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Sample records for annexin a5 microbubbles

  1. Reduction of circulating annexin A5 levels and resistance to annexin A5 anticoagulant activity in women with recurrent spontaneous pregnancy losses.

    NARCIS (Netherlands)

    Rand, J.H.; Arslan, A.A.; Wu, X.X.; Wein, R.; Mulholland, J.; Shah, M.; Heerde, W.L. van; Reutelingsperger, C.P.M.; Lockwood, C.J.; Kuczynski, E.

    2006-01-01

    OBJECTIVE: We investigated whether levels of annexin A5, evidence for resistance to annexin A5 activity, and levels anti-annexin A5 antibodies might be altered in women with a history of recurrent spontaneous pregnancy losses. STUDY DESIGN: These annexin A5 parameters were assayed in 70 nonpregnant

  2. Internalization of annexin A5-functionalized iron oxide particles by apoptotic Jurkat cells

    NARCIS (Netherlands)

    van Tilborg, Geralda A. F.; Geelen, Tessa; Duimel, Hans; Bomans, Paul H. H.; Frederik, Peter M.; Sanders, Honorius M. H. F.; Deckers, Niko M.; Deckers, Roel; Reutelingsperger, Chris P. M.; Strijkers, Gustav J.; Nicolay, Klaas

    2009-01-01

    Apoptosis plays an important role in the etiology of various diseases. Several studies have reported on the use of annexin A5-functionalized iron oxide particles for the detection of apoptosis with MRI, both in vitro and in vivo. The protein annexin A5 binds with high affinity to the phospholipid

  3. [Antiphospholipid antibodies and recurrent abortions: possible pathogenetic role of annexin A5 investigated by confocal microscopy].

    Science.gov (United States)

    Peluso, G; Morrone, G

    2007-06-01

    It has been established that antiphospholipid antibodies (aPL) are associated with recurrent abortions, but the pathophysiologic mechanisms that characterize thrombosis and recurrent pregnancy losses are still not clear.However, it is known that they are associated with the presence of antibodies directed against anionic phospholipids and putative cofactors. In this study the pathogenetic role of annexin A5, a potent anticoagulant cofactor protein for its anticoagulant property in recurrent abortions, was investigated. Endothelial cells of human umbilical veins ''EAHY2936 Line'' in culture were used, incubated with antiphospholipid anticardiolipin (ACA) antibodies purified from plasma of patients with recurrent abortions. The expression of annexin A5 on the cells with ACA was investigated by immunofluorescence and by confocal microscope. The negative control was also carried out: EAHY cells in cultivation medium without ACA. Confocal analysis revealed a uniform distribution of annexin A5 on the cellular membranes in the negative control. Instead, in EAHY cells with ACA, the annexin A5 appears distributed in irregular manners on the cellular membranes (cytoplasmic and nuclear). The results of an irregular ''cluster'' distribution of annexin A5 on the EAHY cells in presence of aPL, and in agreement with the literature, demonstrated that aPL, inhibiting annexin A5 ability to protect anionic phospholipid, promote the coagulation factors to diffuse laterally against phospholipids.

  4. Exhaled breath condensate annexin A5 levels in exercise-induced bronchoconstriction in asthma: A preliminary study.

    Science.gov (United States)

    Tahan, F; Akar, H H; Saraymen, B

    2015-01-01

    The pathogenesis of exercise-induced bronchoconstriction (EIB) in asthma is incompletely understood. The role of exhaled breath condensate (EBC) annexin A5, which is an anti-inflammatory mediator, has not been investigated. The purpose of this study is to evaluate EBC annexin A5 levels in EIB in asthmatic children. Two groups of children were enrolled in this study: asthmatic children with positive (n=11) and negative (n=7) responses to exercise. The levels of pre- and post-exercise EBC annexin A5 were determined with using enzyme-linked immunosorbent assay (ELISA). We observed significant higher pre-exercise EBC annexin A5 levels in the challenge test negative children than in the challenge test positive children (pexercise EBC annexin A5 levels between the groups (p>0.05). Also, no significant difference was observed between pre- and post-exercise EBC annexin A5 levels within each group (p>0.05). There was an inverse correlation between annexin A5 levels and a reduction in forced expiratory volume at one second percent (FEV1%) (p=0.009, r=-0.598). Our preliminary study showed that EBC annexin A5 may have a possible preventive role in EIB in asthma. Annexin A5 and related compounds may provide novel therapeutic approaches to the treatment of EIB in asthma. Copyright © 2014 SEICAP. Published by Elsevier Espana. All rights reserved.

  5. Evaluation of annexin A5 as a biomarker for Alzheimer's disease and Dementia with Lewy bodies

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    Hitoshi eSohma

    2013-04-01

    Full Text Available Background: Alzheimer’s disease (AD differs from other forms of dementia in its relation to amyloid beta-42. Using a cell culture model we previously identified annexin A5 as an AD biomarker. Plasma level of annexin A5 was significantly higher in AD patients compared to that in control. AD has been identified to share a number of clinical and pathological features with Dementia with Lewy bodies (DLB. The present study was done to examine whether or not plasma annexin A5 is a specific marker for AD, when being compared with the levels of DLB patients. As ApoE gene subtype 4 has been noticed as the probable genetic factor for AD, we also examined and compared ApoE genotype in both AD and DLB.Methods: Blood samples were obtained from 150 patients with AD (aged 77.6 ± 6.5 years, 50 patients of DLB (79.4 ± 5.0 and 279 community-dwelling healthy elderly individuals of comparable age and sex (75.6 ± 8.1. All AD patients met NINCDS-ADRDA criteria and all DLB patients were diagnosed as probable DLB according to the latest consensus diagnostic criteria. Quantification was done using the Chemiluminescent Enzyme Immunoassay Technique using the monoclonal antibodies against annexin A5. DNA genotyping of ApoE was performed by distinguishing unique combinations of Hha1 fragments of PCR-amplified genomic DNA products.Results: The plasma level of annexin A5 was significantly higher in AD patients than in the healthy individuals (control (P < 0.0001. The plasma annexin A5 level was also significantly higher in DLB patients than in the control group (P < 0.0001. From the ROC curves, the mean areas under the curve were 0.863 and 0.838 for the AD/control and DLB/control, respectively. The rate of ApoE4 carrier status and the frequency of the e4 allele were significantly higher in AD or DLB than in control and there was no significant difference between AD and DLB.Conclusions: These results suggest that both annexin A5 and ApoE4 are common markers for AD and

  6. Internalization of annexin A5-functionalized iron oxide particles by apoptotic Jurkat cells.

    Science.gov (United States)

    van Tilborg, Geralda A F; Geelen, Tessa; Duimel, Hans; Bomans, Paul H H; Frederik, Peter M; Sanders, Honorius M H F; Deckers, Niko M; Deckers, Roel; Reutelingsperger, Chris P M; Strijkers, Gustav J; Nicolay, Klaas

    2009-01-01

    Apoptosis plays an important role in the etiology of various diseases. Several studies have reported on the use of annexin A5-functionalized iron oxide particles for the detection of apoptosis with MRI, both in vitro and in vivo. The protein annexin A5 binds with high affinity to the phospholipid phosphatidylserine, which is exposed in the outer leaflet of the apoptotic cell membrane. When co-exposed to apoptotic stimuli, this protein was shown to internalize into endocytic vesicles. Therefore in the present study we investigated the possible internalization of commercially available annexin A5-functionalized iron oxide particles (r1 = 34.0 +/- 2.1 and r2 = 205.0 +/- 10.4 mm(-1) s(-1) at 20 MHz), and the effects of their spatial distribution on relaxation rates R2*, R2 and R1. Two different incubation procedures were performed, where (1) Jurkat cells were either incubated with the contrast agent after induction of apoptosis or (2) Jurkat cells were simultaneously incubated with the apoptotic stimulus and the contrast agent. Transmission electron microscopy images and relaxation rates showed that the first incubation strategy mainly resulted in binding of the annexin A5-iron oxide particles to the cell membrane, whereas the second procedure allowed extensive membrane-association as well as a small amount of internalization. Owing to the small extent of internalization, only minor differences were observed between the DeltaR2*/DeltaR2 and DeltaR2/DeltaR1 ratios of cell pellets with membrane-associated or internalized annexin A5 particles. Only the increase in R1 (DeltaR1) appeared to be diminished by the internalization. Internalization of annexin A5-iron oxide particles is also expected to occur in vivo, where the apoptotic stimulus and the contrast agent are simultaneously present. Where the extent of internalization in vivo is similar to that observed in the present study, both T2- and T2*-weighted MR sequences are considered suitable for the detection of these

  7. Development and evaluation of a novel (99mtc-labeled annexin A5 for early detection of response to chemotherapy.

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    Kazuma Ogawa

    Full Text Available (99mTc-HYNIC-annexin A5 can be considered as a benchmark in the field of apoptosis imaging. However, (99mTc-HYNIC-annexin A5 has characteristics of high uptake and long retention in non-target tissues such as kidney and liver. To minimize this problem, we developed a novel (99mTc-labeled annexin A5 using a bis(hydroxamamide derivative [C3(BHam2] as a bifunctional chelating agent, and evaluated its usefulness as an imaging agent for detecting apoptosis. The amino group of C3(BHam2 was converted to a maleimide group, and was coupled to thiol groups of annexin A5 pretreated with 2-iminothiolane. (99mTc labeling was performed by a ligand exchange reaction with (99mTc-glucoheptonate. Biodistribution experiments for both (99mTc-C3(BHam2-annexin A5 and (99mTc-HYNIC-annexin A5 were performed in normal mice. In addition, in tumor-bearing mice, the relationship between the therapeutic effects of chemotherapy (5-FU and the tumor accumulation of (99mTc-C3(BHam2-annexin A5 just after the first treatment of 5-FU was evaluated. (99mTc-C3(BHam2-annexin A5 was prepared with a radiochemical purity of over 95%. In biodistribution experiments, (99mTc-C3(BHam2-annexin A5 had a much lower kidney accumulation of radioactivity than (99mTc-HYNIC-annexin A5. In the organs for metabolism, such as liver and kidney, radioactivity after the injection of (99mTc-HYNIC-annexin A5 was residual for a long time. On the other hand, radioactivity after the injection of (99mTc-C3(BHam2-annexin A5 gradually decreased. In therapeutic experiments, tumor growth in the mice treated with 5-FU was significantly inhibited. Accumulation of (99mTc-C3(BHam2-annexin A5 in tumors significantly increased after 5-FU treatment. The accumulation of radioactivity in tumor correlated positively with the counts of TUNEL-positive cells. These findings suggest that (99mTc-C3(BHam2-annexin A5 may contribute to the efficient detection of apoptotic tumor response after chemotherapy.

  8. Diagnostic value of anti-annexin A5 antibodies in seropositive versus seronegative antiphospholipid syndrome patients

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    Gihan Omar

    2018-04-01

    Full Text Available Background: Current laboratory criteria for antiphospholipid syndrome (APS classification recommend testing positive for antiphospholipid (aPL antibodies. However, there appears to be a subset of patients with classical APS manifestations who test negative. Aim of the work: To analyze the potential clinical usefulness of testing for anti-annexin A5 antibodies in patients with APS and to study the effectiveness of testing for non-criteria aPLs in an attempt to increase the diagnostic yield, particularly in seronegative APS. Patients and methods: 60 APS patients were divided into two groups; 30 seropositive (SP-APS (group I and 30 age and sex matched seronegative (sN-APS testing negative for aPL antibodies. Serum assay for detection of isotypes of anti-annexin A5 antibodies (IgG and IgM were conducted. Results: The mean age of the patients was 32.9 ± 5.8 years, female:male 57:3 and disease duration in SP-APS versus sN-APS (10.17 ± 4.9 years versus 9.6 ± 5.5 years respectively. Secondary APS was present in 16(53.3% patients in group I compared to 3(10% in group II (p < 0.0001. The mean anti-AnxA5 IgG level was 10.7 ± 5.6 U/ml and IgM was 11.2 ± 7.1 U/ml and were comparable between the 2 groups. The obstetric and thrombotic morbidity had no significant differences between SP and sN-APS. The IgG and IgM levels significantly correlated with the pregnancy morbidity, venous and arterial thrombosis events and showed reasonable sensitivities in their prediction (IgG:71.2%,72.8% and 75.8%; IgM: 68%,67.8% and 71.4% respectively and specificities (IgG:75.9%,77.8% and 81.5%; IgM: 70.9%,73.1% and 73.7% respectively. Conclusion: anti-annexinA5 antibodies are promising for detecting obstetric and thrombotic morbidity in both SP- and sN-APS patients. Keywords: Antiphospholipid syndrome, Seropositive APS (SP-APS, Seronegative APS (sN-APS, Anti-annexin A5 antibodies

  9. Quantitative Proteomic Profiling the Molecular Signatures of Annexin A5 in Lung Squamous Carcinoma Cells.

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    Bing Sun

    Full Text Available Lung cancer remains the leading cancer killer around the world. It's crucial to identify newer mechanism-based targets to effectively manage lung cancer. Annexin A5 (ANXA5 is a protein kinase C inhibitory protein and calcium dependent phospholipid-binding protein, which may act as an endogenous regulator of various pathophysiological processes. However, its molecular mechanism in lung cancer remains poorly understood. This study was designed to determine the mechanism of ANXA5 in lung cancer with a hope to obtain useful information to provide a new therapeutic target. We used a stable isotope dimethyl labeling based quantitative proteomic method to identify differentially expressed proteins in NSCLC cell lines after ANXA5 transfection. Out of 314 proteins, we identified 26 and 44 proteins that were down- and up-regulated upon ANXA5 modulation, respectively. The IPA analysis revealed that glycolysis and gluconeogenesis were the predominant pathways modulated by ANXA5. Multiple central nodes, namely HSPA5, FN1, PDIA6, ENO1, ALDOA, JUP and KRT6A appeared to occupy regulatory nodes in the protein-protein networks upon ANXA5 modulation. Taken together, ANXA5 appears to have pleotropic effects, as it modulates multiple key signaling pathways, supporting the potential usefulness of ANXA5 as a potential target in lung cancer. This study might provide a new insight into the mechanism of ANXA5 in lung cancer.

  10. 99mTc-HYNIC-Annexin A5 in Oncology: Evaluating Efficacy of Anti-Cancer Therapies

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    Chris P. Reutelingsperger

    2013-05-01

    Full Text Available Evaluation of efficacy of anti-cancer therapy is currently performed by anatomical imaging (e.g., MRI, CT. Structural changes, if present, become apparent 1–2 months after start of therapy. Cancer patients thus bear the risk to receive an ineffective treatment, whilst clinical trials take a long time to prove therapy response. Both patient and pharmaceutical industry could therefore profit from an early assessment of efficacy of therapy. Diagnostic methods providing information on a functional level, rather than a structural, could present the solution. Recent technological advances in molecular imaging enable in vivo imaging of biological processes. Since most anti-cancer therapies combat tumors by inducing apoptosis, imaging of apoptosis could offer an early assessment of efficacy of therapy. This review focuses on principles of and clinical experience with molecular imaging of apoptosis using Annexin A5, a widely accepted marker for apoptosis detection in vitro and in vivo in animal models. 99mTc-HYNIC-Annexin A5 in combination with SPECT has been probed in clinical studies to assess efficacy of chemo- and radiotherapy within 1–4 days after start of therapy. Annexin A5-based functional imaging of apoptosis shows promise to offer a personalized medicine approach, now primarily used in genome-based medicine, applicable to all cancer patients.

  11. Evaluation of radiolabelled annexin A5 for scintigraphic imaging of cell processes (necrosis/apoptosis) in cardiovascular diseases

    International Nuclear Information System (INIS)

    Sarda-Mantel, L.

    2007-03-01

    Annexin A5, a 35KDa protein, specifically binds with high affinity to phosphatidylserine (P.S.) which is actively redistributed to the external leaflet of plasmic membranes in apoptotic cells and activated platelets. Annexin A5 radiolabelled with 99m Tc( 99m Tc-ANX5) was developed by Strauss (stanford, Usa) to image apoptosis in vivo: tumours cells apoptosis induced by chemo-radiotherapy, ischemia/reperfusion lesions in animals and patients, graft rejection. Additionally, many in vitro data suggest that annexin A5 also stains necrosis (membrane disruption), which occurs in all types of cell death. This preclinical work aimed to evaluate the potential interest of 99m Tc-ANX5 imaging as a clinical tool in cardiovascular diseases. Four studies performed in rat models of myocardial infarction by coronary ligation and ischemia-reperfusion, and in rat models of subacute and acute (isoproterenol-induced) myocarditis show the ability of 99m Tc-ANX5 to detect in vivo cardio myocytes death by apoptosis and necrosis. Another study demonstrates that 99m Tc-ANX5 is highly accurate to evaluate in vivo the biological activity of parietal thrombus in a rat model of elastase-induced abdominal aortic aneurysm. These results suggest that 99m Tc-ANX5 imaging could be used in patients for non invasive diagnosis, prognostic evaluation in acute myocarditis and in various thrombotic cardiovascular diseases. (author)

  12. 99mTc-HYNIC-Annexin A5 in Oncology: Evaluating Efficacy of Anti-Cancer Therapies

    International Nuclear Information System (INIS)

    Schaper, Frédéric L.W.V.J.; Reutelingsperger, Chris P.

    2013-01-01

    Evaluation of efficacy of anti-cancer therapy is currently performed by anatomical imaging (e.g., MRI, CT). Structural changes, if present, become apparent 1–2 months after start of therapy. Cancer patients thus bear the risk to receive an ineffective treatment, whilst clinical trials take a long time to prove therapy response. Both patient and pharmaceutical industry could therefore profit from an early assessment of efficacy of therapy. Diagnostic methods providing information on a functional level, rather than a structural, could present the solution. Recent technological advances in molecular imaging enable in vivo imaging of biological processes. Since most anti-cancer therapies combat tumors by inducing apoptosis, imaging of apoptosis could offer an early assessment of efficacy of therapy. This review focuses on principles of and clinical experience with molecular imaging of apoptosis using Annexin A5, a widely accepted marker for apoptosis detection in vitro and in vivo in animal models. 99m Tc-HYNIC-Annexin A5 in combination with SPECT has been probed in clinical studies to assess efficacy of chemo- and radiotherapy within 1–4 days after start of therapy. Annexin A5-based functional imaging of apoptosis shows promise to offer a personalized medicine approach, now primarily used in genome-based medicine, applicable to all cancer patients

  13. Preclinical Validation of 99mTc–Annexin A5–128 in Experimental Autoimmune Myocarditis and Infective Endocarditis: Comparison with 99mTc–HYNIC–Annexin A5

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    Khadija Benali

    2015-01-01

    Full Text Available Hydrazinonicotinamide–annexin A5 (HYNIC-Anx, a 99m technetium (99mTc-labeled agent targeting phosphatidylserine, proved to be sensitive for the detection of apoptosis and thrombosis but is no longer available for clinical use. A mutant of human annexin designed for direct 99mTc labeling (referred to as Anx A5–128 showed improved binding affinity to phosphatidylserine and is expected to be used in humans. We compared both radiotracers with regard to pharmacokinetics and diagnostic ability in animal models. Biodistribution studies were performed in normal rats. Radiolabeled Anx A5–128 and HYNIC-Anx were compared in cardiovascular settings involving phosphatidylserine expression: experimental autoimmune myocarditis and infective endocarditis. Initial blood clearance was faster for Anx A5–128 than for HYNIC-Anx, and tissue biodistribution was similar overall for both tracers. The diagnostic sensitivity of Anx A5–128 was excellent and comparable to that of HYNIC-Anx. Anx A5–128 showed biodistribution and diagnostic ability similar to those of the HYNIC-Anx derivative, supporting its translation to clinical use.

  14. Comparison of methods for evaluating radiolabelled Annexin A5 uptake in pre-clinical PET oncological studies

    International Nuclear Information System (INIS)

    Grafström, Jonas; Stone-Elander, Sharon

    2014-01-01

    Purpose: The uptakes of radiolabel led AnnexinA5 (AnxA5) and a size-matched control protein in experimental tumours were evaluated by kinetic analyses and compared with standard uptake values (SUVs) to investigate whether the method of analysis may impact on the conclusions that can be drawn. Procedures: PET scans of the 11 C-labelled proteins performed in untreated and doxorubicin-treated mice with head and neck carcinoma xenografts were retrospectively analysed. The appropriateness of using the Logan graphical analyses for reversibly binding radiotracers in these models was evaluated and confirmed. Distribution volume ratios (DVRs) of the regions of interest to reference muscle tissue were compared to those based on the image-derived input function from arterial blood. SUVs were calculated in the same individuals. Results: DVRs based on reference muscle tissue gave results similar to those based on the arterial blood and may be preferred since they are simpler to calculate. In the inter-group comparisons of baseline versus chemotherapy treatment or AnxA5 versus control protein, differences in DVR quantifications had a 20- to 40-fold higher statistical significance than differences in SUVs. As quantified using the control protein, the amount of free ligand in the vascular space of tumours may be large due to enhanced permeability and retention (EPR) contributions at baseline and affected during treatment, which has implications for quantifications of the specifically bound radioligand. Conclusions: These results demonstrate that the quantification method as well as the controls used can be important for interpreting the uptake in tumours of the medium-sized protein ligand AnxA5 and its use in monitoring the effects of therapy on cell death in the tumours. Advances in knowledge and implications for patient care: These results provide additional support for the recognition that more detailed investigations on the effects of the tumour microenvironment on the

  15. Evaluation of the clinical relevance of anti-annexin-A5 antibodies in Chinese patients with antiphospholipid syndrome.

    Science.gov (United States)

    Zhang, Shulan; Wu, Ziyan; Li, Jing; Wen, Xiaoting; Li, Liubing; Zhang, Wen; Zhao, Jiuliang; Zhang, Fengchun; Li, Yongzhe

    2017-02-01

    A hallmark feature of antiphospholipid syndrome (APS) is the presence of antiphospholipid antibodies (aPLs). Few studies have addressed the clinical relevance of anti-annexin A5 antibodies (aANXA5) in Chinese patients with APS. In this study, we evaluated the clinical performance of aANXA5 in the diagnosis of APS. Sera from 313 subjects were tested, including 170 samples from patients with APS, 104 samples from patients with non-APS diseases as disease controls (DC), and 39 healthy controls (HC). Serum IgG and IgM aANXA5 were determined by ELISA. Overall, the levels of both IgG and IgM aANXA5 were significantly increased in patients with primary APS (PAPS) and APS associated to other diseases (APSAOD) compared with DC and HC. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for IgG and IgM aANXA5 in the diagnosis of APS were 33.5 and 15.3, 99.0 and 99.0, 98.3 and 96.3, and 47.7 and 41.7%, respectively. Significant associations between IgG aANXA5 and arterial thrombotic events (OR, 2.60; 95% CI, 1.44-4.71) and between IgG aANXA5 and venous thrombotic events (OR, 2.80; 95% CI, 1.55-5.06) were identified. No correlations were identified between IgG or IgM aANXA5 and obstetric complications. Our data suggest that aANXA5 could serve as a diagnosis biomarker for patients with APS. More importantly, our data highlighted a potential role of IgG aANXA5 in identifying APS patients with high risk of thrombosis.

  16. {sup 99m}Tc-Annexin A5 quantification of apoptotic tumor response: a systematic review and meta-analysis of clinical imaging trials

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    Belhocine, Tarik Z. [Western University, Biomedical Imaging Research Centre (BIRC), London, Ontario (Canada); Blankenberg, Francis G. [Lucile Salter Packard Children' s Hospital, Stanford, Division of Pediatric Radiology, Department of Radiology, Palo Alto, CA (United States); Kartachova, Marina S. [Medical Center Alkmaar, Department of Nuclear Medicine, Alkmaar (Netherlands); Stitt, Larry W. [LW Stitt Statistical Services, London, Ontario (Canada); Vanderheyden, Jean-Luc [JLVMI Consulting LLC, Waukesha, WI (United States); Hoebers, Frank J.P. [Maastricht University Medical Centre, Department of Radiation Oncology (MAASTRO Clinic), GROW School for Oncology and Developmental Biology, Maastricht (Netherlands); Wiele, Christophe van de [University Hospital Ghent, Department of Nuclear Medicine and Radiology, Ghent (Belgium)

    2015-12-15

    {sup 99m}Tc-Annexin A5 has been used as a molecular imaging probe for the visualization, characterization and measurement of apoptosis. In an effort to define the quantitative {sup 99m}Tc-annexin A5 uptake criteria that best predict tumor response to treatment, we performed a systematic review and meta-analysis of the results of all clinical imaging trials found in the literature or publicly available databases. Included in this review were 17 clinical trials investigating quantitative {sup 99m}Tc-annexin A5 (qAnx5) imaging using different parameters in cancer patients before and after the first course of chemotherapy and/or radiation therapy. Qualitative assessment of the clinical studies for diagnostic accuracy was performed using the QUADAS-2 criteria. Of these studies, five prospective single-center clinical trials (92 patients in total) were included in the meta-analysis after exclusion of one multicenter clinical trial due to heterogeneity. Pooled positive predictive values (PPV) and pooled negative predictive values (NPV) (with 95 % CI) were calculated using Meta-Disc software version 1.4. Absolute quantification and/or relative quantification of {sup 99m}Tc-annexin A5 uptake were performed at baseline and after the start of treatment. Various quantitative parameters have been used for the calculation of {sup 99m}Tc-annexin A5 tumor uptake and delta (Δ) tumor changes post-treatment compared to baseline including: tumor-to-background ratio (TBR), ΔTBR, tumor-to-noise ratio, relative tumor ratio (TR), ΔTR, standardized tumor uptake ratio (STU), ΔSTU, maximum count per pixel within the tumor volume (Cmax), Cmax%, absolute ΔU and percentage (ΔU%), maximum ΔU counts, semiquantitative visual scoring, percent injected dose (%ID) and %ID/cm{sup 3}. Clinical trials investigating qAnx5 imaging have included patients with lung cancer, lymphoma, breast cancer, head and neck cancer and other less common tumor types. In two phase I/II single-center clinical trials

  17. Radiolabeling of annexin A5 with {sup 99m}Tc: comparison of HYNIC-Tc vs. iminothiolane-Tc-tricarbonyl conjugates

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    Biechlin, Marie-Laure [Faculte de Medecine Lyon-Sud (EA 3738), 69921 Oullins (France); Radiopharmacie, Centre Hospitalier de Chambery, 73000 Chambery (France)], E-mail: marie.laure.biechlin@ch-chambery.fr; Bonmartin, Alain; Gilly, Francois-Noel [Faculte de Medecine Lyon-Sud (EA 3738), 69921 Oullins (France); Fraysse, Marc [Radiopharmacie, Centre Hospitalier Lyon Sud, 69495 Pierre-Benite (France); Moulinet d' Hardemare, Amaury du [Departement de Chimie Moleculaire, Equipe de Chimie Inorganique Redox et Biomimetique, Universite Joseph Fourier-Grenoble I (DCM UMR 5250, ICMG), 38041 Grenoble cedex 9 (France)], E-mail: amaury.d-hardemare@ujf-grenoble.fr

    2008-08-15

    In the perspective of expanding the use of annexin A5 (anx A5) as radioactive tracer of cell death in vivo, we recently described its radiolabeling with {sup 99m}Tc-tricarbonyl [{sup 99m}Tc(H{sub 2}O){sub 3}(CO){sub 3}]{sup +} via the mercaptobutyrimidyl group (anx A5-SH). The aim of the present article was to compare this new method with the HYNIC strategy (anx A5-HYNIC), recognized at present as the reference for the radiolabeling of proteins with {sup 99m}Tc. Similar radiolabeling yields and better chemical stability were obtained with the [anx A5-SH-{sup 99m}Tc-tricarbonyl] complex. Since the [anx A5-HYNIC-{sup 99m}Tc(tricine){sub 2}] conjugate shows isomeric forms which can affect the biological properties whereas [anx A5-SH-{sup 99m}Tc-tricarbonyl] is less or not prone to such drawback, the latter seems superior to the former. Furthermore, (anx A5-SH) is readily obtained via commercial sources of Traut's reagent whereas (anx A5-HYNIC) is not. The results provide encouraging evidence in the development of anx A5-labeled reagent for apoptose imaging.

  18. Evaluation of radiolabeling of annexin A5 with technetium-99m: influence of the labeling methods on physico-chemical and biological properties of the compounds

    International Nuclear Information System (INIS)

    Santos, Josefina da Silva

    2009-01-01

    Annexin A5 (ANXA5) is an intracellular human protein of 36 kDa with high affinity for membrane-bound phosphatidylserine that is selectively exposed on the surface of cells undergoing apoptosis. Apoptosis is important in normal physiology and innumerous pathologic states. Clinical applications for ANXA5 imaging are being developed in oncology, organ transplantation and cardiovascular diseases. Many strategies to radiolabel the protein have been described, including direct labeling, derivatization through a bifunctional chelating agent (BFC), production of mutated protein or peptide analogs. Several 99 mTc-labeling techniques have been reported using different cores, including [Tc=O] +3 , [Tc]HYNIC, [Tc≡N]+2 and [Tc(CO 3 )] +1 . In this study, we evaluated the influence of 99 mTc cores on biological behavior and physico-chemical properties of radiolabeled annexin. Radiolabeling procedure using [Tc≡N] +2 core was a two-step procedure including the reaction of 99 mTcO4 - with SDH in the presence of SnCl 2 and PDTA to obtain the intermediate 99 mTcN-SDH, and successive addition of ANXA5. The results obtained were not satisfactory, despite the high efficiency in the production of the intermediate. The [Tc=O] +3 core was produced using the ethylene dicysteine (EC) as BFC. TSTU was employed in the derivatization to produce the corresponding hydroxysuccinimide ester. Different ANXA5:EC ratios were studied and all labeling conditions resulted in high radiochemical yield but with differences in lipophilicity, stability, biological distribution and affinity for apoptotic cells. The HYNIC-ANXA5 also produced the labeled protein with high radiochemical yield. The stability of the radiolabeled ANXA5 was evaluated after storing at room temperature, at 2 - 8 degree C and in human serum at 37 degree C. The analysis of these results showed that the 99 mTc-EC-ANXA5 (ratio 10-2) was the most stable compound in all the studied conditions. Partition coefficient assay resulted in

  19. Interaction of annexin A6 with alpha actinin in cardiomyocytes

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    Park Woo J

    2011-01-01

    Full Text Available Abstract Background Annexins are calcium dependent phospholipid binding proteins that are expressed in a wide variety of tissues and implicated in various extra- and intracellular processes. In myocardial tissue, annexins A2, A5 and A6 are particularly abundant, of which the expression levels of annexin A6 has been found to be maximal. Conflicting reports from transgenic mice overexpressing annexin A6 or null mice lacking annexin A6 showed imbalances in intracellular calcium turnover and disturbed cardiac contractility. However, few studies have focussed on the signalling module of annexin A6 in the heart either in normal or in pathological state. Results To identify the putative binding partners of annexin A6 in the heart, ventricular extracts were subjected to glutathione S-transferase (GST- annexin A6 pull down assay and the GST- annexin A6 bound proteins were identified by mass spectrometry. The pull down fractions of ventricular extracts with GST-full length annexin A6 as well as GST-C terminus deleted annexin A6 when immunoblotted with anti sarcomeric alpha (α-actinin antibody showed the presence of α-actinin in the immunoblot which was absent when GST-N terminus deleted annexin A6 was used for pull down. Overexpression of green fluorescent protein (GFP tagged full length annexin A6 showed z-line like appearance in cardiomyocytes whereas GFP-N termimus deleted annexin A6 was mostly localized to the nucleus. Overexpression of GFP-C terminus deleted annexin A6 in cardiomyocytes showed aggregate like appearance in the cytoplasm. Double immunofluorescent staining of cardiomyocytes with anti annexin A6 and anti sarcomeric α-actinin antibodies showed perfect co-localization of these two proteins with annexin A6 appearing like a component of sarcomere. Transient knockdown of annexin A6 in cardiomyocytes by shRNA significantly enhances the contractile functions but does not affect the z-band architecture, as revealed by

  20. Annexin A2 and cancer

    DEFF Research Database (Denmark)

    Christensen, Maria V; Høgdall, Claus K; Jochumsen, Kirsten M

    2018-01-01

    Annexin A2 is a 36-kDa protein interfering with multiple cellular processes especially in cancer progression. The present review aimed to show the relations between Annexin A2 and cancer. A systematic search for studies investigating cancer and Annexin A2 expression was conducted using Pub......Med. Acute lymphoblastic leukaemia, acute promyelocytic leukaemia, clear cell renal cell carcinoma, breast, cervical, colorectal, endometrial, gastric cancer, glioblastoma, hepatocellular carcinoma, lung, multiple myeloma, oesophageal squamous cell carcinoma, ovarian cancer, pancreatic duct adenocarcinoma......, prostate cancer and urothelial carcinoma were evaluated. Annexin A2 expression correlates with resistance to treatment, binding to the bone marrow, histological grade and type, TNM-stage and shortened overall survival. The regulation of Annexin A2 is of interest due to its potential as target for a more...

  1. Shrinking microbubbles with microfluidics: mathematical modelling to control microbubble sizes.

    Science.gov (United States)

    Salari, A; Gnyawali, V; Griffiths, I M; Karshafian, R; Kolios, M C; Tsai, S S H

    2017-11-29

    Microbubbles have applications in industry and life-sciences. In medicine, small encapsulated bubbles (methods are distinguished due to their precise control and ease-of-fabrication. Nevertheless, sub-10 μm diameter bubble generation using microfluidics remains challenging, and typically requires expensive equipment and cumbersome setups. Recently, our group reported a microfluidic platform that shrinks microbubbles to sub-10 μm diameters. The microfluidic platform utilizes a simple microbubble-generating flow-focusing geometry, integrated with a vacuum shrinkage system, to achieve microbubble sizes that are desirable in medicine, and pave the way to eventual clinical uptake of microfluidically generated microbubbles. A theoretical framework is now needed to relate the size of the microbubbles produced and the system's input parameters. In this manuscript, we characterize microbubbles made with various lipid concentrations flowing in solutions that have different interfacial tensions, and monitor the changes in bubble size along the microfluidic channel under various vacuum pressures. We use the physics governing the shrinkage mechanism to develop a mathematical model that predicts the resulting bubble sizes and elucidates the dominant parameters controlling bubble sizes. The model shows a good agreement with the experimental data, predicting the resulting microbubble sizes under different experimental input conditions. We anticipate that the model will find utility in enabling users of the microfluidic platform to engineer bubbles of specific sizes.

  2. Mesenchymal stem cell transplantation enhancement in myocardial infarction rat model under ultrasound combined with nitric oxide microbubbles.

    Directory of Open Access Journals (Sweden)

    Jiayi Tong

    Full Text Available OBJECTIVE: This study evaluated the effects of ultrasound combined with the homemade nitric oxide (NO micro-bubble destruction on the in vitro proliferation, apoptosis, and migration of mesenchymal stem cells (MSCs. Furthermore, we studied whether or not irradiation of the NO micro-bubble combined with bone-marrow derived MSC infusion had a better effect on treating myocardial infarction. The possible mechanism of MSC delivery into the infarcted myocardium was also investigated. METHODS: The murine bone marrow-derived MSCs were isolated, cultured, irradiated, and combined with different concentrations of NO microbubbles. MTT proliferation assay, annexin V-FITC apoptosis detection, migration assay, and RT-PCR were performed 24 h after the irradiation. The NO micro-bubbles was a intravenously injected, followed by the infusion of MSCs, which were labeled by CM-Dil. Myocardium was harvested 48 h later and the distribution of MSCs was observed by laser scanning confocal microscope after frozen sectioning. Echocardiography, histological examination, RT-PCR, and western blotting were performed four weeks after the cell transplantation. RESULTS: Ultrasound combined with 1:70 NO micro-bubbles had no significant impact on the proliferation or apoptosis of MSCs. Transwell chamber findings demonstrated that MSCs migrated more efficiently in group that underwent ultrasound combined with 1:70 NO micro-bubbles. The Real-time PCR results indicated that the expression of CXCR4 was much higher in the group undergoing ultrasound combined with 1:70 NO micro-bubbles. The normalized fluorescence intensity greatly increased in the group of US+NO micro-bubbles and the cardiac function was also markedly improved. Immunohistochemical staining showed that the capillary density was much greater in the group of US+NO micro-bubbles as compared to that of the other groups. RT-PCR and western blotting also revealed a higher SDF-1 and VEGF expression in the group of US

  3. Annexin A2 Heterotetramer: Structure and Function

    Directory of Open Access Journals (Sweden)

    David Waisman

    2013-03-01

    Full Text Available Annexin A2 is a pleiotropic calcium- and anionic phospholipid-binding protein that exists as a monomer and as a heterotetrameric complex with the plasminogen receptor protein, S100A10. Annexin A2 has been proposed to play a key role in many processes including exocytosis, endocytosis, membrane organization, ion channel conductance, and also to link F-actin cytoskeleton to the plasma membrane. Despite an impressive list of potential binding partners and regulatory activities, it was somewhat unexpected that the annexin A2-null mouse should show a relatively benign phenotype. Studies with the annexin A2-null mouse have suggested important functions for annexin A2 and the heterotetramer in fibrinolysis, in the regulation of the LDL receptor and in cellular redox regulation. However, the demonstration that depletion of annexin A2 causes the depletion of several other proteins including S100A10, fascin and affects the expression of at least sixty-one genes has confounded the reports of its function. In this review we will discuss the annexin A2 structure and function and its proposed physiological and pathological roles.

  4. Collective dissolution of microbubbles

    Science.gov (United States)

    Michelin, Sébastien; Guérin, Etienne; Lauga, Eric

    2018-04-01

    A microscopic bubble of soluble gas always dissolves in finite time in an undersaturated fluid. This diffusive process is driven by the difference between the gas concentration near the bubble, whose value is governed by the internal pressure through Henry's law, and the concentration in the far field. The presence of neighboring bubbles can significantly slow down this process by increasing the effective background concentration and reducing the diffusing flux of dissolved gas experienced by each bubble. We develop theoretical modeling of such diffusive shielding process in the case of small microbubbles whose internal pressure is dominated by Laplace pressure. We first use an exact semianalytical solution to capture the case of two bubbles and analyze in detail the shielding effect as a function of the distance between the bubbles and their size ratio. While we also solve exactly for the Stokes flow around the bubble, we show that hydrodynamic effects are mostly negligible except in the case of almost-touching bubbles. In order to tackle the case of multiple bubbles, we then derive and validate two analytical approximate yet generic frameworks, first using the method of reflections and then by proposing a self-consistent continuum description. Using both modeling frameworks, we examine the dissolution of regular one-, two-, and three-dimensional bubble lattices. Bubbles located at the edge of the lattices dissolve first, while innermost bubbles benefit from the diffusive shielding effect, leading to the inward propagation of a dissolution front within the lattice. We show that diffusive shielding leads to severalfold increases in the dissolution time, which grows logarithmically with the number of bubbles in one-dimensional lattices and algebraically in two and three dimensions, scaling respectively as its square root and 2 /3 power. We further illustrate the sensitivity of the dissolution patterns to initial fluctuations in bubble size or arrangement in the case

  5. [Annexins and recurrent pregnancy loss].

    Science.gov (United States)

    Udry, Sebastián; Aranda, Federico; Latino, Omar; Larrañaga, Gabriela de

    2013-01-01

    Recurrent Pregnancy Loss (RPL) affects public health and directly compromises the quality of life of hundreds of women, with a detrimental effect on their physical and mental health. Approximately 50% of RPL are not associated to any of the currently known etiology and will be considered idiopathic. Recently, it has been demonstrated that the expression of annexin 5 (ANXA5), a protein found on the trophoblastic surface, plays a fundamental role in the development of pregnancy due to its immunomodulator and anticoagulant function at the placentary level. Some genetic haplotypes of ANXA5 are associated to alterations in the expression of this gene, such as haplotype M2 which is associated to a decrease in the expression of ANXA5. The presence of this haplotype is related to the following conditions occurring during pregnancy: RPL, foetal intrauterine growth restriction, low child weight at birth, preeclampsia and maternal pulmonary thromboembolism. This review describes the structure, function and genetic expression of ANXA5, as well as its possible implication in RPL.

  6. Microbubble acoustic signatures: bubble deflation

    NARCIS (Netherlands)

    ten Brinke, G.A.; Slump, Cornelis H.

    2006-01-01

    Ultrasound Contrast Agents (UCAs) are used in medical imaging to enhance the visibility of structures, especially blood vessels and the liver. An example application of UCAs is the detection and classification of tumors. The most common UCA consist of microbubbles, which have pronounced non-linear

  7. Annexin-Phospholipid Interactions. Functional Implications

    Directory of Open Access Journals (Sweden)

    Javier Turnay

    2013-01-01

    Full Text Available Annexins constitute an evolutionary conserved multigene protein superfamily characterized by their ability to interact with biological membranes in a calcium dependent manner. They are expressed by all living organisms with the exception of certain unicellular organisms. The vertebrate annexin core is composed of four (eight in annexin A6 homologous domains of around 70 amino acids, with the overall shape of a slightly bent ring surrounding a central hydrophilic pore. Calcium- and phospholipid-binding sites are located on the convex side while the N-terminus links domains I and IV on the concave side. The N-terminus region shows great variability in length and amino acid sequence and it greatly influences protein stability and specific functions of annexins. These proteins interact mainly with acidic phospholipids, such as phosphatidylserine, but differences are found regarding their affinity for lipids and calcium requirements for the interaction. Annexins are involved in a wide range of intra- and extracellular biological processes in vitro, most of them directly related with the conserved ability to bind to phospholipid bilayers: membrane trafficking, membrane-cytoskeleton anchorage, ion channel activity and regulation, as well as antiinflammatory and anticoagulant activities. However, the in vivo physiological functions of annexins are just beginning to be established.

  8. Microbubble dissolution in a multigas environment.

    Science.gov (United States)

    Kwan, James J; Borden, Mark A

    2010-05-04

    Microbubbles occur naturally in the oceans and are used in many industrial and biomedical applications. Here, a theoretical and experimental study was undertaken to determine the fate of a microbubble suddenly suspended in a medium with several gas species as in, for example, the injection of an ultrasound contrast agent into the bloodstream. The model expands on Epstein and Plesset's analysis to include any number of gases. An experimental system was developed which isolates the microbubble in a permeable hollow fiber submerged in a perfusion chamber, allowing rapid exchange of the external aqueous medium. Experimental verification of the model was performed with individual sulfur hexafluoride (SF(6)) microbubbles coated with the soluble surfactant, sodium dodecyl sulfate (SDS). SDS-coated microbubbles suddenly placed in an air-saturated medium initially grew with the influx of O(2) and N(2) and then dissolved under Laplace pressure. SF(6)-filled microbubbles coated with the highly insoluble lipid, dibehenoylphosphatidylcholine, were found to exhibit significantly different behavior owing to a dynamic surface tension. The initial growth phase was diminished, possibly owing to a shell "breakup" tension that exceeded the pure gas/liquid surface tension. Three dissolution regimes were observed: (1) an initial rapid dissolution to the initial diameter followed by (2) steady dissolution with monolayer collapse and finally (3) stabilization below 10 microm diameter. Results indicated that the lipid shell becomes increasingly rigid as the microbubble dissolves, which has important implications on microbubble size distribution, stability, and acoustic properties.

  9. Increased alveolar soluble Annexin V promotes lung inflammation and fibrosis

    OpenAIRE

    Buckley, S.; Shi, W.; Xu, W.; Frey, M.R.; Moats, R.; Pardo, A.; Selman, M.; Warburton, D.

    2015-01-01

    The causes underlying the self-perpetuating nature of idiopathic pulmonary fibrosis (IPF), a progressive and usually lethal disease, remain unknown. We hypothesized that alveolar soluble Annexin V contributes to lung fibrosis, based on the observation that human IPF BALF containing high Annexin V levels promoted fibroblast involvement in alveolar epithelial wound healing that was reduced when Annexin V was depleted from the BALF.

  10. The Microbubble Assisted Bioremediation of Chlorinated Ethenes

    OpenAIRE

    Kaiser, Philip Marc Jr.

    1998-01-01

    This work focused on using a microbubble dispersion to deliver hydrogen and carbon dioxide to anaerobic consortia to stimulate their ability to reductively dehalogenate tetrachloroethylene all the way to ethene and ethane. A continuous flow system, consisting of six anaerobic soil column bioreactors, inoculated with sediments from Virginia Tech's Duck Pond, was used for this study. Two columns received microbubbles containing hydrogen and carbon dioxide, two received sodium propionate, and ...

  11. Biosurfactants for Microbubble Preparation and Application

    OpenAIRE

    Takeo Shiina; Zengshe Liu; Mitsutoshi Nakajima; Qingyi Xu

    2011-01-01

    Biosurfactants can be classified by their chemical composition and their origin. This review briefly describes various classes of biosurfactants based on their origin and introduces a few of the most widely used biosurfactants. The current status and future trends in biosurfactant production are discussed, with an emphasis on those derived from plants. Following a brief introduction of the properties of microbubbles, recent progress in the application of microbubble technology to molecular im...

  12. Nuclear localization of Annexin A7 during murine brain development

    Directory of Open Access Journals (Sweden)

    Noegel Angelika A

    2005-04-01

    Full Text Available Abstract Background Annexin A7 is a member of the annexin protein family, which is characterized by its ability to interact with phospholipids in the presence of Ca2+-ions and which is thought to function in Ca2+-homeostasis. Results from mutant mice showed altered Ca2+-wave propagation in astrocytes. As the appearance and distribution of Annexin A7 during brain development has not been investigated so far, we focused on the distribution of Annexin A7 protein during mouse embryogenesis in the developing central nervous system and in the adult mouse brain. Results Annexin A7 is expressed in cells of the developing brain where a change in its subcellular localization from cytoplasm to nucleus was observed. In the adult CNS, the subcellular distribution of Annexin A7 depends on the cell type. By immunohistochemistry analysis Annexin A7 was detected in the cytosol of undifferentiated cells at embryonic days E5–E8. At E11–E15 the protein is still present in the cytosol of cells predominantly located in the ventricular germinative zone surrounding the lateral ventricle. Later on, at embryonic day E16, Annexin A7 in cells of the intermediate and marginal zone of the neopallium translocates to the nucleus. Neuronal cells of all areas in the adult brain present Annexin A7 in the nucleus, whereas glial fibrillary acidic protein (GFAP-positive astrocytes exhibit both, a cytoplasmic and nuclear staining. The presence of nuclear Annexin A7 was confirmed by extraction of the nucleoplasm from isolated nuclei obtained from neuronal and astroglial cell lines. Conclusion We have demonstrated a translocation of Annexin A7 to nuclei of cells in early murine brain development and the presence of Annexin A7 in nuclei of neuronal cells in the adult animal. The role of Annexin A7 in nuclei of differentiating and mature neuronal cells remains elusive.

  13. Lead-silicate glass optical microbubble resonator

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Pengfei, E-mail: pengfei.wang@dit.ie [Photonics Research Centre, Dublin Institute of Technology, Kevin Street, Dublin 8 (Ireland); Optoelectronics Research Centre, University of Southampton, Southampton SO17 1BJ (United Kingdom); Ward, Jonathan; Yang, Yong; Chormaic, Síle Nic [Light-Matter Interactions Unit, OIST Graduate University, 1919-1 Tancha, Onna-son, Okinawa 904-0495 (Japan); Feng, Xian; Brambilla, Gilberto [Optoelectronics Research Centre, University of Southampton, Southampton SO17 1BJ (United Kingdom); Farrell, Gerald [Photonics Research Centre, Dublin Institute of Technology, Kevin Street, Dublin 8 (Ireland)

    2015-02-09

    Microbubble whispering gallery resonators have the potential to become key components in a variety of active and passive photonic circuit devices by offering a range of significant functionalities. Here, we report on the fabrication, optical characterization, and theoretical analysis of lead-silicate glass and optical microbubble resonators. Evanescent field coupling to the microbubbles was achieved using a 1 μm diameter, silica microfiber at a wavelength of circa 775 nm. High Q-factor modes were efficiently excited in both single-stem and two-stem, lead-silicate glass, and microbubble resonators, with bubble diameters of 38 μm (single-stem) and 48 μm (two-stem). Whispering gallery mode resonances with Q-factors as high as 2.3 × 10{sup 5} (single-stem) and 7 × 10{sup 6} (two-stem) were observed. By exploiting the high-nonlinearity of the lead-silicate glass, this work will act as a catalyst for studying a range of nonlinear optical effects in microbubbles, such as Raman scattering and four-wave mixing, at low optical powers.

  14. Microbubble Distillation for Ethanol-Water Separation

    Directory of Open Access Journals (Sweden)

    Atheer Al-yaqoobi

    2016-01-01

    Full Text Available In the current study, a novel approach for separating ethanol-water mixture by microbubble distillation technology was investigated. Traditional distillation processes require large amounts of energy to raise the liquid to its boiling point to effect removal of volatile components. The concept of microbubble distillation by comparison is to heat the gas phase rather than the liquid phase to achieve separation. The removal of ethanol from the thermally sensitive fermentation broths was taken as a case of study. Consequently the results were then compared with those which could be obtained under equilibrium conditions expected in an “ideal” distillation unit. Microbubble distillation has achieved vapour compositions higher than that which could be obtained under traditional equilibrium conditions. The separation was achieved at liquid temperature significantly less than the boiling point of the mixture. In addition, it was observed that the separation efficiency of the microbubble distillation could be increased by raising the injected air temperature, while the temperature of the liquid mixture increased only moderately. The separation efficiency of microbubble distillation was compared with that of pervaporation for the recovery of bioethanol from the thermally sensitive fermentation broths. The technology could be controlled to give high separation and energy efficiency. This could contribute to improving commercial viability of biofuel production and other coproducts of biorefinery processing.

  15. Acoustically excited encapsulated microbubbles and mitigation of biofouling

    KAUST Repository

    Qamar, Adnan

    2017-08-31

    Provided herein is a universally applicable biofouling mitigation technology using acoustically excited encapsulated microbubbles that disrupt biofilm or biofilm formation. For example, a method of reducing biofilm formation or removing biofilm in a membrane filtration system is provided in which a feed solution comprising encapsulated microbubbles is provided to the membrane under conditions that allow the encapsulated microbubbles to embed in a biofilm. Sonication of the embedded, encapsulated microbubbles disrupts the biofilm. Thus, provided herein is a membrane filtration system for performing the methods and encapsulated microbubbles specifically selected for binding to extracellular polymeric substances (EFS) in a biofilm.

  16. Acoustic Studies on Nanodroplets, Microbubbles and Liposomes

    Science.gov (United States)

    Kumar, Krishna Nandan

    Microbubbles and droplets are nanometer to micron size biocompatible particles which are primarily used for drug delivery and contrast imaging. Our aim is to broaden the use of microbubbles from contrast imaging to other applications such as measuring blood pressure. The other goal is to develop in situ contrast agents (phase shift droplets) which can be used for applications such as cancer tumor imaging. Therefore, the focus is on developing and validating the concept using experimental and theoretical methods. Below is an overview of each of the projects performed on droplets and microbubbles. Phase shift droplets vaporizable by acoustic stimulation offer many advantages over microbubbles as contrast agents due to their higher stability and possibility of smaller sizes. In this study, the acoustic droplet vaporization (ADV) threshold of a suspension of PFP droplets (400-3000nm) was acoustically measured as a function of the excitation frequency by examining the scattered signals, fundamental, sub- and second-harmonic. This work presents the experimental methodology to determine ADV threshold. The threshold increases with frequency: 1.25 MPa at 2.25 MHz, 2.0 MPa at 5 MHz and 2.5 MPa at 10 MHz. The scattered response from droplets was also found to match well with that of independently prepared lipid-coated microbubble suspension in magnitude as well as trends above the threshold value. Additionally, we have employed classical nucleation theory (CNT) to investigate the ADV, specifically the threshold value of the peak negative pressure required for vaporization. The theoretical analysis predicts that the ADV threshold increases with increasing surface tension of the droplet core and frequency of excitation, while it decreases with increasing temperature and droplet size. The predictions are in qualitative agreement with experimental observations. A technique to measure the ambient pressure using microbubbles was developed. Here we are presenting the results of an

  17. The use of microbubbles to target drug delivery

    Directory of Open Access Journals (Sweden)

    Porter Richard

    2004-11-01

    Full Text Available Abstract Ultrasound-mediated microbubbles destruction has been proposed as an innovative method for noninvasive delivering of drugs and genes to different tissues. Microbubbles are used to carry a drug or gene until a specific area of interest is reached, and then ultrasound is used to burst the microbubbles, causing site-specific delivery of the bioactive materials. Furthermore, the ability of albumin-coated microbubbles to adhere to vascular regions with glycocalix damage or endothelial dysfunction is another possible mechanism to deliver drugs even in the absence of ultrasound. This review focuses on the characteristics of microbubbles that give them therapeutic properties and some important aspects of ultrasound parameters that are known to influence microbubble-mediated drug delivery. In addition, current studies involving this novel therapeutical application of microbubbles will be discussed.

  18. Dynamics of micro-bubble sonication inside a phantom vessel

    KAUST Repository

    Qamar, Adnan

    2013-01-10

    A model for sonicated micro-bubble oscillations inside a phantom vessel is proposed. The model is not a variant of conventional Rayleigh-Plesset equation and is obtained from reduced Navier-Stokes equations. The model relates the micro-bubble oscillation dynamics with geometric and acoustic parameters in a consistent manner. It predicts micro-bubble oscillation dynamics as well as micro-bubble fragmentation when compared to the experimental data. For large micro-bubble radius to vessel diameter ratios, predictions are damped, suggesting breakdown of inherent modeling assumptions for these cases. Micro-bubble response with acoustic parameters is consistent with experiments and provides physical insight to the micro-bubble oscillation dynamics.

  19. Radionuclide tumor therapy with ultrasound contrast microbubbles

    NARCIS (Netherlands)

    van Wamel, Annemieke; Bouakaz, Ayache; Bernard, Bert; ten Cate, Folkert; de Jong, N.

    2004-01-01

    Radionuclides have shown to be effective in tumour therapy. However, the side effects determine the maximum deliverable dose. Recently, it has been demonstrated that cells can be permeabilised through sonoporation using ultrasound and contrast microbubbles. The use of sonoporation in treatment of

  20. Nonspherical oscilllations of ultrasound contrast agent microbubbles

    NARCIS (Netherlands)

    Dollet, B.; van der Meer, S.M.; Garbin, V.; Garbin, Valeria; de Jong, N.; Lohse, Detlef; Versluis, Michel

    2008-01-01

    The occurrence of nonspherical oscillations (or surface modes) of coated microbubbles, used as ultrasound contrast agents in medical imaging, is investigated using ultra–high-speed optical imaging. Optical tweezers designed to micromanipulate single bubbles in 3-D are used to trap the bubbles far

  1. Mapping microbubble viscosity using fluorescence lifetime imaging of molecular rotors

    Science.gov (United States)

    Hosny, Neveen A.; Mohamedi, Graciela; Rademeyer, Paul; Owen, Joshua; Wu, Yilei; Tang, Meng-Xing; Eckersley, Robert J.; Stride, Eleanor; Kuimova, Marina K.

    2013-01-01

    Encapsulated microbubbles are well established as highly effective contrast agents for ultrasound imaging. There remain, however, some significant challenges to fully realize the potential of microbubbles in advanced applications such as perfusion mapping, targeted drug delivery, and gene therapy. A key requirement is accurate characterization of the viscoelastic surface properties of the microbubbles, but methods for independent, nondestructive quantification and mapping of these properties are currently lacking. We present here a strategy for performing these measurements that uses a small fluorophore termed a “molecular rotor” embedded in the microbubble surface, whose fluorescence lifetime is directly related to the viscosity of its surroundings. We apply fluorescence lifetime imaging to show that shell viscosities vary widely across the population of the microbubbles and are influenced by the shell composition and the manufacturing process. We also demonstrate that heterogeneous viscosity distributions exist within individual microbubble shells even with a single surfactant component. PMID:23690599

  2. Is annexin 1 a multifunctional protein during stress responses?

    OpenAIRE

    Clark, Greg; Konopka-Postupolska, Dorota; Hennig, Jacek; Roux, Stanley

    2010-01-01

    Accumulating evidence suggest that certain annexins can play a role in abiotic stress responses in plants. We found that for one member of the Arabidopsis thaliana annexin gene family, annexin 1 (AnnAt1), loss-of-function mutants are more sensitive to drought stress and gain-of-function mutants are more tolerant.1 We also found that AnnAt1 is able to regulate accumulation of H2O2 in vivo in Arabidopsis cells based on the observation that the level of ROS accumulation following induction by AB...

  3. A Review of Microbubble and its Applications in Ozonation

    Science.gov (United States)

    Shangguan, Yufei; Yu, Shuili; Gong, Chao; Wang, Yue; Yang, Wangzhen; Hou, Li-an

    2018-03-01

    Ozonation has been demonstrated to be an effective technology for the oxidation of organic matters in water treatment. But the low solubility and low mass transfer efficiency limit the application. Microbubble technology has the potential of enhancing gas-liquid mass transfer efficiency, thus it can be applied in ozonation process. The applications of microbubble ozonation have shown advantages over macro bubble ozonation in mass transfer and reaction rate. Microbubble ozonation will be a promising treatment both in water and wastewater treatment.

  4. Prognostic significance of annexin A2 and annexin A4 expression in patients with cervical cancer

    International Nuclear Information System (INIS)

    Choi, Chel Hun; Chung, Joon-Yong; Chung, Eun Joo; Sears, John D.; Lee, Jeong-Won; Bae, Duk-Soo; Hewitt, Stephen M.

    2016-01-01

    The annexins (ANXs) have diverse roles in tumor development and progression, however, their clinical significance in cervical cancer has not been elucidated. The present study was to investigate the clinical significance of annexin A2 (ANXA2) and annexin A4 (ANXA4) expression in cervical cancer. ANXA2 and ANXA4 immunohistochemical staining were performed on a cervical cancer tissue microarray consisting of 46 normal cervical epithelium samples and 336 cervical cancer cases and compared the data with clinicopathological variables, including the survival of cervical cancer patients. ANXA2 expression was lower in cancer tissue (p = 0.002), whereas ANXA4 staining increased significantly in cancer tissues (p < 0.001). ANXA2 expression was more prominent in squamous cell carcinoma (p < 0.001), whereas ANXA4 was more highly expressed in adeno/adenosquamous carcinoma (p < 0.001). ANXA2 overexpression was positively correlated with advanced cancer phenotypes, whereas ANXA4 expression was associated with resistance to radiation with or without chemotherapy (p = 0.029). Notably, high ANXA2 and ANXA4 expression was significantly associated with shorter disease-free survival (p = 0.004 and p = 0.033, respectively). Multivariate analysis indicated that ANXA2+ (HR = 2.72, p = 0.003) and ANXA2+/ANXA4+ (HR = 2.69, p = 0.039) are independent prognostic factors of disease-free survival in cervical cancer. Furthermore, a random survival forest model using combined ANXA2, ANXA4, and clinical variables resulted in improved predictive power (mean C-index, 0.76) compared to that of clinical-variable-only models (mean C-index, 0.70) (p = 0.006). These findings indicate that detecting ANXA2 and ANXA4 expression may aid the evaluation of cervical carcinoma prognosis. The online version of this article (doi:10.1186/s12885-016-2459-y) contains supplementary material, which is available to authorized users

  5. Evaluation of {sup 99m}Tc-MAG{sub 3}-annexin V: influence of the chelate on in vitro and in vivo properties in mice

    Energy Technology Data Exchange (ETDEWEB)

    Vanderheyden, Jean-Luc [Department of Radiology, University of Massachusetts Medical School, Worcester, MA 01655 (United States)]. E-mail: jlvand@earthlink.net; Liu Guozheng [Department of Radiology, University of Massachusetts Medical School, Worcester, MA 01655 (United States); He, Jiang [Department of Radiology, University of Massachusetts Medical School, Worcester, MA 01655 (United States); Patel, Bhavesh [Department of Radiology, University of Massachusetts Medical School, Worcester, MA 01655 (United States); Tait, Jonathan F. [Department of Laboratory Medicine, Medicine (Medical Genetics) and Pathology, University of Washington, Seattle, WA 98195 (United States); Hnatowich, Donald J. [Department of Radiology, University of Massachusetts Medical School, Worcester, MA 01655 (United States)

    2006-01-15

    We conjugated mercaptoacetyltriglycine (MAG{sub 3}) to rh-annexin V to permit radiolabeling with {sup 99m}Tc in an effort to decrease the high kidney and liver accumulation observed for {sup 99m}Tc-labeled Hynic-annexin V. The 36-kDa protein was conjugated at a 5:1 molar ratio with NHS-MAG{sub 3} in HEPES buffer pH 7.8 at room temperature, then quenched with glycine and purified by dialysis. The biopotency of the resulting MAG{sub 3}-annexin was similar to that of Hynic-annexin as determined by a sensitive red blood cell membrane affinity binding assay and a surface plasmon resonance (SPR) assay. The {sup 99m}Tc radiolabeling of MAG{sub 3}-annexin resulted in radiochemical yields of 90% under mildly basic pH conditions. Biodistribution data in normal mice clearly showed a significant decrease in kidney and liver uptake at 1 h postinjection for the {sup 99m}Tc MAG{sub 3}-annexin compared to the {sup 99m}Tc Hynic-annexin (from 24% ID to 4% ID for the liver, and 45% ID to 15% ID for the kidneys, respectively). Autoradiography of the kidneys showed retention of radioactivity in the collecting tubules following administration of both labeled annexins. The {sup 99m}Tc MAG{sub 3}-annexin biodistribution was also characterized by a lower retention of radioactivity in the whole body, but with small intestine accumulation over fivefold higher than observed with {sup 99m}Tc Hynic-annexin. These findings show a definite improvement in renal and hepatic clearance of the MAG{sub 3} radioligand. However, due to the increased radioactivity uptake in the small intestines, the early in vivo detection of ongoing apoptosis in the lower abdomen might be more difficult with {sup 99m}Tc MAG{sub 3}-annexin. Nevertheless, {sup 99m}Tc MAG{sub 3}-annexin may be an attractive alternative to {sup 99m}Tc Hynic-annexin for the in vivo imaging of phosphatidylserine receptors.

  6. Tunable microbubble generator using electrolysis and ultrasound

    OpenAIRE

    Younes Achaoui; Khaled Metwally; Damien Fouan; Zoubida Hammadi; Roger Morin; Eric Debieu; Cédric Payan; Serge Mensah

    2017-01-01

    This letter reports on a method for producing on demand calibrated bubbles in a non-chemically controlled solution using localized micro-electrolysis and ultrasound. Implementing a feedback loop in the process leads to a point source of stable mono-dispersed microbubbles. This approach overcomes the inertial constraints encountered in microfluidics with the possibility to produce from a single to an array of calibrated bubbles. Moreover, this method avoids the use of additional surfactant tha...

  7. Facilitating Intracellular Drug Delivery by Ultrasound-Activated Microbubbles

    NARCIS (Netherlands)

    Lammertink, BHA

    2017-01-01

    The goal of this thesis was to investigate the combination of ultrasound and microbubbles (USMB) for intracellular delivery of (model) drugs in vitro. We have focused on clinically approved drugs, i.e. cisplatin, and microbubbles, i.e. SonoVue™, to facilitate clinical translation. In addition, model

  8. Light propagation in a turbid medium with insonified microbubbles

    Science.gov (United States)

    Leung, Terence S.; Honeysett, Jack E.; Stride, Eleanor; Deng, Jing

    2013-01-01

    Surfactant stabilized microbubbles are widely used clinical contrast agents for ultrasound imaging. In this work, the light propagation through a turbid medium in the presence of microbubbles has been investigated. Through a series of experiments, it has been found that the optical attenuation is increased when the microbubbles in a turbid medium are insonified by ultrasound. Such microbubble enhanced optical attenuation is a function of both applied ultrasound pressure and microbubble concentration. To understand the mechanisms involved, a Monte Carlo (MC) model has been developed. Under ultrasound exposure, the sizes of microbubbles vary in space and time, and their dynamics are modeled by the Rayleigh-Plesset equation. By using Mie theory, the spatially and temporally varying optical scattering and scattering efficiency of microbubbles are determined based on the bubble sizes and internal refractive indices. The MC model is shown to effectively describe a medium with rapidly changing optical scattering, and the results are validated against both computational results using an N-layered diffusion equation model and experimental results using a clinical microbubble contrast agent (SonoVue).

  9. Optical characterization of individual liposome-loaded microbubbles

    NARCIS (Netherlands)

    Faez, T.; Skachkov, I.; Gelderblom, E.C.; Geers, B.; Lentacker, I.; van der Steen, A.F.W.; Versluis, Michel; de Jong, N.

    2012-01-01

    Newly developed liposome-loaded (LPS) microbubbles are characterized by comparing their oscillating response with standard phospholipid-coated (bare) microbubbles using the ultra-high speed imaging (Brandaris 128) camera. A study of the shell properties indicate nearly the same shell elasticity and

  10. Annexin V and anti-Annexin V antibodies: two interesting aspects in acute myocardial infarction

    Directory of Open Access Journals (Sweden)

    Kholoosi Ensieh

    2009-07-01

    Full Text Available Abstract Background Myocardial infarction is the combined result of environmental factors and personal predispositions. Prothrombotic factors might play an important role in this phenomenon. Annexin V (ANV is a calcium-dependent glycoprotein widely present in various tissues exerting a potent anticoagulant effect in vitro by reducing plaque adhesion and aggregation. Anti-annexin V antibodies (aANVAs are detected in various diseases like rheumatoid arthritis, systemic lupus erythematosus and anti-phospholipid antibody syndrome. The study of ANV in Acute Myocardial Infarction (AMI might shed light on hypercoagulability mechanisms in the pathogenesis of acute coronary syndromes. This study was conducted to investigate the association of plasma ANV, aANVAs and anti-cardiolipin antibodies (aCLAs with AMI. Methods This study recruited 45 patients with the diagnosis of AMI according to WHO criteria in their first 24 hours of admission. 36 matched individuals were studied as the control group with normal coronary artery angiography. Plasma levels of ANV, aANVAs and aCLAs were determined by enzyme-linked immunosorbent assay and the results were compared. Results Plasma ANV levels in the patients with AMI on admission were significantly lower than those in the control group (p = 0.002. Positive test for aANVAs were found to be present in a significant number of our patients (p = 0.004. The studied groups were similar in their rate of patients with positive aCLAs tests. ANV, aANVAs and aCLAs were not correlated with hypertension, diabetes mellitus, hyperlipidemia, sex, age and smoking. Conclusion Our findings suggest that low plasma ANV levels along with positive aANVAs tests in patients with AMI are indicative of hypercoagulable state that is not related to the traditional cardiovascular risk factors.

  11. Immunolocalization of an annexin-like protein in corn

    Science.gov (United States)

    Clark, G. B.; Dauwalder, M.; Roux, S. J.

    1994-08-01

    Although calcium has been proposed to be an important regulatory element in plant gravitropic growth, as yet no specific function of Ca2+ in growth regulation has been discovered. Our recent studies on a Ca2+-binding protein in pea seedlings called p35 indicate that it is a member of the annexin family of proteins and may play a key role in growth regulation through its function in delivering polysaccharides needed for wall construction. We previously reported the isolation of p35 from pea plumules and the production of polyclonal antibodies to it. Immunolocalization analyses of p35 in pea tissues revealed high levels of staining in secretory cell types such as developing vascular cells and outer root cap cells. To test how general was the occurrence and distribution of this annexin-like protein in plant cells we initiated an analysis of annexins in the monocot corn using immunological techniques. Our results indicate the immunochemical properties and localization of corn annexins are very similar to those reported for pea. They are consistent with the postulate that annexins may play a general role in the regulation of the secretion of wall polysaccharides needed for growth, and thus could be an important target of calcium action during gravitropic growth.

  12. Tunable microbubble generator using electrolysis and ultrasound

    Science.gov (United States)

    Achaoui, Younes; Metwally, Khaled; Fouan, Damien; Hammadi, Zoubida; Morin, Roger; Debieu, Eric; Payan, Cédric; Mensah, Serge

    2017-01-01

    This letter reports on a method for producing on demand calibrated bubbles in a non-chemically controlled solution using localized micro-electrolysis and ultrasound. Implementing a feedback loop in the process leads to a point source of stable mono-dispersed microbubbles. This approach overcomes the inertial constraints encountered in microfluidics with the possibility to produce from a single to an array of calibrated bubbles. Moreover, this method avoids the use of additional surfactant that may modify the composition of the host fluid. It impacts across a broad range of scientific domains from bioengineering, sensing to environment.

  13. Tunable microbubble generator using electrolysis and ultrasound

    Directory of Open Access Journals (Sweden)

    Younes Achaoui

    2017-01-01

    Full Text Available This letter reports on a method for producing on demand calibrated bubbles in a non-chemically controlled solution using localized micro-electrolysis and ultrasound. Implementing a feedback loop in the process leads to a point source of stable mono-dispersed microbubbles. This approach overcomes the inertial constraints encountered in microfluidics with the possibility to produce from a single to an array of calibrated bubbles. Moreover, this method avoids the use of additional surfactant that may modify the composition of the host fluid. It impacts across a broad range of scientific domains from bioengineering, sensing to environment.

  14. Minimising microbubble size through oscillation frequency control

    Czech Academy of Sciences Publication Activity Database

    Brittle, S.; Deasi, P.; Ng, W. Ch.; Dunbar, A.; Howell, R.; Tesař, Václav; Zimmerman, W. B.

    2015-01-01

    Roč. 104, December (2015), s. 357-366 ISSN 0263-8762 Institutional support: RVO:61388998 Keywords : microbubbles * process intensification * transfer phenomena Subject RIV: BK - Fluid Dynamics Impact factor: 2.525, year: 2015 http://ac.els-cdn.com/S0263876215002993/1-s2.0-S0263876215002993-main.pdf?_tid=4fca5bdc-9e5f-11e5-85c5-00000aab0f02&acdnat=1449656970_b6957d7afd64592d184a978b367e8e2a

  15. Comprehensive analyses of the annexin gene family in wheat.

    Science.gov (United States)

    Xu, Lei; Tang, Yimiao; Gao, Shiqing; Su, Shichao; Hong, Lin; Wang, Weiwei; Fang, Zhaofeng; Li, Xueyin; Ma, Jinxiu; Quan, Wei; Sun, Hui; Li, Xia; Wang, Yongbo; Liao, Xiangzheng; Gao, Jiangang; Zhang, Fengting; Li, Lei; Zhao, Changping

    2016-05-28

    Annexins are an evolutionarily conserved multigene family of calcium-dependent phospholipid binding proteins that play important roles in stress resistance and plant development. They have been relatively well characterized in model plants Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa), but nothing has been reported in hexaploid bread wheat (Triticum aestivum) and barely (Hordeum vulgare), which are the two most economically important plants. Based on available genomic and transcriptomic data, 25 and 11 putative annexin genes were found through in silico analysis in wheat and barley, respectively. Additionally, eight and 11 annexin genes were identified from the draft genome sequences of Triticum urartu and Aegilops tauschii, progenitor for the A and D genome of wheat, respectively. By phylogenetic analysis, annexins in these four species together with other monocots and eudicots were classified into six different orthologous groups. Pi values of each of Ann1-12 genes among T. aestivum, T. urartu, A. tauschii and H. vulgare species was very low, with the exception of Ann2 and Ann5 genes. Ann2 gene has been under positive selection, but Ann6 and Ann7 have been under purifying selection among the four species in their evolutionary histories. The nucleotide diversities of Ann1-12 genes in the four species were 0.52065, 0.59239, 0.60691 and 0.53421, respectively. No selective pressure was operated on annexin genes in the same species. Gene expression patterns obtained by real-time PCR and re-analyzing the public microarray data revealed differential temporal and spatial regulation of annexin genes in wheat under different abiotic stress conditions such as salinity, drought, cold and abscisic acid. Among those genes, TaAnn10 is specifically expressed in the anther but fails to be induced by low temperature in thermosensitive genic male sterile lines, suggesting that specific down-regulation of TaAnn10 is associated with conditional male sterility in wheat

  16. Current Status and Prospects for Microbubbles in Ultrasound Theranostics

    Science.gov (United States)

    Martin, K. Heath

    2013-01-01

    Encapsulated microbubbles have been developed over the past two decades to provide both improvements in imaging as well as new therapeutic applications. Microbubble contrast agents are used currently for clinical imaging where increased sensitivity to blood flow is required, such as echocardiography. These compressible spheres oscillate in an acoustic field, producing nonlinear responses which can be uniquely distinguished from surrounding tissue, resulting in substantial enhancements in imaging signal-to-noise ratio. Furthermore, with sufficient acoustic energy the oscillation of microbubbles can mediate localized biological effects in tissue including the enhancement of membrane permeability or increased thermal energy deposition. Structurally, microbubbles are comprised of two principal components – an encapsulating shell and an inner gas core. This configuration enables microbubbles to be loaded with drugs or genes for additional therapeutic effect. Application of sufficient ultrasound energy can release this payload, resulting in site-specific delivery. Extensive pre-clinical studies illustrate that combining microbubbles and ultrasound can result in enhanced drug delivery or gene expression at spatially selective sites. Thus, microbbubles can be used for imaging, for therapy, or for both simultaneously. In this sense, microbubbles combined with acoustics may be one of the most universal theranostic tools. PMID:23504911

  17. Photoacoustic technique to measure temperature effects on microbubble viscoelastic properties

    Science.gov (United States)

    Lum, Jordan S.; Stobbe, David M.; Borden, Mark A.; Murray, Todd W.

    2018-03-01

    Phospholipid-coated microbubbles are being developed for several biomedical applications, but little is known about the effect of temperature on the viscoelastic properties of the shell. Here, we report on the use of a photoacoustic technique to study the shell properties of individual microbubbles as a function of temperature. The microbubbles were driven into small-amplitude oscillations by ultrasound waves generated from the absorption of an intensity-modulated infrared laser, and these oscillations were detected by forward-light scattering of a second blue laser. The drive laser modulation frequency was swept to determine the resonant response of 2-4 μm radius microbubbles. Lipid shell elasticity and viscosity were determined by modeling the microbubble response as a linear harmonic oscillator. The results from slow heating showed a linear decrease in elasticity and viscosity between 21 and 53 °C and a corresponding increase in the maximum oscillation amplitude. Rapid heating to 38 °C, on the other hand, showed a transient response in the viscoelastic properties, suggesting shell rupture and reformation during microbubble growth and subsequent dissolution. These effects are important for biomedical applications, which require warming of the microbubbles to body temperature.

  18. Ultrasound-accelerated thrombolysis using microbubbles

    Science.gov (United States)

    Culp, William

    2005-04-01

    Current thrombolytic therapy for ischemic stroke reaches less than 3% of cases and is only moderately successful. Numerous studies have demonstrated increased thrombolytic activity when ultrasound is delivered to clot in the presence of tissue plasminogen activator. Initial human reports using trans-cranial Doppler technology (2 MHz) to deliver continuous ultrasound to intracranial clot are very promising with improved clot lysis and no increase in symptomatic bleeding. However, another human study using low-frequency therapeutic ultrasound resulted in excessive bleeding and was discontinued. Microbubble augmented ultrasound clot lysis has proven successful in several studies ranging from lysis of very small peripheral clots in rabbits to very large clots in dialysis grafts in dogs. Early human studies of thrombosed dialysis grafts show success and no adverse events. Intracranial clot lysis in pigs has been successful with both intra-arterial and intravenous microbubble techniques. Better definition of possible combination therapies and of efficacy and safety is still required in animal models, but human studies should be designed to avoid some of the complications of current therapy. This presentation will review the subject, present recent advances, and define some requirements for successful clot destruction in peripheral vessels and intracranial vessels.

  19. Microbubble generation by piezotransducer for biological studies

    Science.gov (United States)

    Zhu, W.; Alkhazal, M.; Cho, M.; Xiao, S.

    2015-12-01

    Bubbles induced by blast waves or shocks are speculated to be the major cause of damages in biological cells in mild traumatic brain injuries. Microbubble collapse was found to induce noticeable cell detachment from the cell substrate, changes in focal adhesion and biomechanics. To better understand the bubble mechanism, we would like to construct a system, which allows us to clearly differentiate the impact of bubbles from that of shocks. Such a generator needs to be low profile in order to place under a microscope. A piezoelectric transducer system was designed to meet the need. The system uses either a flat or a spherical focusing piezoelectric transducer to produce microbubbles in a cuvette loaded with cell-culture medium. The transducer is placed on the side of the cuvette with its axis lining horizontally. A cover slip is placed on the top of the cuvette. The impact of the waves to the cells is minimized as the cover slip is parallel to the direction of the wave. Only bubbles from the medium reach the cover slip and interact with cells. The effect of bubbles therefore can be separated that of pressure waves. The bubbles collected on a cover slip range in size from 100 μm to 10 μm in radius, but the dominant size is 20-30 μm.

  20. Determination of autoantibodies to annexin XI in systemic autoimmune diseases

    DEFF Research Database (Denmark)

    Jorgensen, C S; Levantino, G; Houen, Gunnar

    2000-01-01

    of 282 sera from patients with systemic rheumatic diseases. The highest number of annexin XI positive sera were found in primary antiphospholipid syndrome (3/17), and in subacute lupus erythematosus (1/6), while lower frequencies of positive sera were found in patients with systemic sclerosis (5...

  1. Annexin II Dependent Mechanism of Breast Cancer Progression

    Science.gov (United States)

    2009-06-01

    microfilament architecture, affecting cellular physiology such as cell-cell interaction, migration and proliferation [37]. Targeted disruption of actin... microfilament assembly has been demonstrated in invasive (MDA-MB231) breast cancer cell death and morphological changes in cell shape [38]. Annexin II has

  2. Regulation of Annexin I in Rheumatoid Synovial Cells by Glucocorticoids and Interleukin-1

    Directory of Open Access Journals (Sweden)

    2006-01-01

    Full Text Available The glucocorticoid (GC-induced antiinflammatory molecule annexin I is expressed in leukocytes and has antiinflammatory effects in animal models of arthritis, but the expression of annexin I in rheumatoid arthritis (RA fibroblast-like synoviocytes (FLS is unknown. We report the constitutive and dexamethasone (DEX-inducible expression of annexin I in RA FLS. DEX increased FLS annexin I protein translocation and mRNA expression. Interleukin (IL-1 β also induced annexin I translocation and mRNA but also increased intracellular protein. DEX and IL-1 had additive effects on annexin I mRNA, but DEX inhibited the inducing effect of IL-1 β on cell surface annexin I. These results indicate that glucocorticoids and IL-1 β upregulate the synthesis and translocation of annexin I in RA FLS, but interdependent signalling pathways are involved.

  3. Theragnostic ultrasound using microbubbles in the treatment of prostate cancer

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hak Jong; Yoon, Young Il; Bae, Yun Jung [Dept. of Radiology, Seoul National University Bundang Hospital, Seongnam (Korea, Republic of)

    2016-08-15

    The use of gas-filled microbubbles in perfusion monitoring as intravascular ultrasound contrast agents has recently become more common. Additionally, microbubbles are employed as carriers of pharmaceutical substances or genes. Microbubbles have great potential to improve the delivery of therapeutic materials into cells and to modify vascular permeability, causing increased extravasation of drugs and drug carriers. Prostate cancer is the most common neoplasm in Europe and America, with an incidence twice to three times that of lung and colorectal cancer. Its incidence is still rising in Asian countries, including Japan and Korea. In this review, we present current strategies regarding the synthesis of microbubbles with targeted ligands on their surfaces, with a focus on prostate cancer.

  4. Real-Time Two-Dimensional Imaging of Microbubble Cavitation

    NARCIS (Netherlands)

    Vignon, F.; Shi, W.T.; Powers, J.E.; Liu, J.; Drvol, L.; Lof, J.; Everbach, C.; Gao, S.; Xie, F.; Porter, T.

    2011-01-01

    Ultrasound cavitation of microbubble contrast agents has a potentialfor therapeutic applications, including sonothrombolysis in acute ischemic stroke. For safety, efficacy, and reproducibility of treatment, it is critical to evaluate the cavitation state (e.g. stable versus inertial forms of

  5. Microbubble-assisted optofluidic control using a photothermal waveguide

    Science.gov (United States)

    Cheng, YuPeng; Yang, JianXin; Li, ZongBao; Zhu, DeBin; Cai, Xiang; Hu, Xiaowen; Huang, Wen; Xing, XiaoBo

    2017-10-01

    A convenient and easily controllable microfluidic system was proposed based on a photothermal device. Here, graphene oxide was assembled on an optical waveguide, which could serve as a miniature heat source to generate a microbubble and to control dynamic behaviors of flow by adjusting optical power at the micrometer scale. Micro/nanoparticles were used to demonstrate the trace of fluid flow around the microbubble, which displayed the ability of the flow to capture, transmit, and rotate particles in thermal convection. Correspondingly, three-dimensional theoretical simulation combining thermodynamics with hydrodynamics analyzed the distribution of the velocity field induced by the microbubble for collection and driving of particles. Furthermore, the photothermal waveguide would be developed into a microbubble-based device in the manipulation or transmission of micro/nanoparticles.

  6. Ultrasound microbubbles for molecular diagnosis, therapy, and theranostics

    NARCIS (Netherlands)

    Kiessling, F.; Fokong, S.; Koczera, P.; Lederle, W.; Lammers, Twan Gerardus Gertudis Maria

    2012-01-01

    Abstract Ultrasound imaging is clinically established for routine screening examinations of breast, abdomen, neck, and other soft tissues, as well as for therapy monitoring. Microbubbles as vascular contrast agents improve the detection and characterization of cancerous lesions, inflammatory

  7. {sup 18}F-labelled annexin V: a PET tracer for apoptosis imaging

    Energy Technology Data Exchange (ETDEWEB)

    Murakami, Yoshihiro; Tatsumi, Mitsuyoshi; Ichise, Rikiya; Nishimura, Shintaro [The Medical and Pharmacological Research Center Foundation, Wo32, 925-0613, Inoyama, Hakui, Ishikawa (Japan); Takamatsu, Hiroyuki; Noda, Akihiro [The Medical and Pharmacological Research Center Foundation, Wo32, 925-0613, Inoyama, Hakui, Ishikawa (Japan); Department of Biotracer Medicine, Kanazawa University Graduate School of Medical Science, 13-1 Takara-machi, 920-0934, Kanazawa, Ishikawa (Japan); Taki, Junichi [Department of Biotracer Medicine, Kanazawa University Graduate School of Medical Science, 13-1 Takara-machi, 920-0934, Kanazawa, Ishikawa (Japan); Tait, Jonathan F. [Department of Laboratory Medicine, Medicine (Medical Genetics) and Pathology, University of Washington, 98195-07110, Seattle, Washington (United States)

    2004-04-01

    Annexin V can be used to detect apoptotic cells in vitro and in vivo, based on its ability to identify extracellular phosphatidylserine, which arises during apoptosis. In the present study, we examined the synthesis of fluorine-18 labelled annexin V as a positron emission tomography tracer for apoptosis imaging. The distribution of [{sup 18}F]annexin V and technetium-99m labelled annexin V, a well-characterised SPET tracer for apoptosis imaging, was compared. [{sup 18}F]annexin V was synthesised using N-succinimidyl 4-[{sup 18}F]fluorobenzoate as an {sup 18}F labelling reagent. Synthesised and purified [{sup 18}F]annexin V was confirmed by SDS-PAGE. In an ex vivo imaging experiment, [{sup 18}F]annexin V was intravenously injected into rats 24 h after the induction of myocardial ischaemia, and accumulation in the left ventricle was examined. [{sup 18}F]annexin V accumulated in the infarct area of the left ventricle, where apoptotic cells were observed. In separate experiments, [{sup 18}F]annexin V or [{sup 99m}Tc]annexin V was intravenously injected into ischaemic or normal animals, and the distribution of the tracers was compared. In ischaemic animals, accumulation of [{sup 18}F]annexin V and [{sup 99m}Tc]annexin V in the infarct area was about threefold higher than in the non-infarct area. Furthermore, the ratio of accumulation in the normal heart to the blood radioactivity was not significantly different between the tracers. In normal animals, however, the uptake of [{sup 18}F]annexin V in the liver, spleen and kidney was much lower than that of [{sup 99m}Tc]annexin V. The low uptake of [{sup 18}F]annexin V in these organs might represent an advantage over [{sup 99m}Tc]annexin V. (orig.)

  8. Mass-transfer properties of microbubbles. 1. Experimental studies.

    Science.gov (United States)

    Bredwell, M D; Worden, R M

    1998-01-01

    Synthesis-gas fermentations have typically been gas-to-liquid mass-transfer-limited due to low solubilities of the gaseous substrates. A potential method to enhance mass-transfer rates is to sparge with microbubble dispersions. Mass-transfer coefficients for microbubble dispersions were measured in a bubble column. Oxygen microbubbles were formed in a dilute Tween 20 solution using a spinning disk apparatus. Axial dispersion coefficients measured for the bubble column ranged from 1.5 to 7.2 cm2/s and were essentially independent of flow rate. A laser-diffraction technique was used to determine the interfacial area per unit gas volume, a. The mass-transfer coefficient, KL, was determined by fitting a plug-flow model to the experimental, steady-state, liquid-phase oxygen-concentration profile. The KL values ranged from 2.9 x 10(-5) to 2.2 x 10(-4) m/s. Volumetric mass-transfer coefficients, KLa, for microbubbles with an average initial diameter of 60 microns ranged from 200 to 1800 h-1. Enhancement of mass transfer using microbubbles was demonstrated for a synthesis-gas fermentation. Butyribacterium methylotrophicum was grown in a continuous, stirred-tank reactor using a tangential filter for total cell recycle. The fermentation KLa values were 14 h-1 for conventional gas sparging through a stainless steel frit and 91 h-1 for microbubble sparging. The Power number of the microbubble generator was determined to be 0.036. Using this value, an incremental power-to-volume ratio to produce microbubbles for a B. methylotrophicum fermentation was estimated to be 0.01 kW/m3 of fermentation capacity.

  9. Effect of electrical potential of microbubbles on ozone dissolution

    Science.gov (United States)

    Hasegawa, H.; Kataoka, H.; Asano, K.

    2009-02-01

    Microbubbles make ozone water generation effective due to the high dissolution rate of gas in contrast to a conventional generating method. Therefore, it is presumable that ozone water generation using microbubbles can be achieved by the low concentration ozone gas. In our previous study, a compact and low power microbubble generator was developed. The microbubbles are generated by the local shear stress in the flow through a pipe with slits. In the present study, in order to investigate the relationship between the electrical potential of the gas-water interface and the cleaning of cloth using ozone microbubbles, two models with different slit angles (θ=30 and 60 deg) were installed. High concentration ozone water is produced for θ = 60 deg in contrast to the θ=30 deg case. When a cloth is washed in the θ=60 deg case, the soiled cloth can be cleaned easily in comparison with the θ=30 deg case, because the zeta potential of microbubbles for θ=60 deg is larger than that for θ=30 deg.

  10. Annexin A1 and A2: roles in retrograde trafficking of Shiga toxin.

    Science.gov (United States)

    Tcatchoff, Lionel; Andersson, Sofia; Utskarpen, Audrun; Klokk, Tove Irene; Skånland, Sigrid S; Pust, Sascha; Gerke, Volker; Sandvig, Kirsten

    2012-01-01

    Annexins constitute a family of calcium and membrane binding proteins. As annexin A1 and A2 have previously been linked to various membrane trafficking events, we initiated this study to investigate the role of these annexins in the uptake and intracellular transport of the bacterial Shiga toxin (Stx) and the plant toxin ricin. Once endocytosed, both toxins are retrogradely transported from endosomes to the Golgi apparatus and the endoplasmic reticulum before being targeted to the cytosol where they inhibit protein synthesis. This study was performed to obtain new information both about toxin transport and the function of annexin A1 and annexin A2. Our data show that depletion of annexin A1 or A2 alters the retrograde transport of Stx but not ricin, without affecting toxin binding or internalization. Knockdown of annexin A1 increases Golgi transport of Stx, whereas knockdown of annexin A2 slightly decreases the same transport step. Interestingly, annexin A1 was found in proximity to cytoplasmic phospholipase A2 (cPLA(2)), and the basal as well as the increased Golgi transport of Stx upon annexin A1 knockdown is dependent on cPLA(2) activity. In conclusion, annexin A1 and A2 have different roles in Stx transport to the trans-Golgi network. The most prominent role is played by annexin A1 which normally works as a negative regulator of retrograde transport from the endosomes to the Golgi network, most likely by complex formation and inhibition of cPLA(2).

  11. Dynamics of microbubble oscillators with delay coupling

    Science.gov (United States)

    Heckman, C. R.; Sah, S. M.; Rand, R. H.

    2010-10-01

    We investigate the stability of the in-phase mode in a system of two delay-coupled bubble oscillators. The bubble oscillator model is based on a 1956 paper by Keller and Kolodner. Delay coupling is due to the time it takes for a signal to travel from one bubble to another through the liquid medium that surrounds them. Using techniques from the theory of differential-delay equations as well as perturbation theory, we show that the equilibrium of the in-phase mode can be made unstable if the delay is long enough and if the coupling strength is large enough, resulting in a Hopf bifurcation. We then employ Lindstedt's method to compute the amplitude of the limit cycle as a function of the time delay. This work is motivated by medical applications involving noninvasive localized drug delivery via microbubbles.

  12. Design and Control of Functional Microbubbles for Medical Applications of Ultrasound

    Science.gov (United States)

    Takagi, Shu; Osaki, Taichi; Ariyoshi, Takuya; Azuma, Takashi; Ichiyanagi, Mitsuhisa; Kinefuchi, Ikuya

    2015-11-01

    Microbubbles are used as a contrast agent for ultrasound diagnosis. It is also expected to be use for the treatment. One of the possible applications is microbubble DDS. For that purpose, microbubbles need to be well-controlled for the generating process and manipulation. In this talk, for the design and control of the functional microbubbles, an experimental study on generation and surface modification of microbubbles are explained. Using a T-junction type microchannel, small bubbles about 5 μm size are successfully generated. For the surface modification, Biotin-coated microbubbles are tried to adhere the Avidin-coated wall. Furthermore, the manipulation of the microbubbles using ultrasound is also discussed. Plane-wave and focused ultrasound is used to manipulate a microbubble and bubble clusters. The experimental results are shown in the presentation. Supported by JSPS KAKENHI Grant Number 15K13865.

  13. Non-linear Response and Viscoelastic Properties of Lipid-Coated Microbubbles: DSPC versus DPPC

    NARCIS (Netherlands)

    van Rooij, T.; Luan, Y.; Renaud, G.; van der Steen, A.F.W.; Versluis, Michel; de Jong, N.; Kooiman, K.

    2015-01-01

    For successful in vivo contrast-enhanced ultrasound imaging (CEUS) and ultrasound molecular imaging, detailed knowledge of stability and acoustical properties of the microbubbles is essential. Here, we compare these aspects of lipid-coated microbubbles that have either

  14. Annexin II-Dependent Mechanism of Breast Cancer Progression

    Science.gov (United States)

    2010-11-01

    nitroaniline that absorbs light at 405 nm. The color absorbancewas measured by an automated 96-well plate reader (Bio-TeK Inc., VT). The colorimetric... autoantibodies and high circulating levels of IL-6 in lung cancer. Proc. Natl. Acad. Sci. U. S. A. 98, 9824–9829. Brownstein, C., et al., 2004. Annexin II...Gladu, J., 2002. Urokinase receptor antibody can reduce tumor volume and detect the presence of occult tumor metastases in vivo. Cancer Res. 62, 2390

  15. Beyond annexin V: fluorescence response of cellular membranes to apoptosis.

    Science.gov (United States)

    Demchenko, Alexander P

    2013-03-01

    Dramatic changes in the structure of cell membranes on apoptosis allow easy, sensitive and non-destructive analysis of this process with the application of fluorescence methods. The strong plasma membrane asymmetry is present in living cells, and its loss on apoptosis is commonly detected with the probes interacting strongly and specifically with phosphatidylserine (PS). This phospholipid becomes exposed to the cell surface, and the application of annexin V labeled with fluorescent dye is presently the most popular tool for its detection. Several methods have been suggested recently that offer important advantages over annexin V assay with the ability to study apoptosis by spectroscopy of cell suspensions, flow cytometry and confocal or two-photon microscopy. The PS exposure marks the integrated changes in the outer leaflet of cell membrane that involve electrostatic potential and hydration, and the attempts are being made to provide direct probing of these changes. This review describes the basic mechanisms underlying the loss of membrane asymmetry during apoptosis and discusses, in comparison with the annexin V-binding assay, the novel fluorescence techniques of detecting apoptosis on cellular membrane level. In more detail we describe the detection method based on smart fluorescent dye F2N12S incorporated into outer leaflet of cell membrane and reporting on apoptotic cell transformation by easily detectable change of the spectral distribution of fluorescent emission. It can be adapted to any assay format.

  16. A novel technology: microfluidic devices for microbubble ultrasound contrast agent generation.

    Science.gov (United States)

    Lin, Hangyu; Chen, Junfang; Chen, Chuanpin

    2016-09-01

    Microbubbles are used as ultrasound contrast agents, which enhance ultrasound imaging techniques. In addition, microbubbles currently show promise in disease therapeutics. Microfluidic devices have increased the ability to produce microbubbles with precise size, and high monodispersity compared to microbubbles created using traditional methods. This paper will review several variations in microfluidic device structures used to produce microbubbles as ultrasound contrast agents. Microfluidic device structures include T-junction, and axisymmetric and asymmetric flow-focusing. These devices have made it possible to produce microbubbles that can enter the vascular space; these microbubbles must be less than 10 μm in diameter and have high monodispersity. For different demands of microbubbles production rate, asymmetric flow-focusing devices were divided into individual and integrated devices. In addition, asymmetric flow-focusing devices can produce double layer and multilayer microbubbles loaded with drug or biological components. Details on the mechanisms of both bubble formation and device structures are provided. Finally, microfluidically produced microbubble acoustic responses, microbubble stability, and microbubble use in ultrasound imaging are discussed.

  17. Condensation phase diagrams for lipid-coated perfluorobutane microbubbles.

    Science.gov (United States)

    Mountford, Paul A; Sirsi, Shashank R; Borden, Mark A

    2014-06-03

    The goal of this study was to explore the thermodynamic conditions necessary to condense aqueous suspensions of lipid-coated gas-filled microbubbles into metastable liquid-filled nanodrops as well as the physicochemical mechanisms involved with this process. Individual perfluorobutane microbubbles and their lipid shells were observed as they were pressurized at 34.5 kPa s(-1) in a microscopic viewing chamber maintained at temperatures ranging from 5 to 75 °C. The microbubbles contracted under pressure, ultimately leading to either full dissolution or microbubble-to-nanodrop condensation. Temperature-pressure phase diagrams conveying condensation and stability transitions were constructed for microbubbles coated with saturated diacylphosphatidylcholine lipids of varying acyl chain length (C16 to C24). The onset of full dissolution was shifted to higher temperatures with the use of longer acyl chain lipids or supersaturated media. Longer chain lipid shells resisted both dissolution of the gas core and mechanical compression through a pronounced wrinkle-to-fold collapse transition. Interestingly, the lipid shell also provided a mechanical resistance to condensation, shifting the vapor-to-liquid transition to higher pressures than for bulk perfluorobutane. This result indicated that the lipid shell can provide a negative apparent surface tension under compression. Overall, the results of this study will aid in the design and formulation of vaporizable fluorocarbon nanodrops for various applications, such as diagnostic ultrasound imaging, targeted drug delivery, and thermal ablation.

  18. Generation of microbubbles from hollow cylindrical ultrasonic horn.

    Science.gov (United States)

    Makuta, Toshinori; Suzuki, Ryodai; Nakao, Takaaki

    2013-01-01

    In this study, we found that microbubbles with diameters of less than 100μm can be easily generated by using a hollow cylindrical ultrasonic horn. Consecutive images of bubbles obtained by using high-speed and high-resolution cameras reveal that a capillary wave is formed on the gas-liquid interface under weak ultrasonic irradiation and that the wave head is detached in the form of bubbles by the fragmentation of the interface as the power of ultrasonic irradiation increases. Moreover, consecutive images of the bubble interface obtained by an ultra-high-speed camera indicate that the breakup of bubbles oscillating harmonically with the ultrasonic irradiation generates many microbubbles that are less than 100μm in diameter. With regard to the orifice diameter of the horn end, we found that its optimum value varies with the ultrasonic power input. When the orifice diameter is small, the capillary wave generated from the horn end easily propagates all over the gas-liquid interface, thereby starting the generation of microbubbles at a lower ultrasonic power input. When the orifice diameter is large, the capillary wave is attenuated because of viscosity and surface tension. Hence, in this case, microbubble generation from the horn requires a higher ultrasonic power input. Furthermore, the maximum yield of microbubbles via primary and secondary bubble generation can be increased by increasing the gas flow rate. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Steering Microbubbles in Physiologically Realistic Flows Using the Bjerknes Force

    Science.gov (United States)

    Clark, Alicia; Aliseda, Alberto

    2017-11-01

    Ultrasound contrast agents (UCAs) are lipid-coated microbubbles that are used to increase contrast in ultrasound imaging due to their ability to scatter sound. Additionally, UCAs can be used in conjunction with ultrasound in medical applications such as targeted drug delivery and thrombolysis. These applications utilize the Bjerknes force, an ultrasound-induced force caused by the phase difference between the incoming ultrasound pressure wave and the microbubble volume oscillations. The dynamics of microbubbles under ultrasound excitation have been studied thoroughly in stagnant fluid baths; however, understanding of the fundamental physics of microbubbles in physiologically realistic flows is lacking. An in vitroexperiment that reproduces the dynamics (Reynolds and Womersley numbers) of a medium-sized blood vessel was used to explore the behavior of microbubbles. Using Lagrangian tracking, the trajectory of each individual bubble was reconstructed using information obtained from high speed imaging. The balance of hydrodynamic forces (lift, drag, added mass, etc.) against the primary Bjerknes force was analyzed. The results show that an increase in ultrasound pulse repetition frequency leads to a linear increase in the Bjerknes force and the increase in the force is quadratic with the amplitude of the excitation.

  20. Marangoni force-driven manipulation of photothermally-induced microbubbles.

    Science.gov (United States)

    Ortega-Mendoza, J G; Sarabia-Alonso, J A; Zaca-Morán, P; Padilla-Vivanco, A; Toxqui-Quitl, C; Rivas-Cambero, I; Ramirez-Ramirez, J; Torres-Hurtado, S A; Ramos-García, R

    2018-03-19

    The generation and manipulation of microbubbles by means of temperature gradients induced by low power laser radiation is presented. A laser beam (λ = 1064 nm) is divided into two equal parts and coupled to two multimode optical fibers. The opposite ends of each fiber are aligned and separated a distance D within an ethanol solution. Previously, silver nanoparticles were photo deposited on the optical fibers ends. Light absorption at the nanoparticles produces a thermal gradient capable of generating a microbubble at the optical fibers end in non-absorbent liquids. The theoretical and experimental studies carried out showed that by switching the thermal gradients, it is possible to generate forces in opposite directions, causing the migration of microbubbles from one fiber optic tip to another. Marangoni force induced by surface tension gradients in the bubble wall is the driving force behind the manipulation of microbubbles. We estimated a maximum Marangoni force of 400nN for a microbubble with a radius of 110 μm.

  1. Microbubble assisted polyhydroxybutyrate production in Escherichia coli.

    Science.gov (United States)

    Inan, Kadriye; Sal, Fulya Ay; Rahman, Asif; Putman, Ryan J; Agblevor, Foster A; Miller, Charles D

    2016-07-09

    One of the potential limitations of large scale aerobic Escherichia coli fermentation is the need for increased dissolved oxygen for culture growth and bioproduct generation. As culture density increases the poor solubility of oxygen in water becomes one of the limiting factors for cell growth and product formation. A potential solution is to use a microbubble dispersion (MBD) generating device to reduce the diameter and increase the surface area of sparged bubbles in the fermentor. In this study, a recombinant E. coli strain was used to produce polyhydroxybutyrate (PHB) under conventional and MBD aerobic fermentation conditions. In conventional fermentation operating at 350 rpm and 0.8 vvm air flow rate, an OD600 of 6.21 and PHB yield of 23 % (dry cell basis) was achieved. MBD fermentation with similar bioreactor operating parameters produced an OD600 of 8.17 and PHB yield of 43 % PHB, which was nearly double that of the conventional fermentation. This study demonstrated that using a MBD generator can increase oxygen mass transfer into the aqueous phase, increasing E. coli growth and bioproduct generation.

  2. Manipulation of Microbubble Clusters Using Focused Ultrasound

    Science.gov (United States)

    Matsuzaki, Hironobu; Osaki, Taichi; Kawaguchi, Kei; Unga, Johan; Ichiyanagi, Mitsuhisa; Azuma, Takashi; Suzuki, Ryo; Maruyama, Kazuo; Takagi, Shu

    2017-11-01

    In recent years, microbubbles (MBs) are expected to be utilized for the ultrasound drug delivery system (DDS). For the MB-DDS, it is important to establish a method of controlling bubbles and bubble clusters using ultrasound field. The objective of this study is to clarify behaviors of bubble clusters with various physical conditions. MBs in the ultrasound field are subjected to the primary Bjerknes force. The force traps MBs at the focal region of the focused ultrasound field. The trapped MBs form a bubble cluster at the region. A bubble cluster continues growing with absorbing surrounding bubbles until it reaches a maximum size beyond which it disappears from the focal region. In the present study, two kinds of MBs are used for the experiment. One is Sonazoid with average diameter of 2.6 um and resonant frequency of 5 MHz. The other is developed by Teikyo Univ., with average diameter of 1.5 um and presumed resonant frequency of 4 MHz. The bubble cluster's behaviors are analyzed using the high-speed camera. Sonazoid clusters have larger critical size than the other in every frequency, and its cluster size is inversely proportional to the ultrasound frequency, while Teikyo-bubble clusters have different tendency. These results are discussed in the presentation.

  3. Modeling Encapsulated Microbubble Dynamics at High Pressure Amplitudes

    Science.gov (United States)

    Heyse, Jan F.; Bose, Sanjeeb; Iaccarino, Gianluca

    2017-11-01

    Encapsulated microbubbles are commonly used in ultrasound contrast imaging and are of growing interest in therapeutic applications where local cavitation creates temporary perforations in cell membranes allowing for enhanced drug delivery. Clinically used microbubbles are encapsulated by a shell commonly consisting of protein, polymer, or phospholipid; the response of these bubbles to externally imposed ultrasound waves is sensitive to the compressibility of the encapsulating shell. Existing models approximate the shell compressibility via an effective surface tension (Marmottant et al. 2005). We present simulations of microbubbles subjected to high amplitude ultrasound waves (on the order of 106 Pa) and compare the results with the experimental measurements of Helfield et al. (2016). Analysis of critical points (corresponding to maximum and minimum expansion) in the governing Rayleigh-Plesset equation is used to make estimates of the parameters used to characterize the effective surface tension of the encapsulating shell. Stanford Graduate Fellowship.

  4. Robust microbubble tracking for super resolution imaging in ultrasound

    DEFF Research Database (Denmark)

    Hansen, Kristoffer B.; Villagómez Hoyos, Carlos Armando; Brasen, Jens Christian

    2016-01-01

    Currently ultrasound resolution is limited by diffraction to approximately half the wavelength of the sound wave employed. In recent years, super resolution imaging techniques have overcome the diffraction limit through the localization and tracking of a sparse set of microbubbles through...... the vasculature. However, this has only been performed on fixated tissue, limiting its clinical application. This paper proposes a technique for making super resolution images on non-fixated tissue by first compensating for tissue movement and then tracking the individual microbubbles. The experiment is performed...... on the kidney of a anesthetized Sprage-Dawley rat by infusing SonoVue at 0.1× original concentration. The algorithm demonstrated in vivo that the motion compensation was capable of removing the movement caused by the mechanical ventilator. The results shows that microbubbles were localized with a higher...

  5. Photothermal generation of microbubbles on plasmonic nanostructures inside microfluidic channels

    Science.gov (United States)

    Li, Jingting; Li, Ming; Santos, Greggy M.; Zhao, Fusheng; Shih, Wei-Chuan

    2016-03-01

    Microbubbles have been utilized as micro-pumps, micro-mixers, micro-valves, micro-robots and surface cleaners. Various generation techniques can be found in the literature, including resistive heating, hydrodynamic methods, illuminating patterned metal films and noble metal nanoparticles of Au or Ag. We present photothermal microbubble generation by irradiating nanoporous gold disk covered microfluidic channels. The size of the microbubble can be controlled by adjusting the laser power. The dynamics of both bubble growth and shrinkage are studied. The advantages of this technique are flexible bubble generation locations, long bubble lifetimes, no need for light-adsorbing dyes, high controllability over bubble size, low power consumption, etc. This technique has the potential to provide new flow control functions in microfluidic devices.

  6. Experimental microbubble generation by sudden pressure drop and fluidics

    Science.gov (United States)

    Franco Gutierrez, Fernando; Figueroa Espinoza, Bernardo; Aguilar Corona, Alicia; Vargas Correa, Jesus; Solorio Diaz, Gildardo

    2014-11-01

    Mass and heat transfer, as well as chemical species in bubbly flow are of importance in environmental and industrial applications. Microbubbles are well suited to these applications due to the large interface contact area and residence time. The objective of this investigation is to build devices to produce microbubbles using two methods: pressure differences and fluidics. Some characteristics, advantages and drawbacks of both methods are briefly discussed, as well as the characterization of the bubbly suspensions in terms of parameters such as the pressure jump and bubble equivalent diameter distribution. The authors acknowledge the support of Consejo Nacional de Ciencia y Tecnología.

  7. Annexin A4 and A6 induce membrane curvature and constriction during cell membrane repair

    DEFF Research Database (Denmark)

    Boye, Theresa Louise; Maeda, Kenji; Pezeshkian, Weria

    2017-01-01

    that annexin A4 binds to artificial membranes and generates curvature force initiated from free edges, whereas annexin A6 induces constriction force. In cells, plasma membrane injury and Ca2+ influx recruit annexin A4 to the vicinity of membrane wound edges where its homo-trimerization leads to membrane......Efficient cell membrane repair mechanisms are essential for maintaining membrane integrity and thus for cell life. Here we show that the Ca2+- and phospholipid-binding proteins annexin A4 and A6 are involved in plasma membrane repair and needed for rapid closure of micron-size holes. We demonstrate...... curvature near the edges. We propose that curvature force is utilized together with annexin A6-mediated constriction force to pull the wound edges together for eventual fusion. We show that annexin A4 can counteract various plasma membrane disruptions including holes of several micrometers indicating...

  8. Annexin A1 and A2: roles in retrograde trafficking of Shiga toxin.

    Directory of Open Access Journals (Sweden)

    Lionel Tcatchoff

    Full Text Available Annexins constitute a family of calcium and membrane binding proteins. As annexin A1 and A2 have previously been linked to various membrane trafficking events, we initiated this study to investigate the role of these annexins in the uptake and intracellular transport of the bacterial Shiga toxin (Stx and the plant toxin ricin. Once endocytosed, both toxins are retrogradely transported from endosomes to the Golgi apparatus and the endoplasmic reticulum before being targeted to the cytosol where they inhibit protein synthesis. This study was performed to obtain new information both about toxin transport and the function of annexin A1 and annexin A2. Our data show that depletion of annexin A1 or A2 alters the retrograde transport of Stx but not ricin, without affecting toxin binding or internalization. Knockdown of annexin A1 increases Golgi transport of Stx, whereas knockdown of annexin A2 slightly decreases the same transport step. Interestingly, annexin A1 was found in proximity to cytoplasmic phospholipase A2 (cPLA(2, and the basal as well as the increased Golgi transport of Stx upon annexin A1 knockdown is dependent on cPLA(2 activity. In conclusion, annexin A1 and A2 have different roles in Stx transport to the trans-Golgi network. The most prominent role is played by annexin A1 which normally works as a negative regulator of retrograde transport from the endosomes to the Golgi network, most likely by complex formation and inhibition of cPLA(2.

  9. Function, expression and localization of annexin A7 in platelets and red blood cells: Insights derived from an annexin A7 mutant mouse

    Directory of Open Access Journals (Sweden)

    Zamparelli Carlotta

    2003-08-01

    Full Text Available Abstract Background Annexin A7 is a Ca2+- and phospholipid-binding protein expressed as a 47 and 51 kDa isoform, which is thought to be involved in membrane fusion processes. Recently the 47 kDa isoform has been identified in erythrocytes where it was proposed to be a key component in the process of the Ca2+-dependent vesicle release, a process with which red blood cells might protect themselves against an attack by for example complement components. Results The role of annexin A7 in red blood cells was addressed in erythrocytes from anxA7-/- mice. Interestingly, the Ca2+-mediated vesiculation process was not impaired. Also, the membrane organization appeared not to be disturbed as assessed using gradient fractionation studies. Instead, lack of annexin A7 led to an altered cell shape and increased osmotic resistance of red blood cells. Annexin A7 was also identified in platelets. In these cells its loss led to a slightly slower aggregation velocity which seems to be compensated by an increased number of platelets. The results appear to rule out an important role of annexin A7 in membrane fusion processes occurring in red blood cells. Instead the protein might be involved in the organization of the membrane cytoskeleton. Red blood cells may represent an appropriate model to study the role of annexin A7 in cellular processes. Conclusion We have demonstrated the presence of both annexin A7 isoforms in red blood cells and the presence of the small isoform in platelets. In both cell types the loss of annexin A7 impairs cellular functions. The defects observed are however not compatible with a crucial role for annexin A7 in membrane fusion processes in these cell types.

  10. Annexin II-binding immunoglobulins in patients with lupus nephritis and their correlation with disease manifestations.

    Science.gov (United States)

    Cheung, Kwok Fan; Yung, Susan; Chau, Mel K M; Yap, Desmond Y H; Chan, Kwok Wah; Lee, Cheuk Kwong; Tang, Colin S O; Chan, Tak Mao

    2017-04-25

    Annexin II on mesangial cell surface mediates the binding of anti-dsDNA antibodies and consequent downstream inflammatory and fibrotic processes. We investigated the clinical relevance of circulating annexin II-binding immunoglobulins (Igs) in patients with severe proliferative lupus nephritis, and renal annexin II expression in relation to progression of nephritis in New Zealand Black and White F1 mice (NZBWF1/J) mice. Annexin II-binding Igs in serum were measured by ELISA. Ultrastructural localization of annexin II was determined by electron microscopy. Seropositivity rates for annexin II-binding IgG and IgM in patients with active lupus nephritis were significantly higher compared with controls (8.9%, 1.3% and 0.9% for annexin II-binding IgG and 11.1%, 4.0% and 1.9% for annexin II-binding IgM for patients with active lupus nephritis, patients with non-lupus renal disease and healthy subjects respectively). In lupus patients, annexin II-binding IgM level was higher at disease flare compared with remission. Annexin II-binding IgG and IgM levels were associated with that of anti-dsDNA and disease activity. Annexin II-binding IgG and IgM levels correlated with histological activity index in lupus nephritis biopsy samples. In NZBWF1/J mice, serum annexin II-binding IgG and IgM levels and glomerular annexin II and p11 expression increased with progression of active nephritis. Annexin II expression was present on mesangial cell surface and in the mesangial matrix, and co-localized with electron-dense deposits along the glomerular basement membrane. Our results show that circulating annexin II-binding IgG and IgM levels are associated with clinical and histological disease activity in proliferative lupus nephritis. The co-localization of annexin II and p11 expression with immune deposition in the kidney suggests pathogenic relevance. © 2017 The Author(s). published by Portland Press Limited on behalf of the Biochemical Society.

  11. Microbubble-mediated ultrasound drug-delivery and therapeutic monitoring.

    Science.gov (United States)

    Sennoga, Charles A; Kanbar, Emma; Auboire, Laurent; Dujardin, Paul-Armand; Fouan, Damien; Escoffre, Jean-Michel; Bouakaz, Ayache

    2017-09-01

    Recent developments in ultrasound imaging and ultrasound contrast agents (UCAs) improved diagnostic confidence in echography and set into motion their combined use as a tool for drug delivery and therapeutic monitoring. Non-invasive, precise and targeted delivery of drug molecules to pathological tissues by employing different mechanisms of drug release is becoming feasible. Areas covered: We sought to describe: the nature and features of UCAs; outline current contrast-specific imaging modes; before describing a variety of strategies for using ultrasound and microbubbles as a drug delivery system. Our expert opinion focusses on results and prospects of using ultrasound and microbubbles as a dual modality for drug delivery and therapeutic monitoring. Expert opinion: Today, ultrasound and microbubbles present a realistic prospect as drug delivery tools that have been demonstrated in a variety of animal models and clinical indications. Besides delivering drugs, ultrasound and microbubbles have demonstrated added value through therapeutic monitoring and assessment. Successful evaluation of the sonoporation mechanism(s), ultrasound parameters, drug type and dose will need to be addressed before translating this technology for clinic use. Ultimately, the development of a strategy for monitoring targeted delivery and its implementation in clinical practice would advance therapeutic treatment to a new qualitative level.

  12. Micromanipulation of endothelial cells: Ultrasound-microbubble-cell interaction

    NARCIS (Netherlands)

    van Wamel, Annemieke; Bouakaz, Ayache; Versluis, Michel; de Jong, N.

    2004-01-01

    Ultrasound (US) in combination with contrast microbubbles has been shown to alter the permeability of cell membranes without affecting cell viability. This permeabilisation feature is used to design new drug delivery systems using US and contrast agents. The underlying mechanisms are still unknown.

  13. Observation of spontaneous combustion of hydrogen and oxygen in microbubbles

    NARCIS (Netherlands)

    Postnikov, A.V.; Uvarov, I.V.; Prokaznikov, A.V.; Svetovoy, Vitaly

    2016-01-01

    Experimental evidence is presented that combustion can ignite at room temperature spontaneously inside microbubbles filled with mixture of hydrogen and oxygen. We perform water electrolysis in a closed microchamber by voltage pulses of alternating polarity at repetition frequencies 100 kHz to pump

  14. Biological in situ characterization of polymeric microbubble contrast agents

    NARCIS (Netherlands)

    Wan, Sha; Egri, Gabriella; Oddo, Letizia; Cerroni, Barbara; Dähne, Lars; Paradossi, Gaio; Salvati, Anna; Lynch, Iseult; Dawson, Kenneth A; Monopoli, Marco P

    Polymeric microbubbles (MBs) are gas filled particles composed of a thin stabilized polymer shell that have been recently developed as valid contrast agents for the combined use of ultrasonography (US), magnetic resonance imaging (MRI) and single photon emission computer tomography (SPECT) imaging.

  15. Acoustic behavior of microbubbles and implications for drug delivery

    NARCIS (Netherlands)

    Kooiman, K.; Vos, H.J.; Versluis, Michel; de Jong, N.

    2014-01-01

    Ultrasound contrast agents are valuable in diagnostic ultrasound imaging, and they increasingly show potential for drug delivery. This review focuses on the acoustic behavior of flexible-coated microbubbles and rigid-coated microcapsules and their contribution to enhanced drug delivery. Phenomena

  16. TOPICAL REVIEW: Ultrasound contrast microbubbles in imaging and therapy: physical principles and engineering

    Science.gov (United States)

    Qin, Shengping; Caskey, Charles F.; Ferrara, Katherine W.

    2009-03-01

    Microbubble contrast agents and the associated imaging systems have developed over the past 25 years, originating with manually-agitated fluids introduced for intra-coronary injection. Over this period, stabilizing shells and low diffusivity gas materials have been incorporated in microbubbles, extending stability in vitro and in vivo. Simultaneously, the interaction of these small gas bubbles with ultrasonic waves has been extensively studied, resulting in models for oscillation and increasingly sophisticated imaging strategies. Early studies recognized that echoes from microbubbles contained frequencies that are multiples of the microbubble resonance frequency. Although individual microbubble contrast agents cannot be resolved—given that their diameter is on the order of microns—nonlinear echoes from these agents are used to map regions of perfused tissue and to estimate the local microvascular flow rate. Such strategies overcome a fundamental limitation of previous ultrasound blood flow strategies; the previous Doppler-based strategies are insensitive to capillary flow. Further, the insonation of resonant bubbles results in interesting physical phenomena that have been widely studied for use in drug and gene delivery. Ultrasound pressure can enhance gas diffusion, rapidly fragment the agent into a set of smaller bubbles or displace the microbubble to a blood vessel wall. Insonation of a microbubble can also produce liquid jets and local shear stress that alter biological membranes and facilitate transport. In this review, we focus on the physical aspects of these agents, exploring microbubble imaging modes, models for microbubble oscillation and the interaction of the microbubble with the endothelium.

  17. Differential Modulation of Annexin I Binding Sites on Monocytes and Neutrophils

    Directory of Open Access Journals (Sweden)

    H. S. Euzger

    1999-01-01

    Full Text Available Specific binding sites for the anti-inflammatory protein annexin I have been detected on the surface of human monocytes and polymorphonuclear leukocytes (PMN. These binding sites are proteinaceous in nature and are sensitive to cleavage by the proteolytic enzymes trypsin, collagenase, elastase and cathepsin G. When monocytes and PMN were isolated independently from peripheral blood, only the monocytes exhibited constitutive annexin I binding. However PMN acquired the capacity to bind annexin I following co-culture with monocytes. PMN incubation with sodium azide, but not protease inhibitors, partially blocked this process. A similar increase in annexin I binding capacity was also detected in PMN following adhesion to endothelial monolayers. We propose that a juxtacrine activation rather than a cleavage-mediated transfer is involved in this process. Removal of annexin I binding sites from monocytes with elastase rendered monocytes functionally insensitive to full length annexin I or to the annexin I-derived pharmacophore, peptide Ac2-26, assessed as suppression of the respiratory burst. These data indicate that the annexin I binding site on phagocytic cells may have an important function in the feedback control of the inflammatory response and their loss through cleavage could potentiate such responses.

  18. Sonodynamically-induced cytotoxicity by rose bengal derivative and microbubbles in isolated sarcoma 180 cells

    Science.gov (United States)

    Sugita, Nami; Hosokawa, Mami; Sunaga, Naoki; Iwase, Yumiko; Yumita, Nagahiko; Ikeda, Toshihiko; Umemura, Shin-ichiro

    2015-07-01

    It is known that the combination of ultrasound and sonodynamic sensitizer (SDS) is effective in noninvasive tumor treatment, referred to as sonodynamic therapy (SDT). Microbubbles have been used in ultrasound therapy as well. The purpose of this paper is to clarify the effect of microbubbles on SDT. Sarcoma 180 cells were suspended in air-saturated phosphate-buffered saline and exposed to ultrasound with the SDS rose bengal derivative (RBD) in standing wave mode in the presence and absence of microbubbles [sonazoid (SZ)]. The ultrasonically induced cytotoxicity with RBD and SZ was about 20 times higher than without either, and about 80% of the SZ microbubbles were destructed by ultrasonic exposure in as short as five seconds. Since microbubbles induce significant cytotoxicity even with short duration, low intensity ultrasound, the application of microbubbles in SDT shows promise in anti-tumor treatment.

  19. Ultrasound modulated optical tomography contrast enhancement with non-linear oscillation of microbubbles.

    Science.gov (United States)

    Ruan, Haowen; Mather, Melissa L; Morgan, Stephen P

    2015-02-01

    Ultrasound modulated optical tomography (USMOT) is an imaging technique used to provide optical functional information inside highly scattering biological tissue. One of the challenges facing this technique is the low image contrast. A contrast enhancement imaging technique based on the non-linear oscillation of microbubbles is demonstrated to improve image contrast. The ultrasound modulated signal was detected using a laser pulse based speckle contrast detection system. Better understanding of the effects of microbubbles on the optical signals was achieved through simultaneous measurement of the ultrasound scattered by the microbubbles. The length of the laser pulse was found to affect the system response of the speckle contrast method with shorter pulses suppressing the fundamental ultrasound modulated optical signal. Using this property, image contrast can be enhanced by detection of the higher harmonic ultrasound modulated optical signals due to nonlinear oscillation and destruction of the microbubbles. Experimental investigations were carried out to demonstrate a doubling in contrast by imaging a scattering phantom containing an embedded silicone tube with microbubbles flowing through it. The contrast enhancement in USMOT resulting from the use of ultrasound microbubbles has been demonstrated. Destruction of the microbubbles was shown to be the dominant effect leading to contrast improvement as shown by simultaneously detecting the ultrasound and speckle contrast signals. Line scans of a microbubble filled silicone tube embedded in a scattering phantom demonstrated experimentally the significant image contrast improvement that can be achieved using microbubbles and demonstrates the potential as a future clinical imaging tool.

  20. Annexin A1 is elevated in patients with COPD and affects lung fibroblast function

    Directory of Open Access Journals (Sweden)

    Lai TW

    2018-02-01

    Full Text Available Tianwen Lai,1,* Yanyu Li,1,* Zongjiong Mai,2 Xiaoxia Wen,1 Yingying Lv,1 Zhanqing Xie,3 Quanchao Lv,1 Min Chen,1 Dong Wu,1 Bin Wu1 1Department of Respiratory and Critical Care Medicine, 2Department of Oncology, 3Department of Thoracic Surgery, The Affiliated Hospital of Guangdong Medical University, Zhanjiang, People’s Republic of China *These authors contributed equally to this work Purpose: Fibrosis in peripheral airways is responsible for airflow limitation in chronic obstructive pulmonary disease (COPD. Annexin A1 modulates several key biological events during inflammation. However, little is known about its role in airway fibrosis in COPD. We investigated whether levels of Annexin A1 were upregulated in patients with COPD, and whether it promoted airway fibrosis.Methods: We quantified serum Annexin A1 levels in never-smokers (n=12, smokers without COPD (n=11, and smokers with COPD (n=22. Correlations between Annexin A1 expression and clinical indicators (eg, lung function were assessed. In vitro, human bronchial epithelial (HBE cells were exposed to cigarette smoke extract (CSE and Annexin A1 expression was assessed. Primary human lung fibroblasts were isolated from patients with COPD and effects of Annexin A1 on fibrotic deposition of lung fibroblasts were evaluated.Results: Serum Annexin A1 was significantly higher in patients with Global Initiative for Chronic Obstructive Lung Disease (GOLD guidelines stage III or IV than in those with GOLD stages I or II (12.8±0.8 ng/mL versus 9.8±0.7 ng/mL; p=0.016. Annexin A1 expression was negatively associated with airflow obstruction (forced expiratory volume in one second % predicted; r=−0.72, p<0.001. In vitro, Annexin A1 was significantly increased in CSE-exposed HBE cells in a time- and concentration-dependent manner. Annexin A1 promoted lung fibroblasts proliferation, migration, differentiation, and collagen deposition via the ERK1/2 and p38 mitogen-activated protein kinase pathways

  1. Micro-Bubble Experiments at the Van de Graaff Accelerator

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Z. J. [Argonne National Lab. (ANL), Argonne, IL (United States); Wardle, Kent E. [Argonne National Lab. (ANL), Argonne, IL (United States); Quigley, K. J. [Argonne National Lab. (ANL), Argonne, IL (United States); Gromov, Roman [Argonne National Lab. (ANL), Argonne, IL (United States); Youker, A. J. [Argonne National Lab. (ANL), Argonne, IL (United States); Makarashvili, Vakhtang [Argonne National Lab. (ANL), Argonne, IL (United States); Bailey, James [Argonne National Lab. (ANL), Argonne, IL (United States); Stepinski, D. C. [Argonne National Lab. (ANL), Argonne, IL (United States); Chemerisov, S. D. [Argonne National Lab. (ANL), Argonne, IL (United States); Vandegrift, G. F. [Argonne National Lab. (ANL), Argonne, IL (United States)

    2015-02-01

    In order to test and verify the experimental designs at the linear accelerator (LINAC), several micro-scale bubble ("micro-bubble") experiments were conducted with the 3-MeV Van de Graaff (VDG) electron accelerator. The experimental setups included a square quartz tube, sodium bisulfate solution with different concentrations, cooling coils, gas chromatography (GC) system, raster magnets, and two high-resolution cameras that were controlled by a LabVIEW program. Different beam currents were applied in the VDG irradiation. Bubble generation (radiolysis), thermal expansion, thermal convection, and radiation damage were observed in the experiments. Photographs, videos, and gas formation (O2 + H2) data were collected. The micro-bubble experiments at VDG indicate that the design of the full-scale bubble experiments at the LINAC is reasonable.

  2. In-plant testing of microbubble column flotation. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, R.H.; Luttrell, G.H.; Adel, G.T.; Mankosa, M.J.

    1991-07-31

    Microbubble column flotation (MCF) was developed at the Virginia Center for Coal and Minerals Processing (VCCMP) for the selective recovery of fine particles. Bench-scale test work conducted at VCCMP, largely under the sponsorship of the U.S. Department of Energy (DOE), showed that the technology worked well for both coal and mineral applications. For the technology to be commercially successful, however, a full-scale demonstration of the MCF technology was deemed necessary. This report summarizes the results of work performed under the DOE project entitled ``In-plant Testing of Microbubble Column Flotation.`` The objectives of this research and development effort were to duplicate the bench-scale performance of the MCF process in a full-scale unit, to verify the scale-up procedure developed in an earlier project, and to demonstrate the applicability of the MCF technology to the coal industry.

  3. In-plant testing of microbubble column flotation

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, R.H.; Luttrell, G.H.; Adel, G.T.; Mankosa, M.J.

    1991-07-31

    Microbubble column flotation (MCF) was developed at the Virginia Center for Coal and Minerals Processing (VCCMP) for the selective recovery of fine particles. Bench-scale test work conducted at VCCMP, largely under the sponsorship of the U.S. Department of Energy (DOE), showed that the technology worked well for both coal and mineral applications. For the technology to be commercially successful, however, a full-scale demonstration of the MCF technology was deemed necessary. This report summarizes the results of work performed under the DOE project entitled In-plant Testing of Microbubble Column Flotation.'' The objectives of this research and development effort were to duplicate the bench-scale performance of the MCF process in a full-scale unit, to verify the scale-up procedure developed in an earlier project, and to demonstrate the applicability of the MCF technology to the coal industry.

  4. A co-flow-focusing monodisperse microbubble generator

    KAUST Repository

    Zhang, Jiaming

    2014-02-14

    We use a simple and inexpensive microfluidic device, which is based on microscope glass slides and two tapered glass capillaries, to produce monodisperse microbubbles. The innermost capillary used for transporting the gas is inserted into the second capillary, with its 2 μm sharp tip aligned with the center of the converging-diverging throat of the second capillary. This configuration provides a small and smooth gas flow rate, and a high velocity gradient at the tube outlet. Highly monodisperse microbubbles with diameters ranging from 3.5 to 60 microns have been successfully produced at a rate of up to 40 kHz. A simple scaling law, which is based on the capillary number and liquid-to-gas flow rate ratio, successfully predicts the bubble size. © 2014 IOP Publishing Ltd.

  5. Microbubble generator excited by fluidic oscillator's third harmonic frequency

    Czech Academy of Sciences Publication Activity Database

    Tesař, Václav

    2014-01-01

    Roč. 92, č. 9 (2014), s. 1603-1615 ISSN 0263-8762 R&D Projects: GA ČR GA13-23046S Institutional support: RVO:61388998 Keywords : fluidic oscillator * microbubble generation * fluidic feedback loop Subject RIV: BK - Fluid Dynamics Impact factor: 2.348, year: 2014 http://dx.doi.org/10.1016/j.cherd.2013.12.004

  6. Microbubbles as drug delivery systems in cerebrovascular diseases.

    Science.gov (United States)

    Spinelli, Mariacarmela; Demitri, Christian; Sannino, Alessandro; Peruzzotti-Jametti, Luca; Bacigaluppi, Marco; Comi, Giancarlo; Corea, Francesco

    2009-11-01

    The field of neurovascular ultrasound is growing rapidly with new applications. While ultrasound contrast agents were initially used to overcome poor transcranial bone windows for identification of cerebral arteries, newgeneration microbubbles in combination with innovative contrast-specific ultrasound techniques now enable potential therapeutic procedures. This article will provide a review of recent and emerging developments along with patents in ultrasound technology and contrast-specific therapeutic techniques for cerebrovascular patients.

  7. Functional comparison of annexin V analogues labeled indirectly and directly with iodine-124

    Energy Technology Data Exchange (ETDEWEB)

    Dekker, Bronwen [CRUK/UMIST Dept. of Radiochemical Targeting and Imaging, Paterson Inst. for Cancer Research, M20 4BX Manchester (United Kingdom)]|[Dept. of Instrumentation and Analytical Science, Univ. of Manchester Inst. for Science and Technology (United Kingdom)]. E-mail: bdekker@picr.man.ac.uk; Keen, Heather [CRUK/UMIST Dept. of Radiochemical Targeting and Imaging, Paterson Inst. for Cancer Research, M20 4BX Manchester (United Kingdom)]|[Dept. of Instrumentation and Analytical Science, Univ. of Manchester Inst. for Science and Technology, M60 1QD Manchester (United Kingdom); Shaw, David [CRUK/UMIST Dept. of Radiochemical Targeting and Imaging, Paterson Inst. for Cancer Research, M20 4BX Manchester (United Kingdom)]|[CRUK Dept. of Immunology, Paterson Inst. for Cancer Research, M20 4BX Manchester (United Kingdom); Disley, Lynn [CRUK/UMIST Dept. of Radiochemical Targeting and Imaging, Paterson Inst. for Cancer Research, M20 4BX Manchester (United Kingdom); Hastings, David; Julyan, Peter [CRUK/UMIST Dept. of Radiochemical Targeting and Imaging, Paterson Inst. for Cancer Research, M20 4BX Manchester (United Kingdom)]|[North Western Medical Physics, Christie Hospital NHS Trust, M20 4BX Manchester (United Kingdom); Hadfield, John [School of Environment and Life Sciences, Univ. of Salford, M5 4WT Manchester (United Kingdom); Reader, Andrew [Dept. of Instrumentation and Analytical Science, Univ. of Manchester Inst. for Science and Technology, M60 1QD Manchester (United Kingdom); Allan, Donald [Physics and Electronic Unit, Paterson Inst. for Cancer Research, M20 4BX Manchester (United Kingdom); Watson, Alastair [Dept. of Medicine, Univ of Liverpool (United Kingdom); Zweit, Jamal [CRUK/UMIST Dept. of Radiochemical Targeting and Imaging, Paterson Inst. for Cancer Research, M20 4BX Manchester (United Kingdom)]|[Dept. of Instrumentation and Analytical Science, Univ. of Manchester Inst. for Science and Technology (United Kingdom)]. E-mail: jzweit@picr.man.ac.uk

    2005-05-01

    We are interested in imaging cell death in vivo using annexin V radiolabeled with {sup 124}I. In this study, [{sup 124}I]4IB-annexin V and [{sup 124}I]4IB-ovalbumin were made using [{sup 124}I]N-hydroxysuccinimidyl-4-iodobenzoate prepared by iododestannylation of N-hydroxysuccinimidyl-4-(tributylstannyl)benzoate. [{sup 124}I]4IB-annexin V binds to phosphatidylserine-coated microtiter plates and apoptotic Jurkat cells and accumulates in hepatic apoptotic lesions in mice treated with anti-Fas antibody, while [{sup 124}I]4IB-ovalbumin does not. In comparison with {sup 124}I-annexin V, [{sup 124}I]4IB-annexin V has a higher rate of binding to phosphatidylserine in vitro, a higher kidney and urine uptake, a lower thyroid and stomach content uptake, greater plasma stability and a lower rate of plasma clearance. Binding of radioactivity to apoptotic cells relative to normal cells in vitro and in vivo appears to be lower for [{sup 124}I]4IB-annexin V than for {sup 124}I-annexin V.

  8. Enhancement of focused jets by using surface microbubbles

    Science.gov (United States)

    Yukisada, Ryosuke; Kiyama, Akihito; Zhang, Xuehua; Tagawa, Yoshiyuki

    2017-11-01

    Focused liquid jets are important for various key technologies, such as material deposition and automated pipetting. It has been challenging to create high speed jets of viscous liquids. Our latest work showed that it is possible to generate viscous jets by applying sudden acceleration to the liquid (Onuki et al., J. J. Multi. Flow, 2015). It was observed that under certain conditions cavitation bubbles form in the liquid, making important contribution to the increment of jet velocity (Kiyama et al., JFM, 2016). The increased velocity depends on the maximum size of expanding bubbles. Thus, for controlling the velocity of focused jets, it is crucial to control the bubble expansion. In this study, we investigate the effects of surface microbubbles on the focused jets. Before the impact is performed, the microbubbles are produced on an inner wall of the liquid container by using water-ethanol exchange technique. We experimentally measure the jet velocity and bubble motion utilizing a high-speed camera. It is found that surface microbubbles expand upon the impact, enhancing the increment of jet velocity under the conditions that do not trigger cavitation inception in the bulk liquid. JSPS KAKENHI Grant Numbers 26709007 and 17H01246.

  9. Microbubbles-Assisted Ultrasound Triggers the Release of Extracellular Vesicles

    Directory of Open Access Journals (Sweden)

    Yuana Yuana

    2017-07-01

    Full Text Available Microbubbles-assisted ultrasound (USMB has shown promise in improving local drug delivery. The formation of transient membrane pores and endocytosis are reported to be enhanced by USMB, and they contribute to cellular drug uptake. Exocytosis also seems to be linked to endocytosis upon USMB treatment. Based on this rationale, we investigated whether USMB triggers exocytosis resulting in the release of extracellular vesicles (EVs. USMB was performed on a monolayer of head-and-neck cancer cells (FaDu with clinically approved microbubbles and commonly used ultrasound parameters. At 2, 4, and 24 h, cells and EV-containing conditioned media from USMB and control conditions (untreated cells, cells treated with microbubbles and ultrasound only were harvested. EVs were measured using flow cytometric immuno-magnetic bead capture assay, immunogold electron microscopy, and western blotting. After USMB, levels of CD9 exposing-EVs significantly increased at 2 and 4 h, whereas levels of CD63 exposing-EVs increased at 2 h. At 24 h, EV levels were comparable to control levels. EVs released after USMB displayed a heterogeneous size distribution profile (30–1200 nm. Typical EV markers CD9, CD63, and alix were enriched in EVs released from USMB-treated FaDu cells. In conclusion, USMB treatment triggers exocytosis leading to the release of EVs from FaDu cells.

  10. Cavitation microstreaming and stress fields created by microbubbles.

    Science.gov (United States)

    Collis, James; Manasseh, Richard; Liovic, Petar; Tho, Paul; Ooi, Andrew; Petkovic-Duran, Karolina; Zhu, Yonggang

    2010-02-01

    Cavitation microstreaming plays a role in the therapeutic action of microbubbles driven by ultrasound, such as the sonoporative and sonothrombolytic phenomena. Microscopic particle-image velocimetry experiments are presented. Results show that many different microstreaming patterns are possible around a microbubble when it is on a surface, albeit for microbubbles much larger than used in clinical practice. Each pattern is associated with a particular oscillation mode of the bubble, and changing between patterns is achieved by changing the sound frequency. Each microstreaming pattern also generates different shear stress and stretch/compression distributions in the vicinity of a bubble on a wall. Analysis of the micro-PIV results also shows that ultrasound-driven microstreaming flows around bubbles are feasible mechanisms for mixing therapeutic agents into the surrounding blood, as well as assisting sonoporative delivery of molecules across cell membranes. Patterns show significant variations around the bubble, suggesting sonoporation may be either enhanced or inhibited in different zones across a cellular surface. Thus, alternating the patterns may result in improved sonoporation and sonothrombolysis. The clear and reproducible delineation of microstreaming patterns based on driving frequency makes frequency-based pattern alternation a feasible alternative to the clinically less desirable practice of increasing sound pressure for equivalent sonoporative or sonothrombolytic effect. Surface divergence is proposed as a measure relevant to sonoporation.

  11. Annexin A1 in primary tumors promotes melanoma dissemination.

    Science.gov (United States)

    Boudhraa, Zied; Rondepierre, Fabien; Ouchchane, Lemlih; Kintossou, Roselyne; Trzeciakiewicz, Anna; Franck, Frederic; Kanitakis, Jean; Labeille, Bruno; Joubert-Zakeyh, Juliette; Bouchon, Bernadette; Perrot, Jean Luc; Mansard, Sandrine; Papon, Janine; Dechelotte, Pierre; Chezal, Jean-Michel; Miot-Noirault, Elisabeth; Bonnet, Mathilde; D'Incan, Michel; Degoul, Françoise

    2014-10-01

    Metastatic melanoma is one of the most aggressive forms of skin cancer and has a poor prognosis. We have previously identified Annexin A1 (ANXA1) as a potential murine melanoma-spreading factor that may modulate cell invasion by binding to formyl peptide receptors (FPRs). Here, we report that (1) in a B16Bl6 spontaneous metastasis model, a siRNA-induced decrease in tumoral ANXA1 expression significantly reduced tumoral MMP2 activity and number of lung metastases; (2) in a retrospective study of 61 patients, metastasis-free survival was inversely related to ANXA1 expression levels in primary tumors (HR 3.15 [1.03-9.69], p = 0.045); (3) in human melanoma cell lines, ANXA1 level was positively correlated with in vitro invasion capacity whereas normal melanocytes contained low ANXA1 levels, and (4) the ANXA1 N-terminal peptide ANXA12-26 stimulated MMP2 activity after interaction with FPRs and significantly stimulated the in vitro invasion of melanomas by acting on FPRs. These findings identify ANXA1 as a proinvasive protein in melanoma that holds promise as a potential prognostic marker and therapeutic target.

  12. Factors that affect the efficiency of antisense oligodeoxyribonucleotide transfection by insonated gas-filled lipid microbubbles

    International Nuclear Information System (INIS)

    Zhao Yingzheng; Lu Cuitao

    2008-01-01

    Objective: To investigate the factors that affect the efficiency of antisense oligodeoxyribonucleotide(AS-ODNs) transfection by insonated gas-filled lipid microbubbles. Methods: Lipid microbubbles filled with two types of gases-air and C 3 F 8 , were prepared respectively. An AS-ODNs sequence HA824 and a breast cancer cell line SK-BR-3 were used to define the various operating variables determining the transfection efficiency of insonated microbubbles. Two mixing methods, three levels of mixing speed, different mixing durations and various ultrasound initiation time after mixing were examined respectively. Transfection efficiency was detected by fluorescence microscopy. Results: C 3 F 8 microbubbles gave higher levels of AS-ODNs transfection efficiency than air microbubbles in all test conditions. Transfection efficiency resulted from mixing method A (incubation of HA824 and microbubbles before mixing cells) did not show significant difference with that of mixing method B (without incubation of HA824 and microbubbles before mixing cells). Mixing speed, duration of mixing and ultrasound initiation time after mixing were central to determining HA824 transfection efficiency in vitro. The optimum parameters for SK-BR-3 cells were found at a mixing speed of 40-50 rpm for 30-60 s with less than 60 s delay before ultrasound. Conclusion: Ultrasound-mediated AS-ODNs transfection enhanced by C 3 F 8 -filled lipid microbubbles represents an effective avenue for AS-ODNs transfer

  13. Annexin A1 influences in breast cancer: Controversies on contributions to tumour, host and immunoediting processes.

    Science.gov (United States)

    Tu, Yan; Johnstone, Cameron N; Stewart, Alastair G

    2017-05-01

    Annexin A1 is a multifunctional protein characterised by its actions in modulating the innate and adaptive immune response. Accumulating evidence of altered annexin A1 expression in many human tumours raises interest in its functional role in cancer biology. In breast cancer, altered annexin A1 expression levels suggest a potential influence on tumorigenic and metastatic processes. However, reports of conflicting results reveal a relationship that is much more complex than first conceptualised. In this review, we explore the diverse actions of annexin A1 on breast tumour cells and various host cell types, including stromal immune and structural cells, particularly in the context of cancer immunoediting. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Oil recovery in the presence of microbubbles in the filtration flow

    Science.gov (United States)

    Mikhailov, D. N.

    2012-05-01

    This paper presents mathematical models for oil-gas flow taking into account the various processes due to the formation of gas micronuclei (microbubbles) in oil: slip of oil relative to the walls of the pore channels (gas lubrication), changes in oil viscosity, and motion of microbubles with respect to oil. We consider examples of oil flow in the near-wellbore zone for the case where a reduction in pressure to the saturation pressure leads to the formation of gas microbubbles and micronuclei and examples of the action of a water-gas mixture in the case where oil foams in the contact area with the injected gas, i.e., a finely dispersed mixture of oil and microbubbles is formed. The behavior of indicator curves for an oil well with the formation of microbubbles is simulated, and the effect of microbubbles on the oil recovery factor in a water-alternating-gas injection process is studied.

  15. Magnetic targeting to enhance microbubble delivery in an occluded microarterial bifurcation.

    Science.gov (United States)

    de Saint Victor, M; Carugo, D; Barnsley, L C; Owen, J; Coussios, C-C; Stride, E

    2017-09-05

    Ultrasound and microbubbles have been shown to accelerate the breakdown of blood clots both in vitro and in vivo. Clinical translation of this technology is still limited, however, in part by inefficient microbubble delivery to the thrombus. This study examines the obstacles to delivery posed by fluid dynamic conditions in occluded vasculature and investigates whether magnetic targeting can improve microbubble delivery. A 2D computational fluid dynamic model of a fully occluded Y-shaped microarterial bifurcation was developed to determine: (i) the fluid dynamic field in the vessel with inlet velocities from 1-100 mm s -1 (corresponding to Reynolds numbers 0.25-25); (ii) the transport dynamics of fibrinolytic drugs; and (iii) the flow behavior of microbubbles with diameters in the clinically-relevant range (0.6-5 µm). In vitro experiments were carried out in a custom-built microfluidic device. The flow field was characterized using tracer particles, and fibrinolytic drug transport was assessed using fluorescence microscopy. Lipid-shelled magnetic microbubbles were fluorescently labelled to determine their spatial distribution within the microvascular model. In both the simulations and experiments, the formation of laminar vortices and an abrupt reduction of fluid velocity were observed in the occluded branch of the bifurcation, severely limiting drug transport towards the occlusion. In the absence of a magnetic field, no microbubbles reached the occlusion, remaining trapped in the first vortex, within 350 µm from the bifurcation center. The number of microbubbles trapped within the vortex decreased as the inlet velocity increased, but was independent of microbubble size. Application of a magnetic field (magnetic flux density of 76 mT, magnetic flux density gradient of 10.90 T m -1 at the centre of the bifurcation) enabled delivery of microbubbles to the occlusion and the number of microbubbles delivered increased with bubble size and with decreasing inlet

  16. Prevalence and clinical significance of pleural microbubbles in computed tomography of thoracic empyema

    International Nuclear Information System (INIS)

    Smolikov, A.; Smolyakov, R.; Riesenberg, K.; Schlaeffer, F.; Borer, A.; Cherniavsky, E.; Gavriel, A.; Gilad, J.

    2006-01-01

    AIM: To determine the prevalence and clinical significance of pleural microbubbles in thoracic empyema. MATERIALS AND METHODS: The charts of 71 consecutive patients with empyema were retrospectively reviewed for relevant demographic, laboratory, microbiological, therapeutic and outcome data. Computed tomography (CT) images were reviewed for various signs of empyema as well as pleural microbubbles. Two patient groups, with and without microbubbles were compared. RESULTS: Mean patient age was 49 years and 72% were males. Microbubbles were detected in 58% of patients. There were no significant differences between patients with and without microbubbles in regard to pleural fluid chemistry. A causative organism was identified in about 75% of cases in both. There was no difference in the rates of pleural thickening and enhancement, increased extra-pleural fat attenuation, air-fluid levels or loculations. Microbubbles were diagnosed after a mean of 7.8 days from admission. Thoracentesis before CT was performed in 90 and 57% of patients with and without microbubbles (p=0.0015), respectively. Patients with microbubbles were more likely to require repeated drainage (65.9 versus 36.7%, p=0.015) and surgical decortication (31.7 versus 6.7%, p=0.011). Mortalities were 9.8 and 6.6% respectively (p=0.53). CONCLUSION: Pleural microbubbles are commonly encountered in CT imaging of empyema but have not been systematically studied to date. Microbubbles may be associated with adverse outcome such as repeated drainage or surgical decortication. The sensitivity and specificity of this finding and its prognostic implications need further assessment

  17. Microbubble enhanced ozonation process for advanced treatment of wastewater produced in acrylic fiber manufacturing industry

    KAUST Repository

    Zheng, Tianlong

    2015-02-02

    This work investigated microbubble-ozonation for the treatment of a refractory wet-spun acrylic fiber wastewater in comparison to macrobubble-ozonation. CODcr, NH3-N, and UV254 of the wastewater were removed by 42%, 21%, and 42%, respectively in the microbubble-ozonation, being 25%, 9%, and 35% higher than the removal rates achieved by macrobubble-ozonation at the same ozone dose. The microbubbles (with average diameter of 45μm) had a high concentration of 3.9×105 counts/mL at a gas flow rate of 0.5L/min. The gas holdup, total ozone mass-transfer coefficient, and average ozone utilization efficiency in the microbubble-ozonation were 6.6, 2.2, and 1.5 times higher than those of the macrobubble-ozonation. Greater generation of hydroxyl radicals and a higher zeta potential of the bubbles were also observed in the microbubble ozonation process. The biodegradability of the wastewater was also significantly improved by microbubble-ozonation, which was ascribed to the enhanced degradation of alkanes, aromatic compounds, and the many other bio-refractory organic compounds in the wastewater. Microbubble-ozonation can thus be a more effective treatment process than traditional macrobubble-ozonation for refractory wastewater produced by the acrylic fiber manufacturing industry.

  18. Sonothrombolysis with BR38 Microbubbles Improves Microvascular Patency in a Rat Model of Stroke.

    Directory of Open Access Journals (Sweden)

    Nadine Schleicher

    Full Text Available Early recanalization of large cerebral vessels in ischemic stroke is associated with improved clinical outcome, however persisting hypoperfusion leads to poor clinical recovery despite large vessel recanalization. Limited experimental sonothrombolysis studies have shown that addition of microbubbles during treatment can improve microvascular patency. We aimed to determine the effect of two different microbubble formulations on microvascular patency in a rat stroke model.We tested BR38 and SonoVue® microbubble-enhanced sonothrombolysis in Wistar rats submitted to 90-minute filament occlusion of the middle cerebral artery. Rats were randomized to treatment (n = 6/group: control, rt-PA, or rt-PA+3-MHz ultrasound insonation with BR38 or SonoVue® at full or 1/3 dose. Treatment duration was 60 minutes, beginning after withdrawal of the filament, and sacrifice was immediately after treatment. Vascular volumes were evaluated with microcomputed tomography.Total vascular volume of the ipsilateral hemisphere was reduced in control and rt-PA groups (p0.1.Microbubble-enhanced sonothrombolysis improves microvascular patency. This effect is not dose- or microbubble formulation-dependent suggesting a class effect of microbubbles promoting microvascular reopening. This study demonstrates that microbubble-enhanced sonothrombolysis may be a therapeutic strategy for patients with persistent hypoperfusion of the ischemic territory.

  19. Comparison of microbubble presence in the right heart during mechanochemical and radiofrequency ablation for varicose veins.

    Science.gov (United States)

    Moon, K H; Dharmarajah, B; Bootun, R; Lim, C S; Lane, Tra; Moore, H M; Sritharan, K; Davies, A H

    2017-07-01

    Objective Mechanochemical ablation is a novel technique for ablation of varicose veins utilising a rotating catheter and liquid sclerosant. Mechanochemical ablation and radiofrequency ablation have no reported neurological side-effect but the rotating mechanism of mechanochemical ablation may produce microbubbles. Air emboli have been implicated as a cause of cerebrovascular events during ultrasound-guided foam sclerotherapy and microbubbles in the heart during ultrasound-guided foam sclerotherapy have been demonstrated. This study investigated the presence of microbubbles in the right heart during varicose vein ablation by mechanochemical abaltion and radiofrequency abaltion. Methods Patients undergoing great saphenous vein ablation by mechanochemical abaltion or radiofrequency ablation were recruited. During the ablative procedure, the presence of microbubbles was assessed using transthoracic echocardiogram. Offline blinded image quantification was performed using International Consensus Criteria grading guidelines. Results From 32 recruited patients, 28 data sets were analysed. Eleven underwent mechanochemical abaltion and 17 underwent radiofrequency abaltion. There were no neurological complications. In total, 39% (11/28) of patients had grade 1 or 2 microbubbles detected. Thirty-six percent (4/11) of mechanochemical abaltion patients and 29% (5/17) of radiofrequency ablation patients had microbubbles with no significant difference between the groups ( p=0.8065). Conclusion A comparable prevalence of microbubbles between mechanochemical abaltion and radiofrequency ablation both of which are lower than that previously reported for ultrasound-guided foam sclerotherapy suggests that mechanochemical abaltion may not confer the same risk of neurological events as ultrasound-guided foam sclerotherapy for treatment of varicose veins.

  20. Advanced treatment of acrylic fiber manufacturing wastewater with a combined microbubble-ozonation/ultraviolet irradiation process

    KAUST Repository

    Zheng, Tianlong

    2015-01-01

    This work investigated the effectiveness of a combination of microbubble-ozonation and ultraviolet (UV) irradiation for the treatment of secondary wastewater effluent of a wet-spun acrylic fiber manufacturing plant. Under reactor condition (ozone dosage of 48 mg L-1, UV fluence rate of 90 mW cm-2, initial pH of 8.0, and reaction time of 120 min), the biodegradability (represented as BOD5/CODcr) of the wastewater improved from 0.18 to 0.47. This improvement in biodegradability is related to the degradation of alkanes, aromatic compounds, and other bio-refractory organic compounds. The combination of microbubble-ozonation and UV irradiation synergistically improved treatment efficiencies by 228%, 29%, and 142% for CODcr, UV254 removal and BOD5/CODcr respectively after 120 min reaction time, as compared with the sum efficiency of microbubble-ozonation alone and UV irradiation alone. Hydroxyl radical production in the microbubble-ozonation/UV process was about 1.8 times higher than the sum production in microbubble-ozonation alone and UV irradiation alone. The ozone decomposition rate in the combined process was about 4.1 times higher than that in microbubble-ozonation alone. The microbubble-ozonation/UV process could be a promising technique for the treatment of bio-refractory organics in the acrylic fiber manufacturing industry. © 2015 Royal Society of Chemistry.

  1. On the relationship between microbubble fragmentation, deflation and broadband superharmonic signal production.

    Science.gov (United States)

    Lindsey, Brooks D; Rojas, Juan D; Dayton, Paul A

    2015-06-01

    Acoustic angiography imaging of microbubble contrast agents uses the superharmonic energy produced from excited microbubbles and enables high-contrast, high-resolution imaging. However, the exact mechanism by which broadband harmonic energy is produced is not fully understood. To elucidate the role of microbubble shell fragmentation in superharmonic signal production, simultaneous optical and acoustic measurements were performed on individual microbubbles at transmit frequencies from 1.75 to 3.75 MHz and pressures near the shell fragmentation threshold for microbubbles of varying diameter. High-amplitude, broadband superharmonic signals were produced with shell fragmentation, whereas weaker signals (approximately 25% of peak amplitude) were observed in the presence of shrinking bubbles. Furthermore, when populations of stationary microbubbles were imaged with a dual-frequency ultrasound imaging system, a sharper decline in image intensity with respect to frame number was observed for 1-μm bubbles than for 4-μm bubbles. Finally, in a study of two rodents, increasing frame rate from 4 to 7 Hz resulted in decreases in mean steady-state image intensity of 27% at 1000 kPa and 29% at 1300 kPa. Although the existence of superharmonic signals when bubbles shrink has the potential to prolong the imaging efficacy of microbubbles, parameters such as frame rate and peak pressure must be balanced with expected re-perfusion rate to maintain adequate contrast during in vivo imaging. Copyright © 2015. Published by Elsevier Inc.

  2. Elevation of plasma membrane permeability upon laser irradiation of extracellular microbubbles.

    Science.gov (United States)

    Zhou, Yu; Zhou, Xi-Yuan; Wang, Zhi-Gang; Zhu, Ye-Feng; Li, Pan

    2010-07-01

    Laser-mediated gene transfection has received much attention as a new method for targeted gene therapy because of the high controllability of laser energy and direction. In this report, we describe a combination laser-microbubble system that enables membrane-impermeable molecules to penetrate cell membranes. The main theories we apply are optical breakdown and photoacoustic generation, which are induced by laser irradiation. Firstly, different types of laser light (Ar-green, Novus Varia poly-wavelength and Nd:YAG laser) were adopted to blast liposome microbubble contrast medium; subsequently, the Nd:YAG laser (1064 nm, 4 ns), which could successfully blast microbubbles, and ultrasound were used in combination to irradiate a mixture of liposome microbubbles and retinoblastoma (Rb) cells. After irradiation, membrane permeability was evaluated by flow cytometric assay using propidium iodide (PI) and fluorescein diacetate (FDA). The proportion of permeabilized resealed cells was affected by changes in the light energy. All of the Nd:YAG laser, Nd:YAG combination laser-microbubble and combination ultrasound-microbubble systems were able to permeabilize the Rb cells. These results suggest that this combination laser-microbubble system is a new means of delivering exogenous materials into living cells.

  3. Degradation of methyl orange using short-wavelength UV irradiation with oxygen microbubbles

    Energy Technology Data Exchange (ETDEWEB)

    Tasaki, Tsutomu; Wada, Tsubasa; Fujimoto, Kanji [JST Innovation Satellite Miyazaki, 1-1 Gakuen Kibanadai Nishi, Miyazaki 889-2192 (Japan); Kai, Shinji; Ohe, Kaoru; Oshima, Tatsuya; Baba, Yoshinari [Department of Applied Chemistry, Faculty of Engineering, University of Miyazaki, 1-1 Gakuen Kibanadai Nishi, Miyazaki 889-2192 (Japan); Kukizaki, Masato [Department of Material Development, Miyazaki Prefecture Industrial Technology Center, 16500-2 Higashi Kaminaka, Sadowara, Miyazaki 880-0303 (Japan)], E-mail: kukizaki@iri.pref.miyazaki.jp

    2009-03-15

    A novel wastewater treatment technique using 8 W low-pressure mercury lamps in the presence of uniform-sized microbubbles (diameter = 5.79 {mu}m) was investigated for the decomposition of methyl orange as a model compound in aqueous solution. Photodegradation experiments were conducted with a BLB black light blue lamp (365 nm), a UV-C germicidal lamp (254 nm) and an ozone lamp (185 nm + 254 nm) both with and without oxygen microbubbles. The results show that the oxygen microbubbles accelerated the decolorization rate of methyl orange under 185 + 254 nm irradiation. In contrast, the microbubbles under 365 and 254 nm irradiation were unaffected on the decolorization of methyl orange. It was found that the pseudo-zero order decolorization reaction constant in microbubble system is 2.1 times higher than that in conventional large bubble system. Total organic carbon (TOC) reduction rate of methyl orange was greatly enhanced by oxygen microbubble under 185 + 254 nm irradiation, however, TOC reduction rate by nitrogen microbubble was much slower than that with 185 + 254 nm irradiation only. Possible reaction mechanisms for the decolorization and mineralization of methyl orange both with oxygen and nitrogen mirobubbles were proposed in this study.

  4. Degradation of methyl orange using short-wavelength UV irradiation with oxygen microbubbles.

    Science.gov (United States)

    Tasaki, Tsutomu; Wada, Tsubasa; Fujimoto, Kanji; Kai, Shinji; Ohe, Kaoru; Oshima, Tatsuya; Baba, Yoshinari; Kukizaki, Masato

    2009-03-15

    A novel wastewater treatment technique using 8 W low-pressure mercury lamps in the presence of uniform-sized microbubbles (diameter = 5.79 microm) was investigated for the decomposition of methyl orange as a model compound in aqueous solution. Photodegradation experiments were conducted with a BLB black light blue lamp (365 nm), a UV-C germicidal lamp (254 nm) and an ozone lamp (185 nm+254 nm) both with and without oxygen microbubbles. The results show that the oxygen microbubbles accelerated the decolorization rate of methyl orange under 185+254 nm irradiation. In contrast, the microbubbles under 365 and 254 nm irradiation were unaffected on the decolorization of methyl orange. It was found that the pseudo-zero order decolorization reaction constant in microbubble system is 2.1 times higher than that in conventional large bubble system. Total organic carbon (TOC) reduction rate of methyl orange was greatly enhanced by oxygen microbubble under 185+254 nm irradiation, however, TOC reduction rate by nitrogen microbubble was much slower than that with 185+254 nm irradiation only. Possible reaction mechanisms for the decolorization and mineralization of methyl orange both with oxygen and nitrogen mirobubbles were proposed in this study.

  5. Study optical properties of biological tissue in the presence of microbubbles

    Science.gov (United States)

    Assadi, Homa; Lee, Vincent; Karshafian, Raffi; Douplik, Alexandre

    2015-03-01

    Optical contrast agents introduce distinct features to induce detectable changes in native tissue properties [1]. In ultrasound imaging, microbubbles (MBs) - a gas-core shell-encapsulated agent - are used clinically as contrast agents. The working hypothesis of this study is that microbubbles can be employed as an intravascular contrast agent in optical imaging systems. Microbubbles can produce a refractive index mismatch which makes it distinguishable from surrounding media. In this work, the interaction of collimated light and microbubbles in a [1] biological phantom solution was investigated. The biological medium was comprised of intralipid and human blood which was constructed to cover the range of soft tissue optical properties. The effect of microbubbles on the optical properties such as reduced scattering and absorption coefficients were considered. Diffuse reflectance (DR) and total transmittance (TT) of a biological phantom solution were measured using a spectroscopic integrating sphere system in the absence and presence of Definity® microbubbles. The optical properties were computed using the inverse adding doubling (IAD) software. The presence of microbubbles increased DR and decreased TT of the phantom. In the presence of MB's (2.5% volume concentration), the reflectance of the phantom increased by 25% in the optical window. There is no absorption event and only scattering happened after light-microbubbles interactions. The reduced scattering coefficient increased significantly (30%) indicating the potential use of MBs as optical contrast agents. In conclusion, reflectance of a media can be enhanced by adding microbubbles to increase scattering properties and more light was detected returning to the surface of tissue.

  6. Enhanced intracellular delivery of a model drug using microbubbles produced by a microfluidic device.

    Science.gov (United States)

    Dixon, Adam J; Dhanaliwala, Ali H; Chen, Johnny L; Hossack, John A

    2013-07-01

    Focal drug delivery to a vessel wall facilitated by intravascular ultrasound and microbubbles holds promise as a potential therapy for atherosclerosis. Conventional methods of microbubble administration result in rapid clearance from the bloodstream and significant drug loss. To address these limitations, we evaluated whether drug delivery could be achieved with transiently stable microbubbles produced in real time and in close proximity to the therapeutic site. Rat aortic smooth muscle cells were placed in a flow chamber designed to simulate physiological flow conditions. A flow-focusing microfluidic device produced 8 μm diameter monodisperse microbubbles within the flow chamber, and ultrasound was applied to enhance uptake of a surrogate drug (calcein). Acoustic pressures up to 300 kPa and flow rates up to 18 mL/s were investigated. Microbubbles generated by the flow-focusing microfluidic device were stabilized with a polyethylene glycol-40 stearate shell and had either a perfluorobutane (PFB) or nitrogen gas core. The gas core composition affected stability, with PFB and nitrogen microbubbles exhibiting half-lives of 40.7 and 18.2 s, respectively. Calcein uptake was observed at lower acoustic pressures with nitrogen microbubbles (100 kPa) than with PFB microbubbles (200 kPa) (p 3). In addition, delivery was observed at all flow rates, with maximal delivery (>70% of cells) occurring at a flow rate of 9 mL/s. These results demonstrate the potential of transiently stable microbubbles produced in real time and in close proximity to the intended therapeutic site for enhancing localized drug delivery. Copyright © 2013 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  7. Bioinspired preparation of alginate nanoparticles using microbubble bursting.

    Science.gov (United States)

    Elsayed, Mohamed; Huang, Jie; Edirisinghe, Mohan

    2015-01-01

    Nanoparticles are considered to be one of the most advanced tools for drug delivery applications. In this research, alginate (a model hydrophilic polymer) nanoparticles 80 to 200 nm in diameter were obtained using microbubble bursting. The natural process of bubble bursting occurs through a number of stages, which consequently produce nano- and microsized droplets via two main production mechanisms, bubble shell disintegration and a jetting process. In this study, nano-sized droplets/particles were obtained by promoting the disintegrating mechanism and suppressing (limiting) the formation of larger microparticles resulting from the jetting mechanism. A T-junction microfluidic device was used to prepare alginate microbubbles with different sizes in a well-controlled manner. The size of the bubbles was varied by controlling two processing parameters, the solution flow rate and the bubbling pressure. Crucially, the bubble size was found to be the determining factor for inducing (or limiting) the bubble shell disintegration mechanism and the size needed to promote this process was influenced by the properties of the solution used for preparing the bubbles, particularly the viscosity. The size of alginate nanoparticles produced via the disintegration mechanism was found to be directly proportional to the viscosity of the alginate solution. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Removal of dimethyl phthalate from water by ozone microbubbles.

    Science.gov (United States)

    Jabesa, Abdisa; Ghosh, Pallab

    2017-08-01

    This work investigates the removal of dimethyl phthalate (DMP) from water using ozone microbubbles in a pilot plant of 20 dm 3 capacity. Experiments were performed under various reaction conditions to examine the effects of the initial concentration of DMP, pH of the medium, ozone generation rate, and the role of H 2 O 2 on the removal of DMP. The DMP present in water was effectively removed by the ozone microbubbles. The removal was effective in neutral and alkaline media. Increase in the initial concentration of the target pollutant negatively affected its removal efficiency. The removal efficiency dramatically increased from 1% to 99% when the ozone generation rate was increased from 0.28 to 1.94 mg s -1 at pH 7. The total organic carbon measurements revealed that a complete mineralization of DMP was achieved within 1.8 ks at the high ozone feed rate. The use of t-butyl alcohol as the hydroxyl radical scavenger confirmed that the reaction between the target organic compound and ·OH radical dominated over its direct reaction with ozone. The reaction between DMP and ozone followed an overall second-order kinetics. The volumetric mass transfer coefficient of ozone in the reacting system and the enhancement factor increased with increasing initial concentration of DMP. Very low values of Hatta number were obtained at all initial concentrations of DMP and pH, which show that the mass transfer resistance was small.

  9. Mass production of monodisperse microbubbles for real applications avoiding microfluidics

    Science.gov (United States)

    Sanchez Quintero, Enrique Jesus; Evangelio, Alvaro; Gordillo, Jose Manuel

    2017-11-01

    In this presentation we report experiments showing the effect on the controlled generation of microbubbles of the pressure gradient imposed by the relative flow of a liquid stream around an airfoil-shaped solid. Taking advantage of the conclusions in, where the local pressure gradient was identified as the mechanism responsible of the generation of microbubbles in microfluidic devices and, with the purpose of overcoming the low production rates associated with these kind of microdevices, we have used the same physical principle but have applied it to a totally different geometry: a rectangular planar wing composed by symmetrical airfoils. The relative velocity field is imposed either submerging the static wing within a flowing hydraulic channel or by rotating the wings within a reservoir containing the otherwise quiescent liquid mass. We provide physical insight on the bubbling process and deduce a scaling law which expresses the diameters of the bubbles formed as a function of the gas flow rate, relative liquid velocity and the angle of attack of the incident flow. In spite of the geometry is totally different, we recover the same results obtained using microfluidic devices but with much higher production rates.

  10. Velocity field measurement in micro-bubble emission boiling

    International Nuclear Information System (INIS)

    Ito, Daisuke; Saito, Yasushi; Natazuka, Jun

    2017-01-01

    Liquid inlet behavior to a heat surface in micro-bubble emission boiling (MEB) was investigated by flow measurement using particle image velocimetry (PIV). Subcooled pool boiling experiments under atmospheric pressure were carried out using a heat surface with a diameter of 10 mm. An upper end of a heater block made of copper was used as the heat surface. Working fluid was the deionized water and the subcooling was varied from 40 K to 70 K. Three K-type thermocouples were installed in the copper block to measure the temperature gradient, and the heat flux and wall superheat were estimated from these temperature data to make a boiling curve. The flow visualization around the heat surface was carried out using a high-speed video camera and a light sheet. The microbubbles generated in the MEB were used as tracer particles and the velocity field was obtained by PIV analysis of the acquired image sequence. As a result, the higher heat fluxes than the critical heat flux could be obtained in the MEB region. In addition, the distribution characteristics of the velocity in MEB region were studied using the PIV results and the location of the stagnation point in the velocity fields was discussed. (author)

  11. AM05-24-002 THE MICRO-BUBBLE GENERATION DUE TO THE VORTEX BREAKDOWN

    OpenAIRE

    山田, 麗徳; 坂入, 信之; 金子, 公久; 京藤, 敏達; 筑波大学システム情報工学研究科; 筑波大学システム情報工学研究科; 筑波大学システム情報工学研究科; 筑波大学システム情報工学研究科

    2005-01-01

    Micro-bubble generation due to a vortex breakdown of high-speed gas-liquid two-phase swirling flow was investigated in the present study. The control of the position and type of the vortex breakdown is crucial for the generation of the micro-bubbles. It greatly depends on the nozzle shape and the air discharge supplied to the nozzle. The air discharge under which micro-bubbles are efficiently generated depends on the pressure in the air vortex core. Numerical analysis was performed to study t...

  12. Characterization of Annexin V Fusion with the Superfolder GFP in Liposomes Binding and Apoptosis Detection.

    Science.gov (United States)

    Abbady, Abdul Qader; Twair, Aya; Ali, Bouthaina; Murad, Hossam

    2017-01-01

    Programed cell death is a critical and unavoidable part of life. One of the most widely used markers for dying cells, by apoptosis or pyroptosis, is the redistribution of phosphatidylserine (PS) from the inner to the outer plasma membrane leaflet. Annexin V protein is a sensitive and specific probe to mark this event because of its high affinity to the exposed PS. Beyond that, annexin V can bind to any PS-containing phospholipid bilayer of almost all tiny forms of membranous vesicles like blood platelets, exosomes, or even nanostructured liposomes. In this work, recombinant human annexin V was produced as a fusion with a highly fluorescent superfolder derivative of the green fluorescent protein ( sf GFP) in Escherichia coli . The fusion protein( sf GFP-ANXV, 64 kDa), annexin V (ANXV, 40 kDa), and sf GFP (27 kDa) were separately produced after cloning their encoding genes in pRSET plasmid, and all proteins were expressed in a soluble form, then purified in high yields because of their N -terminal 6× His tag (~150 mg of pure protein per 1 L culture). Superiority of this fluorescent fusion protein over fluorescein-conjugated annexin V was demonstrated in binding to phospholipids (and their liposomes), prepared from natural sources (soya bean and egg yolk) that have different content of PS, by using different methods including ELISA, dot-blotting, surface plasmon resonance, and flow cytometry. We also applied fluorescent annexin V in the detection of apoptotic cells by flow cytometry and fluorescent microscopy. Interestingly, sf GFP-ANXV fusion was more sensitive to early apoptotic stressed HeLa cells than fluorescein-conjugated-ANXV. This highly expressed and functional sf GFP-ANXV fusion protein provides a promising ready-to-use molecular tool for quantifying liposomes (or similarly exosomes) and detecting apoptosis in cells.

  13. [Progress of ultrasound microbubble contrast technology in the diagnosis and treatment of clinical diseases].

    Science.gov (United States)

    Qiu, Li; Leng, Qian-ying; Luo, Yan

    2014-11-01

    Contrast enhanced ultrasound (CEUS) technology has made important developments over the past decade. It has been applied in the clinical diagnosis for various diseases of multiple organs including liver, kidney, thyroid, breast and so on, which greatly improves the accuracy of ultrasound diagnosis. The emergence of targeted ultrasound microbubble makes ultrasound molecular imaging possible. More than the improvement of ultrasound imaging, microbubble contrast agent also could be an effective drug or gene carrier. Microbubble will rupture under the irradiation effect of local ultrasound, and then the carried drugs or genes will be released to achieve the purpose of targeted therapy. We should pay more attention on the progress of ultrasound microbubble contrast technology in clinical and basic research. It will promote better understanding and clinical applications of this novel medical ultrasound technology.

  14. Assessments of Bubble Dynamics Model and Influential Parameters in Microbubble Drag Reduction

    National Research Council Canada - National Science Library

    Skudarnov, P. V; Lin, C. X

    2006-01-01

    .... The effects of mixture density variation, free stream turbulence intensity, free stream velocity, and surface roughness on the microbubble drag reduction were studied using a single phase model based...

  15. Stabilization and fabrication of microbubbles: applications for medical purposes and functional materials.

    Science.gov (United States)

    Lee, Mina; Lee, Eun Yeol; Lee, Daeyeon; Park, Bum Jun

    2015-03-21

    Microbubbles with diameters ranging from a few micrometers to tens of micrometers have garnered significant attention in various applications including food processing, water treatment, enhanced oil recovery, surface cleaning, medical purposes, and material preparation fields with versatile functionalities. A variety of techniques have been developed to prepare microbubbles, such as ultrasonication, excimer laser ablation, high shear emulsification, membrane emulsification, an inkjet printing method, electrohydrodynamic atomization, template layer-by-layer deposition, and microfluidics. Generated bubbles should be immediately stabilized via the adsorption of stabilizing materials (e.g., surfactants, lipids, proteins, and solid particles) onto the gas-liquid interface to lower the interfacial tension. Such adsorption of stabilizers prevents coalescence between the microbubbles and also suppresses gas dissolution and resulting disproportionation caused by the presence of the Laplace overpressure across the gas-liquid interface. Herein, we comprehensively review three important topics of microbubbles: stabilization, fabrication, and applications.

  16. Dynamic Fluorescence Microscopy of Cellular Uptake of Intercalating Model Drugs by Ultrasound-Activated Microbubbles

    NARCIS (Netherlands)

    Lammertink, B.H.A.; Deckers, R.; Derieppe, M.; De Cock, I.; Lentacker, I.; Storm, G.; Moonen, C. T.W.; Bos, C.

    2017-01-01

    Purpose: The combination of ultrasound and microbubbles can facilitate cellular uptake of (model) drugs via transient permeabilization of the cell membrane. By using fluorescent molecules, this process can be studied conveniently with confocal fluorescence microscopy. This study aimed to investigate

  17. Bevacizumab (Avastin) conjugated microbubbles for anti-VEGF treatment of neovascular age-related macular degeneration

    Science.gov (United States)

    Zhang, Leilei; Xu, Jeff; Huang, Jiwei; Roberts, Cynthia; Xu, Ronald

    2010-02-01

    Bevacizumab (Avastin) has been used as one of the anti-VEGF therapies to manage neovascular age-related macular degeneration (AMD). The drug delivery system for bevacizumab needs to be improved in order to decrease the frequency of injection and reduce the adverse effects. In our study, bevacizumab was conjugated with poly (lactic-co-glycolic acid) (PLGA) microbubbles by activating carboxyl functional groups. The averaged size of microbubbles was estimated 1.055+/-0.258μm, allowing for ultrasound guided drug delivery. The binding efficiency between bevacizumab and microbubbles was evaluated in an enzyme-linked immunosorbent assay plate. The test results demonstrated the potential of using PLGA microbubbles to deliver bevacizumab with imaging guidance.

  18. Intense Microbubbles Mimicking Mobile Thrombus in a Patient with Prosthetic Mitral Valve

    Directory of Open Access Journals (Sweden)

    Fatma Yılmaz Coşkun

    2017-06-01

    Full Text Available Microbubbles have been presumed as gaseous emboli, which originate during mechanical heart valve closure, but are not seen in bioprosthetic valves. In this report, we presented a cluster of microbubbles mimicking mobile thrombus in a patient with mechanical mitral valve prosthesis. A 30-year-old female with a history of implanted mechanical valve at the mitral position underwent a routine examination. She was asymptomatic and her physical examination was unremarkable. Transthoracic echocardiography showed a mobile thrombus-like mass on the ventricular side of the prosthetic mitral valve moving into the left ventricular outflow tract. However, close examination of images indicated that the mass was in fact intense microbubbles mimicking thrombus. Intense mobile microbubbles can be misdiagnosed as a mobile thrombus. We recommend and underscore the importance of detailed echocardiographic examination in case of mobile mass to avoid misdiagnosis in patients with mechanical heart valves.

  19. Subharmonic response from ultrasound contrast microbubbles for noninvasive blood pressure estimation

    Science.gov (United States)

    Katiyar, Amit; Sarkar, Kausik; Forsberg, Flemming

    2010-11-01

    Estimation of local organ-level blood pressure can help in diagnosing and monitoring heart and vascular diseases. Subharmonic signals from ultrasound contrast microbubbles have been proposed as a noninvasive alternative to the current practice of using manometer-tipped catheter. Approximately 10dB linear decrease in subharmonic component with 25 kPa pressure increase (typical blood pressure variation) has been reported for several contrast microbubbles. Here we report a theoretical investigation of the underlying phenomenon. We first study the well established model of a free microbubble to show that reduction of subharmonic with ambient pressure increase occurs only below a certain excitation frequency. Above this critical frequency, subharmonic signal increases with ambient pressure. Furthermore, where it decreases with ambient pressure, the relationship is linear only above certain excitation pressure. The dependence of the critical frequency on bubble radius and possibly bubble size distribution is discussed. We also report similar behavior for several models for encapsulated contrast microbubbles.

  20. Estimating the shell parameters of SonoVue® microbubbles using light scattering

    OpenAIRE

    Tu, Juan; Guan, Jingfeng; Qiu, Yuanyuan; Matula, Thomas J.

    2009-01-01

    Experiments were performed to measure the dynamical response of individual SonoVue® microbubbles subjected to pulsed ultrasound. Three commonly used bubble dynamic models (i.e., Hoff’s, Sarkar’s, and linearized Marmottant’s models) were compared to determine the most appropriate model for fitting to the experimental data. The models were evaluated against published optical microscopy data. The comparison suggests that it is difficult to rank these models for lipid-shelled microbubbles undergo...

  1. Targeted microbubbles: a novel application for the treatment of kidney stones.

    Science.gov (United States)

    Ramaswamy, Krishna; Marx, Vanessa; Laser, Daniel; Kenny, Thomas; Chi, Thomas; Bailey, Michael; Sorensen, Mathew D; Grubbs, Robert H; Stoller, Marshall L

    2015-07-01

    Kidney stone disease is endemic. Extracorporeal shockwave lithotripsy was the first major technological breakthrough where focused shockwaves were used to fragment stones in the kidney or ureter. The shockwaves induced the formation of cavitation bubbles, whose collapse released energy at the stone, and the energy fragmented the kidney stones into pieces small enough to be passed spontaneously. Can the concept of microbubbles be used without the bulky machine? The logical progression was to manufacture these powerful microbubbles ex vivo and inject these bubbles directly into the collecting system. An external source can be used to induce cavitation once the microbubbles are at their target; the key is targeting these microbubbles to specifically bind to kidney stones. Two important observations have been established: (i) bisphosphonates attach to hydroxyapatite crystals with high affinity; and (ii) there is substantial hydroxyapatite in most kidney stones. The microbubbles can be equipped with bisphosphonate tags to specifically target kidney stones. These bubbles will preferentially bind to the stone and not surrounding tissue, reducing collateral damage. Ultrasound or another suitable form of energy is then applied causing the microbubbles to induce cavitation and fragment the stones. This can be used as an adjunct to ureteroscopy or percutaneous lithotripsy to aid in fragmentation. Randall's plaques, which also contain hydroxyapatite crystals, can also be targeted to pre-emptively destroy these stone precursors. Additionally, targeted microbubbles can aid in kidney stone diagnostics by virtue of being used as an adjunct to traditional imaging methods, especially useful in high-risk patient populations. This novel application of targeted microbubble technology not only represents the next frontier in minimally invasive stone surgery, but a platform technology for other areas of medicine. © 2014 The Authors BJU International © 2014 BJU International Published

  2. Guanine nucleotide-binding protein subunit beta-2-like 1, a new Annexin A7 interacting protein

    Energy Technology Data Exchange (ETDEWEB)

    Du, Yue; Meng, Jinyi; Huang, Yuhong; Wu, Jun; Wang, Bo; Ibrahim, Mohammed M.; Tang, Jianwu, E-mail: jianwutdlmedu@163.com

    2014-02-28

    Highlights: • RACK1 formed a complex with Annexin A7. • Depletion of RACK1 inhibited the proliferation, migration and invasion. • RACK1 RNAi abolished RACK1-Annexin A7 interaction. • RACK1-Annexin A7 may play a role in regulating the metastatic potentials. - Abstract: We report for the first time that Guanine nucleotide-binding protein subunit beta-2-like 1 (RACK1) formed a complex with Annexin A7. Hca-F and Hca-P are a pair of syngeneic mouse hepatocarcinoma cell lines established and maintained in our laboratory. Our previous study showed that both Annexin A7 and RACK1 were expressed higher in Hca-F (lymph node metastasis >70%) than Hca-P (lymph node metastasis <30%). Suppression of Annexin A7 expression in Hca-F cells induced decreased migration and invasion ability. In this study, knockdown of RACK1 by RNA interference (RNAi) had the same impact on metastasis potential of Hca-F cells as Annexin A7 down-regulation. Furthermore, by co-immunoprecipitation and double immunofluorescence confocal imaging, we found that RACK1 was in complex with Annexin A7 in control cells, but not in the RACK1-down-regulated cells, indicating the abolishment of RACK1-Annexin A7 interaction in Hca-F cells by RACK1 RNAi. Taken together, these results suggest that RACK1-Annexin A7 interaction may be one of the means by which RACK1 and Annexin A7 influence the metastasis potential of mouse hepatocarcinoma cells in vitro.

  3. Dynamic Interactions between Contrast Agent Microbubbles: High Speed Camera Observations and Simulation Results

    Science.gov (United States)

    Stride, Eleanor; Chetty, Kevin; Eckersley, Robert

    2007-05-01

    The efficacy of coated microbubbles as contrast agents for ultrasound imaging has been well established over the past two decades. More recently, their use as carriers for targeted drug delivery has also become an active area of research. However, the behaviour of microbubbles in an ultrasound field is by no means fully understood. For example, the dynamic interactions between microbubbles have frequently been neglected when considering contrast agent suspensions. In this investigation, high speed camera observations of a commercial contrast agent (SonoVue®) were made under controlled and calibrated acoustic exposure conditions (single 4 cycle (FWHM) Gaussian pulse with 0.5 MHz centre frequency and peak negative pressure <100 kPa). These were compared with numerical simulations of both single and pairs of coated microbubbles corresponding to the experiments. Both the theoretical and experimental results indicate that the dynamic behaviour of a microbubble may be substantially affected by the presence of neighbouring bubbles under certain conditions. This, in turn, may affect the microbubble's acoustic response and its destruction threshold, which has potentially significant implications for both diagnostic and therapeutic applications.

  4. Estimating the shell parameters of SonoVue microbubbles using light scattering.

    Science.gov (United States)

    Tu, Juan; Guan, Jingfeng; Qiu, Yuanyuan; Matula, Thomas J

    2009-12-01

    Experiments were performed to measure the dynamical response of individual SonoVue microbubbles subjected to pulsed ultrasound. Three commonly used bubble dynamic models (i.e., Hoff's, Sarkar's, and linearized Marmottant's models) were compared to determine the most appropriate model for fitting to the experimental data. The models were evaluated against published optical microscopy data. The comparison suggests that it is difficult to rank these models for lipid-shelled microbubbles undergoing small-amplitude oscillations, because under these conditions the shell parameters in these models are closely related. A linearized version of the Marmottant model was used to estimate the shell parameters (i.e., shear modulus and shear viscosity) of SonoVue microbubbles from the experimental light scattering data, as a function of ambient microbubble radius. The SonoVue microbubble shell elasticity and dilatational viscosity increase with ambient bubble radius, in agreement with previously published data. The results suggest that light scattering, used in conjunction with one of several popular bubble dynamics models, is effective at characterizing microbubble response and evaluating shell parameters.

  5. Technique for the Characterization of Phospholipid Microbubbles Coatings by Transmission Electron Microscopy.

    Science.gov (United States)

    Owen, Joshua; Stride, Eleanor

    2015-12-01

    Gas microbubbles stabilized by a surfactant or polymer coating are of considerable clinical interest because of their imaging and drug delivery potential under ultrasound exposure. The utility of microbubbles for a given application is intrinsically linked to their structure and stability. These in turn are highly sensitive to coating composition and fabrication techniques. Various methods including fluorescence and atomic force microscopy have been applied to characterize microbubble properties, but direct observation of coating structure at the nanoscale still poses a considerable challenge. Here we describe a transmission electron microscopy (TEM) technique to observe the surface of microbubbles. Images from a series of phospholipid-coated microbubble systems, including those decorated with nanoparticles, are presented. They indicate that the technique enables visualization of the coating structure, in particular lipid discontinuities and nanoparticle distribution. This information can be used to better understand how microbubble surface structure relates to formulation and/or processing technique and ultimately to functionality. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  6. Particle migration and sorting in microbubble streaming flows

    Science.gov (United States)

    Thameem, Raqeeb; Hilgenfeldt, Sascha

    2016-01-01

    Ultrasonic driving of semicylindrical microbubbles generates strong streaming flows that are robust over a wide range of driving frequencies. We show that in microchannels, these streaming flow patterns can be combined with Poiseuille flows to achieve two distinctive, highly tunable methods for size-sensitive sorting and trapping of particles much smaller than the bubble itself. This method allows higher throughput than typical passive sorting techniques, since it does not require the inclusion of device features on the order of the particle size. We propose a simple mechanism, based on channel and flow geometry, which reliably describes and predicts the sorting behavior observed in experiment. It is also shown that an asymptotic theory that incorporates the device geometry and superimposed channel flow accurately models key flow features such as peak speeds and particle trajectories, provided it is appropriately modified to account for 3D effects caused by the axial confinement of the bubble. PMID:26958103

  7. [Relevance of contrast ultrasound with microbubbles in vascular medecine].

    Science.gov (United States)

    Erdmann, Andreas; Ney, Barbara; Alatri, Adriano; Calanca, Luca; Mazzolai, Lucia

    2016-12-07

    Application of ultrasound contrast media has become a standard in diagnostic imaging in cardiology and in the characterization of focal lesions in multiple organs, especially of the liver. In the past years there was a growing body of evidence for their usefulness in vascular medicine. The development of contrast media, microbubbles with a stabilizing envelope and filled with gaz, small enough to pass through pulmonary capillaries made real-time imaging of organ perfusion possible. Ultrasound contrast media are rapidly eliminated by exhalation and can safely be administered to patients with renal failure. The objective of this review is to describe the basic principles of ultrasound contrast imaging and to inform about vascular applications of contrast ultrasound.

  8. [Impact of instantaneous uniformity of SonoVue microbubbles on binding characteristics of a new contrast agent targeted to choriocarcinoma cells in vitro].

    Science.gov (United States)

    Zhou, Li-Xue; Ding, Hong; Jia, Cai-Xia; Li, Yan; Wei, Qing

    2008-07-01

    At present, the investigation of microbubble contrast agents is a hot spot. Although these contrast agents can increase the ultrasound detection rate of tumor vessels, they lack tissue specificity. This study was to evaluate the impact of instantaneous uniformity of SonoVue microbubbles on binding characteristics, including the adhesion rate and stability, of a new contrast agent targeted to choriocarcinoma cells (JARs) in vitro, in order to establish a foundation to explore targeted ultrasound imaging for localization of tumor cell antigens and increase the early diagnostic rate for tumors. The objects were divided into three groups: the uneven microbubble group (n=10), the uniform microbubble group (n=10) and the tiny microbubble group (n=10). The rosette formation rate was counted. JARs were calculated by flow cytometry (FCM). The shape of the rosette was recorded. The targeted contrast agent was prepared by mixing SonoVue microbubbles of different uniformity with rabbit anti-human chorionic gonadotrophin (HCG) antibody. The binding rates of the contrast agent to JARs before and after PBS rinse were analyzed. The binding rate was significantlylower in the uneven microbubble group (60.4+/-1.5)% than in the uniform microbubble group (84.3+/-5.5)% and the tiny microbubble group (90.6+/-6.8)% (P0.05). The binding rates of the targeted microbubbles labeled with fluorescence to JARs were 72.9%, 81.03% and 88.5% in the uneven microbubble group, the uniform microbubble group and the tiny microbubble group, respectively (PSonoVue microbubbles to JARs is related to instantaneous uniformity of the microbubble, which is determined by the shaking method before preparation. Improving instantaneous uniformity of SonoVue microbubbles may increase the binding rate and stability of targeted microbubbles to JARs, thus to improve the image of JARs.

  9. Nanoparticle coated optical fibers for single microbubble generation

    Science.gov (United States)

    Pimentel-Domínguez, Reinher; Hernández-Cordero, Juan

    2011-09-01

    The study of bubbles and bubbly flows is important in various fields such as physics, chemistry, medicine, geophysics, and even the food industry. A wide variety of mechanical and acoustic techniques have been reported for bubble generation. Although a single bubble may be generated with these techniques, controlling the size and the mean lifetime of the bubble remains a difficult task. Most of the optical methods for generation of microbubbles involve high-power pulsed laser sources focused in absorbing media such as liquids or particle solutions. With these techniques, single micron-sized bubbles can be generated with typical mean lifetimes ranging from nano to microseconds. The main problem with these bubbles is their abrupt implosion: this produces a shock wave that can potentially produce damages on the surroundings. These effects have to be carefully controlled in biological applications and in laser surgery, but thus far, not many options are available to effectively control micron-size bubble growth. In this paper, we present a new technique to generate microbubbles in non-absorbing liquids. In contrast to previous reports, the proposed technique uses low-power and a CW radiation from a laser diode. The laser light is guided through an optical fiber whose output end has been coated with nanostructures. Upon immersing the tip of the fiber in ethanol or water, micron-size bubbles can be readily generated. With this technique, bubble growth can be controlled through adjustments on the laser power. We have obtained micron-sized bubbles with mean lifetimes in the range of seconds. Furthermore, the generated bubbles do not implode, as verified with a high-speed camera and flow visualization techniques.

  10. Annexin A5 Promoter Haplotype M2 Is Not a Risk Factor for Recurrent Pregnancy Loss in Northern Europe

    DEFF Research Database (Denmark)

    Nagirnaja, Liina; Nõmmemees, Diana; Rull, Kristiina

    2015-01-01

    and recurrent pregnancy loss (RPL), however with inconclusive results. STUDY SUBJECTS AND METHODS: A retrospective case-control study combining resequencing and restriction fragment length polymorphism (RFLP) analysis was undertaken in 313 women with unexplained RPL and 214 fertile women from Estonia...... compared to controls both in Estonia (8.1% vs 15.2%, respectively) and Denmark (9.7% vs 12.6%). The high M2 prevalence in fertile controls was consistent with estimations for European and East Asian populations (9.6%-16.0%). CONCLUSIONS: This study cautions to consider the M2 haplotype as a deterministic...

  11. Annexin A1 expression in a pooled breast cancer series : Association with tumor subtypes and prognosis

    NARCIS (Netherlands)

    Sobral-Leite, Marcelo; Wesseling, Jelle; Smit, Vincent T H B M; Nevanlinna, Heli; van Miltenburg, Martine H.; Sanders, Joyce; Hofland, Ingrid; Blows, Fiona M.; Coulson, Penny; Patrycja, Gazinska; Schellens, Jan H M; Fagerholm, Rainer; Heikkilä, Päivi; Aittomäki, Kristiina; Blomqvist, Carl; Provenzano, Elena; Ali, Hamid Raza; Figueroa, Jonine; Sherman, Mark; Lissowska, Jolanta; Mannermaa, Arto; Kataja, Vesa; Kosma, Veli Matti; Hartikainen, Jaana M.; Phillips, Kelly Anne; Couch, Fergus J.; Olson, Janet E.; Vachon, Celine; Visscher, Daniel; Brenner, Hermann; Butterbach, Katja; Arndt, Volker; Holleczek, Bernd; Hooning, Maartje J.; Hollestelle, Antoinette; Martens, John W M; van Deurzen, Carolien H M; van de Water, Bob; Broeks, Annegien; Chang-Claude, Jenny; Chenevix-Trench, Georgia; Easton, Douglas F.; Pharoah, Paul D P; García-Closas, Montserrat; de Graauw, Marjo; Schmidt, Marjanka K.; Aghmesheh, Morteza; Amor, David; Andrews, Lesley; Antill, Yoland; Armitage, Shane; Arnold, Leanne; Balleine, Rosemary; Bankier, Agnes; Bastick, Patti; Beesley, Jonathan; Beilby, John; Bennett, Barbara; Bennett, Ian; Berry, Geoffrey; Blackburn, Anneke; Bogwitz, Michael; Brennan, Meagan; Brown, Melissa; Buckley, Michael; Burgess, Matthew; Burke, Jo; Butow, Phyllis; Byron, Keith; Callen, David; Campbell, Ian; Chauhan, Deepa; Chauhan, Manisha; Christian, Alice; Clarke, Christine; Colley, Alison; Cotton, Dick; Crook, Ashley; Cui, James; Culling, Bronwyn; Cummings, Margaret; Dawson, Sarah Jane; deFazio, Anna; Delatycki, Martin; Dickson, Rebecca; Dixon, Joanne; Dobrovic, Alexander; Dudding, Tracy; Edkins, Ted; Edwards, Stacey; Eisenbruch, Maurice; Farshid, Gelareh; Fawcett, Susan; Fellows, Andrew; Fenton, Georgina; Field, Michael; Firgaira, Frank; Flanagan, James; Fleming, Jean; Fong, Peter; Forbes, John; Fox, Stephen; French, Juliet; Friedlander, Michael; Gaff, Clara; Gardner, Mac; Gattas, Mike; George, Peter; Giles, Graham; Gill, Grantley; Goldblatt, Jack; Greening, Sian; Grist, Scott; Haan, Eric; Hardie, Kate; Harris, Marion; Hart, Stewart; Hayward, Nick; Healey, Sue; Heiniger, Louise; Hopper, John; Humphrey, Evelyn; Hunt, Clare; James, Paul; Jenkins, Mark; Jones, Alison; Kefford, Rick; Kidd, Alexa; Kiely, Belinda; Kirk, Judy; Koehler, Jessica; Kollias, James; Kovalenko, Serguei; Lakhani, Sunil; Leaming, Amanda; Leary, Jennifer; Lim, Jacqueline; Lindeman, Geoff; Lipton, Lara; Lobb, Liz; Mann, Graham; Marsh, Deborah; McLachlan, Sue Anne; Meiser, Bettina; Meldrum, Cliff; Milne, Roger; Mitchell, Gillian; Newman, Beth; Niedermayr, Eveline; Nightingale, Sophie; O'Connell, Shona; O'Loughlin, Imelda; Osborne, Richard; Pachter, Nick; Patterson, Briony; Peters, Lester; Phillips, Kelly; Price, Melanie; Purser, Lynne; Reeve, Tony; Reeve, Jeanne; Richards, Robert; Rickard, Edwina; Robinson, Bridget; Rudzki, Barney; Saleh, Mona; Salisbury, Elizabeth; Sambrook, Joe; Saunders, Christobel; Saunus, Jodi; Sayer, Robyn; Scott, Elizabeth; Scott, Rodney; Scott, Clare; Seshadri, Ram; Sexton, Adrienne; Sharma, Raghwa; Shelling, Andrew; Simpson, Peter; Southey, Melissa; Spurdle, Amanda; Suthers, Graeme; Sykes, Pamela; Tassell, Margaret; Taylor, Donna; Taylor, Jessica; Thierry, Benjamin; Thomas, Susan; Thompson, Ella; Thorne, Heather; Townshend, Sharron; Trainer, Alison; Tran, Lan; Tucker, Kathy; Tyler, Janet; Visvader, Jane; Walker, Logan; Walpole, Ian; Ward, Robin; Waring, Paul; Warner, Bev; Warren, Graham; Williams, Rachael; Wilson, Judy; Winship, Ingrid; Wu, Kathy; Young, Mary Ann; Bowtell, D.; Green, A.; Webb, P.; de Fazio, A.; Gertig, D.

    2015-01-01

    Background: Annexin A1 (ANXA1) is a protein related with the carcinogenesis process and metastasis formation in many tumors. However, little is known about the prognostic value of ANXA1 in breast cancer. The purpose of this study is to evaluate the association between ANXA1 expression, BRCA1/2

  12. "Nonclassical" secretion of annexin A2 to the lumenal side of the enterocyte brush border membrane

    DEFF Research Database (Denmark)

    Danielsen, E Michael; van Deurs, Bo; Hansen, Gert H

    2003-01-01

    side of the microvilli, showing an apical secretion by a "nonclassical" mechanism. In addition, annexin A2 was associated with surface-connected, deep apical tubules in the apical terminal web region and with an underlying pleiomorphic, tubulo-vesicular compartment (subapical compartment...

  13. Evaluation of 18F-SFB-Annexin B1 in detecting apoptosis

    International Nuclear Information System (INIS)

    Zhao Qing; Zhang Yingjian; Wang Fang

    2011-01-01

    Objective: To evaluate 18 F-N- succinimidyl -4-fluorobenzoate (SFB)-Annexin B1 in detecting in vitro and in vivo apoptosis. Methods: Anti-Fas antibody was used to induce apoptosis in Jurkat cells. Apoptosis in Jurkat cells was confirmed by flow cytometer (FCM). Unilateral renal ischemia/reperfusion injury was induced by transient (45 min) ligation of the renal artery in the rabbit. The rabbit was then administrated with 18 F-SFB-Annexin B1 intravenously 24 h later and then imaged by PET/CT at 10, 30, 60, 90, 120 and 240 min postinjection. Apoptosis in kidney was confirmed by terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) assay and HE staining. Results The apoptosis rate induced by anti-Fas antibody was 25.98% (120 min) while that in the control group was only 1.81%. The uptake of 18 F-SFB-Annexin B1 in apoptosis group was greater than that in the control group. PET/CT images at 240 min showed higher uptake in the ligated kidney than the non-ligated kidney. TUNEL assay and HE staining confirmed great amount apoptotic cells in the ligated kidney. Conclusion: 18 F-SFB-Annexin B1 may be potentially useful in detecting apoptosis both in vitro and in vivo. (authors)

  14. Purification and immunolocalization of an annexin-like protein in pea seedlings

    Science.gov (United States)

    Clark, G. B.; Dauwalder, M.; Roux, S. J.

    1992-01-01

    As part of a study to identify potential targets of calcium action in plant cells, a 35-kDa, annexin-like protein was purified from pea (Pisum sativum L.) plumules by a method used to purify animal annexins. This protein, called p35, binds to a phosphatidylserine affinity column in a calcium-dependent manner and binds 45Ca2+ in a dot-blot assay. Preliminary sequence data confirm a relationship for p35 with the annexin family of proteins. Polyclonal antibodies have been raised which recognize p35 in Western and dot blots. Immunofluorescence and immunogold techniques were used to study the distribution and subcellular localization of p35 in pea plumules and roots. The highest levels of immunostain were found in young developing vascular cells producing wall thickenings and in peripheral root-cap cells releasing slime. This localization in cells which are actively involved in secretion is of interest because one function suggested for the animal annexins is involvement in the mediation of exocytosis.

  15. Regulation of annexins following infection like tissue damage – investigated by 2-dimensional gel electrophoresis

    DEFF Research Database (Denmark)

    Wulff, Tune; Nielsen, Michael Engelbrecht

    , internal- and external control were found using a t-test. To investigate numerous proteins in a single study changes in protein abundance were investigated using 2-dimensional gel electrophoresis. Protein of interest were identified using MALDI MS/MS. The results show that both annexin 4 and 5...

  16. Biodistribution, kinetics, and biological fate of SPION microbubbles in the rat

    Directory of Open Access Journals (Sweden)

    Barrefelt A

    2013-08-01

    Full Text Available Åsa Barrefelt,1,2,* Maryam Saghafian,2,* Raoul Kuiper,3 Fei Ye,4 Gabriella Egri,5 Moritz Klickermann,5 Torkel B Brismar,1 Peter Aspelin,1 Mamoun Muhammed,4 Lars Dähne,5 Moustapha Hassan2,6 1Department of Clinical Science, Intervention and Technology, Division of Medical Imaging and Technology, Karolinska Institutet, and Department of Radiology, Karolinska University Hospital-Huddinge, Stockholm, Sweden; 2Experimental Cancer Medicine, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden; 3Karolinska Institute Core Facility for Morphologic Phenotype Analysis, Clinical Research Center, Karolinska University Hospital-Huddinge, Stockholm, Sweden; 4Division of Functional Materials, Department of Materials and Nano Physics, Royal Institute of Technology, Stockholm, Sweden; 5Surflay Nanotec GmbH, Berlin, Germany; 6Clinical Research Center, Karolinska University Hospital-Huddinge, Stockholm, Sweden *These authors contributed equally to this work Background: In the present investigation, we studied the kinetics and biodistribution of a contrast agent consisting of poly(vinyl alcohol (PVA microbubbles containing superparamagnetic iron oxide (SPION trapped between the PVA layers (SPION microbubbles. Methods: The biological fate of SPION microbubbles was determined in Sprague-Dawley rats after intravenous administration. Biodistribution and elimination of the microbubbles were studied in rats using magnetic resonance imaging for a period of 6 weeks. The rats were sacrificed and perfusion-fixated at different time points. The magnetic resonance imaging results obtained were compared with histopathologic findings in different organs. Results: SPION microbubbles could be detected in the liver using magnetic resonance imaging as early as 10 minutes post injection. The maximum signal was detected between 24 hours and one week post injection. Histopathology showed the presence of clustered SPION microbubbles predominantly in the lungs from

  17. Prediction of micro-bubble dissolution characteristics in water and seawater

    Energy Technology Data Exchange (ETDEWEB)

    Kawahara, Akimaro; Sadatomi, Michio; Matsuura, Hidetoshi; Tominaga, Mayo; Noguchi, Masanori [Department of Mechanical System Engineering, Kumamoto University, Kurokami 2-39-1, Kumamoto City 860-8555 (Japan); Matsuyama, Fuminori [Department of Mechanical Engineering, Sasebo National College of Technology (Japan)

    2009-07-15

    This paper is concerned with the prediction of micro-bubble dissolution characteristics in water and seawater when microbubbles are generated by a Sadatomi-type micro-bubble generator (2003) with a spherical body in a flowing liquid tube. In the experiments, in order to know the effects of the salinity on the characteristics, tap water and an artificial seawater with different salt concentrations of 1 and 3 wt% were used as the test liquids. Parameters measured were the Sauter mean diameter of bubbles, d{sub BS}, the void fraction, {alpha}, the rising velocity of bubbles, u{sub G}, the interfacial area concentration, a, the volumetric mass transfer coefficient, K{sub L}a, and the liquid-side mass transfer coefficient, K{sub L}. In the analysis, for predicting {alpha}, K{sub L}a and K{sub L}, some correlations in the literatures were tested against the present data. Furthermore, in order to improve the predictability, new correlations were developed based on the present data. The prediction of K{sub L}a with the new correlation agreed well with Nishino et al.'s [T. Nishino, K. Terasaka, M. Ishida, Application for several micro-bubble generators for gas absorber, in: Proceedings of the Annual Meeting of the Japanese Society for Multiphase Flow, 2006, pp. 276-277 (in Japanese)] and Li and Tsuge's [P. Li, H. Tsuge, Water treatment by induced air flotation using microbubbles, Journal of Chemical Engineering of Japan 39 (2006) 896-903; P. Li, H. Tsuge, Ozone transfer in a new gas-induced contactor with microbubbles, Journal of Chemical Engineering of Japan 39 (2006) 1213-1220] data for different aeration systems using several different micro-bubble generators. (author)

  18. Ultrasonic microbubble contrast agents and the transplant kidney

    Energy Technology Data Exchange (ETDEWEB)

    Kay, D.H., E-mail: davidhkay@doctors.org.u [Department of Radiology, Western Infirmary, Glasgow (United Kingdom); Mazonakis, M.; Geddes, C. [Department of Renal Medicine, Western Infirmary, Glasgow (United Kingdom); Baxter, G. [Department of Radiology, Western Infirmary, Glasgow (United Kingdom)

    2009-11-15

    Aim: To evaluate the potential application of microbubble agents in the immediate post-transplant period, by studying contrast uptake and washout, and to correlate these values with clinical indices, and thus, assess the potential prognostic value of this technique. Materials and methods: The study group comprised 20 consecutive renal transplant patients within 7 days of transplantation. Sonovue was administered as an intravenous bolus with continuous imaging of the transplant kidney at low mechanical index (MI) for 1 min post-injection. These data were analysed off-line by two observers, and time intensity curves (TIC) for the upper, mid, and lower poles constructed. Within each pole, a region of interest (5 mm square) was placed over the cortex, medullary pyramid, and interlobar artery, resulting in a total of nine TIC for each patient. TIC parameters included the arrival time (AT), time to peak (TTP), peak intensity (Max), gradient of the slope (M), and the area under curve (AUC). Results: For both observers there was good agreement for all values measured from the cortex and medulla, but poor interobserver correlation for the vascular values. In addition, there was only agreement for these values in the upper and mid-pole of the transplant with poor agreement for the lower pole values. The mid-pole of the transplant kidney was chosen as the point of measurement for subsequent studies. Mid-pole values were correlated with clinical data and outcome over the 3-month post-transplant period. Renal microbubble perfusion correlated with the transplant estimated glomerular filtration rate (eGFR) at 3 months post-transplantation (p = 0.016). Discussion: In conclusion, this is the first study to confirm reproducibility of the Sonovue TIC data in transplant patients and to quantify regional variation and perfusion. The statistically significant estimates of transplant perfusion may be of future benefit to transplant recipients and potentially utilized as a prognostic tool

  19. Ultrasonic microbubble contrast agents and the transplant kidney

    International Nuclear Information System (INIS)

    Kay, D.H.; Mazonakis, M.; Geddes, C.; Baxter, G.

    2009-01-01

    Aim: To evaluate the potential application of microbubble agents in the immediate post-transplant period, by studying contrast uptake and washout, and to correlate these values with clinical indices, and thus, assess the potential prognostic value of this technique. Materials and methods: The study group comprised 20 consecutive renal transplant patients within 7 days of transplantation. Sonovue was administered as an intravenous bolus with continuous imaging of the transplant kidney at low mechanical index (MI) for 1 min post-injection. These data were analysed off-line by two observers, and time intensity curves (TIC) for the upper, mid, and lower poles constructed. Within each pole, a region of interest (5 mm square) was placed over the cortex, medullary pyramid, and interlobar artery, resulting in a total of nine TIC for each patient. TIC parameters included the arrival time (AT), time to peak (TTP), peak intensity (Max), gradient of the slope (M), and the area under curve (AUC). Results: For both observers there was good agreement for all values measured from the cortex and medulla, but poor interobserver correlation for the vascular values. In addition, there was only agreement for these values in the upper and mid-pole of the transplant with poor agreement for the lower pole values. The mid-pole of the transplant kidney was chosen as the point of measurement for subsequent studies. Mid-pole values were correlated with clinical data and outcome over the 3-month post-transplant period. Renal microbubble perfusion correlated with the transplant estimated glomerular filtration rate (eGFR) at 3 months post-transplantation (p = 0.016). Discussion: In conclusion, this is the first study to confirm reproducibility of the Sonovue TIC data in transplant patients and to quantify regional variation and perfusion. The statistically significant estimates of transplant perfusion may be of future benefit to transplant recipients and potentially utilized as a prognostic tool

  20. Buoyancy-activated cell sorting using targeted biotinylated albumin microbubbles.

    Directory of Open Access Journals (Sweden)

    Yu-Ren Liou

    Full Text Available Cell analysis often requires the isolation of certain cell types. Various isolation methods have been applied to cell sorting, including fluorescence-activated cell sorting and magnetic-activated cell sorting. However, these conventional approaches involve exerting mechanical forces on the cells, thus risking cell damage. In this study we applied a novel isolation method called buoyancy-activated cell sorting, which involves using biotinylated albumin microbubbles (biotin-MBs conjugated with antibodies (i.e., targeted biotin-MBs. Albumin MBs are widely used as contrast agents in ultrasound imaging due to their good biocompatibility and stability. For conjugating antibodies, biotin is conjugated onto the albumin MB shell via covalent bonds and the biotinylated antibodies are conjugated using an avidin-biotin system. The albumin microbubbles had a mean diameter of 2 μm with a polydispersity index of 0.16. For cell separation, the MDA-MB-231 cells are incubated with the targeted biotin-MBs conjugated with anti-CD44 for 10 min, centrifuged at 10 g for 1 min, and then allowed 1 hour at 4 °C for separation. The results indicate that targeted biotin-MBs conjugated with anti-CD44 antibodies can be used to separate MDA-MB-231 breast cancer cells; more than 90% of the cells were collected in the MB layer when the ratio of the MBs to cells was higher than 70:1. Furthermore, we found that the separating efficiency was higher for targeted biotin-MBs than for targeted avidin-incorporated albumin MBs (avidin-MBs, which is the most common way to make targeted albumin MBs. We also demonstrated that the recovery rate of targeted biotin-MBs was up to 88% and the sorting purity was higher than 84% for a a heterogenous cell population containing MDA-MB-231 cells (CD44(+ and MDA-MB-453 cells (CD44-, which are classified as basal-like breast cancer cells and luminal breast cancer cells, respectively. Knowing that the CD44(+ is a commonly used cancer

  1. A study on the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles.

    Science.gov (United States)

    Li, Huiling; Chen, Jinwen; Xu, Xuan; Yang, Ruhao; Xiang, Xudong; Zhang, Dongshan

    2016-02-01

    To study the safety and efficiency of the transfection of antisense oligonucletide into kidney mediated by lipid microbubbles, and to evaluate its potential clinical application. The potential and conditions regarding the transfection self-made lipid microbubbles (CY5)-labeled-oligonucleotide (ODN) or CY5-labeled-ODN connective tissue growth factor (CTGF) into the rat kidney were evaluated. Th e safety was evaluated by HE staining, liver and renal function tests. The transfection efficiency was evaluated by fluorescence microscopy. Th e expression of CTGF was detected by RT-PCR and Western blot. Self-made lipid microbubble and/or ultrasound significantly enhanced the efficiency of gene transfer and expression in the kidney. Especially, 85%-90% of total glomerular could be transfected. CY5-labeled-ODN expression could be observed in glomerular, tubular and interstitial area. Th ere was no significant change in blood tests aft er gene transfer. Levels of LDH in 7 days were decreased compared with that at the fi rst day aft er the transfection (Ptransfection of CTGF-antisense-ODN into kidney. The ultrasound-mediated gene transfer by self-made lipid microbubble could enhance the efficiency of ODN and expression in the rat kidney. Th is self-made lipid microbubbles supplement may be use for transfection of target genes.

  2. Liver haemostasis using microbubble-enhanced ultrasound at a low acoustic intensity.

    Science.gov (United States)

    Zhao, Xiaochen; Li, Lu; Zhao, Hongzhi; Li, Tao; Wu, Shengzheng; Zhong, Yu; Zhao, Yang; Liu, Zheng

    2012-02-01

    To explore the haemostatic effects of microbubble-enhanced ultrasound (MEUS) at a very low acoustic intensity on the bleeding liver of rabbits. Liver incisions made on 20 rabbits were treated with a pulsed therapeutic ultrasound transducer. The transducer was operated at 831 KHz with an acoustic intensity of 0.4 W/cm(2). The treatment was coordinated with intravenous injection of microbubbles. Ultrasound only and sham treatment served as the controls. Visual bleeding score and 10-min bleeding volume were evaluated for haemostatic efficacy. Contrast-enhanced ultrasound (CEUS) was performed to assess the liver perfusion. Nine treated livers were harvested for acute histological examination. Regarding the bleeding incisions made on rabbit livers, the haemorrhage stopped immediately after 2 min of MEUS treatment but bleeding continued in the controls treated by ultrasound or microbubble injection alone. The bleeding scores and the 10-min haemorrhagic volumes dropped significantly in the MEUS group compared with those of the controls (p therapeutic ultrasound during enhancement with intravenous microbubbles • This combined therapy was more effective than ultrasound or intravenous microbubbles alone • More work is required with larger animals before potential human trials.

  3. Effects of ultrasound-combined microbubbles on hippocampal AchE fibers in rats.

    Science.gov (United States)

    Gong, Zi-Li; Luo, Chun-Mei; Wu, Sheng-Zheng; Ran, Hong; Zhu, Jie; Zheng, Jian

    2014-05-01

    To investigate the protective effect of ultrasound-combined microbubbles on hippocampal acetylcholinesterase (AchE) fibers in rats. According to random digits table, 60 SD rats were divided into two groups, marrow stromal cells (MSCs) intracranial transplantation group and MSCs intracranial transplantation + ultrasonic microbubbles group. Marrow stromal cells were cultivated and isolated in vitro; 12 weeks after transplantation, spatial learning and memorizing abilities of rats were assessed by Morris water maze; AchE staining method was used to observe changes in density and appearance of AchE staining positive fibers in hippocampal CA1 region. There was a significant increase in spatial learning and memorizing abilities of rats in MSCs intracranial transplantation + ultrasonic microbubbles group. Hippocampal AchE staining suggested an increase in the density of AchE staining positive fibers in MSCs intracranial transplantation group; the fibers were regular, intact and dense. Density of hippocampal AchE positive fibers was negatively correlated with the escape latent period and was positively correlated with percentage of the time needed to cross each platform quadrant. Better promotion of spatial learning and memorizing abilities of rats in MSCs intracranial transplantation + ultrasonic microbubbles group may be related with the protective effect of ultrasound-combined microbubbles on hippocampal acetylcholine fibers. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  4. Microbubble-based fiber-optic Fabry-Perot pressure sensor for high-temperature application.

    Science.gov (United States)

    Li, Zhe; Jia, Pinggang; Fang, Guocheng; Liang, Hao; Liang, Ting; Liu, Wenyi; Xiong, Jijun

    2018-03-10

    Using arc discharge technology, we fabricated a fiber-optic Fabry-Perot (FP) pressure sensor with a very low temperature coefficient based on a microbubble that can be applied in a high-temperature environment. The thin-walled microbubble can be fabricated by heating the gas-pressurized hollow silica tube (HST) using a commercial fusion splicer. Then, the well-cut single-mode fiber (SMF) was inserted into the microbubble, and they were fused together. Thus, the FP cavity can be formed between the end of the SMF and the inner surface of the microbubble. The diameter of the microbubble can be up to 360 μm with the thickness of the wall being approximately 0.5 μm. Experimental results show that such a sensor has a linear sensitivity of approximately -6.382  nm/MPa, -5.912  nm/MPa at 20°C, and 600°C within the pressure range of 1 MPa. Due to the thermal expansion coefficient of the SMF being slightly larger than that of silica, we can fuse the SMF and the HST with different lengths; thus, the sensor has a very low temperature coefficient of approximately 0.17 pm/°C.

  5. Drag reduction mechanism by microbubble injection within a channel boundary layer

    International Nuclear Information System (INIS)

    Ling Zhen; Hassan, Y.

    2005-01-01

    In this study, the drag reduction due to microbubble injection in the boundary layer of a fully developed turbulent channel flow was investigated. Particle Image Velocimetry (PIV) techniques were taken. The effects of the presence of microbubbles in the boundary layer were assessed. A drag reduction of 38.4% was obtained with void fraction of 4.9%. The algorithms of wavelet auto-correlation maps were applied to the PIV velocity field measurement. Modifications in the wavelet auto-correlation maps due to the presence of microbubbles were studied and compared in three-dimensions. By using 3-D plotting routines and the wavelet auto-correlation maps, it can be deduced from this study that the microbubble injection within the boundary layer increases the turbulent energy of the streamwise velocity components of the large scale (large eddy size, low frequency) range and decreases the energy of the small scale (small eddy size, high frequency) range. The wavelet auto-correlation maps of the normal velocities indicate that the microbubble presence decrease the turbulent energy of normal velocity components for both the large scale (large eddy size, low frequency) and the small scale (small eddy size, high frequency) ranges. (authors)

  6. High-speed optical observations and simulation results of SonoVue microbubbles at low-pressure insonation.

    Science.gov (United States)

    Chetty, Kevin; Stride, Eleanor; Sennoga, Charles A; Hajnal, Joseph V; Eckersley, Robert J

    2008-01-01

    Abstract-Modified Rayleigh-Plesset models are commonly used to characterize the acoustic response of microbubbles under ultrasound exposure. In most instances these models have been parameterized through acoustic measurements taken from bulk suspensions of microbubbles. The aim of this study was to parameterize the Hoff model for the commercial contrast agent SonoVue using optically observed oscillations from individual microbubbles recorded with a high-speed camera. The shell elasticity model term was tuned to fit simulation data to the measured oscillations while the shell viscosity parameter was held constant at 1 Pa??s. The results demonstrate a limited ability of the model to predict the microbubble behavior. The shell elasticity parameter was found to vary proportionally between 10 and 80 MPa with the initial microbubble diameter, implying the viscoelastic shell terms are not a constant property of the shell material. Further analysis using a moving window optimization to probe the microbubble responses suggests that the elasticity of the shell can increase by up to 50% over the course of insonation, particularly for microbubbles oscillating nearer to their resonant frequency. Microbubble oscillations were modeled more successfully by incorporating a varying elasticity term into the model.

  7. Optimization of transfection parameters for ultrasound/SonoVue microbubble-mediated hAng-1 gene delivery in vitro.

    Science.gov (United States)

    Zhou, Qing; Chen, Jin-Ling; Chen, Qian; Wang, Xiao; Deng, Qing; Hu, Bo; Guo, Rui-Qiang

    2012-12-01

    This study aimed to explore the effects of microbubble concentration, gene dosage, cell-microbubble mixing mode and fetal bovine serum (FBS) on gene delivery. 293T cells were transfected with Sonovue microbubbles carrying the hAng-1 gene via ultrasound irradiation. Various ultrasound exposure parameters and microbubble and DNA concentrations were investigated. In addition, FBS and the cell suspension or adherent mode was explored. Transfection efficiency and cell viability were used to determine the optimal transfection parameters. hAng-1 gene transfection efficiency gradually increased with elongation of ultrasound exposure and increasing microbubble concentration. However, if ultrasound irradiation exceeded 1.5 W/cm² and 30 sec or the microbubble concentration was over 20%, hAng-1 gene expression was significantly decreased, coupled with extensive cell death. Gene transfection levels were low under DNA concentrations less than 15 µg/ml. Furthermore, the gene transfer rate was significantly increased under cell suspension mode; FBS had no effect on hAng-1 gene transfection. The integrity of hAng-1 DNA was not affected by ultrasonic irradiation under optimal conditions. The optimal transfection parameters for the hAng-1 gene and Sonovue microbubble were ultrasound exposure of 1.5 W/cm² and 30 sec, 20% microbubbles, 15 µg/ml of DNA and under cell suspension mode.

  8. Dietary flavonoids bind to mono-ubiquitinated annexin A1 in nuclei, and inhibit chemical induced mutagenesis

    International Nuclear Information System (INIS)

    Hirata, Fusao; Harada, Takasuke; Corcoran, George B.; Hirata, Aiko

    2014-01-01

    Highlight: • Nuclear mono-ubiquitinated annexin A1 is involved in DNA damage induced mutagenesis. • Dietary flavonoids bind to and inhibit purified mono-ubiquitinated annexin A1 helicase. • Dietary flavonoids show anti-mutagenic action. • Annexin A1 may serve as a putative target of cancer chemoprevention by flavonoids. - Abstract: In order to investigate the mechanisms of anti-mutagenic action by dietary flavonoids, we investigated if they inhibit mutation of the thymidine kinase (tk) gene in L5178Ytk(±) lymphoma cells. Silibinin, quercetin and genistein suppressed mutation of the tk gene induced in L5178Ytk(±) lymphoma cells by methyl methanesulfonate (MMS) and As 3+ . Flavone and flavonol were less effective. To establish that mutation of the tk gene in L5178Ytk(±) lymphoma cells by MMS and As 3+ is mediated through mono-ubiquitinated annexin A1, L5178Ytk(±) lymphoma cells were treated with annexin A1 anti-sense oligonucleotide. The treatment reduced mRNA as well as protein levels of annexin A1, and suppressed mutation of the tk gene. Nuclear extracts from L5178Ytk(±) lymphoma cells catalyzed translesion DNA synthesis with an oligonucleotide template containing 8-oxo-guanosine in an annexin A1 dependent manner. This translesion DNA synthesis was inhibited by the anti-mutagenic flavonoids, silibinin, quercetin and genistein, in a concentration dependent manner, but only slightly by flavone and flavonol. Because these observations implicate involvement of annexin A1 in mutagenesis, we examined if flavonoids suppress nuclear annexin A1 helicase activity. Silibinin, quercetin and genistein inhibited ssDNA binding, DNA chain annealing and DNA unwinding activities of purified nuclear mono-ubiquitinated annexin A1. Flavone and flavonol were ineffective. The apparent direct binding of anti-mutagenic flavonoids to the annexin A1 molecule was supported by fluorescence quenching. Taken together, these findings illustrate that nuclear annexin A1 may be a novel

  9. Dietary flavonoids bind to mono-ubiquitinated annexin A1 in nuclei, and inhibit chemical induced mutagenesis

    Energy Technology Data Exchange (ETDEWEB)

    Hirata, Fusao, E-mail: fhirata@wayne.edu; Harada, Takasuke; Corcoran, George B.; Hirata, Aiko

    2014-01-15

    Highlight: • Nuclear mono-ubiquitinated annexin A1 is involved in DNA damage induced mutagenesis. • Dietary flavonoids bind to and inhibit purified mono-ubiquitinated annexin A1 helicase. • Dietary flavonoids show anti-mutagenic action. • Annexin A1 may serve as a putative target of cancer chemoprevention by flavonoids. - Abstract: In order to investigate the mechanisms of anti-mutagenic action by dietary flavonoids, we investigated if they inhibit mutation of the thymidine kinase (tk) gene in L5178Ytk(±) lymphoma cells. Silibinin, quercetin and genistein suppressed mutation of the tk gene induced in L5178Ytk(±) lymphoma cells by methyl methanesulfonate (MMS) and As{sup 3+}. Flavone and flavonol were less effective. To establish that mutation of the tk gene in L5178Ytk(±) lymphoma cells by MMS and As{sup 3+} is mediated through mono-ubiquitinated annexin A1, L5178Ytk(±) lymphoma cells were treated with annexin A1 anti-sense oligonucleotide. The treatment reduced mRNA as well as protein levels of annexin A1, and suppressed mutation of the tk gene. Nuclear extracts from L5178Ytk(±) lymphoma cells catalyzed translesion DNA synthesis with an oligonucleotide template containing 8-oxo-guanosine in an annexin A1 dependent manner. This translesion DNA synthesis was inhibited by the anti-mutagenic flavonoids, silibinin, quercetin and genistein, in a concentration dependent manner, but only slightly by flavone and flavonol. Because these observations implicate involvement of annexin A1 in mutagenesis, we examined if flavonoids suppress nuclear annexin A1 helicase activity. Silibinin, quercetin and genistein inhibited ssDNA binding, DNA chain annealing and DNA unwinding activities of purified nuclear mono-ubiquitinated annexin A1. Flavone and flavonol were ineffective. The apparent direct binding of anti-mutagenic flavonoids to the annexin A1 molecule was supported by fluorescence quenching. Taken together, these findings illustrate that nuclear annexin A1 may be

  10. Algae separation from urban landscape water using a high density microbubble layer enhanced by micro-flocculation.

    Science.gov (United States)

    Chen, Shuwen; Xu, Jingcheng; Liu, Jia; Wei, Qiaoling; Li, Guangming; Huang, Xiangfeng

    2014-01-01

    Eutrophication of raw water results in outbreaks of algae, which hinders conventional water treatment. In this study, high density microbubble layers combined with micro-flocculation was adopted to remove algae from urban landscape water, and the effects of pressure, hydraulic loading, microbubble layer height and flocculation dosage on the removal efficiency for algae were studied. The greatest removal efficiency for algae, chemical oxygen demand, nitrogen and phosphorus was obtained at 0.42 MPa with hydraulic loading at 5 m/h and a flocculation dosage of 4 mg/L using a microbubble layer with a height of 130 cm. Moreover, the size, clearance distance and concentration of microbubbles were found to be affected by pressure and the height of the microbubble layer. Based on the study, this method was an alternative for algae separation from urban landscape water and water purification.

  11. Ambient pressure sensitivity of microbubbles investigated through a parameter study

    DEFF Research Database (Denmark)

    Andersen, Klaus Scheldrup; Jensen, Jørgen Arendt

    2009-01-01

    . The behavior of two microbubbles corresponding to two different contrast agents was investigated as a function of driving pulse and ambient overpressure, pov. Simulations of Levovist using a rectangular driving pulse show an almost linear reduction in the subharmonic component as pov is increased. For a 20....... 1999, who found a linear reduction of 9.6 dB. Further simulations of Levovist show that also the shape and the acoustic pressure of the driving pulse are very important factors. The best pressure sensitivity of Levovist was found to be 0.88 dB/kPa. For Sonazoid, a sensitivity of 1.14 dB/kPa has been...... cycles driving pulse, a reduction of 4.6 dB is observed when changing pov from 0 to 25 kPa. Increasing the pulse duration makes the reduction even more clear. For a pulse with 64 cycles, the reduction is 9.9 dB. This simulation is in good correspondence with measurement results presented by Shi et al...

  12. CELL DEATH DIFFERENTIATION IN BLACK HEADED RAMS SPERMATOZOA, USING FLUORESCENT LABELED ANNEXIN V

    Directory of Open Access Journals (Sweden)

    M Ivanova-Kicheva

    2008-11-01

    Full Text Available Double staining kit of Annexin V Cy3.18/6-CFDA was used to investigate the changes in phospholipide asymmetry after treating sperm cells with dexamethasone. The % of spermatozoa with registered translocation of PS in treated with dexamethazone groups at the 10-th min and in control no treated varied from 2.74%±0.65 to 2.30%±0.89. After the 5 hour of incubation these % increased to 39.83±3.33 for the treated group and 23.44±1.12 for the control. It was concluded that Annexin V binding assay is more sensitive in the detection of deterioration in membrane function than other conventional methods such as motility analysis and supravital techniques.

  13. Impact of Annexin A 7 Deficiency on FGF23 Plasma Concentrations

    Directory of Open Access Journals (Sweden)

    Anja T. Umbach

    2016-11-01

    Full Text Available Background/Aims: The release of fibroblast growth factor FGF23, a powerful regulator of 1,25(OH2D3 formation and mineral metabolism, is stimulated by store-operated Ca2+ entry (SOCE, which is accomplished by the pore forming Ca2+ release activated channel protein Orai1. Regulators of Orai1 and thus FGF23 release include serum & glucocorticoid inducible kinase SGK1, a kinase up-regulated by glucocorticosteroids. Some effects of glucocorticoids require the presence of annexin A7, such as suppression of prostaglandin E2 in gastric glands. The present study thus explored whether annexin A7 impacts on FGF23 plasma levels. Methods: Comparisons were made between gene targeted mice lacking functional annexin A7 (Anx7-/- and their wild type littermates (Anx7+/+. Serum C-terminal-FGF23, intact FGF23, 1,25(OH2D3 and PTH concentrations were measured by ELISA or EIA. The serum and urinary phosphate concentrations were measured by colorimetry, the serum Ca2+ concentration and the urinary Ca2+ concentration by flame photometry. Results: Serum C-terminal FGF23 levels and corticosterone levels were significantly higher and serum 1,25(OH2 D3 and PTH levels were significantly lower in Anx7-/- than in Anx7+/+ mice. Water intake was slightly but significantly higher in Anx7-/- mice than in Anx7+/+ mice. No significant difference was observed between Anx7-/- and Anx7+/+ mice in urinary fluid excretion, plasma Ca2+ concentration, plasma phosphate concentration and urinary Ca2+ output. The urinary phosphate output was significantly lower in Anx7-/- mice than in Anx7+/+ mice. Conclusion: Annexin A7 deficiency upregulates FGF23 plasma levels, an effect paralleled by increased corticosterone plasma levels, as well as decreased 1,25(OH2 D3 and PTH plasma levels.

  14. Annexin 1 and Melanocortin Peptide Therapy for Protection Against Ischaemic-Reperfusion Damage in the Heart

    Directory of Open Access Journals (Sweden)

    F.N.E. Gavins

    2006-01-01

    Full Text Available Cardiovascular disease is a major cause of mortality within the western world affecting 2.7 million British people. This review highlights the beneficial effects of naturally occurring hormones and their peptides, in myocardial ischaemic-injury (MI models, a disease pathology in which cytokines and neutrophils play a causal role. Here we discuss two distinct classes of endogenous peptides: the steroid inducible annexin 1 and the melanocortin peptides. Annexin 1 and the melanocortins counteract the most important part of the host inflammatory response, namely, the process of leukocyte extravasation, as well as release of proinflammatory mediators. Their biological effects are mediated via the seven transmembrane G-protein-coupled receptors, the fMLP receptor family (or FPR, and the melanocortin receptors, respectively. Pharmacological analysis has demonstrated that the first 24 amino acids of the N-terminus (termed Ac2-26 are the most active region. Both exogenous annexin 1 and its peptides demonstrate cardioprotectiveness and continuing work is required to understand this annexin 1/FPR relationship fully. The melanocortin peptides are derived from a precursor molecule called the POMC protein. These peptides display potent anti-inflammatory effects in human and animal models of disease. In MI, the MC3R has been demonstrated to play an important role in mediating the protective effects of these peptides. The potential anti-inflammatory role for endogenous peptides in cardiac disease is in its infancy. The inhibition of cell migration and release of cytokines and other soluble mediators appears to play an important role in affording protection in ischaemic injury and thus may lead to potential therapeutic targets.

  15. Micro-bubble morphologies following drop impacts onto a pool surface

    KAUST Repository

    Thoroddsen, Sigurdur T.

    2012-10-01

    When a drop impacts at low velocity onto a pool surface, a hemispheric air layer cushions and can delay direct contact. Herein we use ultra-high-speed video to study the rupture of this layer, to explain the resulting variety of observed distribution of bubbles. The size and distribution of micro-bubbles is determined by the number and location of the primary punctures. Isolated holes lead to the formation of bubble necklaces when the edges of two growing holes meet, whereas bubble nets are produced by regular shedding of micro-bubbles from a sawtooth edge instability. For the most viscous liquids the air film contracts more rapidly than the capillary-viscous velocity through repeated spontaneous ruptures of the edge. From the speed of hole opening and the total volume of micro-bubbles we conclude that the air sheet ruptures when its thickness approaches ?100.

  16. Ultrasound-Stimulated Mutual Interaction Forces between Optically Configured Micro-Bubble Pairs

    Science.gov (United States)

    Prentice, Paul A.; Campbell, Paul A.

    2007-05-01

    The mutual interaction between two oscillating encapsulated microbubbles was investigated using a novel optical trapping arrangement. This approach facilitated the development of an arbitrary, stable, initial spatial configuration for a two-bubble system. Critically, exercising optical control over such a binary bubble system meant that it could be isolated from the resident population of microbubbles during exposure to ultrasound. This ensured that any early stage dynamical evolution of the system was dominated by the mutual interaction of the two bubbles in view, rather than any extraneous influence arising from `crosstalk' with the rest of the bubble population. We observed, using high speed microphotography at 4×105 frames per second, that the action of secondary radiation forces leads to mutual bubble attraction. Phenomena such as coalescence and `bounce' were observed. Estimates of the microbubble [Sonovue™] compressibility could also be made, and tally well with published values obtained for other shelled contrast agents.

  17. Modulation of the molecular arrangement in artificial and biological membranes by phospholipid-shelled microbubbles.

    Science.gov (United States)

    Carugo, Dario; Aron, Miles; Sezgin, Erdinc; Bernardino de la Serna, Jorge; Kuimova, Marina K; Eggeling, Christian; Stride, Eleanor

    2017-01-01

    The transfer of material from phospholipid-coated microbubbles to cell membranes has been hypothesized to play a role in ultrasound-mediated drug delivery. In this study, we employed quantitative fluorescence microscopy techniques to investigate this phenomenon in both artificial and biological membrane bilayers in an acoustofluidic system. The results of the present study provide strong evidence for the transfer of material from microbubble coatings into cell membranes. Our results indicate that transfer of phospholipids alters the organization of molecules in cell membranes, specifically the lipid ordering or packing, which is known to be a key determinant of membrane mechanical properties, protein dynamics, and permeability. We further show that polyethylene-glycol, used in many clinical microbubble formulations, also has a major impact on both membrane lipid ordering and the extent of lipid transfer, and that this occurs even in the absence of ultrasound exposure. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  18. Transit of micro-bubbles through the pulmonary circulation of Thoroughbred horses during exercise.

    Science.gov (United States)

    La Gerche, A; Daffy, J R; Mooney, D J; Forbes, G; Davie, A J

    2013-10-01

    It has been observed that microbubbles may pass through the pulmonary circulation of dogs and humans during exercise. In humans, this phenomenon has been associated with lower pulmonary artery pressures, enhanced right ventricular function and greater exercise capacity. In the exercising Thoroughbred horse, extraordinarily high cardiac outputs exert significant pulmonary vascular stresses. The aim of this study was to determine, using contrast echocardiography, whether Thoroughbred horses performing strenuous exercise developed pulmonary transit of agitated contrast microbubbles (PTAC). At rest, agitated contrast was observed in the right ventricle, but not in the left ventricle. However, post-exercise microbubbles were observed in the left ventricle, confirming the occurrence of PTAC with exercise but not at rest. Further investigation is warranted to investigate whether this phenomenon may be associated with superior physiology and performance measures as has been implicated in other species. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Autoantibodies targeting glomerular annexin A2 identify patients with proliferative lupus nephritis.

    Science.gov (United States)

    Caster, Dawn J; Korte, Erik A; Merchant, Michael L; Klein, Jon B; Wilkey, Daniel W; Rovin, Brad H; Birmingham, Dan J; Harley, John B; Cobb, Beth L; Namjou, Bahram; McLeish, Kenneth R; Powell, David W

    2015-12-01

    Patients with systemic lupus erythematosus (SLE) frequently develop lupus nephritis (LN), a complication frequently leading to end stage kidney disease. Immune complex deposition in the glomerulus is central to the development of LN. Using a targeted proteomic approach, we tested the hypothesis that autoantibodies targeting glomerular antigens contribute to the development of LN. Human podocyte and glomerular proteins were separated by SDS-PAGE and immunoblotted with sera from SLE patients with and without LN. The regions of those gels corresponding to reactive bands observed with sera from LN patients were analyzed using LC-MS/MS. LN reactive bands were seen at approximately 50 kDa in podocyte extracts and between 36 and 50 kDa in glomerular extracts. Those bands were analyzed by LC-MS/MS and 102 overlapping proteins were identified. Bioinformatic analysis determined that 36 of those proteins were membrane associated, including a protein previously suggested to contribute to glomerulonephritis and LN, annexin A2. By ELISA, patients with proliferative LN demonstrated significantly increased antibodies against annexin A2. Proteomic approaches identified multiple candidate antigens for autoantibodies in patients with LN. Serum antibodies against annexin A2 were significantly elevated in subjects with proliferative LN, validating those antibodies as potential biomarkers. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Reversible and irreversible vascular bioeffects induced by ultrasound and microbubbles in chorioallantoic membrane model

    Science.gov (United States)

    Tarapacki, Christine; Kuebler, Wolfgang M.; Tabuchi, Arata; Karshafian, Raffi

    2017-03-01

    Background: The application of ultrasound and microbubbles at therapeutic conditions has been shown to improve delivery of molecules, cause vasoconstriction, modulate blood flow and induce a vascular shut down in in vivo cancerous tissues. The underlying mechanism has been associated with the interaction of ultrasonically-induced microbubble oscillation and cavitation with the blood vessel wall. In this study, the effect of ultrasound and microbubbles on blood flow and vascular architecture was studied using a fertilized chicken egg CAM (chorioallantoic membrane) model. Methods: CAM at day 12 of incubation (Hamburger-Hamilton stage 38-40) were exposed to ultrasound at varying acoustic pressures (160, 240 and 320 kPa peak negative pressure) in the presence of Definity microbubbles and 70 kDa FITC dextran fluorescent molecules. A volume of 50 µL Definity microbubbles were injected into a large anterior vein of the CAM prior to ultrasound exposure. The ultrasound treatment sequence consisted of 5 s exposure at 500 kHz frequency, 8 cycles and 1 kHz pulse repetition frequency with 5 s off for a total exposure of 2 minutes. Fluorescent videos and images of the CAM vasculature were acquired using intravital microscopy prior, during and following the ultrasound exposure. Perfusion was quantified by measuring the length of capillaries in a region of interest using Adobe Illustrator. Results and Discussion: The vascular bioeffects induced by USMB increased with acoustic peak negative pressure. At 160 kPa, no visible differences were observed compared to the control. At 240 kPa, a transient decrease in perfusion with subsequent recovery within 15 minutes was observed, whereas at 320 kPa, the fluorescent images showed an irreversible vascular damage. The study indicates that a potential mechanism for the transient decrease in perfusion may be related to blood coagulation. The results suggest that ultrasound and microbubbles can induce reversible and irreversible vascular

  1. Drag reduction by microbubbles in turbulent flows: the limit of minute bubbles.

    Science.gov (United States)

    L'vov, Victor S; Pomyalov, Anna; Procaccia, Itamar; Tiberkevich, Vasil

    2005-05-06

    Drag reduction by microbubbles is a promising engineering method for improving ship performance. A fundamental theory of the phenomenon is lacking, however, making actual design quite haphazard. We offer here a theory of drag reduction by microbubbles in the limit of very small bubbles, when the effect of the bubbles is mainly to normalize the density and the viscosity of the carrier fluid. The theory culminates with a prediction of the degree of drag reduction given the concentration profile of the bubbles. Comparisons with experiments are discussed and the road ahead is sketched.

  2. Cavitation dynamics and directional microbubble ejection induced by intense femtosecond laser pulses in liquids

    Science.gov (United States)

    Faccio, D.; Tamošauskas, G.; Rubino, E.; Darginavičius, J.; Papazoglou, D. G.; Tzortzakis, S.; Couairon, A.; Dubietis, A.

    2012-09-01

    We study cavitation dynamics when focusing ring-shaped femtosecond laser beams in water. This focusing geometry reduces detrimental nonlinear beam distortions and enhances energy deposition within the medium, localized at the focal spot. We observe remarkable postcollapse dynamics of elongated cavitation bubbles with high-speed ejection of microbubbles out of the laser focal region. Bubbles are ejected along the laser axis in both directions (away and towards the laser). The initial shape of the cavitation bubble is also seen to either enhance or completely suppress jet formation during collapse. In the absence of jetting, microbubble ejection occurs orthogonal to the laser propagation axis.

  3. Cavitation dynamics and directional microbubble ejection induced by intense femtosecond laser pulses in liquids.

    Science.gov (United States)

    Faccio, D; Tamošauskas, G; Rubino, E; Darginavičius, J; Papazoglou, D G; Tzortzakis, S; Couairon, A; Dubietis, A

    2012-09-01

    We study cavitation dynamics when focusing ring-shaped femtosecond laser beams in water. This focusing geometry reduces detrimental nonlinear beam distortions and enhances energy deposition within the medium, localized at the focal spot. We observe remarkable postcollapse dynamics of elongated cavitation bubbles with high-speed ejection of microbubbles out of the laser focal region. Bubbles are ejected along the laser axis in both directions (away and towards the laser). The initial shape of the cavitation bubble is also seen to either enhance or completely suppress jet formation during collapse. In the absence of jetting, microbubble ejection occurs orthogonal to the laser propagation axis.

  4. Laser-induced microbubbles in gold and oxide nanoparticle suspensions: photoacoustic detection

    Science.gov (United States)

    Zhao, Z.; Myllylä, R.

    2010-11-01

    A tunable pulsed laser with nano-second pulse duration is used to generate microbubbles in highly diluted nanoparticle (Au, TiO2 and ZnO) suspensions. The microbubble explosion may produce shock wave which is in-phase detected by a low-frequency piezoelectric transducer. The effects of particle size and category on the threshold laser fluence of shock wave generation and the wave intensity are investigated. The interaction between laser and nanoparticles has significant application in biomedicine such as photothermal diagnostic and therapy, as well as cosmetic or drug delivery in skin.

  5. Carcinogenic heavy metals replace Ca2+ for DNA binding and annealing activities of mono-ubiquitinated annexin A1 homodimer.

    Science.gov (United States)

    Hirata, Aiko; Corcoran, George B; Hirata, Fusao

    2010-10-01

    Mono-ubiquitinated annexin A1 was purified from rat liver nuclei. The homodimer form of mono-ubiquitinated annexin A1 was able to unwind dsDNA in a Mg(2+)- and ATP-dependent manner, and to anneal ssDNA in a Ca(2+)-dependent manner. Phospholipids decreased the concentration of Ca(2+) required for maximal annealing activity. Heavy metals such as As(3+), Cr(6+), Pb(2+) and Cd(2+) substituted for Ca(2+) in the ssDNA binding and annealing activities of annexin A1. While these metals inhibited the unwinding of dsDNA by nuclear annexin A1 in the presence of Mg(2+) and ATP, they enhanced dsDNA-dependent ATPase activity of annexin A1. Heavy metals may have produced dsDNA, a substrate for the DNA unwinding reaction, via the DNA annealing reaction. DNA synthesomes were isolated from L5178Y tk(+/-) mouse lymphoma cells in exponential growth, and were found to contain helicase activities. The As(3+)- or Cr(6+)-induced increases in ssDNA binding activity of DNA synthesomes were reduced by a mono-specific anti-annexin A1 antibody, but not by anti-Ig antibody. Anti-annexin A1 antibody also blocked the inhibitory and stimulatory effects of As(3+) or Cr(6+) towards DNA unwinding and annealing activities of DNA synthesomes. Based on these observations, it can be concluded that the effects of heavy metals on DNA annealing and unwinding activities are mediated, at least in substantial part, through actions of the mono-ubiquitinated annexin A1 homodimer. Copyright (c) 2010 Elsevier Inc. All rights reserved.

  6. Iodine-124 labelled Annexin-V as a potential radiotracer to study apoptosis using positron emission tomography

    Energy Technology Data Exchange (ETDEWEB)

    Glaser, Matthias E-mail: m.glaser@csc.mrc.ac.uk; Collingridge, D.R.; Aboagye, E.O.; Bouchier-Hayes, Lisa; Hutchinson, O. Clyde; Martin, S.J.; Price, Pat; Brady, Frank; Luthra, S.K

    2003-01-01

    Annexin-V is a calcium-dependent protein that binds with high affinity to phosphaditylserine exposed during apoptosis. The aim of this study was to radiolabel annexin-V with iodine-124 for use as a potential probe of apoptosis by positron emission tomography. Annexin-V was radioiodinated directly using the cyclotron-produced positron emitter iodine-124 by the chloramine-T (CAT) method and indirectly by the pre-labelled reagent N-succinimidyl 3-[{sup 124}I]iodobenzoate ([{sup 124}I]m-SIB). Some reaction parameters of the CAT method such as reaction time and pH were optimised to give radiochemical yields of 22.3{+-}2.6% (n=3, gel-filtration). After incubation with [{sup 124}I]m-SIB, radiolabelled annexin-V was obtained in 14% and 25% yield by FPLC and gel-filtration, respectively. The radiochemical purities from direct and indirect labelling were 97.7{+-}1.0% (n=3) and 96.7{+-}2.1% (n=3), respectively. The new radiotracers could be stored for up to four days without significant de-iodination. The biological activity of radiolabelled annexin-V was tested in control and camptothecin-treated (i.e. apoptotic) human leukaemic HL60 cells. A significantly higher (21%) binding in treated cells was observed with [{sup 125}I]m-SIB-annexin-V. The binding of [{sup 125}I]m-SIB labelled annexin-V to camptothecin treated cells was blocked (68%) by a 100-fold excess of unlabelled annexin-V. Abbreviations: Fast protein liquid chromatography (FPLC), Instant thin layer chromatography (ITLC), Sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE), 3-iodobenzoate (m-IBA), N-succinimidyl 3-(trimethylstannyl)benzoate (m-MeATE)

  7. Iodine-124 labelled annexin-V as a potential radiotracer to study apoptosis using positron emission tomography.

    Science.gov (United States)

    Glaser, Matthias; Collingridge, David R; Aboagye, Eric O; Bouchier-Hayes, Lisa; Hutchinson, O Clyde; Martin, Seamus J; Price, Pat; Brady, Frank; Luthra, Sajinder K

    2003-01-01

    Annexin-V is a calcium-dependent protein that binds with high affinity to phosphaditylserine exposed during apoptosis. The aim of this study was to radiolabel annexin-V with iodine-124 for use as a potential probe of apoptosis by positron emission tomography. Annexin-V was radioiodinated directly using the cyclotron-produced positron emitter iodine-124 by the chloramine-T (CAT) method and indirectly by the pre-labelled reagent N-succinimidyl 3-[124I]iodobenzoate ([124I]m-SIB). Some reaction parameters of the CAT method such as reaction time and pH were optimised to give radiochemical yields of 22.3 +/- 2.6%(n = 3, gel-filtration). After incubation with [124I]m-SIB, radiolabelled annexin-V was obtained in 14% and 25% yield by FPLC and gel-filtration, respectively. The radiochemical purities from direct and indirect labelling were 97.7 +/- 1.0%(n = 3) and 96.7 +/- 2.1%(n = 3), respectively. The new radiotracers could be stored for up to four days without significant de-iodination. The biological activity of radiolabelled annexin-V was tested in control and camptothecin-treated (i.e. apoptotic) human leukaemic HL60 cells. A significantly higher (21%) binding in treated cells was observed with [125I]m-SIB-annexin-V. The binding of [125I]m-SIB labelled annexin-V to camptothecin treated cells was blocked (68%) by a 100-fold excess of unlabelled annexin-V. Crown Copyright 2002 Published by Elsevier Science Ltd.

  8. Annexin A2 in amniotic fluid: correlation with histological chorioamnionitis, preterm premature rupture of membranes, and subsequent preterm delivery.

    Science.gov (United States)

    Namba, Fumihiko; Ina, Shihomi; Kitajima, Hiroyuki; Yoshio, Hiroyuki; Mimura, Kazuya; Saito, Shigeru; Yanagihara, Itaru

    2012-01-01

    The aim of this study was to determine whether amniotic fluid levels of annexin A2, a phospholipid-binding protein that is abundant in amnion and regulates fibrin homeostasis, are associated with histological chorioamnionitis, preterm premature rupture of the membranes, and subsequent preterm delivery. Amniotic fluid was obtained from 55 pregnant women with preterm labor and/or preterm premature rupture of the membranes before 32weeks of gestation, and amniotic fluid levels of annexin A2 were measured with a sandwich enzyme-linked immunosorbent assay. Amniotic fluid levels of annexin A2 in patients with histological chorioamnionitis was higher than that in the remainder (P=0.053), whereas amniotic fluid levels of annexin A2 in patients with preterm premature rupture of the membranes was significantly higher than that in the remainder (P=0.002). Amniotic levels of annexin A2 was a fair test (area under receiver-operator characteristic curve=0.679), and amniotic fluid levels of annexin A2>878.2ng/mL had a sensitivity of 68.8%, a specificity of 65.2%, a positive predictive value of 73.3%, and a negative predictive value of 60.0% for predicting delivery within 2weeks after amniotic fluid sampling. Furthermore, the combined use of amniotic fluid cut-off levels of 878.2ng/mL for annexin A2 and 13.3ng/mL for interleukin-8 improved the specificity (91.3%) and the positive predictive value (89.5%). We identified amniotic fluid levels of annexin A2, especially in combination with amniotic fluid levels of interleukin-8, as a novel predictive marker for preterm delivery. © 2011 The Authors. Journal of Obstetrics and Gynaecology Research © 2011 Japan Society of Obstetrics and Gynecology.

  9. Characteristics of phenomenon and sound in microbubble emission boiling

    International Nuclear Information System (INIS)

    Zhu Guangyu; Sun Licheng; Tang Jiguo

    2014-01-01

    Background: Nowadays, the efficient heat transfer technology is required in nuclear energy. Therefore, micro-bubble emission boiling (MEB) is getting more attentions from many researchers due to its extremely high heat-transfer dissipation capability. Purpose: An experimental setup was built up to study the correspondences between the characteristics on the amplitude spectrum of boiling sound in different boiling modes. Methods: The heat element was a copper block heated by four Si-C heaters. The upper of the copper block was a cylinder with the diameter of 10 mm and height of 10 mm. Temperature data were measured by three T-type sheathed thermocouples fitted on the upper of the copper block and recorded by NI acquisition system. The temperature of the heating surface was estimated by extrapolating the temperature distribution. Boiling sound data were acquired by hydrophone and processed by Fourier transform. Bubble behaviors were captured by high-speed video camera with light system. Results: In nucleate boiling region, the boiling was not intensive and as a result, the spectra didn't present any peak. While the MEB fully developed on the heating surface, an obvious peak came into being around the frequency of 300 Hz. This could be explained by analyzing the video data. The periodic expansion and collapse into many extremely small bubbles of the vapor film lead to MEB presenting an obvious characteristic peak in its amplitude spectrum. Conclusion: The boiling mode can be distinguished by its amplitude spectrum. When the MEB fully developed, it presented a characteristic peak in its amplitude spectrum around the frequency between 300-400 Hz. This proved that boiling sound of MEB has a close relation with the behavior of vapor film. (authors)

  10. Annexin A1 N-terminal derived peptide Ac2-26 stimulates fibroblast migration in high glucose conditions.

    Directory of Open Access Journals (Sweden)

    Valentina Bizzarro

    Full Text Available Deficient wound healing in diabetic patients is very frequent, but the cellular and molecular causes are poorly defined. In this study, we have evaluated whether Annexin A1 derived peptide Ac2-26 stimulates fibroblast migration in high glucose conditions. Using normal human skin fibroblasts WS1 in low glucose (LG or high glucose (HG we observed the enrichment of Annexin A1 protein at cell movement structures like lamellipodial extrusions and interestingly, a significant decrease in levels of the protein in HG conditions. The analysis of the translocation of Annexin A1 to cell membrane showed lower levels of Annexin A1 in both membrane pool and supernatants of WS1 cells treated with HG. Wound-healing assays using cell line transfected with Annexin A1 siRNAs indicated a slowing down in migration speed of cells suggesting that Annexin A1 has a role in the migration of WS1 cells. In order to analyze the role of extracellular Annexin A1 in cell migration, we have performed wound-healing assays using Ac2-26 showing that peptide was able to increase fibroblast cell migration in HG conditions. Experiments on the mobilization of intracellular calcium and analysis of p-ERK expression confirmed the activity of the FPR1 following stimulation with the peptide Ac2-26. A wound-healing assay on WS1 cells in the presence of the FPR agonist fMLP, of the FPR antagonist CsH and in the presence of Ac2-26 indicated that Annexin A1 influences fibroblast cell migration under HG conditions acting through FPR receptors whose expression was slightly increased in HG. In conclusion, these data demonstrate that (i Annexin A1 is involved in migration of WS1 cells, through interaction with FPRs; (ii N- terminal peptide of Annexin A1 Ac2-26 is able to stimulate direct migration of WS1 cells in high glucose treatment possibly due to the increased receptor expression observed in hyperglycemia conditions.

  11. PBCA-based polymeric microbubbles for molecular imaging and drug delivery

    NARCIS (Netherlands)

    Koczera, Patrick; Appold, Lia; Shi, Yang; Liu, Mengjiao; Dasgupta, Anshuman; Pathak, Vertika; Ojha, Tarun; Fokong, Stanley; Wu, Zhuojun; Van Zandvoort, Marc; Iranzo, Olga; Kuehne, Alexander J C; Pich, Andrij; Kiessling, Fabian; Lammers, Twan

    2017-01-01

    Microbubbles (MB) are routinely used as contrast agents for ultrasound (US) imaging. We describe different types of targeted and drug-loaded poly(n-butyl cyanoacrylate) (PBCA) MB, and demonstrate their suitability for multiple biomedical applications, including molecular US imaging and US-mediated

  12. Image-guided, targeted and triggered drug delivery to tumors using polymer-based microbubbles.

    NARCIS (Netherlands)

    Fokong, S.; Theek, B.; Koczera, P.; Appold, L.; Resch-Genger, U.; van Zandvoort, M.; Storm, Gerrit; Kiessling, F.; Lammers, Twan Gerardus Gertudis Maria

    2012-01-01

    Abstract Microbubbles (MB) are routinely used contrast agents for functional and molecular ultrasound (US) imaging. In addition, they have been attracting more and more attention for drug delivery purposes, enabling e.g. US-mediated drug delivery across biological barriers and US-induced triggered

  13. Theranastic USPIO-loaded microbubbles for mediating and monitoring blood-brain barrier permeation

    NARCIS (Netherlands)

    Lammers, Twan Gerardus Gertudis Maria; Koczera, Patrick; Fokong, Stanley; Gremse, Felix; Ehling, Josef; Vogt, Michael; Pich, Andrij; Storm, Gerrit; van Zandvoort, Marc; Kiessling, Fabian

    2015-01-01

    Efficient and safe drug delivery across the blood-brain barrier (BBB) remains one of the major challenges of biomedical and (nano-) pharmaceutical research. Here, it is demonstrated that poly(butyl cyanoacrylate)-based microbubbles (MB), carrying ultrasmall superparamagnetic iron oxide (USPIO)

  14. Theranostic USPIO-loaded microbubbles for mediating and monitoring blood-brain barrier permeation

    NARCIS (Netherlands)

    Lammers, Twan; Koczera, Patrick; Fokong, Stanley; Gremse, Felix; Ehling, Josef; Vogt, Michael; Pich, Andrij; Storm, G; Van Zandvoort, Marc; Kiessling, Fabian

    2015-01-01

    Efficient and safe drug delivery across the blood-brain barrier (BBB) remains one of the major challenges of biomedical and (nano-) pharmaceutical research. Here, it is demonstrated that poly(butyl cyanoacrylate)-based microbubbles (MB), carrying ultrasmall superparamagnetic iron oxide (USPIO)

  15. Resonance frequency of microbubbles in small blood vessels: a numerical study

    International Nuclear Information System (INIS)

    Sassaroli, E; Hynynen, K

    2005-01-01

    Microbubbles are currently used as ultrasound contrast agents. Their potential therapeutic applications are also under investigation. This work is designed to provide some insight into the mechanisms of energy absorption and deposition by a preformed gas bubble in the microvasculature to optimize its efficacy. In the linear regime, the most favourable condition for the transfer of energy from an ultrasonic field to a gas bubble occurs when the centre frequency of the ultrasonic field equals the resonance frequency of the bubble. The resonance frequency of gas microbubbles has been investigated up to now mainly in unbounded liquids; however when bubbles are confined in small regions, their resonance frequency is strongly affected by the surrounding boundaries. A parametric study on how the resonance frequency of microbubbles in blood vessels is affected by the bubble radius, vessel radius and the bubble position in the vessel is presented. The resonance frequency decreases below its free value with decreasing vessel radius for vessels smaller than 200-300 μm depending on the bubble size. This model suggests the possibility of using ultrasound in a range of frequencies that are, in general, lower than the ones used now for therapeutic and diagnostic applications of ultrasound (a few MHz). When microbubbles oscillate at their resonance frequency they absorb and therefore emit more energy. This energy may allow specific blood vessels to be targeted for both diagnostic and therapeutic applications of ultrasound

  16. Enhanced gene expression of systemically administered plasmid DNA inthe liver with therapeutic ultrasound and microbubbles

    NARCIS (Netherlands)

    Raju, B.I.; Leyvi, E.; Seip, R.; Sethuraman, S.; Luo, X.; Bird, A.; Li, S.; Koeberl, D.

    2012-01-01

    Ultrasound mediated delivery (USMD) of novel therapeutic agents in the presence of microbubbles is a potentially safe and effective method for gene therapy offering many desired characteristics such as low toxicity, potential for repeated treatment, and organ specificity.In this study we tested the

  17. Advances in ultrasound-targeted microbubble-mediated gene therapy for liver fibrosis

    Directory of Open Access Journals (Sweden)

    Cuiyuan Huang

    2017-07-01

    Full Text Available Hepatic fibrosis develops as a wound-healing scar in response to acute and chronic liver inflammation and can lead to cirrhosis in patients with chronic hepatitis B and C. The condition arises due to increased synthesis and reduced degradation of extracellular matrix (ECM and is a common pathological sequela of chronic liver disease. Excessive deposition of ECM in the liver causes liver dysfunction, ascites, and eventually upper gastrointestinal bleeding as well as a series of complications. However, fibrosis can be reversed before developing into cirrhosis and has thus been the subject of extensive researches particularly at the gene level. Currently, therapeutic genes are imported into the damaged liver to delay or prevent the development of liver fibrosis by regulating the expression of exogenous genes. One technique of gene delivery uses ultrasound targeting of microbubbles combined with therapeutic genes where the time and intensity of the ultrasound can control the release process. Ultrasound irradiation of microbubbles in the vicinity of cells changes the permeability of the cell membrane by its cavitation effect and enhances gene transfection. In this paper, recent progress in the field is reviewed with emphasis on the following aspects: the types of ultrasound microbubbles, the construction of an ultrasound-mediated gene delivery system, the mechanism of ultrasound microbubble–mediated gene transfer and the application of ultrasound microbubbles in the treatment of liver fibrosis.

  18. Decontamination System Development of Radioative Activated Carbon using Micro-bubbles

    Energy Technology Data Exchange (ETDEWEB)

    Jeon, Jong seon; Kim, Wi soo [NESS, Daejeon (Korea, Republic of); Han, Byoung sub. [Enesys Co., Daejeon (Korea, Republic of)

    2016-10-15

    This study was aimed to develop a decontamination system by applying such technical characteristics that minimizes a generation of secondary wastes while decontaminating radiation wastes. The radioactive activated carbon is removed from the end-of-life air cleaning filter in replacement or decommission of nuclear power plant or nuclear facility. By removing radioactive activated carbon, the filter would be classified as a low radioactive contaminant. And thus the amount of radioactive wastes and the treatment cost would be decreased. We are in development of the activated carbon cleaning technique by utilizing micro-bubbles, which improve efficiency and minimize damage of activated carbon. The purpose of using micro-bubbles is to decontamination carbon micropore, which is difficult to access, by principle of cavitation phenomenon generated in collapse of micro-bubbles. In this study, we introduced the micro-bubble decontamination system developed to decontaminate activated carbon. For further researches, we will determine carbon weight change and the decontamination rate under the experimental conditions such as temperature and pH.

  19. Microbubble-induced detachment of coadhering oral bacteria from salivary pellicles

    NARCIS (Netherlands)

    Sharma, PK; Gibcus, MJ; van der Mei, HC; Busscher, HJ

    The presence and maturity of the salivary pellicle influences microbial adhesion and its tenacity in the oral cavity, posing a challenge to different plaque-control systems. Some plaque-control systems rely on surface-tension forces arising from passing microbubbles sprayed over the pellicle.

  20. Ultrasound-targeted microbubble destruction enhances naked plasmid DNA transfection in rabbit Achilles tendons in vivo.

    Science.gov (United States)

    Qiu, L; Zhang, L; Wang, L; Jiang, Y; Luo, Y; Peng, Y; Lin, L

    2012-07-01

    The study was to investigate the probability of increasing the transfection of the gene in tendons by ultrasound-targeted microbubble destruction (UTMD), and to search for the most suitable transfection conditions. A mixture of microbubbles and enhanced green fluorescent protein (EGFP) plasmids was injected into rabbit Achilles tendons by different administration routes and the tendons were ultrasound pulse by different ultrasonic conditions in order to determine the most appropriate conditions. Then, the rabbits were divided into four groups: (1) ultrasound + microbubbles + plasmid; (2) ultrasound+ plasmid; (3) microbubble + plasmid; (4) plasmid only. EGFP expression in the tendons and other tissues, and the damage to tendon and paratenon were all observed. The results showed that EGFP expression in the tendon was higher by ultrasound pulse with 2 W cm(-2) of output intensity and a 20% duty cycle for 10 min. Local injection was determined to be the better administration route. Among the four groups, EGFP expression in Group 1 was higher than that in other groups. EGFP expression was highest on seventh day, then it gradually decrease over time, and lasted more than 56 days. EGFP expression was not found in other tissues. There was no obvious injury caused by UTMD. Under suitable conditions, it is feasible to use UTMD as a safe and effective gene transfection therapy for tendon injuries.

  1. Increase of intracellular cisplatin levels and radiosensitization by ultrasound in combination with microbubbles

    NARCIS (Netherlands)

    Lammertink, Bart H A; Bos, Clemens; van der Wurff-Jacobs, Kim M.; Storm, G; Moonen, Chrit T.; Deckers, Roel

    2016-01-01

    The possibility to enhance drug delivery by using ultrasound in combination with microbubbles (USMB) is extensively studied. So far, these studies have focused on the delivery and efficacy of a single drug, e.g. in chemotherapy. In this study, we investigated the intracellular delivery of cisplatin

  2. Robust Whispering-Gallery-Mode Microbubble Lasers from Colloidal Quantum Dots.

    Science.gov (United States)

    Wang, Yue; Ta, Van Duong; Leck, Kheng Swee; Tan, Beng Hau Ian; Wang, Zeng; He, Tingchao; Ohl, Claus-Dieter; Demir, Hilmi Volkan; Sun, Handong

    2017-04-12

    Microlasers hold great promise for the development of photonics and optoelectronics. Among the discovered optical gain materials, colloidal quantum dots (CQDs) have been recognized as the most appealing candidate due to the facile emission tunability and solution processability. However, to date, it is still challenging to develop CQD-based microlasers with low cost yet high performance. Moreover, the poor long-term stability of CQDs remains to be the most critical issue, which may block their laser aspirations. Herein, we developed a unique but generic approach to forming a novel type of a whispering-gallery-mode (WGM) microbubble laser from the hybrid CQD/poly(methyl methacrylate) (PMMA) nanocomposites. The formation mechanism of the microbubbles was unraveled by recording the drying process of the nanocomposite droplets. Interestingly, these microbubbles naturally serve as the high-quality WGM laser resonators. By simply changing the CQDs, the lasing emission can be tuned across the whole visible spectral range. Importantly, these microbubble lasers exhibit unprecedented long-term stability (over one year), sufficient for practical applications. As a proof-of-concept, the potential of water vapor sensing was demonstrated. Our results represent a significant advance in microlasers based on the advantageous CQDs and may offer new possibilities for photonics and optoelectronics.

  3. Magnetic stents retain nanoparticle-bound antirestenotic drugs transported by lipid microbubbles.

    Science.gov (United States)

    Räthel, T; Mannell, H; Pircher, J; Gleich, B; Pohl, U; Krötz, F

    2012-05-01

    Coating coronary stents with antirestenotic drugs revolutionized interventional cardiology. We developed a system for post-hoc drug delivery to uncoated stents. We coupled rapamycin or a chemically similar fluorescent dye to superparamagnetic nanoparticles. The antiproliferative activity of rapamycin coupled to nanoparticles was confirmed in vitro in primary porcine vascular cells. The particles were then incorporated into lipid based microbubbles. Commercially available stents were made magnetizable by nickel plating and used to induce strong field gradients in order to capture magnetic microbubbles from flowing liquids when placed in an external magnetic field. Nanoparticle bound Rapamycin dose dependently inhibited cell proliferation in vitro. Magnetic microcbubbles carrying coated nanoparticles were caught by magnets placed external to a flow-through tube. Plating commercial stents with nickel resulted in increased deposition at stent struts and allowed for widely increased distance of external magnets. Deposition depended on circulation time and velocity and distance of magnets. Deposited microbubbles were destroyed by ultrasound and delivered their cargo to targeted sites. Drugs can be incorporated into nanoparticle loaded microbubbles and thus be delivered to magnetizable stents from circulating fluids by applying external magnetic fields. This technology could allow for post-hoc drug coating of already implanted vascular stents.

  4. Immunohistochemistry for annexin A10 can distinguish sporadic from Lynch syndrome-associated microsatellite-unstable colorectal carcinoma.

    Science.gov (United States)

    Pai, Reetesh K; Shadrach, Bonnie L; Carver, Paula; Heald, Brandie; Moline, Jessica; Church, James; Kalady, Matthew F; Burke, Carol A; Plesec, Thomas P; Lai, Keith K; Gonzalo, David H; Pai, Rish K

    2014-04-01

    Differentiating sporadic microsatellite-unstable colorectal carcinoma due to MLH1 promoter hypermethylation from Lynch syndrome (LS)-associated tumors due to mutations in mismatch-repair proteins is time consuming, cost intensive, and requires advanced laboratory testing. A mutation in BRAF has been shown to be highly specific for sporadic tumors; however, a significant proportion of sporadic microsatellite-unstable tumors lack BRAF mutations. MLH1 promoter methylation analysis is subsequently used to differentiate LS and sporadic tumors, but both tests require specialized laboratories and are costly. Through previous gene expression profiling of serrated polyps, we identified annexin A10 as a protein highly expressed in sessile serrated adenomas/polyps. As these polyps give rise to the majority of sporadic microsatellite-unstable tumors, we evaluated the ability of annexin A10 expression to discriminate between LS and sporadic tumors. A marked increase in annexin A10 mRNA was observed in sporadic microsatellite-unstable tumors compared with LS tumors (378-fold increase, Pimmunohistochemistry, annexin A10 was expressed in 23/53 (43%) BRAF-mutated and 9/22 (41%) BRAF wild-type sporadic tumors. In contrast, only 3/56 (5%) LS tumors were positive for annexin A10 (Pimmunohistochemistry. Only 1/28 (4%) LS tumors with loss of MLH1 was positive for annexin A10. This patient did not have a deleterious MLH1 mutation but rather germline promoter hypermethylation of MLH1. On the basis of these results, immunohistochemistry for annexin A10 may be a useful marker to distinguish sporadic from LS-associated microsatellite-unstable colon cancer.

  5. Anti-annexin antibodies, cholesterol levels and disability in multiple sclerosis.

    Science.gov (United States)

    Mandoj, Chiara; Renna, Rosaria; Plantone, Domenico; Sperduti, Isabella; Cigliana, Giovanni; Conti, Laura; Koudriavtseva, Tatiana

    2015-10-08

    So far, no studies have been conducted to evaluate possible correlations between lipid/lipoprotein levels and the anti-phospholipid antibody (aPL) positivity in multiple sclerosis (MS). In this cross-sectional study, we aimed to investigate the relationships between serum lipid profile and aPL positivity rates in MS patients, and their possible differences among secondary-progressive MS (SPMS) patients, relapsing-remitting MS patients in remission (REM) and in relapse (REL). We included 16 SPMS, 58 REM and 26 REL. Their sera were tested for aPL (anti-cardiolipin, anti-β2glycoproteinI, anti-prothrombin, anti-annexinV), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG) and lipoprotein(a) levels. High TC levels were more frequent in SPMS patients than other groups (p=0.05). The REL had significantly higher rates of positivity for anti-β2glycoproteinI IgM (p<0.0001), anti-prothrombin IgG and IgM (both p=0.05) than the other groups. A significant positive correlation was found between age and both TC and LDL-C, disability and both TC and LDL-C, disease duration and LDL-C. TC levels were significantly higher (p=0.007) in anti-annexinV-IgG positive patients. The anti-annexinV-IgG positivity significantly associated with high levels of TC (p=0.002) and LDL-C (p=0.03). Our results support the hypothesis that both thrombogenic and neurodegenerative mechanisms associated with an abnormal cholesterol homeostasis might contribute to MS progression. Our study may have interesting practical implications, which could potentially open new therapeutic approaches in the context of appropriately designed clinical trials. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  6. Annexin A2 Mediates the Localization of Measles Virus Matrix Protein at the Plasma Membrane.

    Science.gov (United States)

    Koga, Ritsuko; Kubota, Marie; Hashiguchi, Takao; Yanagi, Yusuke; Ohno, Shinji

    2018-02-28

    Annexins are a family of structurally related proteins that bind negatively charged membrane phospholipids in a Ca 2+ -dependent manner. Annexin A2 (AnxA2), a member of the family, has been implicated in a variety of cellular functions including the organization of membrane domains, vesicular trafficking and cell-cell adhesion. AnxA2 generally forms the heterotetrameric complex with a small Ca 2+ -binding protein S100A10. Measles virus (MV), a member of the family Paramyxoviridae , is an enveloped virus with a nonsegmented negative strand RNA genome. Knockdown of AnxA2 greatly reduced MV growth in cells, without affecting its entry and viral RNA production. In MV-infected, AnxA2-knockdown cells, the expression level of the matrix (M) protein, but not other viral proteins, was reduced compared with that in control cells, and the distribution of the M protein at the plasma membrane was decreased. The M protein lines the inner surface of the envelope and plays an important role in virus assembly by connecting the nucleocapsid to the envelope proteins. The M protein bound to AnxA2 independently of AnxA2's phosphorylation or its association with S100A10, and was co-localized with AnxA2 within cells. Truncation of the N-terminal 10 amino acid residues, but not the N-terminal 5 residues, compromised the ability of the M protein to interact with AnxA2 and localize at the plasma membrane. These results indicate that AnxA2 mediates the localization of the MV M protein at the plasma membrane by interacting with its N-terminal region (especially residues at positions 6-10), thereby aiding in MV assembly. IMPORTANCE Measles virus (MV) is an important human pathogen, still claiming ∼ 100,000 lives per year despite the presence of effective vaccines, and causes occasional outbreaks even in developed countries. Replication of viruses largely relies on the functions of host cells. Our study revealed that the reduction of the host protein annexin A2 compromises the replication of

  7. Study of 99Tcm-annexin V distribution in inferior vena cava thrombus models of rabbits

    International Nuclear Information System (INIS)

    Wu Dayong; Zhang Wenyan; Bian Yanzhu; Hu Yujing

    2013-01-01

    To study 99 Tc m -Annexin V distribution in inferior vena cava thrombus models of rabbits and uptake of 99 Tc m -Annexin V in fresh and old venous thrombus. Rabbits (n=15) were randomly grouped into 3 groups (the fresh thrombus group, old thrombus group, and control group). The rabbits of two thrombus groups developed inferior vena cava thrombus models by operations. The control group received sham operation. The fresh thrombus group and control group rabbits were injected 99 Tc m -Annexin V after operating 1 d; the old thrombus group 14 d. After 1 h all rabbits were killed by injecting overdose pentobarbital sodium. The thrombus (or the inferior vena cava about 3 cm below inferior pole of right kidney level in the control group rabbits), blood, thrombus area inferior vena cava, head lateral inferior vena cava (except the control group), thigh muscle, stomach, myocardium, pulmonary, liver, kidney, spleen, bone and small intestine were obtained from all group rabbits. The ex tissue and blood were weighed and measured by a Well-type detector. The percentage of the injected dose per gram of ex tissue (or blood) was calculated by the above data. The thrombus to blood, thrombus area inferior vena cava, head lateral inferior vena cava and thigh muscle ratios were calculated by percentage of the injected dose per gram of ex tissue (or blood). The test was used to compare the fresh thrombus group and old thrombus group by SPSS 17.0. The percentage of' the injected dose per gram of thrombi (0.01894± 0.002 16% ID/g) in the fresh thrombus group was higher than the old thrombus group (0.00473±0.001 28% ID/g), P<0.05. The thrombus to blood, thrombus area inferior vena cava, head lateral inferior vena cava and muscle ratios (3.42±1.06, 26.32±13.60, 31.23 ±16.00, 111.62±52.23) in the fresh thrombus group were higher than the old thrombus group (0.98±0.09, 5.12±2.01, 6.25±2.38, 21.82±5.93), P<0.05 for all. All the thrombi of the fresh thrombus group were confirmed

  8. Annexin A2 complexes with S100 proteins: structure, function and pharmacological manipulation

    OpenAIRE

    Liu, Yidong; Myrvang, Helene K; Dekker, Lodewijk V

    2014-01-01

    Annexin A2 (AnxA2) was originally identified as a substrate of the pp60v-src oncoprotein in transformed chicken embryonic fibroblasts. It is an abundant protein that associates with biological membranes as well as the actin cytoskeleton, and has been implicated in intracellular vesicle fusion, the organization of membrane domains, lipid rafts and membrane-cytoskeleton contacts. In addition to an intracellular role, AnxA2 has been reported to participate in processes localized to the cell surf...

  9. Advance of apoptosis imaging with radiolabeled annexin V in tumor research

    International Nuclear Information System (INIS)

    Huang Daijuan

    2003-01-01

    One of the most important reasons that cause tumor is decrease or complete absence of apoptosis of tumor cells. Conversely successful anti-tumor therapy is correlated with the introduction of apoptosis into tumor cells. Radiolabeled annexin V is used to image in vivo the phosphatidylserine (PS) that explode on the outer surface of cell membrane after apoptosis so that apoptosis can be detected on the early stage. This imaging method can be introduced into the research of tumor in order to help direct the choose of tumor therapy, inspect the effect and evaluate the prognosis

  10. Identifying low density lipoprotein cholesterol associated variants in the Annexin A2 (ANXA2) gene

    DEFF Research Database (Denmark)

    Fairoozy, Roaa Hani; Cooper, Jackie; White, Jon

    2017-01-01

    Background and aims: Annexin-A2 (AnxA2) is an endogenous inhibitor of proprotein convertase subtilisin/kexin type-9 (PCSK9). The repeat-one (R1) domain of AnxA2 binds to PCSK9, blocking its ability to promote degradation of low-density lipoprotein cholesterol-receptors (LDL-R) and thereby regulat...... plasma LDL-C levels, and thus implicate this protein as a potential therapeutic target for LDL-C lowering. (C) 2017 The Authors. Published by Elsevier Ireland Ltd....

  11. Nanobody-coupled microbubbles as novel molecular tracer.

    Science.gov (United States)

    Hernot, Sophie; Unnikrishnan, Sunil; Du, Zhongmin; Shevchenko, Talent; Cosyns, Bernard; Broisat, Alexis; Toczek, Jakub; Caveliers, Vicky; Muyldermans, Serge; Lahoutte, Tony; Klibanov, Alexander L; Devoogdt, Nick

    2012-03-10

    Camelid-derived single-domain antibody-fragments (~15kDa), called nanobodies, are a new class of molecular tracers that are routinely identified with nanomolar affinity for their target and that are easily tailored for molecular imaging and drug delivery applications. We hypothesized that they are well-suited for the design of targeted microbubbles (μBs) and aimed to develop and characterize eGFP- and VCAM-1-targeted μBs. Anti-eGFP (cAbGFP4) and anti-VCAM-1 (cAbVCAM1-5) nanobodies were site-specifically biotinylated in bacteria. This metabolic biotinylation method yielded functional nanobodies with one biotin located at a distant site of the antigen-binding region of the molecule. The biotinylated nanobodies were coupled to biotinylated lipid μBs via streptavidin-biotin bridging. The ability of μB-cAbGFP4 to recognize eGFP was tested as proof-of-principle by fluorescent microscopy and confirmed the specific binding of eGFP to μB-cAbGFP4. Dynamic flow chamber studies demonstrated the ability of μB-cAbVCAM1-5 to bind VCAM-1 in fast flow (up to 5 dynes/cm(2)). In vivo targeting studies were performed in MC38 tumor-bearing mice (n=4). μB-cAbVCAM1-5 or control μB-cAbGFP4 were injected intravenously and imaged using a contrast-specific ultrasound imaging mode. The echo intensity in the tumor was measured 10min post-injection. μB-cAbVCAM1-5 showed an enhanced signal compared to control μBs (p<0.05). Using metabolic and site-specific biotinylation of nanobodies, a method to develop nanobody-coupled μBs was described. The application of VCAM-1-targeted μBs as novel molecular ultrasound contrast agent was demonstrated both in vitro and in vivo. Copyright © 2011 Elsevier B.V. All rights reserved.

  12. I-124 labeled recombinant human annexin V produced by E. coli for apoptosis image using small animal PET

    Energy Technology Data Exchange (ETDEWEB)

    Jung, J. H.; Lee, I. S.; Woo, S. K.; Woo, G. S.; Chung, W. S.; Kang, J. H.; Cheon, G. J.; Choi, C. W.; Urn, S. M. [Korea Institute of Radiological and Medical Sciences, Daejeon (Korea, Republic of)

    2007-07-01

    Annexin V labeled with radioisotope and optical probe has been used to detect apoptosis. To evaluate annexin V as a multimodal apoptosis imaging agent, large-scale preparation of Annexin V (AV) is preliminary. The aim of this study is to produce and purify recombinant human Annexin V (rh-AV) in E. coli system and radiolabeled rh-AV evaluate in vitro and in vivo apoptosis model system. Annexin V cDNA was obtained from human placenta and rh-AV cloning vector used fusion E. coli vector. Expression vector was based on the E. coli pET system. Induction of rh-AV was used Isopropyl--D-thiogalactoside (IPTG) and purification was used TALON metal affinity resin and T7 - Taq. Purification yield confirmed through SDS-PAGE. In camptothecin (0, 50, 100 uM) induced Jurkat T cell apoptosis model, AV-PI flow cytometry analysis and in vitro binding assay of I-124 labeled rh - AV were performed and compared. Small animal PET images of I-124 labeled rh-AV were obtained in Fas-mediated hepatic apoptosis model. Optimum expression condition was at 37, 250 rpm, 8 hr in 2X YT media including 1mM IPTG, Through two step purification process, rh-AV confirmed about 35 Kd single band by SDS-PAGE. As camptothecin concentration increasing, annexin V-FITC positive % increased in flow cytometry analysis and uptake of I-124 labeled rh-AV also increased. Annexin V-FITC positive % was correlated with and uptake of I-124 labeled rh-AV (R{sup 2}=0.99). In Fas-mediated hepatic apoptosis model, I-124 labeled rh-AV was selectively localized in liver region in PET image. Recombinant Human annexin V was produced by E. coli system and purified using two step affinity chromatography. Radiolabeled rh-AV was useful for the evaluation of apoptosis in vitro and in vivo model. Recombinant human annexin V could be used as apoptosis imaging agent with various radiolabel and optical probe.

  13. Targeted microbubbles for imaging tumor angiogenesis: assessment of whole-body biodistribution with dynamic micro-PET in mice

    DEFF Research Database (Denmark)

    Willmann, Jürgen K; Cheng, Zhen; Davis, Corrine

    2008-01-01

    To evaluate in vivo whole-body biodistribution of microbubbles (MBs) targeted to tumor angiogenesis-related vascular endothelial growth factor (VEGF) receptor 2 (VEGFR2) by using dynamic micro-positron emission tomography (PET) in living mice....

  14. Imaging myocardial ischemia and reperfusion injury via Cy5.5 Annexin V

    Energy Technology Data Exchange (ETDEWEB)

    Tian, Rong [Sichuan Univ., Chengdu (China); Pan, Dong Feng [Univ. of Virginia, VA (United States)

    2012-09-15

    The aim of this article is to present the results of an imaging study of myocardial apoptosis induced by ischemia/reperfusion injury. Twenty nude mice were randomly divided into an experimental group, myocardial apoptosis was induced by ligation of the left anterior descending coronary artery (LAD)for 30 min. This was followed by reperfusion for 90 min. In the control group, the heart was exposed for the same length of time as in the experimental group. Cy5.5 annexin V (25{mu}g)was injected into both sets of mice after the onset of reperfusion. At 90 min post injection, the mice were imaged. The region of interest (ROI)was obtained, and the fluorescence intensity of the ROI was quantified. The animals were sacrificed, and myocardial apoptosis was assayed by TUNEL assay. Fluorescence intensity in the ischemia/reperfusion hearts was significantly higher than that in the control group (P<0.05). In the TUNEL assay, more apoptotic cells were observed in the experimental group than in the control group, correlating with imaging results. Fluorescence imaging of Cy5.5 annexin V in a mouse model of myocardial ischemia/reperfusion can be used in vivo as a noninvasive means of detecting ischemia/reperfusion induced apoptotic cells in the heart.

  15. Usage of CO2 microbubbles as flow-tracing contrast media in X-ray dynamic imaging of blood flows.

    Science.gov (United States)

    Lee, Sang Joon; Park, Han Wook; Jung, Sung Yong

    2014-09-01

    X-ray imaging techniques have been employed to visualize various biofluid flow phenomena in a non-destructive manner. X-ray particle image velocimetry (PIV) was developed to measure velocity fields of blood flows to obtain hemodynamic information. A time-resolved X-ray PIV technique that is capable of measuring the velocity fields of blood flows under real physiological conditions was recently developed. However, technical limitations still remained in the measurement of blood flows with high image contrast and sufficient biocapability. In this study, CO2 microbubbles as flow-tracing contrast media for X-ray PIV measurements of biofluid flows was developed. Human serum albumin and CO2 gas were mechanically agitated to fabricate CO2 microbubbles. The optimal fabricating conditions of CO2 microbubbles were found by comparing the size and amount of microbubbles fabricated under various operating conditions. The average size and quantity of CO2 microbubbles were measured by using a synchrotron X-ray imaging technique with a high spatial resolution. The quantity and size of the fabricated microbubbles decrease with increasing speed and operation time of the mechanical agitation. The feasibility of CO2 microbubbles as a flow-tracing contrast media was checked for a 40% hematocrit blood flow. Particle images of the blood flow were consecutively captured by the time-resolved X-ray PIV system to obtain velocity field information of the flow. The experimental results were compared with a theoretically amassed velocity profile. Results show that the CO2 microbubbles can be used as effective flow-tracing contrast media in X-ray PIV experiments.

  16. TREATMENT OF MICROVASCULAR MICRO-EMBOLIZATION USING MICROBUBBLES AND LONG-TONE-BURST ULTRASOUND: AN IN VIVO STUDY

    Science.gov (United States)

    Pacella, John J.; Brands, Judith; Schnatz, Frederick G.; Black, John J.; Chen, Xucai; Villanueva, Flordeliza S.

    2015-01-01

    Despite epicardial coronary artery reperfusion by percutaneous coronary intervention, distal micro-embolization into the coronary microcirculation limits myocardial salvage during acute myocardial infarction. Thrombolysis using ultrasound and microbubbles (sonothrombolysis) is an approach that induces microbubble oscillations to cause clot disruption and restore perfusion. We sought to determine whether this technique could restore impaired tissue perfusion caused by thrombotic microvascular obstruction. In 16 rats, an imaging transducer was placed on the biceps femoris muscle, perpendicular to a single-element 1-MHz treatment transducer. Ultrasound contrast perfusion imaging was performed at baseline and after micro-embolization. Therapeutic ultrasound (5000 cycles, pulse repetition frequency = 5 0.33 Hz, 1.5 MPa) was delivered to nine rats for two 10-min sessions during intra-arterial infusion of lipid-encapsulated microbubbles; seven control rats received no ultrasound–microbubble therapy. Ultrasound contrast perfusion imaging was repeated after each treatment or control period, and microvascular volume was measured as peak video intensity. There was a 90% decrease in video intensity after micro-embolization (from 8.6 ± 4.8 to 0.7 ± 0.8 dB, p ultrasound–microbubble sessions were respectively followed by video intensity increases of 5.8 ± 5.1 and 8.7 ± 5.7 dB (p ultrasound with microbubbles has a therapeutic effect on microvascular perfusion and may be a valuable adjunct to reperfusion therapy for acute myocardial infarction. PMID:25542487

  17. The effect of microbubbles on gas-liquid mass transfer coefficient and degradation rate of COD in wastewater treatment.

    Science.gov (United States)

    Yao, Kangning; Chi, Yong; Wang, Fei; Yan, Jianhua; Ni, Mingjiang; Cen, Kefa

    2016-01-01

    A commonly used aeration device at present has the disadvantages of low mass transfer rate because the generated bubbles are several millimeters in diameter which are much bigger than microbubbles. Therefore, the effect of a microbubble on gas-liquid mass transfer and wastewater treatment process was investigated. To evaluate the effect of each bubble type, the volumetric mass transfer coefficients for microbubbles and conventional bubbles were determined. The volumetric mass transfer coefficient was 0.02905 s(-1) and 0.02191 s(-1) at a gas flow rate of 0.67 L min(-1) in tap water for microbubbles and conventional bubbles, respectively. The degradation rate of simulated municipal wastewater was also investigated, using aerobic activated sludge and ozone. Compared with the conventional bubble generator, the chemical oxygen demand (COD) removal rate was 2.04, 5.9, 3.26 times higher than those of the conventional bubble contactor at the same initial COD concentration of COD 200 mg L(-1), 400 mg L(-1), and 600 mg L(-1), while aerobic activated sludge was used. For the ozonation process, the rate of COD removal using microbubble generator was 2.38, 2.51, 2.89 times of those of the conventional bubble generator. Based on the results, the effect of initial COD concentration on the specific COD degradation rate were discussed in different systems. Thus, the results revealed that microbubbles could enhance mass transfer in wastewater treatment and be an effective method to improve the degradation of wastewater.

  18. Mechanical and dynamic characteristics of encapsulated microbubbles coupled by magnetic nanoparticles as multifunctional imaging and drug delivery agents

    Science.gov (United States)

    Guo, Gepu; Lu, Lu; Yin, Leilei; Tu, Juan; Guo, Xiasheng; Wu, Junru; Xu, Di; Zhang, Dong

    2014-11-01

    Development of magnetic encapsulated microbubble agents that can integrate multiple diagnostic and therapeutic functions is a key focus in both biomedical engineering and nanotechnology and one which will have far-reaching impact on medical diagnosis and therapies. However, properly designing multifunctional agents that can satisfy particular diagnostic/therapeutic requirements has been recognized as rather challenging, because there is a lack of comprehensive understanding of how the integration of magnetic nanoparticles to microbubble encapsulating shells affects their mechanical properties and dynamic performance in ultrasound imaging and drug delivery. Here, a multifunctional imaging contrast and in-situ gene/drug delivery agent was synthesized by coupling super paramagnetic iron oxide nanoparticles (SPIOs) into albumin-shelled microbubbles. Systematical studies were performed to investigate the SPIO-concentration-dependence of microbubble mechanical properties, acoustic scattering response, inertial cavitation activity and ultrasound-facilitated gene transfection effect. These demonstrated that, with the increasing SPIO concentration, the microbubble mean diameter and shell stiffness increased and ultrasound scattering response and inertial cavitation activity could be significantly enhanced. However, an optimized ultrasound-facilitated vascular endothelial growth factor transfection outcome would be achieved by adopting magnetic albumin-shelled microbubbles with an appropriate SPIO concentration of 114.7 µg ml-1. The current results would provide helpful guidance for future development of multifunctional agents and further optimization of their diagnostic/therapeutic performance in clinic.

  19. Enhancement of aerobic biodegradation in an oxygen-limiting environment using a saponin-based microbubble suspension

    International Nuclear Information System (INIS)

    Choi, Yong Ju; Kim, Young-Jin; Nam, Kyoungphile

    2009-01-01

    This study investigated the ability of a saponin-based microbubble suspension to enhance aerobic biodegradation of phenanthrene by subsurface delivery. As the microbubble suspension flowed through a sand column pressure buildup and release was repeatedly observed, which delivered oxygen to the less permeable regions. Burkholderia cepacia RPH1, a phenanthrene-degrading bacterium, was mainly transported in a suspended form in the microbubble suspension. When three pore volumes of the microbubble suspension containing B. cepacia RPH1 was introduced into a column contaminated with phenanthrene (100 mg/kg), the oxygen content declined to 5% from an initial value of 20% within 5 days and correspondingly, 34.4% of initial phenanthrene was removed in 8 days. The addition of two further three pore volumes enhanced the biodegradation efficiency by a factor of 2.2. Our data suggest that a saponin-based microbubble suspension could be a potential carrier for enhancing the aerobic biodegradation under an oxygen-limiting environment. - Microbubble suspension can enhance the phenanthrene biodegradation under an oxygen-limiting condition.

  20. Influencing factors on microbubble ozonation treatment of acid red 3R wastewater

    Directory of Open Access Journals (Sweden)

    Yurong YA

    2017-08-01

    Full Text Available The microbubble ozonation was used to treat acid red 3R wastewater in order to investigate the influencing factors on its performance. The effects of ozone dose, initial acid red 3R concentration and activated carbon on the performance of microbubble ozonation treatment of acid red 3R wastewater are investigated. The decolorization rate, TOC removal rate, pH variation and ozone utilization efficiency in the microbubble ozonation treatment are compared under different treatment conditions. The results indicate that when increasing ozone dose or decreasing initial acid red 3R concentration, both decolorization rate and TOC removal rate of acid red 3R wastewater increase, but ozone utilization efficiency decreases. The coal-based activated carbon shows strong catalytic activity for microbubble ozonation, which could enhance the decolorization rate and TOC removal rate of acid red 3R wastewater. The better performance of microbubble ozonation treatment is achieved when the ozone dose is 48.3 mg/min and the initial acid red 3R mass concentration is 100 mg/L. Under these conditions, the decolorization efficiency reaches to 100% after treatment for 30 min, the TOC removal efficiency reaches to 78.0% after treatment for 120 min, the reaction rate constant of TOC removal is 0.015 min-1 and the ozone utilization efficiency is higher than 99%. With addition of the coal-based activated carbon of 5 g/L, the decolorization efficiency reaches to 100% after treatment for 15 min, the TOC removal efficiency reaches to 91.2% after treatment for 120 min and the reaction rate constant of TOC removal increases to 0037 min-1.The accumulation and following degradation of intermediate products of small molecule organic acid happens during treatment process, and as a result, the solution pH decreases initially and then increases. Therefore, the optimization of influencing factors for microbubble ozonation could increase both contaminant removal

  1. Intracellular delivery of peptides and siRNAs using microbubble enhanced focused ultrasound

    Science.gov (United States)

    Kinoshita, Manabu; Hynynen, Kullervo

    2006-05-01

    Bioactive substances such as peptides and nucleic acid based agents have attracted great attention for the next generation drug for various diseases. However, the greatest challenge for using these bioactive substances is the development of their delivery system, especially the method for delivering these substances through the cell membrane. With the advancement of ultrasound and ultrasound contrast agent technology, it has become possible to transiently change the permeability of the cell membrane. Moreover, using a focused ultrasound transducer, it is possible to narrow and focus the ultrasound energy within a small target, avoiding damage to the surrounding tissue. In this research we have searched the possibility of delivering the Bak BH3 peptide, the death domain of the Bc1-2 family of proteins, or the short interfering RNA (siRNA) targeting the enhanced green fluorescent protein (EGFP) using microbubble-enhanced focused ultrasound in an in vitro setting. Using a 1.696 MHz focused ultrasound and a microbubble ultrasound contrast agent OPTISON®, we first tested the stability of BH3 peptide under microbubble-enhanced focused ultrasound exposure and proved that the peptide is stable under these circumstances. Next, we have tested the cell-killing effect of the intracellularly delivered Bak BH3 peptide in HeLa and BJAB cell line and observed a statistically enhanced cell death in BJAB cells but not in HeLa cells, leading to the conclusion that intracellularly delivered BH3 peptide by microbubble-enhanced ultrasound can exert its cell killing effect in some cells. We also investigated if we can silence the EGFP expression in the cell by delivering siRNA targeting the EGFP in both transient and stable EGFP expression cell line. Using a 1.653 MHz focused ultrasound and OPTISON®, in both cases, intracellularly delivered siRNA by microbubble-enhanced ultrasound was able to knock down the EGFP expression, which demonstrates the feasibility of using this novel method

  2. Diagnostic and therapeutic research on ultrasound microbubble/nanobubble contrast agents (Review).

    Science.gov (United States)

    Ma, Jing; Xu, Chang Song; Gao, Feng; Chen, Ming; Li, Fan; Du, Lian Fang

    2015-09-01

    The contrast enhanced imaging function of ultrasound contrast agents (UCAs) has been extensively investigated using physical acoustic signatures. It has a number of novel applications, including tissue‑specific molecular imaging and multi‑modal imaging. In addition there are numerous other therapeutic applications of UCAs, for example as vehicles for drug or gene delivery. These uses are discussed, as well as the acoustically‑induced biological effects, including ultrasound targeted microbubble destruction (UTMD). This review also explores the considerations for the safe use of UCA from an acoustic standpoint. The scope of the application of UCA has markedly expanded in recent years, and it is a rapidly growing field of medical research. The current article reviews recent advances in the diagnostic and therapeutic applications of ultrasound microbubble/nanobubble contrast agents.

  3. Cross correlation coefficients of turbulent boundary layer with micro-bubble injection

    International Nuclear Information System (INIS)

    Claudia del Carmen Gutierrez-Torres; Yassin A Hassan; Jose Alfredo Jimenez-Bernal

    2005-01-01

    Full text of publication follows: Injection of micro-bubbles within the turbulent boundary layer has been investigated for a several years as a method to achieve drag reduction. However, the physical mechanism of this phenomenon is not fully understood yet. Experiments in a channel flow for single phase (water) and two phase (water and micro-bubbles) flows under different void fraction conditions are reported for a Reynolds number of 5128. Particle Image Velocimetry technique is used to measure instantaneous velocity fields. Consequently the cross-correlation coefficient Ruv can be calculated along the stream-wise direction for various different y + positions and along the normal direction for the fluctuating components of the velocity obtained from the instantaneous velocity fields. The experiments were carried out in a rectangular acrylic channel, whose dimensions are 4.8 m length, 20.6 cm wide and 5.6 cm height. Water was driven trough the channel by gravity from a tank, which was located 3 m above the channel. Then, water was conducted to a lower tank; from which water was pumped to the upper thank forming a closed loop. Upper tank's water level was kept constant through the tests to ensure constant flow rate trough the channel. The velocity field in the x-y plane was obtained by particle image velocimetry (PIV) at 3.15 m downstream from the channel inlet. A Nd:YAG laser with a wavelength of 532 nm (green light) and power of 350 mJ per pulse is utilized. The particles used for seeding have a diameter that goes from 6-9 μm with a specific gravity almost identical to water s specific gravity. The laser light scattered from the seeding particles was recorded using a CCD Kodak Megaplus camera, Model ES 1.0, 1008 x 1018 pixels. The viewing area was 1.28 cm 2 and was located close to the channel wall. The system recorded 30 velocity fields per second. Each velocity field was obtained from a pair of consecutive images capturing the second image of the pair 1 ms after

  4. Measurement of real pulsatile blood flow using X-ray PIV technique with CO2 microbubbles.

    Science.gov (United States)

    Park, Hanwook; Yeom, Eunseop; Seo, Seung-Jun; Lim, Jae-Hong; Lee, Sang-Joon

    2015-03-06

    Synchrotron X-ray imaging technique has been used to investigate biofluid flows in a non-destructive manner. This study aims to investigate the feasibility of the X-ray PIV technique with CO2 microbubbles as flow tracer for measurement of pulsatile blood flows under in vivo conditions. The traceability of CO2 microbubbles in a pulsatile flow was demonstrated through in vitro experiment. A rat extracorporeal bypass loop was used by connecting a tube between the abdominal aorta and jugular vein of a rat to obtain hemodynamic information of actual pulsatile blood flows without changing the hemorheological properties. The decrease in image contrast of the surrounding tissue was also investigated for in vivo applications of the proposed technique. This technique could be used to accurately measure whole velocity field information of real pulsatile blood flows and has strong potential for hemodynamic diagnosis of cardiovascular diseases.

  5. Measurement of real pulsatile blood flow using X-ray PIV technique with CO2 microbubbles

    Science.gov (United States)

    Park, Hanwook; Yeom, Eunseop; Seo, Seung-Jun; Lim, Jae-Hong; Lee, Sang-Joon

    2015-01-01

    Synchrotron X-ray imaging technique has been used to investigate biofluid flows in a non-destructive manner. This study aims to investigate the feasibility of the X-ray PIV technique with CO2 microbubbles as flow tracer for measurement of pulsatile blood flows under in vivo conditions. The traceability of CO2 microbubbles in a pulsatile flow was demonstrated through in vitro experiment. A rat extracorporeal bypass loop was used by connecting a tube between the abdominal aorta and jugular vein of a rat to obtain hemodynamic information of actual pulsatile blood flows without changing the hemorheological properties. The decrease in image contrast of the surrounding tissue was also investigated for in vivo applications of the proposed technique. This technique could be used to accurately measure whole velocity field information of real pulsatile blood flows and has strong potential for hemodynamic diagnosis of cardiovascular diseases. PMID:25744850

  6. Ultrasound Improves the Delivery and Therapeutic Effect of Nanoparticle-Stabilized Microbubbles in Breast Cancer Xenografts

    OpenAIRE

    Snipstad, Sofie; Berg, Sigrid; Mørch, Ýrr Asbjørg; Bjørkøy, Astrid; Sulheim, Einar; Hansen, Rune; Grimstad, Ingeborg; van Wamel, Annemieke; Maaland, Astri Fjelde; Torp, Sverre Helge; Davies, Ruth Catharina de Lange

    2017-01-01

    Compared with conventional chemotherapy, encapsulation of drugs in nanoparticles can improve efficacy and reduce toxicity. However, delivery of nanoparticles is often insufficient and heterogeneous because of various biological barriers and uneven tumor perfusion. We investigated a unique multifunctional drug delivery system consisting of microbubbles stabilized by polymeric nanoparticles (NPMBs), enabling ultrasound-mediated drug delivery. The aim was to examine mechanisms of ultrasound-medi...

  7. On the design and simulation of an airlift loop bioreactorwith microbubble generation by fluidic oscillation

    Czech Academy of Sciences Publication Activity Database

    Zimmerman, W. B.; Tesař, Václav; Hewakandamby, B.N.; Bandulasena, H.C.H.; Omotowa, O.A.

    2009-01-01

    Roč. 87, C3 (2009), s. 215-227 ISSN 0960-3085 Institutional research plan: CEZ:AV0Z20760514 Keywords : microbubbles * fluidic oscillators * transport phenomena Subject RIV: BK - Fluid Dynamics Impact factor: 0.952, year: 2009 http://apps.isiknowledge.com/full_record.do?product=UA&search_mode=GeneralSearch&qid=1&SID=V2FDdpCMohHOjGaLDMi&page=1&doc=3&colname=WOS

  8. Acoustic detection of microbubble formation induced by enhanced optical breakdown of silver/dendrimer nanocomposites

    International Nuclear Information System (INIS)

    Milas, Susanne M.; Ye, Jing Yong; Norris, Theodore B.; Balogh, Lajos P.; Baker, James R. Jr.; Hollman, Kyle W.; Emelianov, Stanislav; O'Donnell, Matthew

    2003-01-01

    We utilize a real-time acoustic technique, based on pulse-echo measurements to detect formation of microbubbles in an aqueous solution of a silver/dendrimer nanocomposite (DNC). Wave-field plots of successive recordings illustrate the generation and behavior of bubbles created by the optical breakdown process. A significant threshold reduction is achieved with DNC particles compared to its host dendrimer, enabling a diverse field of low-threshold breakdown applications

  9. Emerging functions as host cell factors - an encyclopedia of annexin-pathogen interactions.

    Science.gov (United States)

    Kuehnl, Alexander; Musiol, Agnes; Raabe, Carsten A; Rescher, Ursula

    2016-10-01

    Emerging infectious diseases and drug-resistant infectious agents call for the development of innovative antimicrobial strategies. With pathogenicity now considered to arise from the complex and bi-directional interplay between a microbe and the host, host cell factor targeting has emerged as a promising approach that might overcome the limitations of classical antimicrobial drug development and could open up novel and efficient therapeutic strategies. Interaction with and modulation of host cell membranes is a recurrent theme in the host-microbe relationship. In this review, we provide an overview of what is currently known about the role of the Ca2+ dependent, membrane-binding annexin protein family in pathogen-host interactions, and discuss their emerging functions as host cell derived auxiliary proteins in microbe-host interactions and host cell targets.

  10. Technetium 99m-labeled annexin v scintigraphy of platelet activation in vegetations of experimental endocarditis

    Energy Technology Data Exchange (ETDEWEB)

    Rouzet, F.; Sarda-Mantel, L.; Le Guludec, D. [Nucl Med Serv, Grp Hosp Bichat Claude Bernard, AP-HP, Paris (France); Rouzet, F.; Sarda-Mantel, L.; LeGuludec, D. [Univ Denis Diderot Paris 7, UMR S773, Paris (France); Rouzet, F.; Sarda-Mantel, L.; Le Guludec, D. [INSERM, U773, Paris (France); Hernandez, M.D.; Louedec, L.; Michel, J.B. [Univ Paris 07, CHU Xavier Bichat, INSERM, U698, Paris (France); Hervatin, F. [CEA, DSV, DRM, SHFJ, Orsay (France); Lefort, A.; Fantin, B. [Univ Denis Diderot Paris 7, EA 3964, Paris (France); Duval, X. [Univ Denis Diderot Paris 7, INSERM, CIC 007, Paris (France); Duval, X. [Univ Denis Diderot Paris 7, AP-HP, Grp Hosp Bichat Claude Bernard, Ctr Invest Clin, Paris (France); Hernandez, M.D. [Univ Guadalajara, DeptPathol, Guadalajara 44430, Jalisco (Mexico)

    2008-07-01

    Background: The pathophysiology of infective endocarditis involves a pathogen/host tissue interaction, leading to formation of infected thrombotic vegetations. Annexin V is a ligand of phosphatidyl-serines exposed by activated platelets and apoptotic cells. Because vegetations are platelet-fibrin clots in which platelet pro-aggregant activity is enhanced by bacterial colonization, we investigated the ability of annexin V labeled with technetium {sup 99m}Tc ({sup 99m}Tc-ANX) to provide functional imaging of these vegetations in experimental models of infective endocarditis. This ability was assessed in rabbits and rats because of the different interest of these 2 species in preclinical analysis. Methods and Results: Non-bacterial thrombotic endocarditis was induced with the use of a catheter left indwelling through the aortic or tricuspid valve, and animals were injected with either a bacterial inoculum or saline. Scintigraphic investigations were performed 5 days later and showed a higher {sup 99m}Tc-ANX uptake by vegetations in infected versus non-infected animals (ratio,1.3 for in vivo acquisitions and 2 for autoradiography; P {<=} 0.0001 for all), whereas no significant uptake was present in controls. Right-sided endocarditis was associated with pulmonary uptake foci corresponding to emboli. Histological analysis of vegetations showed a specific uptake of {sup 99m}Tc-ANX at the interface between circulating blood and vegetation. In parallel, underlying myocardial tissue showed myocyte apoptosis and mucoid degeneration, without extracellular matrix degradation at this stage. Conclusions: {sup 99m}Tc-ANX is suitable for functional imaging of platelet-fibrin vegetations in endocarditis, as well as embolic events. {sup 99m}Tc-ANX uptake reflects mainly platelet activation in the luminal layer of vegetations. This uptake is enhanced by bacterial colonization. (authors)

  11. Technetium 99m-labeled annexin v scintigraphy of platelet activation in vegetations of experimental endocarditis

    International Nuclear Information System (INIS)

    Rouzet, F.; Sarda-Mantel, L.; Le Guludec, D.; Rouzet, F.; Sarda-Mantel, L.; LeGuludec, D.; Rouzet, F.; Sarda-Mantel, L.; Le Guludec, D.; Hernandez, M.D.; Louedec, L.; Michel, J.B.; Hervatin, F.; Lefort, A.; Fantin, B.; Duval, X.; Duval, X.; Hernandez, M.D.

    2008-01-01

    Background: The pathophysiology of infective endocarditis involves a pathogen/host tissue interaction, leading to formation of infected thrombotic vegetations. Annexin V is a ligand of phosphatidyl-serines exposed by activated platelets and apoptotic cells. Because vegetations are platelet-fibrin clots in which platelet pro-aggregant activity is enhanced by bacterial colonization, we investigated the ability of annexin V labeled with technetium 99m Tc ( 99m Tc-ANX) to provide functional imaging of these vegetations in experimental models of infective endocarditis. This ability was assessed in rabbits and rats because of the different interest of these 2 species in preclinical analysis. Methods and Results: Non-bacterial thrombotic endocarditis was induced with the use of a catheter left indwelling through the aortic or tricuspid valve, and animals were injected with either a bacterial inoculum or saline. Scintigraphic investigations were performed 5 days later and showed a higher 99m Tc-ANX uptake by vegetations in infected versus non-infected animals (ratio,1.3 for in vivo acquisitions and 2 for autoradiography; P ≤ 0.0001 for all), whereas no significant uptake was present in controls. Right-sided endocarditis was associated with pulmonary uptake foci corresponding to emboli. Histological analysis of vegetations showed a specific uptake of 99m Tc-ANX at the interface between circulating blood and vegetation. In parallel, underlying myocardial tissue showed myocyte apoptosis and mucoid degeneration, without extracellular matrix degradation at this stage. Conclusions: 99m Tc-ANX is suitable for functional imaging of platelet-fibrin vegetations in endocarditis, as well as embolic events. 99m Tc-ANX uptake reflects mainly platelet activation in the luminal layer of vegetations. This uptake is enhanced by bacterial colonization. (authors)

  12. Annexin A7 deficiency potentiates cardiac NFAT activity promoting hypertrophic signaling

    Energy Technology Data Exchange (ETDEWEB)

    Voelkl, Jakob; Alesutan, Ioana; Pakladok, Tatsiana; Viereck, Robert; Feger, Martina; Mia, Sobuj [Department of Physiology, University of Tübingen, Tübingen (Germany); Schönberger, Tanja [Department of Cardiology and Cardiovascular Medicine, University of Tübingen, Tübingen (Germany); Noegel, Angelika A. [Center for Biochemistry, Institute of Biochemistry I, University of Cologne, Köln (Germany); Gawaz, Meinrad [Department of Cardiology and Cardiovascular Medicine, University of Tübingen, Tübingen (Germany); Lang, Florian, E-mail: florian.lang@uni-tuebingen.de [Department of Physiology, University of Tübingen, Tübingen (Germany)

    2014-02-28

    Highlights: • Cardiac Anxa7 expression was up-regulated following TAC. • The hypertrophic response following TAC was augmented in Anxa7-deficient mice. • Silencing of Anxa7 increased indicators of HL-1 cardiomyocytes hypertrophy. • Silencing of Anxa7 induced Nfatc1 nuclear translocation. • Silencing of Anxa7 enhanced NFAT-dependent transcriptional activity. - Abstract: Annexin A7 (Anxa7) is a cytoskeletal protein interacting with Ca{sup 2+} signaling which in turn is a crucial factor for cardiac remodeling following cardiac injury. The present study explored whether Anxa7 participates in the regulation of cardiac stress signaling. To this end, mice lacking functional Anxa7 (anxa7{sup −/−}) and wild-type mice (anxa7{sup +/+}) were investigated following pressure overload by transverse aortic constriction (TAC). In addition, HL-1 cardiomyocytes were silenced with Anxa7 siRNA and treated with isoproterenol. Transcript levels were determined by quantitative RT-PCR, transcriptional activity by luciferase reporter assay and protein abundance by Western blotting and confocal microscopy. As a result, TAC treatment increased the mRNA and protein levels of Anxa7 in wild-type mice. Moreover, TAC increased heart weight to body weight ratio and the cardiac mRNA levels of αSka, Nppb, Col1a1, Col3a1 and Rcan1, effects more pronounced in anxa7{sup −/−} mice than in anxa7{sup +/+} mice. Silencing of Anxa7 in HL-1 cardiomyocytes significantly increased nuclear localization of Nfatc1. Furthermore, Anxa7 silencing increased NFAT-dependent transcriptional activity as well as αSka, Nppb, and Rcan1 mRNA levels both, under control conditions and following β-adrenergic stimulation by isoproterenol. These observations point to an important role of annexin A7 in the regulation of cardiac NFAT activity and hypertrophic response following cardiac stress conditions.

  13. Preparation of monodisperse microbubbles using an integrated embedded capillary T-junction with electrohydrodynamic focusing.

    Science.gov (United States)

    Parhizkar, Maryam; Stride, Eleanor; Edirisinghe, Mohan

    2014-07-21

    This work investigates the generation of monodisperse microbubbles using a microfluidic setup combined with electrohydrodynamic processing. A basic T-junction microfluidic device was modified by applying an electrical potential difference across the outlet channel. A model glycerol air system was selected for the experiments. In order to investigate the influence of the electric field strength on bubble formation, the applied voltage was increased systematically up to 21 kV. The effect of solution viscosity and electrical conductivity was also investigated. It was found that with increasing electrical potential difference, the size of the microbubbles reduced to ~25% of the capillary diameter whilst their size distribution remained narrow (polydispersity index ~1%). A critical value of 12 kV was found above which no further significant reduction in the size of the microbubbles was observed. The findings suggest that the size of the bubbles formed in the T-junction (i.e. in the absence of the electric field) is strongly influenced by the viscosity of the solution. The eventual size of bubbles produced by the composite device, however, was only weakly dependent upon viscosity. Further experiments, in which the solution electrical conductivity was varied by the addition of a salt indicated that this had a much stronger influence upon bubble size.

  14. Drug perfusion enhancement in tissue model by steady streaming induced by oscillating microbubbles.

    Science.gov (United States)

    Oh, Jin Sun; Kwon, Yong Seok; Lee, Kyung Ho; Jeong, Woowon; Chung, Sang Kug; Rhee, Kyehan

    2014-01-01

    Drug delivery into neurological tissue is challenging because of the low tissue permeability. Ultrasound incorporating microbubbles has been applied to enhance drug delivery into these tissues, but the effects of a streaming flow by microbubble oscillation on drug perfusion have not been elucidated. In order to clarify the physical effects of steady streaming on drug delivery, an experimental study on dye perfusion into a tissue model was performed using microbubbles excited by acoustic waves. The surface concentration and penetration length of the drug were increased by 12% and 13%, respectively, with streaming flow. The mass of dye perfused into a tissue phantom for 30s was increased by about 20% in the phantom with oscillating bubbles. A computational model that considers fluid structure interaction for streaming flow fields induced by oscillating bubbles was developed, and mass transfer of the drug into the porous tissue model was analyzed. The computed flow fields agreed with the theoretical solutions, and the dye concentration distribution in the tissue agreed well with the experimental data. The computational results showed that steady streaming with a streaming velocity of a few millimeters per second promotes mass transfer into a tissue. © 2013 Published by Elsevier Ltd.

  15. Floating Hydrogel with Self-Generating Micro-Bubbles for Intravesical Instillation

    Directory of Open Access Journals (Sweden)

    Tingsheng Lin

    2016-12-01

    Full Text Available Intravesical instillation is the main therapy for bladder cancer and interstitial cystitis. However, most drug solutions are eliminated from bladder after the first voiding of urine. To solve this problem, we proposed a floating hydrogel with self-generating micro-bubbles as a new delivery system. It floated in urine, avoiding the urinary obstruction and bladder irritation that ordinary hydrogels caused. In this study, we abandoned traditional gas-producing method like chemical decomposition of NaHCO3, and used the foamability of Poloxamer 407 (P407 instead. Through simple shaking (just like shaking SonoVue for contrast-enhanced ultrasound in clinical, the P407 solution will “lock” many micro-bubbles and float in urine as quickly and steadily as other gas producing materials. In vivo release experiments showed that drug was released continually from hydrogel for 10 h during the erosion process. Thus, the residence time of drug in bladder was prolonged and drug efficacy was improved. In vivo efficacy study using rabbit acute bladder injury model showed that prolonged drug residence time in bladder increased the efficiency of heparin in the protection of bladder mucosal permeability. Therefore, our floating hydrogel system with self-generating micro-bubbles was single-component, simply prepared and efficacy enhancing, successfully exempting users from worries on safety and clinical efficiency from bench to bedside.

  16. Microbubble-Mediated Ultrasound Enhances the Lethal Effect of Gentamicin on Planktonic Escherichia coli

    Directory of Open Access Journals (Sweden)

    Han-Xiao Zhu

    2014-01-01

    Full Text Available Previous research has found that low-intensity ultrasound enhanced the lethal effect of gentamicin on planktonic E. coli. We aimed to further investigate whether microbubble-mediated low-intensity ultrasound could further enhance the antimicrobial efficacy of gentamicin. The planktonic E. coli (ATCC 25922 was distributed to four different interventions: control (GCON, microbubble only (GMB, ultrasound only (GUS, and microbubble-mediated ultrasound (GMUS. Ultrasound was applied with 100 mW/cm2 (average intensity and 46.5 KHz, which presented no bactericidal activity. After 12 h, plate counting was used to estimate the number of bacteria, and bacterial micromorphology was observed with transmission electron microscope. The results showed that the viable counts of E. coli in GMUS were decreased by 1.01 to 1.42 log10 CFU/mL compared with GUS (P<0.01. The minimal inhibitory concentration (MIC of gentamicin against E. coli was 1 μg/mL in the GMUS and GUS groups, lower than that in the GCON and GMB groups (2 μg/mL. Transmission electron microscopy (TEM images exhibited more destruction and higher thickness of bacterial cell membranes in the GMUS than those in other groups. The reason might be the increased permeability of cell membranes for gentamicin caused by acoustic cavitation.

  17. Acoustic microstreaming due to an ultrasound contrast microbubble near a wall

    Science.gov (United States)

    Mobadersany, Nima; Sarkar, Kausik

    2017-11-01

    In an ultrasound field, in addition to the sinusoidal motion of fluid particles, particles experience a steady streaming velocity due to nonlinear second order effects. Here, we have simulated the microstreaming flow near a plane rigid wall caused by the pulsations of contrast microbubbles. Although these microbubbles were initially developed as a contrast enhancing agents for ultrasound imaging, they generate additional therapeutic effects that can be harnessed for targeted drug delivery or blood brain barrier (BBB) opening. The microbubbles have a gas core coated with a stabilizing layer of lipids or proteins. We use analytical models as well as boundary element (BEM) simulation to simulate the flow around these bubbles implementing interfacial rheology models for the coating. The microstreaming flow is characterized by two wall bounded vortices. The size of the vortices decreases with the decrease of the separation from the wall. The vortex-induced shear stress is simulated and analyzed as a function of excitation parameters and geometry. These microstreaming shear stress plays a critical role in increasing the membrane permeability facilitating drug delivery or rupturing biological tissues.

  18. Combined effect of ultrasound/SonoVue microbubble on CD4(+)CD25(+) regulatory T cells viability and optimized parameters for its transfection.

    Science.gov (United States)

    Shi, Chunying; Zhang, Yu; Yang, Haichao; Dong, Tianxiu; Chen, Yaodong; Xu, Yutong; Yang, Xiuhua

    2015-09-01

    The purpose of this study was to investigate the combined effect of ultrasound and SonoVue microbubble on CD4(+)CD25(+) regulatory T cells (Tregs) viability and to explore the appropriate parameters for Tregs transfection. Tregs were separated from peripheral venous blood of patients with hepatocellular carcinoma and seeded in 96-well plates. The optimal ultrasound exposure time and optimal SonoVue microbubble concentration for Tregs were measured by mechanical index (MI) of 1.2 or 1.4, exposure time of 0, 30, 60, 90, 120, 150, 180s, and 0, 10, 20, 30, 40, 50μL/100μL microbubble per well, respectively. In addition, the combined effect of ultrasound and microbubble on Tregs viability was evaluated according to the following parameters: MI 1.2/1.4+exposure time of 120, 150, 180s+0, 10, 20, 30, 40, 50μL/100μL microbubble per well. Tregs viability investigations were performed in order to explore the optimal transfection condition. The efficiency of plasmid transfer was determined by detection of luciferase activity on the microscopic examinations. The proliferation of Tregs could be promoted by ultrasound exposures, while being decreased with the increasing concentration of microbubbles. Under the current experimental conditions, the optimal ultrasound parameters were MI=1.4 and exposure time=150/180s. The optimal microbubble concentration was 10μL/100μL. Compared with treatment with ultrasound or microbubbles alone, the transfection efficiency of Tregs improved 50% by combining ultrasound and microbubble. The results indicate that both ultrasound and microbubble could affect the Tregs proliferation and the optimal Treg transfection rate was obtained by treating with 10% microbubbles and ultrasound exposure for 150/180s under ultrasound MI of 1.4. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Characterization of a novel annexin gene from cotton (Gossypium hirsutum cv CRI 35) and antioxidative role of its recombinant protein.

    Science.gov (United States)

    Zhou, Lu; Duan, Jin; Wang, Xiao-Ming; Zhang, Heng-Mu; Duan, Ming-Xing; Liu, Jin-Yuan

    2011-05-01

    Plant annexins represent a multigene family involved in cellular elongation and development. A cDNA encoding a novel annexin was isolated from a cotton (Gossypium hirsutum) fiber cDNA library and designated GhAnx1. This gene encodes a 316 amino acid protein with a theoretical molecular mass of 36.06 kDa and a theoretical pI of 6.19. At the amino acid level, it shares high sequence similarity and has evolutionary relationships with annexins from higher plants. The purified recombinant protein expressed in Escherichia coli was used to investigate its physicochemical properties. Circular dichroism spectrum analyses showed a positive peak rising to the maximum at 196 nm and a broad negative band rounding 215 nm, suggesting that the GhAnx1 protein was prominently α-helical. The fluorescence measurements indicated that it could bind to Ca(2+) in vitro. These results demonstrated that GhAnx1 was a typical annexin protein in cotton. A bioassay experiment was conducted to analyze its potential function and showed that E. coli cells expressing GhAnx1 were protected from tert-butyl hydroperoxide (tBH) stress, suggesting that it had a potential antioxidative role. Northern blot analyses revealed that GhAnx1 was highly expressed in fibers, especially during the elongation stage, suggesting that it might be important for fiber elongation. © 2011 Institute of Botany, Chinese Academy of Sciences.

  20. Potato Annexin STANN1 Promotes Drought Tolerance and Mitigates Light Stress in Transgenic Solanum tuberosum L. Plants

    Czech Academy of Sciences Publication Activity Database

    Szalonek, M.; Sierpien, B.; Rymaszewski, W.; Gieczewska, K.; Vaňková, Radomíra; Dobrev, Petre; Szczesny, P.; Marczewski, W.; Krusiewicz, D.; Strzelczyk-Zyta, D.; Konopka-Postupolska, D.

    2015-01-01

    Roč. 10, č. 7 (2015), e0132683 E-ISSN 1932-6203 Institutional support: RVO:61389030 Keywords : MEDIATED OXIDATIVE STRESS * VIOLAXANTHIN DE-EPOXIDASE * BRASSICA-JUNCEA ANNEXIN-3 Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.057, year: 2015

  1. Suppression of annexin A2 by prostaglandin E₂ impairs phagocytic ability of peritoneal macrophages in women with endometriosis.

    Science.gov (United States)

    Wu, Meng-Hsing; Chuang, Pei-Chin; Lin, Yiu-Juian; Tsai, Shaw-Jenq

    2013-04-01

    Is annexin A2 involved in the reduced phagocytic ability of macrophages in endometriosis? Data from women with endometriosis and a murine model of the disease show that expression of annexin A2 in peritoneal macrophages is inhibited by prostaglandin E2 (PGE2) and this impairs the phagocytic ability of macrophages. Endometriosis is a chronic inflammatory disease that recruits many immune cells, especially macrophages, to the peritoneal cavity. The phagocytic ability of peritoneal macrophages isolated from women with endometriosis is reduced. A laboratory study. Thirty-five patients (20 with and 15 without endometriosis) of reproductive age with normal menstrual cycles were recruited. Peritoneal macrophages isolated from women with or without endometriosis were cultured and treated with vehicle, PGE2 and different EP receptor agonists, and the expression of annexin A2 was quantified by RT-PCR and western blotting. Annexin A2 was knocked down (by small interfering RNA) in normal macrophages or overexpressed (by treatment with recombinant protein) in endometriotic macrophages and their phagocytic ability was measured by flow cytometry. Peritoneal macrophages were isolated from a mouse model of endometriosis and treated with PGE2 or cyclo-oxygenase (COX) inhibitors, and annexin A2 mRNA was quantified. Levels of annexin A2 were markedly reduced in peritoneal macrophages from women with endometriosis versus controls (mRNA: P endometriosis versus control) via the EP2/EP4 receptor-dependent signaling pathway. Treatment with PGE2 or knockdown of annexin A2 inhibited the phagocytic ability of macrophages (P peritoneal macrophages were markedly reduced in mice treated with PGE2 (P peritoneal macrophages (P peritoneal cells from patients with endometriosis or that their endometriotic fluid contains increased amounts of PGE2 when compared with control subjects. Inhibiting PGE2 signaling, in order to restore or enhance the phagocytic capability of macrophages, may represent a new

  2. Microbubble-mediated ultrasound therapy: a review of its potential in cancer treatment

    Directory of Open Access Journals (Sweden)

    Ibsen S

    2013-05-01

    Full Text Available Stuart Ibsen,1 Carolyn E Schutt,2 Sadik Esener31Moores Cancer Center, University of California at San Diego, La Jolla, CA, USA; 2Department of Bioengineering, University of California at San Diego, La Jolla, CA, USA; 3Department of Nanoengineering, Moores Cancer Center, University of California at San Diego, La Jolla, CA, USAAbstract: The inherently toxic nature of chemotherapy drugs is essential for them to kill cancer cells but is also the source of the detrimental side effects experienced by patients. One strategy to reduce these side effects is to limit the healthy tissue exposure by encapsulating the drugs in a vehicle that demonstrates a very low leak rate in circulation while simultaneously having the potential for rapid release once inside the tumor. Designing a vehicle with these two opposing properties is the major challenge in the field of drug delivery. A triggering event is required to change the vehicle from its stable circulating state to its unstable release state. A unique mechanical actuation type trigger is possible by harnessing the size changes that occur when microbubbles interact with ultrasound. These mechanical actuations can burst liposomes and cell membranes alike allowing for rapid drug release and facilitating delivery into nearby cells. The tight focusing ability of the ultrasound to just a few cubic millimeters allows for precise control over the tissue location where the microbubbles destabilize the vehicles. This allows the ultrasound to highlight the tumor tissue and cause rapid drug release from any carrier present. Different vehicle designs have been demonstrated from carrying drug on just the surface of the microbubble itself to encapsulating the microbubble along with the drug within a liposome. In the future, nanoparticles may extend the circulation half-life of these ultrasound triggerable drug-delivery vehicles by acting as nucleation sites of ultrasound-induced mechanical actuation. In addition to the

  3. Potato Annexin STANN1 Promotes Drought Tolerance and Mitigates Light Stress in Transgenic Solanum tuberosum L. Plants.

    Science.gov (United States)

    Szalonek, Michal; Sierpien, Barbara; Rymaszewski, Wojciech; Gieczewska, Katarzyna; Garstka, Maciej; Lichocka, Malgorzata; Sass, Laszlo; Paul, Kenny; Vass, Imre; Vankova, Radomira; Dobrev, Peter; Szczesny, Pawel; Marczewski, Waldemar; Krusiewicz, Dominika; Strzelczyk-Zyta, Danuta; Hennig, Jacek; Konopka-Postupolska, Dorota

    2015-01-01

    Annexins are a family of calcium- and membrane-binding proteins that are important for plant tolerance to adverse environmental conditions. Annexins function to counteract oxidative stress, maintain cell redox homeostasis, and enhance drought tolerance. In the present study, an endogenous annexin, STANN1, was overexpressed to determine whether crop yields could be improved in potato (Solanum tuberosum L.) during drought. Nine potential potato annexins were identified and their expression characterized in response to drought treatment. STANN1 mRNA was constitutively expressed at a high level and drought treatment strongly increased transcription levels. Therefore, STANN1 was selected for overexpression analysis. Under drought conditions, transgenic potato plants ectopically expressing STANN1 were more tolerant to water deficit in the root zone, preserved more water in green tissues, maintained chloroplast functions, and had higher accumulation of chlorophyll b and xanthophylls (especially zeaxanthin) than wild type (WT). Drought-induced reductions in the maximum efficiency and the electron transport rate of photosystem II (PSII), as well as the quantum yield of photosynthesis, were less pronounced in transgenic plants overexpressing STANN1 than in the WT. This conferred more efficient non-photochemical energy dissipation in the outer antennae of PSII and probably more efficient protection of reaction centers against photooxidative damage in transgenic plants under drought conditions. Consequently, these plants were able to maintain effective photosynthesis during drought, which resulted in greater productivity than WT plants despite water scarcity. Although the mechanisms underlying this stress protection are not yet clear, annexin-mediated photoprotection is probably linked to protection against light-induced oxidative stress.

  4. Potato Annexin STANN1 Promotes Drought Tolerance and Mitigates Light Stress in Transgenic Solanum tuberosum L. Plants

    Science.gov (United States)

    Szalonek, Michal; Sierpien, Barbara; Rymaszewski, Wojciech; Gieczewska, Katarzyna; Garstka, Maciej; Lichocka, Malgorzata; Sass, Laszlo; Paul, Kenny; Vass, Imre; Vankova, Radomira; Dobrev, Peter; Szczesny, Pawel; Marczewski, Waldemar; Krusiewicz, Dominika; Strzelczyk-Zyta, Danuta; Hennig, Jacek; Konopka-Postupolska, Dorota

    2015-01-01

    Annexins are a family of calcium- and membrane-binding proteins that are important for plant tolerance to adverse environmental conditions. Annexins function to counteract oxidative stress, maintain cell redox homeostasis, and enhance drought tolerance. In the present study, an endogenous annexin, STANN1, was overexpressed to determine whether crop yields could be improved in potato (Solanum tuberosum L.) during drought. Nine potential potato annexins were identified and their expression characterized in response to drought treatment. STANN1 mRNA was constitutively expressed at a high level and drought treatment strongly increased transcription levels. Therefore, STANN1 was selected for overexpression analysis. Under drought conditions, transgenic potato plants ectopically expressing STANN1 were more tolerant to water deficit in the root zone, preserved more water in green tissues, maintained chloroplast functions, and had higher accumulation of chlorophyll b and xanthophylls (especially zeaxanthin) than wild type (WT). Drought-induced reductions in the maximum efficiency and the electron transport rate of photosystem II (PSII), as well as the quantum yield of photosynthesis, were less pronounced in transgenic plants overexpressing STANN1 than in the WT. This conferred more efficient non-photochemical energy dissipation in the outer antennae of PSII and probably more efficient protection of reaction centers against photooxidative damage in transgenic plants under drought conditions. Consequently, these plants were able to maintain effective photosynthesis during drought, which resulted in greater productivity than WT plants despite water scarcity. Although the mechanisms underlying this stress protection are not yet clear, annexin-mediated photoprotection is probably linked to protection against light-induced oxidative stress. PMID:26172952

  5. Potato Annexin STANN1 Promotes Drought Tolerance and Mitigates Light Stress in Transgenic Solanum tuberosum L. Plants.

    Directory of Open Access Journals (Sweden)

    Michal Szalonek

    Full Text Available Annexins are a family of calcium- and membrane-binding proteins that are important for plant tolerance to adverse environmental conditions. Annexins function to counteract oxidative stress, maintain cell redox homeostasis, and enhance drought tolerance. In the present study, an endogenous annexin, STANN1, was overexpressed to determine whether crop yields could be improved in potato (Solanum tuberosum L. during drought. Nine potential potato annexins were identified and their expression characterized in response to drought treatment. STANN1 mRNA was constitutively expressed at a high level and drought treatment strongly increased transcription levels. Therefore, STANN1 was selected for overexpression analysis. Under drought conditions, transgenic potato plants ectopically expressing STANN1 were more tolerant to water deficit in the root zone, preserved more water in green tissues, maintained chloroplast functions, and had higher accumulation of chlorophyll b and xanthophylls (especially zeaxanthin than wild type (WT. Drought-induced reductions in the maximum efficiency and the electron transport rate of photosystem II (PSII, as well as the quantum yield of photosynthesis, were less pronounced in transgenic plants overexpressing STANN1 than in the WT. This conferred more efficient non-photochemical energy dissipation in the outer antennae of PSII and probably more efficient protection of reaction centers against photooxidative damage in transgenic plants under drought conditions. Consequently, these plants were able to maintain effective photosynthesis during drought, which resulted in greater productivity than WT plants despite water scarcity. Although the mechanisms underlying this stress protection are not yet clear, annexin-mediated photoprotection is probably linked to protection against light-induced oxidative stress.

  6. Universal phase and force diagrams for a microbubble or pendant drop in static fluid on a surface

    Science.gov (United States)

    Wei, P. S.; Hsiao, C. C.; Chen, K. Y.

    2008-01-01

    Dimensionless three-dimensional universal phase and lift force diagrams of a microbubble (or pendant drop) in static liquid on a solid surface (or orifice) are presented in this work. Microbubble dynamics has been found to play a vital role in mass, momentum, energy, and concentration transfer rates in contemporary micro- and nanosciences and technologies. In this study, dimensionless phase and force diagrams are introduced by utilizing the analytical solutions of the microbubble shape reported in the literature. It shows that phase and force diagrams can be universally specified by two dimensionless independent parameters, Bond number, and contact angle (or base radius). Based on the presence of an inflection point or neck on the microbubble surface, each diagram exhibits three regions. Growth, detachment, and entrapment of a microbubble can be described by path lines in three regions. The corresponding universal total lift forces include hydrostatic buoyancy, difference in gas, and hydrostatic pressures at the base, capillary pressure, as well as surface tension induced by the variation of circumference, which has not been treated in the literature so far. In the absence of viscous stress and Marangoni force, the total lift force equals surface tension induced by the variation of circumference. The latter can be an attaching or lifting force, depending on whether the state in the distinct regions and contact angle is less than or greater than a critical angle. The critical angle, which is slightly less than the inclination angle at the inflection point, is decreased with increasing Bond number.

  7. Value of amniotic fluid IL-8 and Annexin A2 in prediction of preterm delivery in preterm labor and preterm premature rupture of membranes.

    Science.gov (United States)

    Jia, Xiaohui

    2014-01-01

    To investigate the clinical significance and value in the prediction of preterm delivery of combined amniotic fluid IL-8 and Annexin A2 levels in preterm premature rupture of membranes (PPROM) and preterm labor (PTL). Sixty pregnant women at < 32 gestational weeks who developed PTL were divided into a PPROM group and a non-PPROM group. Ten normal pregnant women served as a control group. IL-8 and Annexin A2 levels were measured in amniotic fluid samples from each patient. Amniotic fluid IL-8 and Annexin-A2 levels in PTL (PPROM and non-PPROM groups) were significantly higher than those of the controls (p < 0.05). The PPROM group displayed higher amniotic fluid Annexin-A2 levels than did the non-PPROM group, with a statistically significant difference (p < 0.05). The PPROM group showed higher amniotic fluid IL-8 levels than did the non-PPROM group; however, this was statistically insignificant (p = 0.56). Combined detection of amniotic fluid IL-8 and Annexin-A2 in the prediction of preterm delivery within 2 weeks of measurement showed sensitivity of 81.25%, specificity of 88.89% and PPV of 92.86%. Amniotic fluid IL-8 and Annexin-A2 levels are associated with the occurrence of PPROM and PTL. Combined detection of IL-8 and Annexin-A2 levels in identifying preterm delivery within 2 weeks in PTL and PPROM is of possible clinical and predictive value.

  8. Highly sensitive temperature sensor based on cascaded polymer-microbubble cavities by employing a subtraction between reciprocal thermal responses.

    Science.gov (United States)

    Cao, Kunjian; Liu, Yi; Qu, Shiliang

    2016-09-05

    A miniature, robust, and highly sensitive optical fiber temperature sensor based on cascaded polymer-microbubble cavities was fabricated by polymer-filling and subsequent heat-curing process. The expansion of polymer cavity results in the compression of microbubble cavity when the sensor is heated. We demodulated the interference spectrum by means of the fast-Fourier transform (FFT) and signal filtering. Since the thermal response of the polymer cavity is positive and that of the microbubble cavity is negative, a high sensitivity of the temperature sensor is achieved by a subtraction between the two reciprocal thermal responses. Experimental results show that the sensitivity of the temperature sensor is as high as 5.013 nm/°C in the measurement range between 20 °C and 55 °C. Meanwhile, such a sensor has potential for mass production, owing to the simple, nontoxic, and cost-effective process of fabrication.

  9. The multiphase flow system used in exploiting depleted reservoirs: water-based Micro-bubble drilling fluid

    Science.gov (United States)

    Li-hui, Zheng; Xiao-qing, He; Li-xia, Fu; Xiang-chun, Wang

    2009-02-01

    Water-based micro-bubble drilling fluid, which is used to exploit depleted reservoirs, is a complicated multiphase flow system that is composed of gas, water, oil, polymer, surfactants and solids. The gas phase is separate from bulk water by two layers and three membranes. They are "surface tension reducing membrane", "high viscosity layer", "high viscosity fixing membrane", "compatibility enhancing membrane" and "concentration transition layer of liner high polymer (LHP) & surfactants" from every gas phase centre to the bulk water. "Surface tension reducing membrane", "high viscosity layer" and "high viscosity fixing membrane" bond closely to pack air forming "air-bag", "compatibility enhancing membrane" and "concentration transition layer of LHP & surfactants" absorb outside "air-bag" to form "incompact zone". From another point of view, "air-bag" and "incompact zone" compose micro-bubble. Dynamic changes of "incompact zone" enable micro-bubble to exist lonely or aggregate together, and lead the whole fluid, which can wet both hydrophilic and hydrophobic surface, to possess very high viscosity at an extremely low shear rate but to possess good fluidity at a higher shear rate. When the water-based micro-bubble drilling fluid encounters leakage zones, it will automatically regulate the sizes and shapes of the bubbles according to the slot width of fracture, the height of cavern as well as the aperture of openings, or seal them by making use of high viscosity of the system at a very low shear rate. Measurements of the rheological parameters indicate that water-based micro-bubble drilling fluid has very high plastic viscosity, yield point, initial gel, final gel and high ratio of yield point and plastic viscosity. All of these properties make the multiphase flow system meet the requirements of petroleum drilling industry. Research on interface between gas and bulk water of this multiphase flow system can provide us with information of synthesizing effective agents to

  10. Treatment of microvascular micro-embolization using microbubbles and long-tone-burst ultrasound: an in vivo study.

    Science.gov (United States)

    Pacella, John J; Brands, Judith; Schnatz, Frederick G; Black, John J; Chen, Xucai; Villanueva, Flordeliza S

    2015-02-01

    Despite epicardial coronary artery reperfusion by percutaneous coronary intervention, distal micro-embolization into the coronary microcirculation limits myocardial salvage during acute myocardial infarction. Thrombolysis using ultrasound and microbubbles (sonothrombolysis) is an approach that induces microbubble oscillations to cause clot disruption and restore perfusion. We sought to determine whether this technique could restore impaired tissue perfusion caused by thrombotic microvascular obstruction. In 16 rats, an imaging transducer was placed on the biceps femoris muscle, perpendicular to a single-element 1-MHz treatment transducer. Ultrasound contrast perfusion imaging was performed at baseline and after micro-embolization. Therapeutic ultrasound (5000 cycles, pulse repetition frequency = 0.33 Hz, 1.5 MPa) was delivered to nine rats for two 10-min sessions during intra-arterial infusion of lipid-encapsulated microbubbles; seven control rats received no ultrasound-microbubble therapy. Ultrasound contrast perfusion imaging was repeated after each treatment or control period, and microvascular volume was measured as peak video intensity. There was a 90% decrease in video intensity after micro-embolization (from 8.6 ± 4.8 to 0.7 ± 0.8 dB, p ultrasound-microbubble sessions were respectively followed by video intensity increases of 5.8 ± 5.1 and 8.7 ± 5.7 dB (p ultrasound with microbubbles has a therapeutic effect on microvascular perfusion and may be a valuable adjunct to reperfusion therapy for acute myocardial infarction. Copyright © 2015 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  11. The dynamic behavior of microbubbles during long ultrasound tone-burst excitation: mechanistic insights into ultrasound-microbubble mediated therapeutics using high-speed imaging and cavitation detection

    Science.gov (United States)

    Pacella, John J.; Villanueva, Flordeliza S.

    2015-01-01

    Ultrasound (US)-microbubble (MB) mediated therapies have been shown to restore perfusion and enhance drug/gene delivery. Due to the presumption that MBs do not persist during long US exposure under high acoustic pressures, most schemes utilize short US pulses when a high US pressure is employed. However, we recently observed an enhanced thrombolytic effect using long US pulses at high acoustic pressures. Therefore we explored the fate of MBs during long tone-burst exposures (5 ms) at various acoustic pressures and MB concentrations via direct high-speed optical observation and passive cavitation detection. MBs first underwent stable or inertial cavitation depending on the acoustic pressure, and then formed gas-filled clusters that continued to oscillate, break up, and form new clusters. Cavitation detection confirmed continued, albeit diminishing acoustic activity throughout the 5-ms US excitation. These data suggest that persisting cavitation activity during long tone-bursts may confer additional therapeutic effects. PMID:26603628

  12. Integrative Modeling Reveals Annexin A2-mediated Epigenetic Control of Mesenchymal Glioblastoma.

    Science.gov (United States)

    Kling, Teresia; Ferrarese, Roberto; Ó hAilín, Darren; Johansson, Patrik; Heiland, Dieter Henrik; Dai, Fangping; Vasilikos, Ioannis; Weyerbrock, Astrid; Jörnsten, Rebecka; Carro, Maria Stella; Nelander, Sven

    2016-10-01

    Glioblastomas are characterized by transcriptionally distinct subtypes, but despite possible clinical relevance, their regulation remains poorly understood. The commonly used molecular classification systems for GBM all identify a subtype with high expression of mesenchymal marker transcripts, strongly associated with invasive growth. We used a comprehensive data-driven network modeling technique (augmented sparse inverse covariance selection, aSICS) to define separate genomic, epigenetic, and transcriptional regulators of glioblastoma subtypes. Our model identified Annexin A2 (ANXA2) as a novel methylation-controlled positive regulator of the mesenchymal subtype. Subsequent evaluation in two independent cohorts established ANXA2 expression as a prognostic factor that is dependent on ANXA2 promoter methylation. ANXA2 knockdown in primary glioblastoma stem cell-like cultures suppressed known mesenchymal master regulators, and abrogated cell proliferation and invasion. Our results place ANXA2 at the apex of a regulatory cascade that determines glioblastoma mesenchymal transformation and validate aSICS as a general methodology to uncover regulators of cancer subtypes. Copyright © 2016. Published by Elsevier B.V.

  13. Integrative Modeling Reveals Annexin A2-mediated Epigenetic Control of Mesenchymal Glioblastoma

    Directory of Open Access Journals (Sweden)

    Teresia Kling

    2016-10-01

    Full Text Available Glioblastomas are characterized by transcriptionally distinct subtypes, but despite possible clinical relevance, their regulation remains poorly understood. The commonly used molecular classification systems for GBM all identify a subtype with high expression of mesenchymal marker transcripts, strongly associated with invasive growth. We used a comprehensive data-driven network modeling technique (augmented sparse inverse covariance selection, aSICS to define separate genomic, epigenetic, and transcriptional regulators of glioblastoma subtypes. Our model identified Annexin A2 (ANXA2 as a novel methylation-controlled positive regulator of the mesenchymal subtype. Subsequent evaluation in two independent cohorts established ANXA2 expression as a prognostic factor that is dependent on ANXA2 promoter methylation. ANXA2 knockdown in primary glioblastoma stem cell-like cultures suppressed known mesenchymal master regulators, and abrogated cell proliferation and invasion. Our results place ANXA2 at the apex of a regulatory cascade that determines glioblastoma mesenchymal transformation and validate aSICS as a general methodology to uncover regulators of cancer subtypes.

  14. Annexin A2 system in human biology: cell surface and beyond.

    Science.gov (United States)

    Luo, Min; Hajjar, Katherine A

    2013-06-01

    Annexin A2 (A2) is a multicompartmental, multifunctional protein that orchestrates a growing spectrum of biologic processes. At the endothelial cell surface, A2 and S100A10 (p11) form a heterotetramer, which accelerates tissue plasminogen activator-dependent activation of the fibrinolytic protease, plasmin. In antiphospholipid syndrome, anti-A2 antibodies are associated with clinical thrombosis, whereas overexpression of A2 in acute promyelocytic leukemia promotes hyperfibrinolytic bleeding. A2 is upregulated in hypoxia, and mice deficient in A2 are resistant to oxygen-induced retinal neovascularization, suggesting a role for A2 in human retinal vascular proliferation. In solid malignancies, the (A2•p11)(2) tetramer may promote cancer cell invasion, whereas in multiple myeloma A2 enables malignant plasmacyte growth and predicts prognosis. In the central nervous system, the p11 enables membrane insertion of serotonin receptors that govern mood. In the peripheral nervous system, p11 directs sodium channels to the plasma membrane, enabling pain perception. In cerebral cortex neurons, A2 stabilizes the microtubule-associated tau protein, which, when mutated, is associated with frontotemporal dementia. In inflammatory dendritic cells, A2 maintains late endosomal/lysosomal membrane integrity, thus modulating inflammasome activation and cytokine secretion in a model of aseptic arthritis. Together, these findings suggest an emerging, multifaceted role for A2 in human health and disease. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

  15. Annexin II mRNA expression in bovine oocytes during follicular development

    Directory of Open Access Journals (Sweden)

    Luis Fabiano Santos da Costa

    2006-01-01

    Full Text Available We investigated the expression of calcium-dependent phospholipid binding protein annexin-II (Ann-II messenger RNA (mRNA during preantral follicle development and in oocytes from antral follicles of different diameters ( 8 mm. The action of retinol on Ann-II mRNA expression in mature oocytes was also examined. Only oocytes from secondary preantral follicles expressed Ann-II mRNA and at the germinal vesicle stage expression by oocytes from follicles larger than 8 mm was significantly higher (p < 0.05 compared with oocytes from follicles smaller than 3 mm or between 5 and 8 mm. Ann-II mRNA expression by metaphase II oocytes from follicles larger than 8 mm was significantly higher (p < 0.05 than that from oocytes from follicles smaller than 3 mm, with oocytes from both these size-classes showing similar levels of Ann-II mRNA expression as oocytes recovered from 5-8 mm follicles. In the presence of retinol, Ann-II mRNA expression was higher than when retinol was absent (p < 0.05. Our data indicate that Ann-II mRNA expression is highest in competent oocytes and that retinol increases Ann-II mRNA and may be involved in the regulation of oocyte competence by decreasing the translation and/or degradation of Ann-II mRNA.

  16. Annexin A10 expression correlates with serrated pathway features in colorectal carcinoma with microsatellite instability.

    Science.gov (United States)

    Kim, Jung Ho; Rhee, Ye-Young; Kim, Kyung-Ju; Cho, Nam-Yun; Lee, Hye Seung; Kang, Gyeong Hoon

    2014-12-01

    Annexin A10 (ANXA10) has recently been identified as a marker of sessile serrated adenomas/polyps of the colorectum. Although the serrated neoplasia pathway is thought to be involved in the majority of microsatellite instability-high (MSI-H) sporadic colorectal carcinomas (CRCs), the clinicopathological implications of ANXA10 expression in CRC are unknown. Here, we evaluated ANXA10 expression status in 168 MSI-H CRCs by immunohistochemistry. Among 168 MSI-H CRCs, nuclear staining for ANXA10 in tumor cells revealed 28 cases (17%) with ANXA10-positive (ANXA10+) tumors. Most of the ANXA10+ tumors were located in the proximal colon (96%, p < 0.001). The ANXA10+ phenotype in MSI-H CRC was significantly associated with female gender (68%, p = 0.016), CpG island methylator phenotype-high (CIMP-H) (68%, p < 0.001), MLH1 promoter hypermethylation (61%, p < 0.001), loss of MLH1 expression (82%, p = 0.019), and wild-type KRAS status (96%, p = 0.023). Survival analysis revealed no prognostic significance of ANXA10 expression in MSI-H CRC. In conclusion, ANXA10+ MSI-H colon carcinomas are characterized by serrated pathway features, including proximal location, female predominance, and high frequencies of CIMP-H status and MLH1 methylation. © 2014 APMIS. Published by John Wiley & Sons Ltd.

  17. Proteins Annexin A2 and PSA in Prostate Cancer Biopsies Do Not Predict Biochemical Failure.

    Science.gov (United States)

    Lamb, David S; Sondhauss, Sven; Dunne, Jonathan C; Woods, Lisa; Delahunt, Brett; Ferguson, Peter; Murray, Judith; Nacey, John N; Denham, James W; Jordan, T William

    2017-12-01

    We previously reported the use of mass spectrometry and western blotting to identify proteins from tumour regions of formalin-fixed paraffin-embedded biopsies from 16 men who presented with apparently localized prostate cancer, and found that annexin A2 (ANXA2) appeared to be a better predictor of subsequent biochemical failure than prostate-specific antigen (PSA). In this follow-up study, ANXA2 and PSA were measured using western blotting of proteins extracted from biopsies from 37 men from a subsequent prostate cancer trial. No significant differences in ANXA2 and PSA levels were observed between men with and without biochemical failure. The statistical effect sizes were small, d=0.116 for ANXA2, and 0.266 for PSA. ANXA2 and PSA proteins measured from biopsy tumour regions are unlikely to be good biomarkers for prediction of the clinical outcome of prostate cancer presenting with apparently localized disease. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

  18. Microbubble-based enhancement of radiation effect: Role of cell membrane ceramide metabolism.

    Directory of Open Access Journals (Sweden)

    Azza Al-Mahrouki

    Full Text Available Ultrasound (US stimulated microbubbles (MB is a new treatment approach that sensitizes cancer cells to radiation (XRT. The molecular pathways in this response remain unelucidated, however, previous data has supported a role for cell membrane-metabolism related pathways including an up regulation of UDP glycosyltransferase 8 (UGT8, which catalyzes the transfer of galactose to ceramide, a lipid that is associated with the induction of apoptotic signalling. In this study, the role of UGT8 in responses of prostate tumours to ultrasound-stimulated microbubble radiation enhancement therapy is investigated. Experiments were carried out with cells in vitro and tumours in vivo in which UGT8 levels had been up regulated or down regulated. Genetically modified PC3 cells were treated with XRT, US+MB, or a combination of XRT+US+MB. An increase in the immunolabelling of ceramide was observed in cells where UGT8 was down-regulated as opposed to cells where UGT8 was either not regulated or was up-regulated. Clonogenic assays have revealed a decreased level of cellular survival with the down-regulation of UGT8. Xenograft tumours generated from stably transfected PC3 cells were also treated with US+MB, XRT or US+MB+XRT. Histology demonstrated more cellular damage in tumours with down-regulated UGT8 in comparison with control tumours. In contrast, tumours with up-regulated UGT8 had less damage than control tumours. Power Doppler imaging indicated a reduction in the vascular index with UGT8 down-regulation and photoacoustic imaging revealed a reduction in oxygen saturation. This was contrary to when UGT8 was up regulated. The down regulation of UGT8 led to the accumulation of ceramide resulting in more cell death signalling and therefore, a greater enhancement of radiation effect when vascular disruption takes place through the use of ultrasound-stimulated microbubbles.

  19. Vascular gene transfer and drug delivery in vitro using low-frequency ultrasound and microbubbles

    Science.gov (United States)

    Yang, Hong; Liu, Zhong-hua; Liu, Yi-yao; Lou, Chang-chun; Ren, Zheng-long; Miyoshi, Hirokazu

    2010-01-01

    Aim: To determine the effects of ultrasound exposure in combination with a microbubble contrast agent (SonoVue) on the cellular uptake and delivery of drugs/genes into human umbilical vein endothelial cells (HUVECs) as well as their biological effects on migration. Methods: HUVECs in suspension were exposed to pulsed ultrasound with a 10% duty cycle in combination with various concentrations of a microbubble contrast agent (SonoVue) using a digital sonifier at a frequency of 20 kHz and an intensity of 3.77 W/cm2 on the surface of a horn tip. Cell culture inserts were used to determine the cell migration ability. Results: Exposure to pulsed ultrasound resulted in enhanced green fluorescent protein (EGFP) gene transfection efficiencies ranging from 0.2% to 2%. The transfection efficiency of HUVECs was approximately 3-fold higher in the presence of SonoVue than in its absence at the effective exposure time of 6 s. For drug delivery to HUVECs using ultrasound, the delivery efficiencies of a low-molecular-weight model drug (TO-PRO®−1, MW 645.38) were significantly higher when compared to drug delivery without ultrasound, with a maximum efficiency of approximately 34%. However, the delivery efficiencies of a high-molecular-weight model drug (Dextran-Rhodamine B, MW 70 000) were low, with a maximum delivery efficiency of nearly 0.5%, and gene transfection results were similarly poor. The migration ability of HUVECs exposed to ultrasound was also lower than that of the control (no exposure). Conclusion: The use of low-frequency and low-energy ultrasound in combination with microbubbles could be a potent physical method of increasing drug/gene delivery efficiency. This technique is a promising nonviral approach that can be used in cardiovascular disease therapy. PMID:20348943

  20. An algorithm for sensing venous oxygenation using ultrasound-modulated light enhanced by microbubbles

    Science.gov (United States)

    Honeysett, Jack E.; Stride, Eleanor; Deng, Jing; Leung, Terence S.

    2012-02-01

    Near-infrared spectroscopy (NIRS) can provide an estimate of the mean oxygen saturation in tissue. This technique is limited by optical scattering, which reduces the spatial resolution of the measurement, and by absorption, which makes the measurement insensitive to oxygenation changes in larger deep blood vessels relative to that in the superficial tissue. Acousto-optic (AO) techniques which combine focused ultrasound (US) with diffuse light have been shown to improve the spatial resolution as a result of US-modulation of the light signal, however this technique still suffers from low signal-to-noise when detecting a signal from regions of high optical absorption. Combining an US contrast agent with this hybrid technique has been proposed to amplify an AO signal. Microbubbles are a clinical contrast agent used in diagnostic US for their ability to resonate in a sound field: in this work we also make use of their optical scattering properties (modelled using Mie theory). A perturbation Monte Carlo (pMC) model of light transport in a highly absorbing blood vessel containing microbubbles surrounded by tissue is used to calculate the AO signal detected on the top surface of the tissue. An algorithm based on the modified Beer-Lambert law is derived which expresses intravenous oxygen saturation in terms of an AO signal. This is used to determine the oxygen saturation in the blood vessel from a dual wavelength microbubble-contrast AO measurement. Applying this algorithm to the simulation data shows that the venous oxygen saturation is accurately recovered, and this measurement is robust to changes in the oxygenation of the superficial tissue layer.

  1. Quantitative ultrasound characterization of tumor cell death: ultrasound-stimulated microbubbles for radiation enhancement.

    Directory of Open Access Journals (Sweden)

    Hyunjung Christina Kim

    Full Text Available The aim of this study was to assess the efficacy of quantitative ultrasound imaging in characterizing cancer cell death caused by enhanced radiation treatments. This investigation focused on developing this ultrasound modality as an imaging-based non-invasive method that can be used to monitor therapeutic ultrasound and radiation effects. High-frequency (25 MHz ultrasound was used to image tumor responses caused by ultrasound-stimulated microbubbles in combination with radiation. Human prostate xenografts grown in severe combined immunodeficiency (SCID mice were treated using 8, 80, or 1000 µL/kg of microbubbles stimulated with ultrasound at 250, 570, or 750 kPa, and exposed to 0, 2, or 8 Gy of radiation. Tumors were imaged prior to treatment and 24 hours after treatment. Spectral analysis of images acquired from treated tumors revealed overall increases in ultrasound backscatter intensity and the spectral intercept parameter. The increase in backscatter intensity compared to the control ranged from 1.9±1.6 dB for the clinical imaging dose of microbubbles (8 µL/kg, 250 kPa, 2 Gy to 7.0±4.1 dB for the most extreme treatment condition (1000 µL/kg, 750 kPa, 8 Gy. In parallel, in situ end-labelling (ISEL staining, ceramide, and cyclophilin A staining demonstrated increases in cell death due to DNA fragmentation, ceramide-mediated apoptosis, and release of cyclophilin A as a result of cell membrane permeabilization, respectively. Quantitative ultrasound results indicated changes that paralleled increases in cell death observed from histology analyses supporting its use for non-invasive monitoring of cancer treatment outcomes.

  2. Dynamic manipulation of the subharmonic scattering of phospholipid-coated microbubbles

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    Faez, Telli; Renaud, Guillaume; De Jong, Nico [Biomedical Engineering Thoraxcenter, Erasmus Medical Center, PO Box 2040, 3000 CA Rotterdam (Netherlands); Defontaine, Marielle; Calle, Samuel, E-mail: t.faez@erasmusmc.nl [INSERM U930-CNRS ERL3106, Universite Francois Rabelais, UFR Medecine, 10 bd Tonnelle, 37000 Tours (France)

    2011-10-07

    In this paper, the influence of a dynamic variation in the ambient pressure on the subharmonic response of phospholipid-coated microbubbles was investigated. The ambient pressure in water was modulated by a 2.5 kHz acoustic wave with a peak amplitude of 15 kPa. We investigated the fundamental and subharmonic emissions at two driving frequencies: 5 and 10 MHz. The modulation of the bubble radius induced by the dynamic variation in the liquid ambient pressure subsequently causes modulations of the scattered acoustic pressure at the fundamental and subharmonic frequencies (half the fundamental frequency). As a first result, we measured that the variation in the ambient pressure of 15 kPa can modulate the subharmonic amplitude up to 10 dB as compared to the static atmospheric pressure condition. As a second result, we noticed that the relative subharmonic amplitude modulation as a function of the LF acoustic pressure was symmetrical for the 5 MHz driving frequency but asymmetric for 10 MHz. In the latter case, the subharmonic amplitude was more enhanced for an ambient overpressure than reduced for an ambient depression of the same amplitude likely due to the buckling of the lipid shell. However, the fundamental amplitude was symmetrically modulated during bubble compression and expansion. Moreover, subharmonic and fundamental amplitude modulations were found to be either in phase or out of phase with the low-frequency acoustic pressure. Numerical simulations showed that this behavior can be obtained depending on the bubbles' diameter. The highest subharmonic amplitude was measured when microbubbles were insonified at 10 MHz. This fact together with the asymmetry observed in the subharmonic modulation suggests that smaller bubbles with a buckling shell are excited at 10 MHz compared to 5 MHz. These results present new potentials for in vitro characterization of contrast agent microbubbles and possibly a new imaging modality.

  3. First Pregnancy, Somatic and Psychological Status of a 4-Year-Old Child Born following Annexin V TESA Sperm Separation

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    Krzysztof Lukaszuk

    2015-10-01

    Full Text Available Introduction - Sperm DNA integrity is a crucial paternal factor affecting fertilization and pregnancy rates, as well as embryo development. Case - The present case report describes the successful pregnancy after testicular sperm aspiration (TESA combined with intracytoplasmic sperm injection (ICSI (TESA-ICSI in a couple where the male presented high sperm DNA fragmentation. In order to sort damaged sperm presenting DNA fragmentation, magnetic activated cell sorting (MACS with annexin V microbeads (MACS Miltenyi Biotec, Teterow, Germany was used. Conclusion - The authors present the first description of a successful medical case using TESA-ICSI annexin V sperm sorting. Additionally, a follow-up of the child at the age of 4 years old was done.

  4. Scaling behavior of microbubbles rising in water-saturated porous media

    Science.gov (United States)

    Kong, X.; Ma, Y.; Scheuermann, A.; Bringemeier, D.; Galindo-Torres, S. A.; Saar, M. O.; Li, L.

    2015-12-01

    Gas transport in the form of discrete microbubbles in saturated porous media is of importance in a number of processes relevant to many geo-environmental and engineering systems such as bubbling of greenhouse gases in river and sea beds, hydrocarbon gas migration in coal cleats and rock fractures, and air sparging for remediation of soil contaminated with volatile organic compounds. Under the assumption of no or minor volume expansion during gravity-driven migration, the transport of a single microbubble can be well described using various drag force models. However, not enough attention has been paid to the collective behavior of microbubbles during their ascend as a plume through the saturated porous medium, involving dynamic interactions between individual bubbles, bubbles and the ambient fluid, as well as bubbles and the solid matrix. With our quasi-2D, lab-scale microbubble migration experiments, where bubbles are continuously released from a diffuser at the bottom of a porous bed of hydrated gel beads, we establish a scaling relationship between the gas (bubble) release rate and various characteristic parameters of the bubble plume, such as plume tip velocity, plume width, and breakthrough time of the plume front. We find that the characteristic width of the bubble plume varies as a power of both the gas release rate and the bed thickness, with exponents of 0.2 and 0.4, respectively. Moreover, the characteristic breakthrough time also scales with both the gas release rate and the bed thickness with power-law exponents of -0.4 and 1.2, respectively. The mean pore-water velocity of the circulating ambient water also follows a power-law relationship with the gas release rate being an exponent of 0.6 of the gas release rate. This can be quantitatively proven using a simplified momentum exchange model together with the above power-law exponents for the bubble plume. These analyses on the experimental results are carried out on the basis of non

  5. Cesium decontamination using a microbubble-treated aqueous solution of sodium metasilicate

    International Nuclear Information System (INIS)

    Ueda, Yoshikatsu; Tokuda, Yomei; Goto, Hiroshi

    2015-01-01

    The remediation of materials contaminated with radioactive materials, such as 137 Cs, is important. Here, we report the effectiveness of an aqueous Sodium Metasilicate prepared via a microbubble Crushing process (SMC) for the removal of radioactive 137 Cs from granule conglomerates and nonwoven cloth. A 137 Cs removal ratio of 78% was achieved for the nonwoven cloth sample, and multiple washings at low SMC concentrations were effective. In addition, the volume of the waste solution could be reduced by neutralizing the SMC and using gelation to remove the radioactive material. The decontamination achieved using this process was more efficient than that with sodium hydroxide, even for granule conglomerates. (author)

  6. Resonance Frequency of Optical Microbubble Resonators: Direct Measurements and Mitigation of Fluctuations

    Directory of Open Access Journals (Sweden)

    Alessandro Cosci

    2016-08-01

    Full Text Available This work shows the improvements in the sensing capabilities and precision of an Optical Microbubble Resonator due to the introduction of an encaging poly(methyl methacrylate (PMMA box. A frequency fluctuation parameter σ was defined as a score of resonance stability and was evaluated in the presence and absence of the encaging system and in the case of air- or water-filling of the cavity. Furthermore, the noise interference introduced by the peristaltic and the syringe pumping system was studied. The measurements showed a reduction of σ in the presence of the encaging PMMA box and when the syringe pump was used as flowing system.

  7. Domains I and IV of Annexin A2 Affect the Formation and Integrity of In Vitro Capillary-Like Networks

    OpenAIRE

    Raddum, Aase M.; Evensen, Lasse; Holl?s, Hanne; Grindheim, Ann Kari; Lorens, James B.; Vedeler, Anni

    2013-01-01

    Annexin A2 (AnxA2) is a widely expressed multifunctional protein found in different cellular compartments. In spite of lacking a hydrophobic signal peptide, AnxA2 is found at the cell surface of endothelial cells, indicative of a role in angiogenesis. Increased extracellular levels of AnxA2 in tumours correlate with neoangiogenesis, metastasis and poor prognosis. We hypothesised that extracellular AnxA2 may contribute to angiogenesis by affecting endothelial cell-cell interactions and motilit...

  8. Evaluation of adenosine preconditioning with 99mTc-His10-annexin V in a porcine model of myocardium ischemia and reperfusion injury: preliminary study

    International Nuclear Information System (INIS)

    Ye Fei; Fang Wei; Wang Feng; Hua Zichun; Wang Zizheng; Yang Xiang

    2011-01-01

    Purpose: The goal of this study was to evaluate the feasibility of 99m Tc-His 10 -annexin V for the detection of acute myocardial cell death and to assess the effect of adenosine preconditioning in a porcine model of myocardium ischemia and reperfusion injury (RI). Materials and Methods: 99m Tc-His 10 -annexin V was prepared by one-step direct labeling, and RCP and radiostability were tested. The binding of 99m Tc-His 10 -annexin V to apoptosis was validated in vitro using camptothecin-induced Jurkat cells. In vivo biodistribution was determined in mice by the dissection method. Ischemia of 20-30 min was induced by balloon occlusion of the epicardial coronary artery of the porcine model (n=14). Adenosine was infused intravenously in six pigs before coronary occlusion. 99m Tc-His 10 -annexin V (n=12) was injected intravenously at 1 h after reperfusion. SPECT/CT was acquired at 3 h postinjection. Myocardial perfusion imaging (MPI) with 99m Tc-MIBI was also performed 1 day after His 10 -annexin V imaging. Cardiac tissues were analyzed postmortem using hematoxylin-and-eosin and TUNEL staining. Caspase-3 activity was measured to confirm the presence of apoptosis. Results: 99m Tc-His 10 -annexin V had a RCP >98% and high stability 2 h after radiolabeling; it could bind to apoptotic cells with high affinity. Biodistribution of 99m Tc-His 10 -annexin V showed a predominant uptake in the kidney and relatively low uptake in the myocardium, liver and gastrointestinal tract; rapid clearance from blood and kidney was observed. In the untreated group, intense uptake of His 10 -annexin V was visualized in the defect which was shown in MPI, whereas in the adenosine group a mild uptake of 99m Tc-His 10 -annexin was found in the risk area which showed no defects in the 99m Tc-MIBI image. TUNEL staining and activated caspase-3 confirmed the ongoing apoptosis in RI. Adenosine preconditioning significantly diminished the level of apoptosis. Uptake of His 10 -annexin V in RI correlated

  9. The Annexin a2 Promotes Development in Arthritis through Neovascularization by Amplification Hedgehog Pathway.

    Science.gov (United States)

    Yi, Jun; Zhu, Yan; Jia, Yin; Jiang, Hongdie; Zheng, Xin; Liu, Dejing; Gao, Shunxiang; Sun, Mingjuan; Hu, Bo; Jiao, Binghua; Wang, Lianghua; Wang, Kaihui

    2016-01-01

    The neovascularization network of pannus formation plays a crucial role in the development of rheumatoid arthritis (RA). Annexin a2 (Axna2) is an important mediating agent that induces angiogenesis in vascular diseases. The correlation between Axna2 and pannus formation has not been studied. Here, we provided evidence that compared to osteoarthritis (OA) patients and healthy people, the expression of Axna2 and Axna2 receptor (Axna2R) were up-regulated in patients with RA. Joint swelling, inflammation and neovascularization were increased significantly in mice with collagen-induced arthritis (CIA) that were exogenously added Axna2. Cell experiments showed that Axna2 promoted HUVEC proliferation by binding Axna2R, and could activate Hedgehog (HH) signaling and up-regulate the expression of Ihh and Gli. Besides, expression of Ihh, Patched (Ptc), Smoothened (Smo) and Gli and matrix metalloproteinase-2 (MMP-2), vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2), angiogenic growth factor of HH signaling downstream, were down-regulated after inhibition of expression Axna2R on HUVEC. Together, our research definitely observed that over-expression of Axna2 could promote the development of CIA, especially during the process of pannus formation for the first time. Meanwhile, Axna2 depended on combining Axna2R to activate and enlarge HH signaling and the expression of its downstream VEGF, Ang-2 and MMP-2 to promote HUVEC proliferation, and eventually caused to angiogenesis. Therefore, the role of Axna2 is instructive for understanding the development of RA, suppress the effect of Axna2 might provide a new potential measure for treatment of RA.

  10. The Annexin a2 Promotes Development in Arthritis through Neovascularization by Amplification Hedgehog Pathway.

    Directory of Open Access Journals (Sweden)

    Jun Yi

    Full Text Available The neovascularization network of pannus formation plays a crucial role in the development of rheumatoid arthritis (RA. Annexin a2 (Axna2 is an important mediating agent that induces angiogenesis in vascular diseases. The correlation between Axna2 and pannus formation has not been studied. Here, we provided evidence that compared to osteoarthritis (OA patients and healthy people, the expression of Axna2 and Axna2 receptor (Axna2R were up-regulated in patients with RA. Joint swelling, inflammation and neovascularization were increased significantly in mice with collagen-induced arthritis (CIA that were exogenously added Axna2. Cell experiments showed that Axna2 promoted HUVEC proliferation by binding Axna2R, and could activate Hedgehog (HH signaling and up-regulate the expression of Ihh and Gli. Besides, expression of Ihh, Patched (Ptc, Smoothened (Smo and Gli and matrix metalloproteinase-2 (MMP-2, vascular endothelial growth factor (VEGF and angiopoietin-2 (Ang-2, angiogenic growth factor of HH signaling downstream, were down-regulated after inhibition of expression Axna2R on HUVEC. Together, our research definitely observed that over-expression of Axna2 could promote the development of CIA, especially during the process of pannus formation for the first time. Meanwhile, Axna2 depended on combining Axna2R to activate and enlarge HH signaling and the expression of its downstream VEGF, Ang-2 and MMP-2 to promote HUVEC proliferation, and eventually caused to angiogenesis. Therefore, the role of Axna2 is instructive for understanding the development of RA, suppress the effect of Axna2 might provide a new potential measure for treatment of RA.

  11. Annexin A1 induces skeletal muscle cell migration acting through formyl peptide receptors.

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    Valentina Bizzarro

    Full Text Available Annexin A1 (ANXA1, lipocortin-1 is a glucocorticoid-regulated 37-kDa protein, so called since its main property is to bind (i.e. to annex to cellular membranes in a Ca(2+-dependent manner. Although ANXA1 has predominantly been studied in the context of immune responses and cancer, the protein can affect a larger variety of biological phenomena, including cell proliferation and migration. Our previous results show that endogenous ANXA1 positively modulates myoblast cell differentiation by promoting migration of satellite cells and, consequently, skeletal muscle differentiation. In this work, we have evaluated the hypothesis that ANXA1 is able to exert effects on myoblast cell migration acting through formyl peptide receptors (FPRs following changes in its subcellular localization as in other cell types and tissues. The analysis of the subcellular localization of ANXA1 in C2C12 myoblasts during myogenic differentiation showed an interesting increase of extracellular ANXA1 starting from the initial phases of skeletal muscle cell differentiation. The investigation of intracellular Ca(2+ perturbation following exogenous administration of the ANXA1 N-terminal derived peptide Ac2-26 established the engagement of the FPRs which expression in C2C12 cells was assessed by qualitative PCR. Wound healing assay experiments showed that Ac2-26 peptide is able to increase migration of C2C12 skeletal muscle cells and to induce cell surface translocation and secretion of ANXA1. Our results suggest a role for ANXA1 as a highly versatile component in the signaling chains triggered by the proper calcium perturbation that takes place during active migration and differentiation or membrane repair since the protein is strongly redistributed onto the plasma membranes after an rapid increase of intracellular levels of Ca(2+. These properties indicate that ANXA1 may be involved in a novel repair mechanism for skeletal muscle and may have therapeutic implications with

  12. Serial changes in plasma annexin A1 and cortisol levels in sepsis patients.

    Science.gov (United States)

    Tsai, Wen-Hui; Li, I-Ting; Yu, Yuan-Bin; Hsu, Hui-Chi; Shih, Chung-Hung

    2014-02-28

    Annexin A1 (AnxA1), originally identified as a glucocorticoid-regulated protein, is an impor- tant endogenous anti-inflammatory mediator during the resolution phase of inflammation, and its cir- culating level has been rarely studied in sepsis patients. Glucocorticoid has been extensively used in treating patients with sepsis. However, it is unclear whether endogenous cortisol or exogenous glucocor- ticoid contributes to the regulation of AnxA1 levels in peripheral blood of sepsis patients. The aim of this study was to investigate: [1] serial changes over time in the plasma levels of AnxA1 and cortisol in sepsis patients; and [2] prognostic value of AnxA1 level in the survival of sepsis patients. Fifty-eight adult sepsis patients admitted to an intensive care unit (ICU) were enrolled. The plasma levels of cortisol and AnxA1 were determined by specific enzyme-link immunosorbent assay. Results show that the median daily levels of cortisol at the 1st, 3rd, 5th and 7th day after admission to ICU were signifi- cantly elevated over the cortisol level of the control subjects. However, the AnxA1 level was elevated in only thirty-three patients (56%) over the observation period. There was no significant correlation between cortisol levels and AnxA1 levels. Further analysis indicated that steroid treatment resulted in significant elevation of the cortisol level over time, but did not affect the AnxA1 level. AnxA1 levels were also not statistically different between surviving and non-surviving patients. In conclusions, the circu- lating level of AnxA1 is elevated in a subgroup of sepsis patients, and the AnxA1 level does not correlate with the cortisol level in the peripheral blood of sepsis patients.

  13. Serum annexin A2 levels in acute brucellosis and brucellar spondylodiscitis.

    Science.gov (United States)

    Aktug Demir, N; Kolgelier, S; Sumer, S; Inkaya, A C; Ozcimen, S; Demir, L S; Ural, O; Arpaci, A

    2014-10-01

    Brucellosis is a chronic granulomatous infection and may present with various clinical manifestations. Brucellar spondylodiscitis symptoms are initially subtle and nonspecific. Annexin A2 (ANXA2) is involved in various biological functions, including osteoclast formation, bone resorption, and cell growth regulation. In this study, we aimed to determine the clinical significance of serum ANXA2 levels in acute brucellosis and brucellar spondylodiscitis. This prospective study included 96 acute brucellosis patients and 51 healthy controls. Acute brucellosis was diagnosed by a 1/160 or higher titer in a standard tube agglutination (STA) test or a four-fold increase in titers between two STA tests performed two weeks apart in the presence of clinical symptoms within the last eight weeks and/or growth of Brucella spp. in appropriately prepared culture media. ANXA2 levels were determined with an enzyme-linked immunosorbent assay (ELISA). Forty (41.7 %) of 96 acute brucellosis patients were male and 56 (58.3 %) were female. Serum ANXA2 levels were elevated in patients compared to healthy controls (p = 0.001). Eighteen of 96 (18.7 %) acute brucellosis patients had brucellar spondylodiscitis. The serum ANXA2 levels of patients with brucellar spondylodiscitis were higher than those of patients with acute disease without brucellar spondylodiscitis (p = 0.001). ANXA2, C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) values were elevated in the brucellar spondylodiscitis group compared to patients without brucellar spondylodiscitis. Serum ANXA2 measurement together with ESR and CRP is thought to be indicative in the diagnosis of brucellar spondylodiscitis, a common complication of brucellosis.

  14. Annexin A2 is a natural extrahepatic inhibitor of the PCSK9-induced LDL receptor degradation.

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    Nabil G Seidah

    Full Text Available Proprotein convertase subtilisin/kexin-9 (PCSK9 enhances the degradation of hepatic low-density lipoprotein receptor (LDLR. Deletion of PCSK9, and loss-of-function mutants in humans result in lower levels of circulating LDL-cholesterol and a strong protection against coronary heart disease. Accordingly, the quest for PCSK9 inhibitors has major clinical implications. We have previously identified annexin A2 (AnxA2 as an endogenous binding partner and functional inhibitor of PCSK9. Herein, we studied the relevance of AnxA2 in PCSK9 inhibition and lipid metabolism in vivo. Plasma analyses of AnxA2(-/- mice revealed: i a ∼1.4-fold increase in LDL-cholesterol without significant changes in VLDLs or HDLs, and ii a ∼2-fold increase in circulating PCSK9 levels. Western blotting and immunohistochemistry of AnxA2(-/- tissues revealed that the LDLR was decreased by ∼50% in extrahepatic tissues, such as adrenals and colon. We also show that AnxA2-derived synthetic peptides block the PCSK9≡LDLR interaction in vitro, and adenoviral overexpression of AnxA2 in mouse liver increases LDLR protein levels in vivo. These results suggest that AnxA2 acts as an endogenous regulator of LDLR degradation, mostly in extrahepatic tissues. Finally, we identified an AnxA2 coding polymorphism, V98L, that correlates with lower circulating levels of PCSK9 thereby extending our results on the physiological role of AnxA2 in humans.

  15. HPV16 E6 regulates annexin 1 (ANXA1) protein expression in cervical carcinoma cell lines

    International Nuclear Information System (INIS)

    Calmon, Marilia Freitas; Sichero, Laura; Boccardo, Enrique; Villa, Luisa Lina; Rahal, Paula

    2016-01-01

    Annexin 1 (ANXA1) is a substrate for E6AP mediated ubiquitylation. It has been hypothesized that HPV 16 E6 protein redirects E6AP away from ANXA1, increasing its stability and possibly contributing to viral pathogenesis. We analyzed ANXA1 expression in HPV-positive and negative cervical carcinoma-derived cells, in cells expressing HPV-16 oncogenes and in cells transduced with shRNA targeting E6AP. We observed that ANXA1 protein expression increased in HPV-16-positive tumor cells, in keratinocytes expressing HPV-16 E6wt (wild-type) or E6/E7 and C33 cells expressing HPV-16 E6wt. ANXA1 protein expression decreased in cells transfected with E6 Dicer-substrate RNAs (DsiRNA) and C33 cells cotransduced with HPV-16 E6wt and E6AP shRNA. Moreover, colony number and proliferation rate decreased in HPV16-positive cells transduced with ANXA1 shRNA. We observed that in cells infected with HPV16, the E6 binds to E6AP to degrade p53 and upregulate ANXA1. We suggest that ANXA1 may play a role in HPV-mediated carcinogenesis. - Highlights: • ANXA1 upregulation requires the presence of E6 and E6AP and is dependent on E6 integrity. • E6 binds to E6AP to degrade p53 and upregulate ANXA1 in cells infected with HPV16. • ANXA1 plays a role in cell proliferation in HPV-positive cervical cells.

  16. HPV16 E6 regulates annexin 1 (ANXA1) protein expression in cervical carcinoma cell lines

    Energy Technology Data Exchange (ETDEWEB)

    Calmon, Marilia Freitas [Department of Biology, Institute of Bioscience, Language and Exact Science, São Paulo State University, São Jose do Rio Preto (Brazil); Sichero, Laura [Molecular Biology Laboratory, Centre for Translational Research in Oncology, Instituto do Câncer do Estado de São Paulo (ICESP), São Paulo (Brazil); Boccardo, Enrique [Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo., São Paulo (Brazil); Villa, Luisa Lina [Department of Radiology and Oncology, Faculdade de Medicina, Universidade de São Paulo, São Paulo (Brazil); Rahal, Paula, E-mail: rahalp@yahoo.com.br [Department of Biology, Institute of Bioscience, Language and Exact Science, São Paulo State University, São Jose do Rio Preto (Brazil)

    2016-09-15

    Annexin 1 (ANXA1) is a substrate for E6AP mediated ubiquitylation. It has been hypothesized that HPV 16 E6 protein redirects E6AP away from ANXA1, increasing its stability and possibly contributing to viral pathogenesis. We analyzed ANXA1 expression in HPV-positive and negative cervical carcinoma-derived cells, in cells expressing HPV-16 oncogenes and in cells transduced with shRNA targeting E6AP. We observed that ANXA1 protein expression increased in HPV-16-positive tumor cells, in keratinocytes expressing HPV-16 E6wt (wild-type) or E6/E7 and C33 cells expressing HPV-16 E6wt. ANXA1 protein expression decreased in cells transfected with E6 Dicer-substrate RNAs (DsiRNA) and C33 cells cotransduced with HPV-16 E6wt and E6AP shRNA. Moreover, colony number and proliferation rate decreased in HPV16-positive cells transduced with ANXA1 shRNA. We observed that in cells infected with HPV16, the E6 binds to E6AP to degrade p53 and upregulate ANXA1. We suggest that ANXA1 may play a role in HPV-mediated carcinogenesis. - Highlights: • ANXA1 upregulation requires the presence of E6 and E6AP and is dependent on E6 integrity. • E6 binds to E6AP to degrade p53 and upregulate ANXA1 in cells infected with HPV16. • ANXA1 plays a role in cell proliferation in HPV-positive cervical cells.

  17. Annexin A2 complexes with S100 proteins: structure, function and pharmacological manipulation.

    Science.gov (United States)

    Liu, Yidong; Myrvang, Helene K; Dekker, Lodewijk V

    2015-04-01

    Annexin A2 (AnxA2) was originally identified as a substrate of the pp60v-src oncoprotein in transformed chicken embryonic fibroblasts. It is an abundant protein that associates with biological membranes as well as the actin cytoskeleton, and has been implicated in intracellular vesicle fusion, the organization of membrane domains, lipid rafts and membrane-cytoskeleton contacts. In addition to an intracellular role, AnxA2 has been reported to participate in processes localized to the cell surface including extracellular protease regulation and cell-cell interactions. There are many reports showing that AnxA2 is differentially expressed between normal and malignant tissue and potentially involved in tumour progression. An important aspect of AnxA2 function relates to its interaction with small Ca(2+) -dependent adaptor proteins called S100 proteins, which is the topic of this review. The interaction between AnxA2 and S100A10 has been very well characterized historically; more recently, other S100 proteins have been shown to interact with AnxA2 as well. The biochemical evidence for the occurrence of these protein interactions will be discussed, as well as their function. Recent studies aiming to generate inhibitors of S100 protein interactions will be described and the potential of these inhibitors to further our understanding of AnxA2 S100 protein interactions will be discussed. © 2014 The Authors. British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of The British Pharmacological Society.

  18. Proresolving protein Annexin A1: The role in type 2 diabetes mellitus and obesity.

    Science.gov (United States)

    Pietrani, Nathalia T; Ferreira, Cláudia N; Rodrigues, Kathryna F; Perucci, Luiza O; Carneiro, Fernanda S; Bosco, Adriana A; Oliveira, Marina C; Pereira, Solange S; Teixeira, Antônio L; Alvarez-Leite, Jacqueline I; Ferreira, Adaliene V; Sousa, Lirlândia P; Gomes, Karina B

    2018-04-17

    Annexin A1 (AnxA1) is a protein involved in inflammation resolution that might be altered in obesity-associated type 2 diabetes mellitus (DM), which is a chronic inflammatory disease. The aim of this study was to evaluate AnxA1 serum levels in individuals with and without DM stratified according to the body mass index (BMI), and the dynamic of AnxA1 expression in adipose tissue from humans with obesity and non-obesity. Serum samples were obtained from 41 patients with DM (lean, overweight and obese) and 40 controls, and adipose tissue samples were obtained from 16 individuals with obesity (with or without DM), and 15 controls. DM patients showed similar AnxA1 serum levels when compared to controls. However, when the individuals were stratified according to BMI, AnxA1 levels were higher in individuals with obesity than lean or overweight, and in overweight compared to lean individuals. Moreover, AnxA1 was correlated positively with IL-6 levels. AnxA1 levels were also positively correlated with BMI, waist circumference and waist-to-hip ratio. Furthermore, higher levels of cleaved AnxA1 were observed in adipose tissue from individuals with obesity, independently of DM status. Enhanced levels of AnxA1 in serum of individuals with obesity suggest an attempt to counter-regulate the systemic inflammation process in this disease. However, the higher levels of cleaved AnxA1 in the adipose tissue of individuals with obesity could compromise its anti-inflammatory and proresolving actions, locally. Considering our data, AnxA1 cleavage in the adipose tissue, despite increased serum levels of this protein, and consequently the failure in inflammation resolution, suggests an important pathophysiological mechanism involved in inflammatory status observed in obesity. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  19. Dysregulation of anti-inflammatory annexin A1 expression in progressive Crohns Disease.

    Science.gov (United States)

    Sena, Angela; Grishina, Irina; Thai, Anne; Goulart, Larissa; Macal, Monica; Fenton, Anne; Li, Jay; Prindiville, Thomas; Oliani, Sonia Maria; Dandekar, Satya; Goulart, Luiz; Sankaran-Walters, Sumathi

    2013-01-01

    Development of inflammatory bowel disease (IBD) involves the interplay of environmental and genetic factors with the host immune system. Mechanisms contributing to immune dysregulation in IBD are not fully defined. Development of novel therapeutic strategies is focused on controlling aberrant immune response in IBD. Current IBD therapy utilizes a combination of immunomodulators and biologics to suppress pro-inflammatory effectors of IBD. However, the role of immunomodulatory factors such as annexin A1 (ANXA1) is not well understood. The goal of this study was to examine the association between ANXA1 and IBD, and the effects of anti-TNF-α, Infliximab (IFX), therapy on ANXA1 expression. ANXA1 and TNF-α transcript levels in PBMC were measured by RT PCR. Clinical follow up included the administration of serial ibdQs. ANXA1 expression in the gut mucosa was measured by IHC. Plasma ANXA1 levels were measured by ELISA. We found that the reduction in ANXA1 protein levels in plasma coincided with a decrease in the ANXA1 mRNA expression in peripheral blood of IBD patients. ANXA1 expression is upregulated during IFX therapy in patients with a successful intervention but not in clinical non-responders. The IFX therapy also modified the cellular immune activation in the peripheral blood of IBD patients. Decreased expression of ANXA1 was detected in the colonic mucosa of IBD patients with incomplete resolution of inflammation during continuous therapy, which correlated with increased levels of TNF-α transcripts. Gut mucosal epithelial barrier disruption was evident by increased plasma bacterial 16S levels. Loss of ANXA1 expression may support inflammation during IBD and can serve as a biomarker of disease progression. Changes in ANXA1 levels may be predictive of therapeutic efficacy.

  20. Characterization of Different Microbubbles in Assisting Focused Ultrasound-Induced Blood-Brain Barrier Opening

    Science.gov (United States)

    Wu, Sheng-Kai; Chu, Po-Chun; Chai, Wen-Yen; Kang, Shih-Tsung; Tsai, Chih-Hung; Fan, Ching-Hsiang; Yeh, Chih-Kuang; Liu, Hao-Li

    2017-04-01

    Microbubbles (MBs) serve as a critical catalyst to amplify local cavitation in CNS capillary lumen to facilitate focused ultrasound (FUS) to transiently open the blood-brain barrier (BBB). However, limited understanding is available regarding the effect of different microbubbles to induce BBB opening. The aim of this study is to characterize different MBs on their effect in FUS-induced BBB opening. Three MBs, SonoVue, Definity, and USphere, were tested, with 0.4-MHz FUS exposure at 0.62-1.38 of mechanical index (MI) on rats. Evans blue, dynamic contrast-enhanced (DCE) MRI and small-animal ultrasound imaging were used as surrogates to allow molecule-penetrated quantification, BBB-opened observation, and MBs circulation/persistence. Cavitation activity was measured via the passive cavitation detection (PCD) setup to correlate with the exposure level and the histological effect. Under given and identical MB concentrations, the three MBs induced similar and equivalent BBB-opening effects and persistence. In addition, a treatment paradigm by adapting exposure time is proposed to compensate MB decay to retain the persistence of BBB-opening efficiency in multiple FUS exposures. The results potentially improve understanding of the equivalence among MBs in focused ultrasound CNS drug delivery, and provide an effective strategy for securing persistence in this treatment modality.

  1. Nonlinear radial oscillations of encapsulated microbubbles subject to ultrasound: the effect of membrane constitutive law.

    Science.gov (United States)

    Tsiglifis, Kostas; Pelekasis, Nikos A

    2008-06-01

    The nonlinear radial oscillations of bubbles that are encapsulated in an elastic shell are investigated numerically subject to three different constitutive laws describing the viscoelastic properties of the shell: the Mooney-Rivlin (MR), the Skalak (SK), and the Kelvin-Voigt (KV) models are used in order to describe strain-softening, strain-hardening and small displacement (Hookean) behavior of the shell material, respectively. Due to the isotropic nature of the acoustic disturbances, the area dilatation modulus is the important parameter. When the membrane is strain softening (MR) the resonance frequency decreases with increasing sound amplitude, whereas the opposite happens when the membrane is strain hardening (SK). As the amplitude of the acoustic disturbance increases the total scattering cross section of a microbubble with a SK membrane tends to decrease, whereas that of a KV or a MR membrane tends to increase. The importance of strain-softening behavior in the abrupt onset of volume pulsations, that is often observed with small insonated microbubbles at moderately large sound amplitudes, is discussed.

  2. Mechanism of Microbubble Growth at Mitral Mechanical Heart Valve (MHV) Closure

    Science.gov (United States)

    Rambod, Edmond; Beizaie, Masoud; Shusser, Michael; Gharib, Morteza

    1999-11-01

    The growth mechanism of microbubbles at mitral MHV closure has been experimentally studied. In the heart, some of the tiny bubbles grow explosively and form larger and persistent bubbles. An experimental set-up was designed to allow the passage of micron-size bubbles through an 80 micron-wide slot, simulating a typical gap between the housing ring and the occluders in MHV. The bubbles were generated using an air-liquid dispenser and were delivered to the system via a 250 micron-diameter hypedermic needle positioned vertically near the slot. A solenoid valve was used to deliver a 10cc volume of liquid in 25ms time through the slot. High-speed imaging was used to study the impact of flow through the slot on bubble growth. The velocity of liquid through the slot was assessed to be in the range of 12-15 m/s. Our observations confirmed the rapid and drastic growth of microbubbles following their passage through the narrow slot, due to pressure drop. Vortices, which were induced by flow separation on the downstream of the slot, caused the grown bubbles to shatter and form more stable bubbles.

  3. Power cavitation-guided blood-brain barrier opening with focused ultrasound and microbubbles

    Science.gov (United States)

    Burgess, M. T.; Apostolakis, I.; Konofagou, E. E.

    2018-03-01

    Image-guided monitoring of microbubble-based focused ultrasound (FUS) therapies relies on the accurate localization of FUS-stimulated microbubble activity (i.e. acoustic cavitation). Passive cavitation imaging with ultrasound arrays can achieve this, but with insufficient spatial resolution. In this study, we address this limitation and perform high-resolution monitoring of acoustic cavitation-mediated blood-brain barrier (BBB) opening with a new technique called power cavitation imaging. By synchronizing the FUS transmit and passive receive acquisition, high-resolution passive cavitation imaging was achieved by using delay and sum beamforming with absolute time delays. Since the axial image resolution is now dependent on the duration of the received acoustic cavitation emission, short pulses of FUS were used to limit its duration. Image sets were acquired at high-frame rates for calculation of power cavitation images analogous to power Doppler imaging. Power cavitation imaging displays the mean intensity of acoustic cavitation over time and was correlated with areas of acoustic cavitation-induced BBB opening. Power cavitation-guided BBB opening with FUS could constitute a standalone system that may not require MRI guidance during the procedure. The same technique can be used for other acoustic cavitation-based FUS therapies, for both safety and guidance.

  4. The Role of 99mTc-Annexin V Apoptosis Scintigraphy in Visualizing Early Stage Glucocorticoid-Induced Femoral Head Osteonecrosis in the Rabbit

    Directory of Open Access Journals (Sweden)

    Xiaolong Wang

    2016-01-01

    Full Text Available Objective. To validate the ability of 99mTc-Annexin V to visualize early stage of glucocorticoid-induced femoral head necrosis by comparing with 99mTc-MDP bone scanning. Methods. Femoral head necrosis was induced in adult New Zealand white rabbits by intramuscular injection of methylprednisolone. 99mTc-Annexin scintigraphy and 99mTc-MDP scans were performed before and 5, 6, and 8 weeks after methylprednisolone administration. Rabbits were sacrificed at various time points and conducted for TUNEL and H&E staining. Results. All methylprednisolone treated animals developed femoral head necrosis; at 8 weeks postinjection, destruction of bone structure was evident in H&E staining, and apoptosis was confirmed by the TUNEL assay. This was matched by 99mTc-Annexin V images, which showed a significant increase in signal over baseline. Serial 99mTc-Annexin V scans revealed that increased 99mTc-Annexin V uptake could be observed in 5 weeks. In contrast, there was no effect on 99mTc-MDP signal until 8 weeks. The TUNEL assay revealed that bone cell apoptosis occurred at 5 weeks. Conclusion. 99mTc-Annexin V is superior to 99mTc-MDP for the early detection of glucocorticoid-induced femoral head necrosis in the rabbit and may be a better strategy for the early detection of glucocorticoid-induced femoral head necrosis in patients.

  5. The Role of (99m)Tc-Annexin V Apoptosis Scintigraphy in Visualizing Early Stage Glucocorticoid-Induced Femoral Head Osteonecrosis in the Rabbit.

    Science.gov (United States)

    Wang, Xiaolong; Liu, Yu; Wang, Xuemei; Liu, Rui; Li, Jianbo; Zhang, Guoliang; Li, Qiang; Wang, Lei; Bai, Zhigang; Zhao, Jianmin

    2016-01-01

    To validate the ability of (99m)Tc-Annexin V to visualize early stage of glucocorticoid-induced femoral head necrosis by comparing with (99m)Tc-MDP bone scanning. Femoral head necrosis was induced in adult New Zealand white rabbits by intramuscular injection of methylprednisolone. (99m)Tc-Annexin scintigraphy and (99m)Tc-MDP scans were performed before and 5, 6, and 8 weeks after methylprednisolone administration. Rabbits were sacrificed at various time points and conducted for TUNEL and H&E staining. All methylprednisolone treated animals developed femoral head necrosis; at 8 weeks postinjection, destruction of bone structure was evident in H&E staining, and apoptosis was confirmed by the TUNEL assay. This was matched by (99m)Tc-Annexin V images, which showed a significant increase in signal over baseline. Serial (99m)Tc-Annexin V scans revealed that increased (99m)Tc-Annexin V uptake could be observed in 5 weeks. In contrast, there was no effect on (99m)Tc-MDP signal until 8 weeks. The TUNEL assay revealed that bone cell apoptosis occurred at 5 weeks. (99m)Tc-Annexin V is superior to (99m)Tc-MDP for the early detection of glucocorticoid-induced femoral head necrosis in the rabbit and may be a better strategy for the early detection of glucocorticoid-induced femoral head necrosis in patients.

  6. Apoptotic abscess imaging with {sup 99m}Tc-HYNIC-rh-Annexin-V

    Energy Technology Data Exchange (ETDEWEB)

    Penn, David L.; Kim, Christopher; Zhang, Kaijun; Mukherjee, Archana; Devakumar, Devadhas; Jungkind, Donald [Department of Radiology, Thomas Jefferson University, Philadelphia, PA 19107 (United States); Thakur, Mathew L. [Department of Radiology, Thomas Jefferson University, Philadelphia, PA 19107 (United States)], E-mail: mathew.thakur@jefferson.edu

    2010-01-15

    Abscess formation causes systemic and localized up-regulation of neutrophil [polymorphonuclear leukocytes (PMNs)] signaling pathways. In the abscess, following bacterial ingestion or PMN activation by inflammatory mediators, PMN apoptosis is elevated and leads to the externalization of phosphatidylserine. Annexin-V (AnxV) has been shown to have high affinity to externalized phosphatidylserine. We hypothesized that {sup 99m}Tc-AnxV will target high densities of apoptotic PMNs and image abscesses. AnxV, conjugated with hydrazinenicaotinamide (HYNIC), was labeled with reduced {sup 99m}TcO{sub 4}{sup -} and its purity was determined by instant thin-layer chromatography. Apoptosis was induced in isolated human PMNs by incubation in 2% saline for 17 and 22 h at 37 deg. C. PMNs were then incubated with {sup 99m}Tc-HYNIC-AnxV and associated {sup 99m}Tc was determined. Abscesses were induced in mice by intramuscular injection of bacteria or turpentine. Following intravenous administration of {sup 99m}Tc-HYNIC-AnxV, mice were imaged and tissue distribution studied at 4 and 24 h. Radiochemical purity of {sup 99m}Tc-HYNIC-AnxV was 84.9{+-}8.11%. At 17 h, {sup 99m}Tc-HYNIC-AnxV bound to apoptotic PMNs was 71.6{+-}0.01% and 48.6{+-}0.01% for experimental and control cells, respectively (P=.002). At 22 h, experimental cells retained 74.9{+-}0.02% and control cells retained 47.2{+-}0.02% (P=.005). {sup 99m}Tc-HYNIC-AnxV associated with bacterial abscesses was 1.25{+-}0.09 and 3.75{+-}0.83 percent injected dose per gram (%ID/g) at 4 and 24 h compared to turpentine abscesses which was 1.02{+-}0.16 and 0.72{+-}0.17 %ID/g at 4 (P{<=}.05) and 24 h (P{<=}.01). {sup 99m}Tc-HYNIC-AnxV represents a minimally invasive and promising agent to image and potentially distinguish between infectious and inflammatory abscesses.

  7. Annexin A2 and zinc finger transcription factor Snail expression in glioma tissue and the regulating effect of corresponding siRNA on glioma cells

    Directory of Open Access Journals (Sweden)

    Zheng-Hai Deng

    2016-12-01

    Full Text Available Objective: To study the Annexin A2 and zinc finger transcription factor Snail expression in glioma tissue and the regulating effect of corresponding siRNA on glioma cells. Methods: Glioma and peri-tumor tissue were collected to determine AnnexinA2 and Snail expression; glioma cell lines U373-MG were cultured and transfected with AnnexinA2, Snail and NC siRNA, and then the cell viability, number of migrating and invading cells as well as the expression levels of proliferation and epithelial-mesenchymal transition genes were detected. Results: AnnexinA2 and Snail mRNA levels in glioma tissues were significantly higher than those in peri-tumor tissues; cell viability as well as Ras, Raf, MEK and ERK mRNA levels of AnnexinA2-siRNA group was significantly lower than those of NC-siRNA group, and the migrating cell number and invading cell number as well as E-cadherin, N-cadherin, Vimentin and α-SMA mRNA levels were not significantly different from those of NC-siRNA group; migrating cell number and invading cell number as well as N-cadherin, Vimentin and α-SMA mRNA levels of Snail-siRNA group were significantly lower than those of NC-siRNA group, E-cadherin mRNA level was significantly higher than that of NC-siRNA group, and the cell viability as well as Ras, Raf, MEK and ERK mRNA levels were not significantly different from those of NC-siRNA group. Conclusions: AnnexinA2 and Snail expression levels significantly increase in glioma tissues, highly expressed AnnexinA2 can promote cell proliferation and highly expressed Snail can promote epithelial-mesenchymal transition.

  8. New markers of pancreatic cancer identified through differential gene expression analyses: claudin 18 and annexin A8.

    Science.gov (United States)

    Karanjawala, Zarir E; Illei, Peter B; Ashfaq, Raheela; Infante, Jeffrey R; Murphy, Kathleen; Pandey, Akhilesh; Schulick, Richard; Winter, Jordan; Sharma, Rajni; Maitra, Anirban; Goggins, Michael; Hruban, Ralph H

    2008-02-01

    New markers to distinguish benign reactive glands from infiltrating ductal adenocarcinoma of the pancreas are needed. The gene expression patterns of 24 surgically resected primary infiltrating ductal adenocarcinomas of the pancreas were compared with 18 non-neoplastic samples using the Affymetrix U133 Plus 2.0 Arrays and the Gene Logic GeneExpress Software System. Gene fragments from 4 genes (annexin A8, claudin 18, CXCL5, and S100 A2) were selected from the fragments found to be highly expressed in infiltrating adenocarcinomas when compared with normal tissues. The protein expression of these genes was examined using immunohistochemical labeling of tissue microarrays. Claudin 18 labeled infiltrating carcinomas in a membranous pattern. When compared with normal and reactive ducts, claudin 18 was overexpressed, at least focally, in 159 of 166 evaluable carcinomas (96%). Strong and diffuse claudin 18 overexpression was most often seen in well-differentiated carcinomas (P=0.02). Claudin 18 was overexpressed in 51 of 52 cases (98%) of pancreatic intraepithelial neoplasia. Annexin A8 was at least focally overexpressed in 149 of 154 evaluable infiltrating carcinomas (97%). S100 A2 was at least focally overexpressed in 118 of 154 evaluable infiltrating carcinomas (77%). Non-neoplastic glands also frequently expressed S100 A2 diminishing its potential diagnostic utility. Immunolabeling with antibodies directed against CXCL5 did not reveal any significant differences in protein expression between infiltrating adenocarcinomas and normal pancreatic ducts. Claudin 18 and annexin A8 are frequently highly overexpressed in infiltrating ductal adenocarcinomas when compared with normal reactive ducts, suggesting a role for these molecules in pancreatic ductal adenocarcinomas. Furthermore, these may serve as diagnostic markers, as screening tests and as therapeutic targets.

  9. The S100A10 subunit of the annexin A2 heterotetramer facilitates L2-mediated human papillomavirus infection.

    Science.gov (United States)

    Woodham, Andrew W; Da Silva, Diane M; Skeate, Joseph G; Raff, Adam B; Ambroso, Mark R; Brand, Heike E; Isas, J Mario; Langen, Ralf; Kast, W Martin

    2012-01-01

    Mucosotropic, high-risk human papillomaviruses (HPV) are sexually transmitted viruses that are causally associated with the development of cervical cancer. The most common high-risk genotype, HPV16, is an obligatory intracellular virus that must gain entry into host epithelial cells and deliver its double stranded DNA to the nucleus. HPV capsid proteins play a vital role in these steps. Despite the critical nature of these capsid protein-host cell interactions, the precise cellular components necessary for HPV16 infection of epithelial cells remains unknown. Several neutralizing epitopes have been identified for the HPV16 L2 minor capsid protein that can inhibit infection after initial attachment of the virus to the cell surface, which suggests an L2-specific secondary receptor or cofactor is required for infection, but so far no specific L2-receptor has been identified. Here, we demonstrate that the annexin A2 heterotetramer (A2t) contributes to HPV16 infection and co-immunoprecipitates with HPV16 particles on the surface of epithelial cells in an L2-dependent manner. Inhibiting A2t with an endogenous annexin A2 ligand, secretory leukocyte protease inhibitor (SLPI), or with an annexin A2 antibody significantly reduces HPV16 infection. With electron paramagnetic resonance, we demonstrate that a previously identified neutralizing epitope of L2 (aa 108-120) specifically interacts with the S100A10 subunit of A2t. Additionally, mutation of this L2 region significantly reduces binding to A2t and HPV16 pseudovirus infection. Furthermore, downregulation of A2t with shRNA significantly decreases capsid internalization and infection by HPV16. Taken together, these findings indicate that A2t contributes to HPV16 internalization and infection of epithelial cells and this interaction is dependent on the presence of the L2 minor capsid protein.

  10. The S100A10 subunit of the annexin A2 heterotetramer facilitates L2-mediated human papillomavirus infection.

    Directory of Open Access Journals (Sweden)

    Andrew W Woodham

    Full Text Available Mucosotropic, high-risk human papillomaviruses (HPV are sexually transmitted viruses that are causally associated with the development of cervical cancer. The most common high-risk genotype, HPV16, is an obligatory intracellular virus that must gain entry into host epithelial cells and deliver its double stranded DNA to the nucleus. HPV capsid proteins play a vital role in these steps. Despite the critical nature of these capsid protein-host cell interactions, the precise cellular components necessary for HPV16 infection of epithelial cells remains unknown. Several neutralizing epitopes have been identified for the HPV16 L2 minor capsid protein that can inhibit infection after initial attachment of the virus to the cell surface, which suggests an L2-specific secondary receptor or cofactor is required for infection, but so far no specific L2-receptor has been identified. Here, we demonstrate that the annexin A2 heterotetramer (A2t contributes to HPV16 infection and co-immunoprecipitates with HPV16 particles on the surface of epithelial cells in an L2-dependent manner. Inhibiting A2t with an endogenous annexin A2 ligand, secretory leukocyte protease inhibitor (SLPI, or with an annexin A2 antibody significantly reduces HPV16 infection. With electron paramagnetic resonance, we demonstrate that a previously identified neutralizing epitope of L2 (aa 108-120 specifically interacts with the S100A10 subunit of A2t. Additionally, mutation of this L2 region significantly reduces binding to A2t and HPV16 pseudovirus infection. Furthermore, downregulation of A2t with shRNA significantly decreases capsid internalization and infection by HPV16. Taken together, these findings indicate that A2t contributes to HPV16 internalization and infection of epithelial cells and this interaction is dependent on the presence of the L2 minor capsid protein.

  11. Porous Polymeric Films from Microbubbles Generated Using a T-Junction Microfluidic Device.

    Science.gov (United States)

    Elsayed, M; Kothandaraman, A; Edirisinghe, M; Huang, J

    2016-12-20

    In this work, a simple microfluidic junction with a T geometry and coarse (200 μm diameter) capillaries was used to generate monodisperse microbubbles with an alginate polymer shell. Subsequently, these bubbles were used to prepare porous alginate films with good control over the pore structure. The lack of pore size, shape, and surface control in scalable forming of polymeric films is a major application-limiting drawback at present. Controlling the thinning process of the shell of the bubbles to tune the surface of the resulting structures was also explored. Films were prepared with nanopatterned surfaces by controlling the thinning of the bubble shell, with the aid of surfactants, to induce efficient bursting (fragmentation) of bubbles to generate nanodroplets, which become embedded within the film surface. This novel feature greatly expands and enhances the use of hydrophilic polymers in a wide range of biomedical applications, particularly in drug delivery and tissue engineering, such as studying cellular responses to different morphological surfaces.

  12. Scalable and reusable micro-bubble removal method to flatten large-area 2D materials

    Science.gov (United States)

    Pham, Phi H. Q.; Quach, Nhi V.; Li, Jinfeng; Burke, Peter J.

    2018-04-01

    Bubbles generated during electro-delamination and chemical etch during large-area two-dimensional (2D) material transfer has been shown to cause rippling, and consequently, results in tears and wrinkles in the transferred film. Here, we demonstrate a scalable and reusable method to remove surface adhered micro-bubbles by using hydrophobic surfaces modified by self-assembled monolayers (SAMs). Bubble removal allows the 2D film to flatten out and prevents the formation of defects. Electrical characterization was used to verify improved transfer quality and was confirmed by increased field-effect mobility and decreased sheet resistance. Raman spectroscopy was also used to validate enhanced electrical quality following transfer. The bubble removal method can be applied to an assortment of 2D materials using diverse hydrophobic SAM variants. Our studies can be integrated into large scale applications and will lead to improved large-area 2D electronics in general.

  13. A model for an acoustically driven microbubble inside a rigid tube

    KAUST Repository

    Qamar, Adnan

    2014-09-10

    A theoretical framework to model the dynamics of acoustically driven microbubble inside a rigid tube is presented. The proposed model is not a variant of the conventional Rayleigh-Plesset category of models. It is derived from the reduced Navier-Stokes equation and is coupled with the evolving flow field solution inside the tube by a similarity transformation approach. The results are computed, and compared with experiments available in literature, for the initial bubble radius of Ro=1.5μm and 2μm for the tube diameter of D=12μm and 200μm with the acoustic parameters as utilized in the experiments. Results compare quite well with the existing experimental data. When compared to our earlier basic model, better agreement on a larger tube diameter is obtained with the proposed coupled model. The model also predicts, accurately, bubble fragmentation in terms of acoustic and geometric parameters.

  14. Numerical investigation of microbubble formation in liquid-liquid impact events

    Science.gov (United States)

    Mirjalili, Seyedshahabaddin; Mani, Ali

    2012-11-01

    A numerical study of the problem of a droplet impacting another layer of the same liquid is performed with the primary motivation of understanding the steps that lead to the formation of multiple micro-bubbles in the Mesler entrainment mechanism. Simulations start before impact, where a thin gas layer is present and are continued to stages after impact, taking care of topological changes, and finally depicting the formation of the chandelier-like pattern of small bubbles observed in Mesler entrainment. A two dimensional boundary element approach similar to the work of M. Mani, Mandre and Brenner (JFM, vol. 647, p. 163, 2010) is undertaken with the appropriate assumption of inviscid, incompressible potential flow in the liquid bodies, thin structure lubrication flow in the gas layer, with modifications to allow for large interface deflection and topological changes assuming uniform pressure in the bubbles. Supported by the Office of Naval Research.

  15. Generation of emulsion droplets and micro-bubbles in microfluidic devices

    KAUST Repository

    Zhang, Jiaming

    2016-04-01

    Droplet-based microfluidic devices have become a preferred versatile platform for various fields in physics, chemistry and biology to manipulate small amounts of liquid samples. In addition to microdroplets, microbubbles are also needed for various pro- cesses in the food, healthcare and cosmetic industries. Polydimethylsiloxane (PDMS) soft lithography, the mainstay for fabricating microfluidic devices, usually requires the usage of expensive apparatus and a complex manufacturing procedure. In ad- dition, current methods have the limited capabilities for fabrication of microfluidic devices within three dimensional (3D) structures. Novel methods for fabrication of droplet-based microfluidic devices for the generation microdroplets and microbubbles are therefore of great interest in current research. In this thesis, we have developed several simple, rapid and low-cost methods for fabrication of microfluidic devices, especially for generation of microdroplets and mi- crobubbles. We first report an inexpensive full-glass microfluidic devices with as- sembly of glass capillaries, for generating monodisperse multiple emulsions. Different types of devices have been designed and tested and the experimental results demon- strated the robust capability of preparing monodisperse single, double, triple and multi-component emulsions. Second, we propose a similar full-glass device for generation of microbubbles, but with assembly of a much smaller nozzle of a glass capillary. Highly monodisperse microbubbles with diameter range from 3.5 to 60 microns have been successfully produced, at rates up to 40 kHz. A simple scaling law based on the capillary number and liquid-to-gas flow rate ratio, successfully predicts the bubble size. Recently, the emergent 3D printing technology provides an attractive fabrication technique, due to its simplicity and low cost. A handful of studies have already demonstrated droplet production through 3D-printed microfluidic devices. However, two

  16. Advantages of the phosphatidylserine-recognizing peptide PSP1 for molecular imaging of tumor apoptosis compared with annexin V.

    Directory of Open Access Journals (Sweden)

    Soyoun Kim

    Full Text Available A number of peptide-based indicators have been identified and reported as potential apoptosis probes, offering great promise for early assessment of therapeutic efficacy in several types of cancer. Direct comparison of the newly developed probes with previously used ones would be an important step in assessing possible applications. Here, we compared the newly identified peptide-based phosphatidylserine (PS indicator PSP1 (CLSYYPSYC with annexin V, a common probe for molecular imaging of apoptotic cells, with respect to PS binding kinetics, apoptotic cell-targeting ability, and the efficacy of homing to apoptotic tumor cells in a mouse model after treatment with the anticancer agent camptothecin. Our results indicate that PSP1 efficiently targeted apoptotic cells and generated apoptosis/tumor-specific signals after cancer treatment in the animal model, whereas a similar dose of annexin V showed weak signals. The formation of a stable complex of PSP1 with PS might be one reason for the efficient in vivo targeting. We suggest that PSP1 has potential advantages for in vivo apoptotic cell imaging and could serve as a platform for the development of de novo peptide-based probes for apoptosis.

  17. Role of lipoxygenases and lipoxin A(4)/annexin-1 receptor in gastric protection induced by 20% ethanol or sodium salicylate in rats.

    Science.gov (United States)

    Peskar, Brigitta M; Ehrlich, Karlheinz; Schuligoi, Rufina; Peskar, Bernhard A

    2009-01-01

    The role of cyclooxygenases and prostaglandins in experimental models of gastroprotection is well established. We investigated the effects of the 5-lipoxygenase inhibitor A63162, the 12-lipoxygenase inhibitor baicalein and the 15-lipoxygenase inhibitor PD146176 as well as the nonspecific lipoxin A(4)/annexin-1 antagonist Boc1 on adaptive protection induced by 20% ethanol against 70% ethanol, and on protection induced by sodium salicylate against the mucosal-damage-aggravating effects of celecoxib and dexamethasone during local ischemia-reperfusion in rats. It was found that both types of gastroprotection were antagonized by the lipoxygenase inhibitors and the lipoxin A(4)/annexin-1 antagonist in doses that have no direct damaging effect on gastric mucosa. The results suggest that not only cyclooxygenases, but also active lipoxygenases and, possibly, annexin-1 are required for these types of gastroprotection to occur. Copyright 2009 S. Karger AG, Basel.

  18. Annexin A2 and its downstream IL-6 and HB-EGF as secretory biomarkers in the differential diagnosis of Her-2 negative breast cancer.

    Science.gov (United States)

    Shetty, Praveenkumar; Patil, Vidya S; Mohan, Rajashekar; D'souza, Leonard Clinton; Bargale, Anil; Patil, Basavaraj R; Dinesh, U S; Haridas, Vikram; Kulkarni, Shrirang P

    2017-07-01

    Background AnnexinA2 (AnxA2) membrane deposition has a critical role in HB-EGF shedding as well as IL-6 secretion in breast cancer cells. This autocrine cycle has a major role in cancer cell proliferation, migration and metastasis. The objective of the study is to demonstrate annexinA2-mediated autocrine regulation via HB-EGF and IL-6 in Her-2 negative breast cancer progression. Methods Secretory annexinA2, HB-EGF and IL-6 were analysed in the peripheral blood sample of Her-2 negative ( n = 20) and positive breast cancer patients ( n = 16). Simultaneously, tissue expression was analysed by immunohistochemistry. The membrane deposition of these secretory ligands and their autocrine regulation was demonstrated using triple-negative breast cancer cell line model. Results Annexina2 and HB-EGF expression are inversely correlated with Her-2, whereas IL-6 expression is seen in both Her-2 negative and positive breast cancer cells. RNA interference studies and upregulation of annexinA2 proved that annexinA2 is the upstream of this autocrine pathway. Abundant soluble serum annexinA2 is secreted in Her-2 negative breast cancer (359.28 ± 63.73 ng/mL) compared with normal (286.10 ± 70.04 ng/mL, P breast cancer phenotypes as compared with normal ( P breast cancer tissues, increased secretion compared with normal cells, and their major role in the regulation of EGFR downstream signalling makes these molecules as a potential tissue and serum biomarker and an excellent therapeutic target in Her-2 negative breast cancer.

  19. Therapeutic effects of microbubble added to combined high-intensity focused ultrasound and chemotherapy in a pancreatic cancer xenograft model

    International Nuclear Information System (INIS)

    Yu, Mi Hye; Lee, Jae Young; Kim, Bo Ram; Park, Eun Joo; Kim, Hoe Suk; Han, Joon Koo; Kim, Hae Ri; Choi, Byung Ihn

    2016-01-01

    To investigate whether high-intensity focused ultrasound (HIFU) combined with microbubbles enhances the therapeutic effects of chemotherapy. A pancreatic cancer xenograft model was established using BALB/c nude mice and luciferase-expressing human pancreatic cancer cells. Mice were randomly assigned to five groups according to treatment: control (n = 10), gemcitabine alone (GEM; n = 12), HIFU with microbubbles (HIFU + MB, n = 11), combined HIFU and gemcitabine (HIGEM; n = 12), and HIGEM + MB (n = 13). After three weekly treatments, apoptosis rates were evaluated using the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay in two mice per group. Tumor volume and bioluminescence were monitored using high-resolution 3D ultrasound imaging and in vivo bioluminescence imaging for eight weeks in the remaining mice. The HIGEM + MB group showed significantly higher apoptosis rates than the other groups (p < 0.05) and exhibited the slowest tumor growth. From week 5, the tumor-volume-ratio relative to the baseline tumor volume was significantly lower in the HIGEM + MB group than in the control, GEM, and HIFU + MB groups (p < 0.05). Despite visible distinction, the HIGEM and HIGEM + MB groups showed no significant differences. High-intensity focused ultrasound combined with microbubbles enhances the therapeutic effects of gemcitabine chemotherapy in a pancreatic cancer xenograft model

  20. Therapeutic Effects of Microbubbles Added to Combined High-Intensity Focused Ultrasound and Chemotherapy in a Pancreatic Cancer Xenograft Model

    International Nuclear Information System (INIS)

    Yu, Mi Hye; Lee, Jae Young; Kim, Hae Ri; Kim, Bo Ram; Park, Eun-Joo; Kim, Hoe Suk; Han, Joon Koo; Choi, Byung Ihn

    2016-01-01

    To investigate whether high-intensity focused ultrasound (HIFU) combined with microbubbles enhances the therapeutic effects of chemotherapy. A pancreatic cancer xenograft model was established using BALB/c nude mice and luciferase-expressing human pancreatic cancer cells. Mice were randomly assigned to five groups according to treatment: control (n = 10), gemcitabine alone (GEM; n = 12), HIFU with microbubbles (HIFU + MB, n = 11), combined HIFU and gemcitabine (HIGEM; n = 12), and HIGEM + MB (n = 13). After three weekly treatments, apoptosis rates were evaluated using the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay in two mice per group. Tumor volume and bioluminescence were monitored using high-resolution 3D ultrasound imaging and in vivo bioluminescence imaging for eight weeks in the remaining mice. The HIGEM + MB group showed significantly higher apoptosis rates than the other groups (p < 0.05) and exhibited the slowest tumor growth. From week 5, the tumor-volume-ratio relative to the baseline tumor volume was significantly lower in the HIGEM + MB group than in the control, GEM, and HIFU + MB groups (p < 0.05). Despite visible distinction, the HIGEM and HIGEM + MB groups showed no significant differences. High-intensity focused ultrasound combined with microbubbles enhances the therapeutic effects of gemcitabine chemotherapy in a pancreatic cancer xenograft model

  1. Therapeutic effects of microbubble added to combined high-intensity focused ultrasound and chemotherapy in a pancreatic cancer xenograft model

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Mi Hye [Dept. of Radiology, Konkuk University Medical Center, Seoul (Korea, Republic of); Lee, Jae Young; Kim, Bo Ram; Park, Eun Joo; Kim, Hoe Suk; Han, Joon Koo [Dept. of Radiology, Seoul National University Hospital, Seoul (Korea, Republic of); Kim, Hae Ri [Dept. of Pre-Dentistry, Gangneung-Wonju National University College of Dentistry, Gangneung (Korea, Republic of); Choi, Byung Ihn [Dept. of Radiology, Chung-Ang University Hospital, Seoul (Korea, Republic of)

    2016-09-15

    To investigate whether high-intensity focused ultrasound (HIFU) combined with microbubbles enhances the therapeutic effects of chemotherapy. A pancreatic cancer xenograft model was established using BALB/c nude mice and luciferase-expressing human pancreatic cancer cells. Mice were randomly assigned to five groups according to treatment: control (n = 10), gemcitabine alone (GEM; n = 12), HIFU with microbubbles (HIFU + MB, n = 11), combined HIFU and gemcitabine (HIGEM; n = 12), and HIGEM + MB (n = 13). After three weekly treatments, apoptosis rates were evaluated using the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay in two mice per group. Tumor volume and bioluminescence were monitored using high-resolution 3D ultrasound imaging and in vivo bioluminescence imaging for eight weeks in the remaining mice. The HIGEM + MB group showed significantly higher apoptosis rates than the other groups (p < 0.05) and exhibited the slowest tumor growth. From week 5, the tumor-volume-ratio relative to the baseline tumor volume was significantly lower in the HIGEM + MB group than in the control, GEM, and HIFU + MB groups (p < 0.05). Despite visible distinction, the HIGEM and HIGEM + MB groups showed no significant differences. High-intensity focused ultrasound combined with microbubbles enhances the therapeutic effects of gemcitabine chemotherapy in a pancreatic cancer xenograft model.

  2. Therapeutic Effects of Microbubbles Added to Combined High-Intensity Focused Ultrasound and Chemotherapy in a Pancreatic Cancer Xenograft Model

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Mi Hye [Department of Radiology, Konkuk University Medical Center, Seoul 05030 (Korea, Republic of); Lee, Jae Young [Department of Radiology, Seoul National University Hospital, Seoul 03080 (Korea, Republic of); Kim, Hae Ri [Department of Pre-Dentistry, Gangneung-Wonju National University College of Dentistry, Gangneung 25457 (Korea, Republic of); Kim, Bo Ram; Park, Eun-Joo; Kim, Hoe Suk; Han, Joon Koo [Department of Radiology, Seoul National University Hospital, Seoul 03080 (Korea, Republic of); Choi, Byung Ihn [Department of Radiology, Chung-Ang University Hospital, Seoul 06973 (Korea, Republic of)

    2016-11-01

    To investigate whether high-intensity focused ultrasound (HIFU) combined with microbubbles enhances the therapeutic effects of chemotherapy. A pancreatic cancer xenograft model was established using BALB/c nude mice and luciferase-expressing human pancreatic cancer cells. Mice were randomly assigned to five groups according to treatment: control (n = 10), gemcitabine alone (GEM; n = 12), HIFU with microbubbles (HIFU + MB, n = 11), combined HIFU and gemcitabine (HIGEM; n = 12), and HIGEM + MB (n = 13). After three weekly treatments, apoptosis rates were evaluated using the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay in two mice per group. Tumor volume and bioluminescence were monitored using high-resolution 3D ultrasound imaging and in vivo bioluminescence imaging for eight weeks in the remaining mice. The HIGEM + MB group showed significantly higher apoptosis rates than the other groups (p < 0.05) and exhibited the slowest tumor growth. From week 5, the tumor-volume-ratio relative to the baseline tumor volume was significantly lower in the HIGEM + MB group than in the control, GEM, and HIFU + MB groups (p < 0.05). Despite visible distinction, the HIGEM and HIGEM + MB groups showed no significant differences. High-intensity focused ultrasound combined with microbubbles enhances the therapeutic effects of gemcitabine chemotherapy in a pancreatic cancer xenograft model.

  3. Yeast two-hybrid screening of proteins interacting with plasmin receptor subunit: C-terminal fragment of annexin A2.

    Science.gov (United States)

    Li, Qun; Laumonnier, Yves; Syrovets, Tatiana; Simmet, Thomas

    2011-11-01

    To identify proteins that interact with the C-terminal fragment of annexin A2 (A2IC), generated by plasmin cleavage of the plasmin receptor, a heterotetramer (AA2t) containing annexin A2. The gene that encodes the A2IC fragment was obtained from PCR-amplified cDNA isolated from human monocytes, and was ligated into the pBTM116 vector using a DNA ligation kit. The resultant plasmid (pBTM116-A2IC) was sequenced with an ABI PRISM 310 Genetic Analyzer. The expression of an A2IC bait protein fused with a LexA-DNA binding domain (BD) was determined using Western blot analysis. The identification of proteins that interact with A2IC and are encoded in a human monocyte cDNA library was performed using yeast two-hybrid screening. The DNA sequences of the relevant cDNAs were determined using an ABI PRISM BigDye terminator cycle sequencing ready reaction kit. Nucleotide sequence databases were searched for homologous sequences using BLAST search analysis (http://www.ncbi.nlm.nih.gov). Confirmation of the interaction between the protein LexA-A2IC and each of cathepsin S and SNX17 was conducted using a small-scale yeast transformation and X-gal assay. The yeast transformed with plasmids encoding the bait proteins were screened with a human monocyte cDNA library by reconstituting full-length transcription factors containing the GAL4-active domain (GAL4-AD) as the prey in a yeast two-hybrid approach. After screening 1×10(7) clones, 23 independent β-Gal-positive clones were identified. Sequence analysis and a database search revealed that 15 of these positive clones matched eight different proteins (SNX17, ProCathepsin S, RPS2, ZBTB4, OGDH, CCDC32, PAPD4, and actin which was already known to interact with annexin A2). A2IC A2IC interacts with various proteins to form protein complexes, which may contribute to the molecular mechanism of monocyte activation induced by plasmin. The yeast two-hybrid system is an efficient approach for investigating protein interactions.

  4. ANNEXIN A1 N-TERMINAL DERIVED PEPTIDE AC2-26 EXERTS CHEMOKINETIC EFFECTS ON HUMAN NEUTROPHILS

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    Jesmond eDalli

    2012-02-01

    Full Text Available It is postulated that peptides derived from the N-terminal region of Annexin A1, a glucocorticoid-regulated 37-kDa protein, could act as biomimetics of the parent protein. However, recent evidence, amongst which the ability to interact with distinct receptors other then that described for Annexin A1, suggest that these peptides might fulfil other functions at variance to those reported for the parent protein. Here we tested the ability of peptide Ac2-26 to induce chemotaxis of human neutrophils, showing that this peptide can elicit responses comparable to those produced by the canonical activator formyl-Met-Leu-Phe (or FMLP. However, whilst disruption of the chemical gradient abolished the FMLP response, addition of peptide Ac2-26 in the top well of the chemotaxis chamber did not affect (10 µM or augmented (at 30 µM the neutrophil locomotion to the bottom well, as elicited by 10 µM peptide Ac2-26. Intriguingly, the sole addition of peptide Ac2-26 in the top wells produced a marked migration of neutrophils. A similar behaviour was observed when human primary monocytes were used. Thus, peptide Ac2-26 is a genuine chemokinetic agent towards human blood leukocytes.Neutralization strategies indicated that engagement of either the GPCR termed FPR1 or its cognate receptor FPR2/ALX was sufficient to sustain peptide Ac2-26 induced neutrophil migration. Similarly, application of pharmacological inhibitors showed that cell locomotion to peptide Ac2-26 was mediated primarily by the ERK, but not the JNK and p38 pathways.In conclusion, we report here novel in vitro properties for peptide Ac2-26, promoting neutrophil and monocyte chemokinesis, a process that may contribute to accelerate the resolution phase of inflammation. Here we postulate that the generation Annexin A1 N-terminal peptides at the site of inflammation may expedite the egress of migrated leukocytes thus promoting the return to homeostasis.

  5. Evaluation of adenosine preconditioning with {sup 99m}Tc-His{sub 10}-annexin V in a porcine model of myocardium ischemia and reperfusion injury: preliminary study

    Energy Technology Data Exchange (ETDEWEB)

    Ye Fei [Department of Cardiology, Nanjing First Hospital Affiliated to Nanjing Medical University, 68 Changle Road, Nanjing 210006 (China); Fang Wei [Cardiovascular Institute and Fuwai Hospital, No. 167 Bei-Li-Shi-Lu, Beijing 100037 (China); Wang Feng, E-mail: fengwang1972cn@gmail.co [Department of Nuclear Medicine, Nanjing First Hospital Affiliated to Nanjing Medical University, 68 Changle Road, Nanjing 210006 (China); State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, College of Life Sciences, Nanjing University, 22 Hankou Road, Nanjing 210093 (China); Hua Zichun [State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, College of Life Sciences, Nanjing University, 22 Hankou Road, Nanjing 210093 (China); Wang Zizheng [Department of Nuclear Medicine, Nanjing First Hospital Affiliated to Nanjing Medical University, 68 Changle Road, Nanjing 210006 (China); Yang Xiang [State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, College of Life Sciences, Nanjing University, 22 Hankou Road, Nanjing 210093 (China)

    2011-05-15

    Purpose: The goal of this study was to evaluate the feasibility of {sup 99m}Tc-His{sub 10}-annexin V for the detection of acute myocardial cell death and to assess the effect of adenosine preconditioning in a porcine model of myocardium ischemia and reperfusion injury (RI). Materials and Methods: {sup 99m}Tc-His{sub 10}-annexin V was prepared by one-step direct labeling, and RCP and radiostability were tested. The binding of {sup 99m}Tc-His{sub 10}-annexin V to apoptosis was validated in vitro using camptothecin-induced Jurkat cells. In vivo biodistribution was determined in mice by the dissection method. Ischemia of 20-30 min was induced by balloon occlusion of the epicardial coronary artery of the porcine model (n=14). Adenosine was infused intravenously in six pigs before coronary occlusion. {sup 99m}Tc-His{sub 10}-annexin V (n=12) was injected intravenously at 1 h after reperfusion. SPECT/CT was acquired at 3 h postinjection. Myocardial perfusion imaging (MPI) with {sup 99m}Tc-MIBI was also performed 1 day after His{sub 10}-annexin V imaging. Cardiac tissues were analyzed postmortem using hematoxylin-and-eosin and TUNEL staining. Caspase-3 activity was measured to confirm the presence of apoptosis. Results: {sup 99m}Tc-His{sub 10}-annexin V had a RCP >98% and high stability 2 h after radiolabeling; it could bind to apoptotic cells with high affinity. Biodistribution of {sup 99m}Tc-His{sub 10}-annexin V showed a predominant uptake in the kidney and relatively low uptake in the myocardium, liver and gastrointestinal tract; rapid clearance from blood and kidney was observed. In the untreated group, intense uptake of His{sub 10}-annexin V was visualized in the defect which was shown in MPI, whereas in the adenosine group a mild uptake of {sup 99m}Tc-His{sub 10}-annexin was found in the risk area which showed no defects in the {sup 99m}Tc-MIBI image. TUNEL staining and activated caspase-3 confirmed the ongoing apoptosis in RI. Adenosine preconditioning significantly

  6. Combination Low Dose Tissue Type-Plasminogen Activator plus Annexin A2 for Improving Thrombolytic Stroke Therapy

    Directory of Open Access Journals (Sweden)

    Yinghua eJiang

    2015-10-01

    Full Text Available Risk of hemorrhagic transformation, incomplete reperfusion, neurotoxicity and a short treatment time window comprise major challenges for tissue plasminogen activator (tPA thrombolytic stroke therapy. Improving tPA therapy has become one of the highest priorities in the stroke field. This mini review article focuses on our recent efforts aimed at evaluating a novel combination approach of low-dose tPA plus recombinant annexin A2 (rA2, a tPA and plasminogen co-receptor, that might enhance tPA thrombolytic efficacy, while reducing its associated complications related to intracerebral hemorrhagic (ICH transformation. Results of our experimental studies using a focal embolic stroke model in rats support the feasibility of the combination approach and suggest the potential for successful clinical translation.

  7. Evaluation of microbubbles as contrast agents for ultrasonography and magnetic resonance imaging.

    Directory of Open Access Journals (Sweden)

    Ling Li

    Full Text Available BACKGROUND: Microbubbles (MBs can serve as an ultrasound contrast agent, and has the potential for magnetic resonance imaging (MRI. Due to the relatively low effect of MBs on MRI, it is necessary to develop new MBs that are more suitable for MRI. In this study, we evaluate the properties of SonoVue® and custom-made Fe(3O(4-nanoparticle-embedded microbubbles (Fe(3O(4-MBs in terms of contrast agents for ultrsonography (US and MRI. METHODOLOGY/PRINCIPAL FINDINGS: A total of 20 HepG2 subcutaneous-tumor-bearing nude mice were randomly assigned to 2 groups (i.e., n = 10 mice each group, one for US test and the other for MRI test. Within each group, two tests were performed for each mouse. The contrast agent for the first test is SonoVue®, and the second is Fe(3O(4-MBs. US was performed using a Technos(MPX US system (Esaote, Italy with a contrast-tuned imaging (CnTI™ mode. MRI was performed using a 7.0T Micro-MRI (PharmaScan, Bruker Biospin GmbH, Germany with an EPI-T(2* sequence. The data of signal-to-noise ratio (SNR from the region-of-interest of each US and MR image was calculated by ImageJ (National Institute of Health, USA. In group 1, enhancement of SonoVue® was significantly higher than Fe(3O(4-MBs on US (P0.05. The SNR analysis of the enhancement process reveals a strong negative correlation in both cases (i.e., SonoVue® r = -0.733, Fe(3O(4-MBs r = -0.903, with P<0.05. CONCLUSIONS: It might be important to change the Fe(3O(4-MBs' shell structure and/or the imagining strategy of US to improve the imaging quality of Fe(3O(4-MBs on US. As an intriguing prospect that can be detected by US and MRI, MBs are worthy of further study.

  8. Ultrasound sonication with microbubbles disrupts blood vessels and enhances tumor treatments of anticancer nanodrug

    Directory of Open Access Journals (Sweden)

    Lin CY

    2012-04-01

    Full Text Available Chung-Yin Lin1*, Hsiao-Ching Tseng1*, Heng-Ruei Shiu1, Ming-Fang Wu2, Cheng-Ying Chou3, Win-Li Lin1,41Institute of Biomedical Engineering, 2Laboratory Animal Center, 3Department of Bio-Industrial Mechatronics Engineering, National Taiwan University, Taipei, Taiwan; 4Division of Medical Engineering Research, National Health Research Institutes, Miaoli, Taiwan*These authors contributed equally to this workAbstract: Ultrasound (US sonication with microbubbles (MBs has the potential to disrupt blood vessels and enhance the delivery of drugs into the sonicated tissues. In this study, mouse ear tumors were employed to investigate the therapeutic effects of US, MBs, and pegylated liposomal doxorubicin (PLD on tumors. Tumors started to receive treatments when they grew up to about 15 mm3 (early stage with injection of PLD 10 mg/kg, or up to 50 mm3 (medium stage with PLD 6 (or 4 mg/kg. Experiments included the control, PLD alone, PLD + MBs + US, US alone, and MBs + US groups. The procedure for the PLD + MBs + US group was that PLD was injected first, MB (SonoVue injection followed, and then US was immediately sonicated on the tumor. The results showed that: (1 US sonication with MBs was always able to produce a further hindrance to tumor growth for both early and medium-stage tumors; (2 for the medium-stage tumors, 6 mg/kg PLD alone was able to inhibit their growth, while it did not work for 4 mg/kg PLD alone; (3 with the application of MBs + US, 4 mg/kg PLD was able to inhibit the growth of medium-stage tumors; (4 for early stage tumors after the first treatment with a high dose of PLD alone (10 mg/kg, the tumor size still increased for several days and then decreased (a biphasic pattern; (5 MBs + US alone was able to hinder the growth of early stage tumors, but unable to hinder that of medium stage tumors. The results of histological examinations and blood perfusion measurements indicated that the application of MBs + US disrupts the tumor blood

  9. Lack of annexin 1 results in an increase in corticotroph number in male but not female mice.

    Science.gov (United States)

    Morris, J F; Omer, S; Davies, E; Wang, E; John, C; Afzal, T; Wain, S; Buckingham, J C; Flower, R J; Christian, H C

    2006-11-01

    Annexin 1 (ANXA1) is a member of the annexin family of phospholipid- and calcium-binding proteins with a well demonstrated role in early delayed (30 min to 3 h) inhibitory feedback of glucocorticoids in the pituitary. We have examined corticotrophs in wild-type and ANXA1 knockout mice to determine the effects of lack of ANXA1 in male and female animals. Anterior pituitary tissue from ANXA1 wild-type, heterozygote and null mice was fixed and examined (i) by confocal immunocytochemistry to determine the number of corticotrophs and (ii) by electron microscopy to examine the size, secretory granule population and secretory machinery of corticotrophs. No differences in these parameters were detected in female mice. In male ANXA1 null mice, there were approximately four-fold more corticotrophs than in wild-type animals. However, the corticotrophs in ANXA1 null mice were smaller and had reduced numbers of secretory granules (the reduction in granules paralleled the reduction in cell size). No differences in the numerical density of folliculo-stellate, gonadotroph, lactotroph or somatotroph cells were detected in male ANXA1 null mice. Plasma corticosterone, adrenocorticotrophic hormone (ACTH) and pituitary pro-opiomelanocortin mRNA were unchanged but pituitary ACTH content was increased in male ANXA1 null mice. Interleukin (IL)-6 pituitary content was significantly elevated in male and reduced in female ANXA1 null mice compared to wild-type. In conclusion, these data indicate that ANXA1 deficiency is associated with gender-specific changes in corticotroph number and structure, via direct actions of ANXA1 and/or indirect changes in factors such as IL-6.

  10. Spermatozoa with numerical chromosomal abnormalities are more prone to be retained by Annexin V-MACS columns.

    Science.gov (United States)

    Esbert, M; Godo, A; Soares, S R; Florensa, M; Amorós, D; Ballesteros, A; Vidal, F

    2017-07-01

    Colloidal super-paramagnetic microbeads conjugated with annexin V are effective for separating apoptotic spermatozoa by MACS as a result of the high affinity of annexin V for externalized PS molecules. The effectiveness of the procedure in reducing the percentage of sperm with fragmented DNA and abnormal morphology has also been reported. However, it is still unknown if it could decrease the percentage of aneuploid spermatozoa. The objective of our prospective study, performed on 16 males with abnormal FISH on spermatozoa, was to assess if MACS columns were useful tools to retain spermatozoa carrying chromosomal abnormalities in semen samples processed after density gradient centrifugation (DGC). The pellet obtained after DGC was subjected to MACS, and sperm FISH analyses were performed both in the eluded fraction and in the fraction retained in the column. The observed frequencies of disomy and nullisomy 13, 18, and 21, X and Y, as well as the diploidy rates in the MACS eluded fraction and the fraction retained in the MACS column were recorded. We observed that the frequencies of aneuploidies in the eluded fraction were lower than in the fraction retained in the MACS column (0.59% vs. 0.75%; p = 0.010). DGC determined a significant reduction in sperm concentration (z-ratio = 2.83; p = 0.005) and a significant increase in sperm progressive motility (z-ratio = -3.5; p spermatozoa carrying chromosomal abnormalities. Furthermore, the performance of DGC and MACS on semen samples leads to an enrichment of progressive motility. © 2017 American Society of Andrology and European Academy of Andrology.

  11. The stable microbubble test for determining continuous positive airway pressure (CPAP) success in very preterm infants receiving nasal CPAP from birth.

    Science.gov (United States)

    Bhatia, Risha; Morley, Colin J; Argus, Brenda; Tingay, David G; Donath, Susan; Davis, Peter G

    2013-01-01

    Very preterm infants can be treated with nasal continuous positive airway pressure (CPAP) from birth, but some fail. A rapid test, such as the stable microbubble test (SMT) on gastric aspirate, may identify those who can be managed successfully using CPAP. To determine if SMT can identify soon after birth, very preterm infants who may be successfully managed on CPAP alone. Stable microbubbles (diameter CPAP from birth. Infants failed CPAP if intubated at CPAP in the delivery room at a median (interquartile range) pressure 7 (6-8) cmH2O and FiO2 0.25 (0.21-0.3). Gastric aspirates were taken at a median (interquartile range) age of 0.5 (0.3-0.6) hours. The best cut-off point for predicting CPAP success or failure was a SMT count of 8 microbubbles/mm(2). The area under the receiver operating characteristic curve was 0.8 (95% CI 0.7-0.9). A SMT count ≥8 microbubbles/mm(2) had a sensitivity of 53%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 60% for predicting CPAP success. Infants treated with CPAP from birth, who had SMT counts ≥8 microbubbles/mm(2) on their gastric aspirate, did not fail CPAP. Copyright © 2013 S. Karger AG, Basel.

  12. Cytoplasmic capes are nuclear envelope intrusions that are enriched in endosomal proteins and depend upon βH-spectrin and Annexin B9.

    Science.gov (United States)

    Wu, Juan; Bakerink, Katelyn J; Evangelista, Meagan E; Thomas, Graham H

    2014-01-01

    It is increasingly recognized that non-erythroid spectrins have roles remote from the plasma membrane, notably in endomembrane trafficking. The large spectrin isoform, βH, partners with Annexin B9 to modulate endosomal processing of internalized proteins. This modulation is focused on the early endosome through multivesicular body steps of endocytic processing and loss of either protein appears to cause a traffic jam before removal of ubiquitin at the multivesicular body. We previously reported that βH/Annexin B9 influenced EGF receptor signaling. While investigating this effect we noticed that mSptiz, the membrane bound precursor of the secreted EGF receptor ligand sSpitz, is located in striking intrusions of the nuclear membrane. Here we characterize these structures and identify them as 'cytoplasmic capes', which were previously identified in old ultrastructural studies and probably coincide with recently recognized sites of non-nuclear-pore RNA export. We show that cytoplasmic capes contain multiple endosomal markers and that their existence is dependent upon βH and Annexin B9. Diminution of these structures does not lead to a change in mSpitz processing. These results extend the endosomal influence of βH and its partner Annexin B9 to this unusual compartment at the nuclear envelope.

  13. Cytoplasmic capes are nuclear envelope intrusions that are enriched in endosomal proteins and depend upon βH-spectrin and Annexin B9.

    Directory of Open Access Journals (Sweden)

    Juan Wu

    Full Text Available It is increasingly recognized that non-erythroid spectrins have roles remote from the plasma membrane, notably in endomembrane trafficking. The large spectrin isoform, βH, partners with Annexin B9 to modulate endosomal processing of internalized proteins. This modulation is focused on the early endosome through multivesicular body steps of endocytic processing and loss of either protein appears to cause a traffic jam before removal of ubiquitin at the multivesicular body. We previously reported that βH/Annexin B9 influenced EGF receptor signaling. While investigating this effect we noticed that mSptiz, the membrane bound precursor of the secreted EGF receptor ligand sSpitz, is located in striking intrusions of the nuclear membrane. Here we characterize these structures and identify them as 'cytoplasmic capes', which were previously identified in old ultrastructural studies and probably coincide with recently recognized sites of non-nuclear-pore RNA export. We show that cytoplasmic capes contain multiple endosomal markers and that their existence is dependent upon βH and Annexin B9. Diminution of these structures does not lead to a change in mSpitz processing. These results extend the endosomal influence of βH and its partner Annexin B9 to this unusual compartment at the nuclear envelope.

  14. Mapping of treatment-induced apoptosis in normal structures: 99mTc-Hynic-rh-annexin V SPECT and CT image fusion

    NARCIS (Netherlands)

    Kartachova, Marina S.; Valdés Olmos, Renato A.; Haas, Rick L. M.; Hoebers, Frank J. P.; van den Brekel, Michiel W.; van Zandwijk, Nico; van Herk, Marcel; Verheij, Marcel

    2006-01-01

    PURPOSE: The purpose of this study was to map treatment-induced (99m)Tc-Hynic-rh-annexin V uptake in normal tissues using co-registration of SPECT and CT. METHODS: Nineteen patients (11 male, 8 female, mean age 57 years) with various malignant tumours (12 lymphomas, four non-small cell lung cancers

  15. Detection of focal hypoxic-ischemic injury and neuronal stress in a rodent model of unilateral MCA occlusion/reperfusion using radiolabeled annexin V

    Energy Technology Data Exchange (ETDEWEB)

    Mari, Carina; Goris, Michael L. [Division of Nuclear Medicine, Department of Radiology, Stanford University Hospital, CA 94305, Stanford (United States); Karabiyikoglu, Murat; Yenari, Midori Anne [Departments of Neurosurgery and Neurology, Stanford University Hospital, CA 94305, Stanford (United States); Tait, Jonathan F. [Department of Laboratory Medicine, University of Washington Medical Center, WA 98195-7110, Seattle (United States); Blankenberg, Francis G. [Division of Nuclear Medicine, Department of Radiology, Stanford University Hospital, CA 94305, Stanford (United States); Division of Pediatric Radiology, Department of Radiology, Stanford University, Lucile Salter Packard Children' s Hospital, 725 Welch Road, Room 1673, CA 94305, Palo Alto (United States)

    2004-05-01

    In this study we wished to determine whether technetium-99m annexin V, an in vivo marker of cellular injury and death, could be used to noninvasively monitor neuronal injury following focal middle cerebral artery (MCA) occlusion/reperfusion injury. Sixteen adult male Sprague-Dawley rats (along with four controls) underwent left (unilateral) MCA intraluminal beaded thread occlusion for 2 h followed by reperfusion. One hour following tail vein injection of 5-10 mCi of {sup 99m}Tc-annexin V, animals underwent either single-photon emission computerized tomography (SPECT) or autoradiography followed by immunohistochemical analyses. There was abnormal, bilateral, multifocal uptake of {sup 99m}Tc-annexin V in each cerebral hemisphere as seen by both SPECT and autoradiography at 4 h and 1, 3, and 7 days after initiation of occlusion. The average maximal annexin V uptake at 4 h was 310%{+-}85% and 365%{+-}151% above control values (P<0.006) within the right and left hemispheres, respectively, peaking on day 3 with values of 925%{+-}734% and 1,194%{+-}643% (P<0.03) that decreased by day 7 to 489%{+-}233% and 785%{+-}225% (P<0.01). Total lesional volume of the left hemisphere was 226%, 261%, and 451% (P<0.03) larger than the right at 4, 24, and 72 h after injury, respectively. Annexin V localized to the cytoplasm of injured neurons ipsilateral to the site of injury as well as to otherwise normal-appearing neurons of the contralateral hemisphere as confirmed by dual fluorescent microscopy. It is concluded that there is abnormal bilateral, multifocal annexin V uptake, greater on the left than on the right side, within 4 h of unilateral left MCA ischemic injury and that the uptake peaks at 3 days and decreases by 7 days after injury. This pattern suggests that neuronal stress may play a role in the response of the brain to focal injury and be responsible for annexin V uptake outside the region of ischemic insult. (orig.)

  16. Fabricating multifunctional microbubbles and nanobubbles for concurrent ultrasound and photoacoustic imaging

    Science.gov (United States)

    Qin, Ruogu; Xu, Jeff; Xu, Ronald; Kim, Chulhong; Wang, Lihong V.

    2010-02-01

    Background: Clinical ultrasound (US) uses ultrasonic scattering contrast to characterize subcutaneous anatomic structures. Photoacoustic (PA) imaging detects the functional properties of thick biological tissue with high optical contrast. In the case of image-guided cancer ablation therapy, simultaneous US and PA imaging can be useful for intraoperative assessment of tumor boundaries and ablation margins. In this regard, accurate co-registration between imaging modalities and high sensitivity to cancer cells are important. Methods: We synthesized poly-lactic-co-glycolic acid (PLGA) microbubbles (MBs) and nanobubbles (NBs) encapsulating India ink or indocyanine green (ICG). Multiple tumor simulators were fabricated by entrapping ink MBs or NBs at various concentrations in gelatin phantoms for simultaneous US and PA imaging. MBs and NBs were also conjugated with CC49 antibody to target TAG-72, a human glycoprotein complex expressed in many epithelial-derived cancers. Results: Accurate co-registration and intensity correlation were observed in US and PA images of MB and NB tumor simulators. MBs and NBs conjugating with CC49 effectively bound with over-expressed TAG-72 in LS174T colon cancer cell cultures. ICG was also encapsulated in MBs and NBs for the potential to integrate US, PA, and fluorescence imaging. Conclusions: Multifunctional MBs and NBs can be potentially used as a general contrast agent for multimodal intraoperative imaging of tumor boundaries and therapeutic margins.

  17. Delivery of dopamine transporter tracer (PE2I) through blood brain barrier with ultrasound and microbubbles

    Science.gov (United States)

    Serrière, Sophie; Escoffre, Jean-Michel; Bodard, Sylvie; Novell, Anthony; Vergote, Jackie; Vercouillie, Johnny; Thiéry, Jean-Claude; Chalon, Sylvie; Bouakaz, Ayache

    2012-10-01

    The blood-brain barrier plays a major role in controlling the delivery of therapeutic and imaging agents to the brain. The aim of this study was to investigate the use of ultrasound and microbubbles to increase its delivery through the BBB and by determining the optimal experimental conditions that achieve a transient and safe BBB disruption. First, we established the ultrasound conditions that achieved a transient BBB disruption in rats using a non-permeant marker, Evans blue. Hence SonoVue® (450 μL/kg) and Evans blue (100 mg/kg) were intravenously administered. BBB leakage was obtained using ultrasound insonation through the rat skull at 1.6 MPa, PRF 1 Hz, duty cycle 12%, burst 10 ms during 120 sec. BBB disruption was observed in all treated animals (N=4) by histological analysis. The same experimental conditions were applied to enhance brain uptake of PE2I. Biological samples were analyzed using a scintillation counter apparatus. The results showed 50% and 20% increase of 125I-PE2I uptake in the striatum and cerebral cortex, respectively, in the treated rats (N=5) versus control (N=4). Similar enhancements were observed using SonoVue® at half concentration. This innovative method provides a great potential for intracerebral delivery of molecular ligands that could be used for the therapy of brain diseases.

  18. Dual-functionalized nanoparticles loaded microbubbles for enhancement of drug uptake.

    Science.gov (United States)

    Li, Yingjia; Zhang, Xia; Luo, Wanxian; Wang, Dongxiao; Yang, Li; Wang, Jianguo; Zhang, Li; Zhang, Shiyu; Luo, Shuyi; Wang, Ying

    2018-07-01

    The application of microbubble (MB)-assisted ultrasound (US) can combine the advantages of real-time imaging and targeted drug delivery. However, the drug loading capacity of MB is limited restricting its application in antitumor procedure. In contrast, nanoparticles (NPs) can carry drugs more efficiently, but adverse side effect induced by unspecific accumulation can not be ignored. Herein, we developed a dual-functionalized NP loaded MB to investigate its potential feasibility for tumor-targeted drug delivery. Firstly, we prepared NPs using heparin as backbone. Targeting ligand folate and cell-penetrating ligand Tat peptide were conjugated to the backbone to deliver paclitaxel (H-F-Tat-P NPs). Subsequently, the dual-functionalized NPs were incorporated with MBs via avidin-biotin linkage to fabricate H-F-Tat-P NPs loaded MBs (NPs-loaded MBs). The combined strategy can take profit of dual functionalities from NPs and sonoporation effect from MBs triggered by US. The prepared NPs have been characterized. The excellent cellular uptake of NPs were qualitative and quantitative analysis by flow cytometry and confocal microscope, the results indicated that it was attributed to not only dual functionalities but also US effect. Foremost, the NPs-loaded MBs combined with US exhibited significant cytotoxicity on both folate receptor (FR) overexpressing and deficiency cells. The combination of dual-functionalized NPs and MBs with US is expected to be a promising strategy for targeted anticancer drug delivery and ultrasound imaging simultaneously. Copyright © 2018 Elsevier B.V. All rights reserved.

  19. Thrombin-Activatable Microbubbles as Potential Ultrasound Contrast Agents for the Detection of Acute Thrombosis.

    Science.gov (United States)

    Lux, Jacques; Vezeridis, Alexander M; Hoyt, Kenneth; Adams, Stephen R; Armstrong, Amanda M; Sirsi, Shashank R; Mattrey, Robert F

    2017-11-01

    Acute deep vein thrombosis (DVT) is the formation of a blood clot in the deep veins of the body that can lead to fatal pulmonary embolism. Acute DVT is difficult to distinguish from chronic DVT by ultrasound (US), the imaging modality of choice, and is therefore treated aggressively with anticoagulants, which can lead to internal bleeding. Here we demonstrate that conjugating perfluorobutane-filled (PFB-filled) microbubbles (MBs) with thrombin-sensitive activatable cell-penetrating peptides (ACPPs) could lead to the development of contrast agents that detect acute thrombosis with US imaging. Successful conjugation of ACPP to PFB-filled MBs was confirmed by fluorescence microscopy and flow cytometry. Fluorescein-labeled ACPP was used to evaluate the efficiency of thrombin-triggered cleavage by measuring the mean fluorescence intensity of ACPP-labeled MBs (ACPP-MBs) before and after incubation at 37 °C with thrombin. Lastly, control MBs and ACPP-MBs were infused through a tube containing a clot, and US contrast enhancement was measured with or without the presence of a thrombin inhibitor after washing the clot with saline. With thrombin activity, 91.7 ± 14.2% of the signal was retained after ACPP-MB infusion and washing, whereas only 16.7 ± 4% of the signal was retained when infusing ACPP-MBs in the presence of hirudin, a potent thrombin inhibitor.

  20. Transition process leading to microbubble emission boiling on horizontal circular heated surface in subcooled pool

    Science.gov (United States)

    Ueno, Ichiro; Ando, Jun; Horiuchi, Kazuna; Saiki, Takahito; Kaneko, Toshihiro

    2016-11-01

    Microbubble emission boiling (MEB) produces a higher heat flux than critical heat flux (CHF) and therefore has been investigated in terms of its heat transfer characteristics as well as the conditions under which MEB occurs. Its physical mechanism, however, is not yet clearly understood. We carried out a series of experiments to examine boiling on horizontal circular heated surfaces of 5 mm and of 10 mm in diameter, in a subcooled pool, paying close attention to the transition process to MEB. High-speed observation results show that, in the MEB regime, the growth, condensation, and collapse of the vapor bubbles occur within a very short time. In addition, a number of fine bubbles are emitted from the collapse of the vapor bubbles. By tracking these tiny bubbles, we clearly visualize that the collapse of the vapor bubbles drives the liquid near the bubbles towards the heated surface, such that the convection field around the vapor bubbles under MEB significantly differs from that under nucleate boiling. Moreover, the axial temperature gradient in a heated block (quasi-heat flux) indicates a clear difference between nucleate boiling and MEB. A combination of quasi-heat flux and the measurement of the behavior of the vapor bubbles allows us to discuss the transition to MEB. This work was financially supported by the 45th Research Grant in Natural Sciences from The Mitsubishi Foundation (2014 - 2015), and by Research Grant for Boiler and Pressurized Vessels from The Japan Boiler Association (2016).

  1. Isolation of Low Abundance Proteins and Cells Using Buoyant Glass Microbubble Chromatography

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    Steingrimur Stefansson

    2013-01-01

    Full Text Available Conventional protein affinity chromatography relies on highly porous resins that have large surface areas. These properties are ideal for fast flow separation of proteins from biological samples with maximum yields, but these properties can also lead to increased nonspecific protein binding. In certain applications where the purity of an isolated protein is more important than the yield, using a glass solid phase could be advantageous as glass is nonporous and hydrophilic and has a low surface area and low nonspecific protein binding. As a proof of principle, we used protein A-conjugated hollow glass microbubbles to isolate fluorescently labeled neurofilament heavy chain spiked into serum and compared them to protein A Sepharose and protein A magnetic beads (Dynabeads using an anti-neurofilament protein antibody. As expected, a greater volume of glass bubbles was required to match the binding capacity of the magnetic beads and Sepharose resins. On the other hand, nonspecific protein binding to glass bubbles was greatly reduced compared to the other resins. Additionally, since the glass bubbles are buoyant and transparent, they are well suited for isolating cells from biological samples and staining them in situ.

  2. Ultrasound Targeted Microbubble Destruction Stimulates Cellular Endocytosis in Facilitation of Adeno-Associated Virus Delivery

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    Lian-Fang Du

    2013-05-01

    Full Text Available The generally accepted mechanism for ultrasound targeted microbubble destruction (UTMD to enhance drug and gene delivery is through sonoporation. However, passive uptake of adeno-associated virus (AAV into cells following sonoporation does not adequately explain observations of enhanced transduction by UTMD. This study investigated alternative mechanisms of UTMD enhancement in AAV delivery. UTMD significantly enhanced transduction efficiency of AAV in a dose-dependent manner. UTMD stimulated a persistent uptake of AAV into the cytoplasm and nucleus. This phenomenon occurred over several hours, suggesting that some viral particles are endocytosed by cells rather than exclusively passing through pores created by sonoporation. Additionally, UTMD enhanced clathrin expression and accumulation at the plasma membrane suggesting greater clathrin-mediated endocytosis following UTMD. Transmission electron microscopy (TEM revealed that UTMD stimulated formation of clathrin-coated pits (CPs and uncoated pits (nCPs. Furthermore, inhibition of clathrin-mediated endocytosis partially blocked the enhancement of AAV uptake following UTMD. The results of this study implicate endocytosis as a mechanism that contributes to UTMD-enhanced AAV delivery.

  3. Effects of Microbubble Size on Ultrasound-Mediated Gene Transfection in Auditory Cells

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    Ai-Ho Liao

    2014-01-01

    Full Text Available Gene therapy for sensorineural hearing loss has recently been used to insert genes encoding functional proteins to preserve, protect, or even regenerate hair cells in the inner ear. Our previous study demonstrated a microbubble- (MB-facilitated ultrasound (US technique for delivering therapeutic medication to the inner ear. The present study investigated whether MB-US techniques help to enhance the efficiency of gene transfection by means of cationic liposomes on HEI-OC1 auditory cells and whether MBs of different sizes affect such efficiency. Our results demonstrated that the size of MBs was proportional to the concentration of albumin or dextrose. At a constant US power density, using 0.66, 1.32, and 2.83 μm albumin-shelled MBs increased the transfection rate as compared to the control by 30.6%, 54.1%, and 84.7%, respectively; likewise, using 1.39, 2.12, and 3.47 μm albumin-dextrose-shelled MBs increased the transfection rates by 15.9%, 34.3%, and 82.7%, respectively. The results indicate that MB-US is an effective technique to facilitate gene transfer on auditory cells in vitro. Such size-dependent MB oscillation behavior in the presence of US plays a role in enhancing gene transfer, and by manipulating the concentration of albumin or dextrose, MBs of different sizes can be produced.

  4. Blood-brain barrier disruption induced by diagnostic ultrasound combined with microbubbles in mice.

    Science.gov (United States)

    Zhao, Bingxia; Chen, Yihan; Liu, Jinfeng; Zhang, Li; Wang, Jing; Yang, Yali; Lv, Qing; Xie, Mingxing

    2018-01-12

    To investigate the effects of the microbubble (MB) dose, mechanism index (MI) and sonication duration on blood-brain barrier (BBB) disruption induced by diagnostic ultrasound combined with MBs as well as to investigate the potential molecular mechanism. The extent of BBB disruption increased with MB dose, MI and sonication duration. A relatively larger extent of BBB disruption associated with minimal tissue damage was achieved by an appropriate MB dose and ultrasound exposure parameters with diagnostic ultrasound. Decreased expression of ZO-1, occludin and claudin-5 were correlated with disruption of the BBB, as confirmed by paracellular passage of the tracer lanthanum nitrate into the brain parenchyma after BBB disruption. These findings indicated that this technique is a promising tool for promoting brain delivery of diagnostic and therapeutic agents in the diagnosis and treatment of brain diseases. The extent of BBB disruption was qualitatively assessed by Evans blue (EB) staining and quantitatively analyzed by an EB extravasation measurement. A histological examination was performed to evaluate tissue damage. Expression of tight junction (TJ) related proteins ZO-1, occludin and claudin-5 was determined by western blotting analysis and immunohistofluorescence. Transmission electron microscopy was performed to observe ultrastructure changes of TJs after BBB disruption.

  5. Cardioprotective effects of erythropoietin in rats subjected to ischemia-reperfusion injury: assessment of infarct size with 99mTc-annexin V.

    Science.gov (United States)

    Doue, Tomoki; Ohtsuki, Katsuichi; Ogawa, Kazuma; Ueda, Masashi; Azuma, Akihiro; Saji, Hideo; Strauss, Harry W; Matsubara, Hiroaki

    2008-10-01

    Administration of erythropoietin (EPO) during or immediately after myocardial ischemia can reduce subsequent myocardial apoptosis, a key phenomenon in myocardial ischemia-reperfusion injury. In this study, we assessed the effect of EPO on (99m)Tc-annexin V myocardial uptake and whether the accumulation of (99m)Tc-annexin V can predict cardiac remodeling and functional deterioration. Eighteen rats with left coronary artery (LCA) occlusion were randomized to receive either an intravenous injection of EPO (EPO group) or saline (nontherapy [nT] group) immediately after release of the occlusion. After 20 min of LCA occlusion and 30 min of reperfusion, the rats were injected with (99m)Tc-annexin V. One hour after (99m)Tc-annexin V injection, the LCA was reoccluded and (201)Tl was injected intravenously, and the rats were sacrificed 1 min later. The heart was removed and sectioned, and dual-tracer autoradiography was performed to evaluate the distribution of the area at risk (defined on the thallium autoradiograph) and the area of apoptosis (defined on the annexin autoradiograph). Adjacent histologic specimens had deoxyuridine triphosphate nick-end labeling (TUNEL) staining to confirm the presence of apoptosis and were compared with autoradiography. Another 16 rats were randomized to EPO and nT groups and underwent echocardiography immediately after release of the LCA occlusion and at 2 and 4 wk after surgery. The areas of (99m)Tc-annexin V accumulation in the EPO group were smaller than those in the nT group, though the (201)Tl defect areas of these 2 groups were comparable (area ratio, 0.318 +/- 0.038 vs. 0.843 +/- 0.051, P < 0.001, for annexin and 24.8 +/- 2.1 vs. 25.9 +/- 2.6 mm(2), P = NS, for thallium). (99m)Tc-annexin V accumulation correlated with the density of TUNEL-positive cells (r = 0.886, P < 0.001). In the nT group, left ventricular end-diastolic dimension (Dd) increased from baseline at 2 wk by 34.7% +/- 3.8% and remained stable at 34.9% +/- 5.0% at 4 wk

  6. Apo And Calcium-Bound Crystal Structures of Alpha-11 Giardin, An Unusual Annexin From 'Giardia Lamblia'

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    Pathuri, P.; Nguyen, E.T.; Svard, S.G.; Luecke, H.; /UC, Irvine /Uppsala U. /Karolinska Inst.

    2007-07-12

    Alpha-11 giardin is a member of the multi-gene alpha-giardin family in the intestinal protozoan, Giardia lamblia. This gene family shares an ancestry with the annexin super family, whose common characteristic is calcium-dependent binding to membranes that contain acidic phospholipids. Several alpha giardins are highly expressed during parasite-induced diarrhea in humans. Despite being a member of a large family of proteins, little is known about the function and cellular localization of alpha-11 giardin, although giardins are often associated with the cytoskeleton. It has been shown that Giardia exhibits high levels of alpha-11 giardin mRNA transcript throughout its life cycle; however, constitutive over-expression of this protein is lethal to the parasite. Determining the three-dimensional structure of an alpha-giardin is essential to identifying functional domains shared in the alpha-giardin family. Here we report the crystal structures of the apo and Ca{sup 2+}-bound forms of alpha-11 giardin, the first alpha giardin to be characterized structurally. Crystals of apo and Ca{sup 2+}-bound alpha-11 giardin diffracted to 1.1 angstroms and 2.93 angstroms, respectively. The crystal structure of selenium-substituted apo alpha-11 giardin reveals a planar array of four tandem repeats of predominantly {alpha}-helical domains, reminiscent of previously determined annexin structures, making this the highest-resolution structure of an annexin to date. The apo alpha-11 giardin structure also reveals a hydrophobic core formed between repeats I/IV and II/III, a region typically hydrophilic in other annexins. Surprisingly, the Ca{sup 2+}-bound structure contains only a single calcium ion, located in the DE loop of repeat I and coordinated differently from the two types of calcium sites observed in previous annexin structures. The apo and Ca{sup 2+}-bound alpha-11 giardin structures assume overall similar conformations; however, Ca2+-bound alpha-11 giardin crystallized in a lower

  7. Carcinogenic heavy metals, As3+ and Cr6+, increase affinity of nuclear mono-ubiquitinated annexin A1 for DNA containing 8-oxo-guanosine, and promote translesion DNA synthesis.

    Science.gov (United States)

    Hirata, Aiko; Corcoran, George B; Hirata, Fusao

    2011-04-15

    To elucidate the biological roles of mono-ubiquitinated annexin A1 in nuclei, we investigated the interaction of purified nuclear mono-ubiquitinated annexin A1 with intact and oxidatively damaged DNA. We synthesized the 80mer 5'-GTCCACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCA-3' (P0G), and four additional 80mers, each with a selected single G in position 14, 30, 37 or 48 replaced by 8-oxo-guanosine (8-oxo-G) to model DNA damaged at a specific site by oxidation. Nuclear mono-ubiquitinated annexin A1 was able to bind oligonucleotides containing 8-oxo-G at specific positions, and able to anneal damaged oligonucleotide DNA to M13mp18 in the presence of Ca(2+) or heavy metals such as As(3+) and Cr(6+). M13mp18/8-oxo-G-oligonucleotide duplexes were unwound by nuclear annexin A1 in the presence of Mg(2+) and ATP. The binding affinity of nuclear annexin A1 for ssDNA was higher for oxidatively damaged oligonucleotides than for the undamaged oligonucleotide P0G, whereas the maximal binding was not significantly changed. The carcinogenic heavy metals, As(3+) and Cr(6+), increased the affinity of mono-ubiquitinated annexin A1 for oxidatively damaged oligonucleotides. Nuclear mono-ubiquitinated annexin A1 stimulated translesion DNA synthesis by Pol β. Nuclear extracts of L5178Y tk(+/-) lymphoma cells also promoted translesion DNA synthesis in the presence of the heavy metals As(3+) and Cr(6+). This DNA synthesis was inhibited by anti-annexin A1 antibody. These observations do not prove but provide strong evidence for the hypothesis that nuclear mono-ubiquitinated annexin A1 is involved in heavy metal promoted translesion DNA synthesis, thereby exhibiting the capacity to increase the introduction of mutations into DNA. Copyright © 2011 Elsevier Inc. All rights reserved.

  8. Microbubble-enhanced ultrasound exposure improves gene transfer in vascular endothelial cells

    Science.gov (United States)

    Nie, Fang; Xu, Hui-Xiong; Tang, Qing; Lu, Ming-De

    2006-01-01

    AIM: To explore the effects of ultrasound exposure combined with microbubble contrast agent (SonoVue) on the permeability of the cellular membrane and on the expression of plasmid DNA encoding enhanced green fluorescent protein (pEGFP) transfer into human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs with fluorescein isothiocyanate-dextran (FD500) and HUVECs with pEGFP were exposed to continuous wave (1.9 MHz, 80.0 mW/cm2) for 5 min, with or without a SonoVue. The percentage of FD500 taken by the HUVECs and the transient expression rate of pEGFP in the HUVECs were examined by fluorescence microscopy and flow cytometry, respectively. RESULTS: The percentage of FD500-positive HUVECs in the group of ultrasound exposure combined with SonoVue was significantly higher than that of the group of ultrasound exposure alone (24.0% ± 5.5% vs 66.6% ± 4.1%, P SonoVue (16.1% ± 1.9% vs 1.5% ± 0.2%, P SonoVue (94.1% ± 2.3% vs 91.1% ± 4.1%). CONCLUSION: The cell membrane permeability of HUVECs and the transfection efficiency of pEGFP into HUVECs exposed to ultrasound are significantly increased after addition of an ultrasound contrast agent without obvious damage to the survival of HUVECs. This non-invasive gene transfer method may be a useful tool for clinical gene therapy of hepatic tumors. PMID:17167842

  9. Myocardial regeneration in adriamycin cardiomyopathy by nuclear expression of GLP1 using ultrasound targeted microbubble destruction

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Shuyuan [Baylor Research Institute, Baylor University Medical Center, 3812 Elm Street, Dallas, TX (United States); Chen, Jiaxi [The University of Texas Southwestern Medical Center at Dallas, Medical School, 5235 Harry Hine Blvd., Dallas, TX (United States); Huang, Pintong [Department of Ultrasonography, The 2nd Affiliated Hospital of Zhejiang University College of Medicine, Hangzhou, Zhejiang Province (China); Meng, Xing-Li; Clayton, Sandra; Shen, Jin-Song [Baylor Research Institute, Baylor University Medical Center, 3812 Elm Street, Dallas, TX (United States); Grayburn, Paul A., E-mail: paulgr@baylorhealth.edu [Baylor Research Institute, Baylor University Medical Center, 3812 Elm Street, Dallas, TX (United States); Department of Internal Medicine, Division of Cardiology, Baylor Heart and Vascular Institute, Baylor University Medical Center, 621 N. Hall St, Suite H030, Dallas, TX (United States)

    2015-03-20

    Recently GLP-1 was found to have cardioprotective effects independent of those attributable to tight glycemic control. Methods and results: We employed ultrasound targeted microbubble destruction (UTMD) to deliver piggybac transposon plasmids encoding the GLP-1 gene with a nuclear localizing signal to rat hearts with adriamycin cardiomyopathy. After a single UTMD treatment, overexpression of transgenic GLP-1 was found in nuclei of rat heart cells with evidence that transfected cardiac cells had undergone proliferation. UTMD-GLP-1 gene therapy restored LV mass, fractional shortening index, and LV posterior wall diameter to nearly normal. Nuclear overexpression of GLP-1 by inducing phosphorylation of FoxO1-S256 and translocation of FoxO1 from the nucleus to the cytoplasm significantly inactivated FoxO1 and activated the expression of cyclin D1 in nuclei of cardiac muscle cells. Reversal of adriamycin cardiomyopathy appeared to be mediated by dedifferentiation and proliferation of nuclear FoxO1-positive cardiac muscle cells with evidence of embryonic stem cell markers (OCT4, Nanog, SOX2 and c-kit), cardiac early differentiation markers (NKX2.5 and ISL-1) and cellular proliferation markers (BrdU and PHH3) after UTMD with GLP-1 gene therapy. Conclusions: Intranuclear myocardial delivery of the GLP-1gene can reverse established adriamycin cardiomyopathy by stimulating myocardial regeneration. - Highlights: • The activation of nuclear FoxO1 in cardiac muscle cells associated with adriamycin cardiomyopathy. • Myocardial nuclear GLP-1 stimulates myocardial regeneration and reverses adriamycin cardiomyopathy. • The process of myocardial regeneration associated with dedifferentiation and proliferation.

  10. Microbubble mediated thrombus dissolution with diagnostic ultrasound for the treatment of chronic venous thrombi.

    Directory of Open Access Journals (Sweden)

    Shelby Kutty

    Full Text Available Central venous catheter (CVC thrombi result in significant morbidity in children, and currently available treatments are associated with significant risk. We sought to investigate the therapeutic efficacy of microbubble (MB enhanced sonothrombolysis for aged CVC associated thrombi in vivo.A model of chronic indwelling CVC in the low superior vena cava with thrombus in situ was established after feasibility and safety testing in 7 pigs; and subsequently applied for repeated, sonothrombolytic treatments in 9 pigs (total 24 treatments. Baseline intracardiac echocardiography (ICE, 10.5F, Siemens, fluoroscopy and saline flushing confirmed the absence of any pre-existing CVC thrombus. A thrombus was then allowed to form and age over 24 hours. The created thrombus was localized and measured by ICE, and transthoracic image guided high mechanical index (MI two-dimensional US treatments (1.1-1.7 MI; iE33, Philips applied intermittently whenever intravenously infused MBs (3% MRX-801; NuVox were visualized near the thrombus (n = 10; Group A. Control pigs (n = 10; Group B received US without MB. All treatments were randomized. Post-treatment thrombus area by ICE planimetry was compared with pre-treatment measurements. Thrombus area measurements before and after treatment were 0.22 and 0.10 cm(2 respectively in Group A; compared to 0.24 and 0.21 cm(2 in Group B (p  = 0.0003. Effectiveness of longer duration US and MB thrombolytic treatments were studied (n = 4, which suggested that near complete thrombus dissolution is possible. No pulmonary emboli, alterations in oxygen saturation, or hemodynamics occurred with either treatment.Guided high MI diagnostic US+systemic MB facilitates reduction of aged CVC associated thrombi in vivo. MB enhanced sonothrombolytic therapy may be a non-invasive safe alternative to thrombolytic agents in treating thrombotic CVC occlusions.

  11. Ultrasound Microbubble Treatment Enhances Clathrin-Mediated Endocytosis and Fluid-Phase Uptake through Distinct Mechanisms.

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    Farnaz Fekri

    Full Text Available Drug delivery to tumors is limited by several factors, including drug permeability of the target cell plasma membrane. Ultrasound in combination with microbubbles (USMB is a promising strategy to overcome these limitations. USMB treatment elicits enhanced cellular uptake of materials such as drugs, in part as a result of sheer stress and formation of transient membrane pores. Pores formed upon USMB treatment are rapidly resealed, suggesting that other processes such as enhanced endocytosis may contribute to the enhanced material uptake by cells upon USMB treatment. How USMB regulates endocytic processes remains incompletely understood. Cells constitutively utilize several distinct mechanisms of endocytosis, including clathrin-mediated endocytosis (CME for the internalization of receptor-bound macromolecules such as Transferrin Receptor (TfR, and distinct mechanism(s that mediate the majority of fluid-phase endocytosis. Tracking the abundance of TfR on the cell surface and the internalization of its ligand transferrin revealed that USMB acutely enhances the rate of CME. Total internal reflection fluorescence microscopy experiments revealed that USMB treatment altered the assembly of clathrin-coated pits, the basic structural units of CME. In addition, the rate of fluid-phase endocytosis was enhanced, but with delayed onset upon USMB treatment relative to the enhancement of CME, suggesting that the two processes are distinctly regulated by USMB. Indeed, vacuolin-1 or desipramine treatment prevented the enhancement of CME but not of fluid phase endocytosis upon USMB, suggesting that lysosome exocytosis and acid sphingomyelinase, respectively, are required for the regulation of CME but not fluid phase endocytosis upon USMB treatment. These results indicate that USMB enhances both CME and fluid phase endocytosis through distinct signaling mechanisms, and suggest that strategies for potentiating the enhancement of endocytosis upon USMB treatment may

  12. Annexin A8 controls leukocyte recruitment to activated endothelial cells via cell surface delivery of CD63

    Science.gov (United States)

    Poeter, Michaela; Brandherm, Ines; Rossaint, Jan; Rosso, Gonzalo; Shahin, Victor; Skryabin, Boris V.; Zarbock, Alexander; Gerke, Volker; Rescher, Ursula

    2014-04-01

    To enable leukocyte adhesion to activated endothelium, the leukocyte receptor P-selectin is released from Weibel-Palade bodies (WPB) to the endothelial cell surface where it is stabilized by CD63. Here we report that loss of annexin A8 (anxA8) in human umbilical vein endothelial cells (HUVEC) strongly decreases cell surface presentation of CD63 and P-selectin, with a concomitant reduction in leukocyte rolling and adhesion. We confirm the compromised leukocyte adhesiveness in inflammatory-activated endothelial venules of anxA8-deficient mice. We find that WPB of anxA8-deficient HUVEC contain less CD63, and that this is caused by improper transport of CD63 from late multivesicular endosomes to WPB, with CD63 being retained in intraluminal vesicles. Consequently, reduced CD63 cell surface levels are seen following WPB exocytosis, resulting in enhanced P-selectin re-internalization. Our data support a model in which anxA8 affects leukocyte recruitment to activated endothelial cells by supplying WPB with sufficient amounts of the P-selectin regulator CD63.

  13. Upregulation of annexin A1 expression by butyrate in human melanoma cells induces invasion by inhibiting E-cadherin expression.

    Science.gov (United States)

    Shin, Jimin; Song, In-Sung; Pak, Jhang Ho; Jang, Sung-Wuk

    2016-11-01

    Epithelial to mesenchymal transition (EMT) is a critical step in the metastasis of epithelial cancer cells. Butyrate, which is produced from dietary fiber by colonic bacterial fermentation, has been reported to influence EMT. However, some studies have reported that butyrate promotes EMT, while others have reported an inhibitory effect. To clarify these controversial results, it is necessary to elucidate the mechanism by which butyrate can influence EMT. In this study, we examined the potential role of annexin A1 (ANXA1), which was previously reported to promote EMT in breast cancer cells, as a mediator of EMT regulation by butyrate. We found that ANXA1 mRNA and protein were expressed in highly invasive melanoma cell lines (A2058 and A375), but not in SK-MEL-5 cells, which are less invasive. We also showed that butyrate induced ANXA1 mRNA and protein expression and promoted EMT-related cell invasion in SK-MEL-5 cells. Downregulation of ANXA1 expression using specific small interfering RNAs in butyrate-treated SK-MEL-5 cells resulted in increased expression of the epithelial marker E-cadherin and decreased cell invasion. Moreover, overexpressing ANXA1 decreased the expression of the E-cadherin. Collectively, these results indicate that butyrate induces the expression of ANXA1 in human melanoma cells, which then promotes invasion through activating the EMT signaling pathway.

  14. Study of the Annexin A1 and Its Associations with Carcinoembryonic Antigen and Mismatch Repair Proteins in Colorectal Cancer.

    Science.gov (United States)

    Ydy, Lenuce Ribeiro Aziz; do Espírito Santo, Gilmar Ferreira; de Menezes, Ivana; Martins, Michelle Santos; Ignotti, Eliane; Damazo, Amílcar Sabino

    2016-03-01

    Annexin-A1 (ANXA1) has been implicated in various tumor types, but few studies have investigated its involvement in colorectal cancer. The study aimed to analyze ANXA1 expression in the normal margin and colorectal tumor tissues of 104 patients who underwent surgery for colorectal cancer and to associate the ANXA1 expression with predictive clinicopathological variables. Hematoxylin-eosin and immunohistochemical staining were used for the analysis. ANXA1 expression was higher in colorectal cancer than in normal margin tissue (p = 0.0001). However, no differences were observed when we analyzed the ANXA1 expression in colon and rectal tumors (p = 0.830). Also, this protein positivity was associated with increased carcinoembryonic antigen levels (p = 0.004). Our data in the DNA-mismatch repair proteins expression was in accordance to the literature. And their positivity was not associated with ANXA1 presence in colorectal cancer. The high incidence of ANXA1 positive expression in colorectal cancer and its association with carcinoembryonic antigen levels might indicate the importance of this protein in the colorectal cancer biology.

  15. Crystallization of the Membrane-Associated Annexin B1: Roles of Additive Screen, Dynamic Light Scattering, and Bioactivity Assay

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    Ding, F.; Xu, Y; Azzi, A; Zhu, D; Rehse, D; Chen, C; Sun, S; Lin, S

    2010-01-01

    Annexin B1 (AnxB1) is a calcium-dependent phospholipid binding protein from Taenia solium cysticercus and has been reported to possess anticoagulant activity, to inhibit phospholipase A{sub 2}, and to regulate membrane transport. Native AnxB1 and its selenomethionyl derivative have been overproduced in Escherichia coli and purified. The results of dynamic light scattering analysis showed that Hepes buffer combined with low concentration salts (NaCl or CaCl{sub 2}) was beneficial for preventing aggregation and for AnxB1 stabilization in the storage. After the additive screen, crystals have been yielded in the presence of guanidine hydrochloride (Gn-HCl). We determined that a low concentration of Gn-HCl significantly delayed clotting time and increased anticoagulant activity. Analysis of the crystal showed that in the presence of Gn-HCl, AnxB1 crystallizes in orthorhombic space group, which is modified from the cubic space group for crystals grown in the absence of Gn-HCl. A high quality data set (at 1.9 {angstrom}) has been collected successfully for crystals of L-selenomethionine labeled protein in the presence of Gn-HCl, to solve the structure with the single anomalous dispersion method (SAD). The unit cell parameters are a = 102.35 {angstrom}, b = 103.59 {angstrom}, c = 114.60 {angstrom}, {alpha} = {beta} = {gamma} = 90.00{sup o}.

  16. Investigation of a Potential Scintigraphic Tracer for Imaging Apoptosis: Radioiodinated Annexin V-Kunitz Protease Inhibitor Fusion Protein

    Directory of Open Access Journals (Sweden)

    Mei-Hsiu Liao

    2011-01-01

    Full Text Available Radiolabeled annexin V (ANV has been widely used for imaging cell apoptosis. Recently, a novel ANV-Kunitz-type protease inhibitor fusion protein, ANV-6L15, was found to be a promising probe for improved apoptosis detection based on its higher affinity to phosphatidylserine (PS compared to native ANV. The present paper investigates the feasibility of apoptosis detection using radioiodinated ANV-6L15. Native ANV and ANV-6L15 were labeled with iodine-123 and iodine-125 using Iodogen method. The binding between the radioiodinated proteins and erythrocyte ghosts or chemical-induced apoptotic cells was examined. ANV-6L15 can be radioiodinated with high yield (40%−60% and excellent radiochemical purity (>95%. 123I-ANV-6L15 exhibited a higher binding ratio to erythrocyte ghosts and apoptotic cells compared to 123I-ANV. The biodistribution of 123I-ANV-6L15 in mice was also characterized. 123I-ANV-6L15 was rapidly cleared from the blood. High uptake in the liver and the kidneys may limit the evaluation of apoptosis in abdominal regions. Our data suggest that radiolabled ANV-6L15 may be a better scintigraphic tracer than native ANV for apoptosis detection.

  17. Specific localization of the annexin II heterotetramer in brain lipid raft fractions and its changes in spatial learning.

    Science.gov (United States)

    Zhao, Wei-Qin; Waisman, David M; Grimaldi, Maurizio

    2004-08-01

    Annexin-II (AII) is a Ca(2+)-dependent phospholipid-binding protein that is present in both intracellular and extracellular compartments. In the present study AII immunoreactivity was found in a subpopulation of neurons in specific brain regions, including the cerebral cortex and the surface of hippocampal pyramidal neurons from adult rats. AII from synaptic membranes was detected by immunoblotting as multiple species containing the monomer (AII36) and heterotetramer (AIIt). AIIt was resistant to beta-mercaptoethanol and dithiothreitol in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but was completely reduced to monomers (36 kDa) by two-dimensional electrophoresis. AIIt resided exclusively in the detergent-resistant lipid rafts concentrated in neuronal dendrites, and its recruitment to those structures was enhanced by antibody cross-link. AII abundantly distributed on the outer leaflet of neuronal membranes and between spaces of neurons appeared to be neuronal adhesive. The formation of AIIt required synthesis of sphingolipids and cholesterol, and its stability depended on Ca2+. Increases in neuronal activities such as depolarization and learning were shown to promote formation of AIIt. Our results suggest that, via a dynamic association with dendritic lipid rafts, AII may play a role in synaptic signal transduction and remodeling. This probably involves focal adhesion and interactions with actin that are associated with brain development and memory consolidation.

  18. Microbubbles as a scattering contrast agent for grating-based x-ray dark-field imaging.

    Science.gov (United States)

    Velroyen, A; Bech, M; Malecki, A; Tapfer, A; Yaroshenko, A; Ingrisch, M; Cyran, C C; Auweter, S D; Nikolaou, K; Reiser, M; Pfeiffer, F

    2013-02-21

    In clinically established-absorption-based-biomedical x-ray imaging, contrast agents with high atomic numbers (e.g. iodine) are commonly used for contrast enhancement. The development of novel x-ray contrast modalities such as phase contrast and dark-field contrast opens up the possible use of alternative contrast media in x-ray imaging. We investigate using ultrasound contrast agents, which unlike iodine-based contrast agents can also be administered to patients with renal impairment and thyroid dysfunction, for application with a recently developed novel x-ray dark-field imaging modality. To produce contrast from these microbubble-based contrast agents, our method exploits ultra-small-angle coherent x-ray scattering. Such scattering dark-field x-ray images can be obtained with a grating-based x-ray imaging setup, together with refraction-based differential phase-contrast and the conventional attenuation contrast images. In this work we specifically show that ultrasound contrast agents based on microbubbles can be used to produce strongly enhanced dark-field contrast, with superior contrast-to-noise ratio compared to the attenuation signal. We also demonstrate that this method works well with an x-ray tube-based setup and that the relative contrast gain even increases when the pixel size is increased from tenths of microns to clinically compatible detector resolutions about up to a millimetre.

  19. Numerical investigation of air film breakup and micro-bubble formation in liquid-liquid impact events

    Science.gov (United States)

    Mirjalili, Seyedshahabaddin; Mani, Ali

    2013-11-01

    Experimental evidence shows that micro-bubbles can be generated when a droplet of the size of a few millimeters impacts a layer of the same liquid with a velocity of a few meters per second. This phenomenon, also known as Mesler entrainment is cumbersome to numerically simulate due to the small time and length scales involved. In order to gain a better understanding of the relevant scales, parameters, and regions, 2-D boundary element simulations inspired by M. Mani, Mandre, Brenner (JFM, vol. 647, p. 143, 2010) were performed. By developing treatments for topological changes, these simulations are extended to after impact events and finally depict the formation of micro-bubbles of sizes similar to entrapped bubbles in Mesler entrainment. Compressibility effects on final bubble size are discussed, and the requirements for a resolved CFD calculation are obtained. Thereafter, a 2-D two-phase flow calculation using a diffuse interface model is undertaken and based on grid-converged results, the statistics of the bubbles are examined and compared with available experimental data. Supported by the Office of Naval Research.

  20. Degradation of textile dyes Remazol Brilliant Blue using plasma electrolysis method with the addition of microbubble and Fe2+ ion

    Science.gov (United States)

    Zainah, Saksono, Nelson

    2017-11-01

    Dye waste is a liquid waste that mostly generated from the textile industry and is very dangerous for the environment. Plasma electrolysis method is an effective method in degrading dye waste because of its ability to produce radical OH in large quantities. This study aims to test the ability of plasma electrolysis method to degrade one of the textile dyes, Remazol Brilliant Blue, with the addition of Fe2+ ion and microbubble. The dye waste degredation reached 99.74% for 180 minutes with the addition of 40 mg/L of Fe2+ ion as a result of fenton reaction. The addition of microbubble will also increase OH radical production by up to 4.8% and be able to reduce energy consumption by 11.3%. The COD value decreased until 20.56 mg/L and has fulfilled the Government standard of 50 mg/L. In addition, the dye waste concentration decreased significantly from 150 mg/L to 0.388 mg/L. Maximum conditions are obtained by using 0.02 M Na2SO4, 700 Volt operating voltage, and 1 cm anode depth.

  1. Ex Vivo Perfusion-Simulation Measurements of Microbubbles as a Scattering Contrast Agent for Grating-Based X-Ray Dark-Field Imaging.

    Directory of Open Access Journals (Sweden)

    Astrid Velroyen

    Full Text Available The investigation of dedicated contrast agents for x-ray dark-field imaging, which exploits small-angle scattering at microstructures for contrast generation, is of strong interest in analogy to the common clinical use of high-atomic number contrast media in conventional attenuation-based imaging, since dark-field imaging has proven to provide complementary information. Therefore, agents consisting of gas bubbles, as used in ultrasound imaging for example, are of particular interest. In this work, we investigate an experimental contrast agent based on microbubbles consisting of a polyvinyl-alcohol shell with an iron oxide coating, which was originally developed for multimodal imaging and drug delivery. Its performance as a possible contrast medium for small-animal angiography was examined using a mouse carcass to realistically consider attenuating and scattering background signal. Subtraction images of dark field, phase contrast and attenuation were acquired for a concentration series of 100%, 10% and 1.3% to mimic different stages of dilution in the contrast agent in the blood vessel system. The images were compared to the gold-standard iodine-based contrast agent Solutrast, showing a good contrast improvement by microbubbles in dark-field imaging. This study proves the feasibility of microbubble-based dark-field contrast-enhancement in presence of scattering and attenuating mouse body structures like bone and fur. Therefore, it suggests a strong potential of the use of polymer-based microbubbles for small-animal dark-field angiography.

  2. Modeling complicated rheological behaviors in encapsulating shells of lipid-coated microbubbles accounting for nonlinear changes of both shell viscosity and elasticity

    Science.gov (United States)

    Li, Qian; Matula, Thomas J.; Tu, Juan; Guo, Xiasheng; Zhang, Dong

    2013-02-01

    It has been accepted that the dynamic responses of ultrasound contrast agent (UCA) microbubbles will be significantly affected by the encapsulating shell properties (e.g., shell elasticity and viscosity). In this work, a new model is proposed to describe the complicated rheological behaviors in an encapsulating shell of UCA microbubbles by applying the nonlinear ‘Cross law’ to the shell viscous term in the Marmottant model. The proposed new model was verified by fitting the dynamic responses of UCAs measured with either a high-speed optical imaging system or a light scattering system. The comparison results between the measured radius-time curves and the numerical simulations demonstrate that the ‘compression-only’ behavior of UCAs can be successfully simulated with the new model. Then, the shell elastic and viscous coefficients of SonoVue microbubbles were evaluated based on the new model simulations, and compared to the results obtained from some existing UCA models. The results confirm the capability of the current model for reducing the dependence of bubble shell parameters on the initial bubble radius, which indicates that the current model might be more comprehensive to describe the complex rheological nature (e.g., ‘shear-thinning’ and ‘strain-softening’) in encapsulating shells of UCA microbubbles by taking into account the nonlinear changes of both shell elasticity and shell viscosity.

  3. Assessment of portal venous system patency in the liver transplant candidate: A prospective study comparing ultrasound, microbubble-enhanced colour Doppler ultrasound, with arteriography and surgery

    International Nuclear Information System (INIS)

    Marshall, M.M.; Beese, R.C.; Muiesan, P.; Sarma, D.I.; O'Grady, J.; Sidhu, P.S.

    2002-01-01

    AIM: To determine the role of microbubble-enhanced colour Doppler ultrasound (CDUS) in assessing portal venous patency prior to liver transplantation. MATERIALS AND METHODS: Over a 2-year period, all patients with chronic liver disease undergoing routine pre-transplant CDUS examination in whom the portal venous system was inadequately demonstrated were recruited to the study. CDUS was performed in 368 patients and 33 patients (9%) were recruited. A repeat CDUS examination following an intravenous bolus injection of the microbubble contrast agent Levovist[reg] (Schering Healthcare AG, Berlin, Germany) was performed. Diagnostic confidence was recorded on a free linear analogue scale for both examinations. Findings were compared with indirect portography and surgery. RESULTS: Of the 33 patients with sub-optimal baseline examinations, improvement in portal vein visualization was achieved in 31 patients (94%). Median diagnostic confidence increased from 50% (interquartile range 30-60) to 90% (interquartile range 75-98) (P < 0.001) following administration of Levovist[reg]. Overall accuracy of portal vein assessment using microbubble-enhanced CDUS in 15 patients in whom a definitive diagnosis was made within 2 months was 87%. CONCLUSION: Microbubble-enhanced CDUS is a simple, inexpensive adjunct to standard pre liver transplant screening of the portal vein. It is particularly helpful in patients with end-stage cirrhosis who are at high risk of portal vein thrombosis and in whom the conventional examination is sub-optimal.Marshall, M.M. et al. (2002)

  4. A low molecular weight zinc{sup 2+}-dipicolylamine-based probe detects apoptosis during tumour treatment better than an annexin V-based probe

    Energy Technology Data Exchange (ETDEWEB)

    Palmowski, Karin [RWTH-Aachen University, Department of Experimental Molecular Imaging, Aachen (Germany); University of Heidelberg, Department of Pneumology, Thoraxklinik Heidelberg, Heidelberg (Germany); Rix, Anne; Lederle, Wiltrud; Kiessling, Fabian [RWTH-Aachen University, Department of Experimental Molecular Imaging, Aachen (Germany); Behrendt, Florian F. [RWTH-Aachen University, Department of Nuclear Medicine, Aachen (Germany); Mottaghy, Felix M. [RWTH-Aachen University, Department of Nuclear Medicine, Aachen (Germany); Maastricht University Medical Center, Department of Nuclear Medicine, Maastricht (Netherlands); Gray, Brian D. [Molecular Targeting Technologies, Inc., West Chester, PA (United States); Pak, Koon Y. [University Medical Center Heidelberg, Academic Radiology Baden-Baden, Diagnostic and Interventional Radiology, Heidelberg (Germany); Palmowski, Moritz [RWTH-Aachen University, Department of Experimental Molecular Imaging, Aachen (Germany); RWTH-Aachen University, Department of Nuclear Medicine, Aachen (Germany); University Medical Center Heidelberg, Academic Radiology Baden-Baden, Diagnostic and Interventional Radiology, Heidelberg (Germany)

    2014-02-15

    Molecular imaging of apoptosis is frequently discussed for monitoring cancer therapies. Here, we compare the low molecular weight phosphatidylserine-targeting ligand zinc{sup 2+}-dipicolylamine (Zn{sup 2+}-DPA) with the established but reasonably larger protein annexin V. Molecular apoptosis imaging with the fluorescently labelled probes annexin V (750 nm, 36 kDa) and Zn{sup 2+}-DPA (794 nm, 1.84 kDa) was performed in tumour-bearing mice (A431). Three animal groups were investigated: untreated controls and treated tumours after 1 or 4 days of anti-angiogenic therapy (SU11248). Additionally, μPET with {sup 18} F-FDG was performed. Imaging data were displayed as tumour-to-muscle ratio (TMR) and validated by quantitative immunohistochemistry. Compared with untreated control tumours, TUNEL staining indicated significant apoptosis after 1 day (P < 0.05) and 4 days (P < 0.01) of treatment. Concordantly, Zn{sup 2+}-DPA uptake increased significantly after 1 day (P < 0.05) and 4 days (P < 0.01). Surprisingly, annexin V failed to detect significant differences between control and treated animals. Contrary to the increasing uptake of Zn{sup 2+}-DPA, {sup 18} F-FDG tumour uptake decreased significantly at days 1 (P < 0.05) and 4 (P < 0.01). Increase in apoptosis during anti-angiogenic therapy was detected significantly better with the low molecular weight probe Zn{sup 2+}-DPA than with the annexin V-based probe. Additionally, significant treatment effects were detectable as early using Zn{sup 2+}-DPA as with measurements of the glucose metabolism using {sup 18} F-FDG. (orig.)

  5. Ultrasound-targeted microbubble destruction (UTMD assisted delivery of shRNA against PHD2 into H9C2 cells.

    Directory of Open Access Journals (Sweden)

    Li Zhang

    Full Text Available Gene therapy has great potential for human diseases. Development of efficient delivery systems is critical to its clinical translation. Recent studies have shown that microbubbles in combination with ultrasound (US can be used to facilitate gene delivery. An aim of this study is to investigate whether the combination of US-targeted microbubble destruction (UTMD and polyethylenimine (PEI (UTMD/PEI can mediate even greater gene transfection efficiency than UTMD alone and to optimize ultrasonic irradiation parameters. Another aim of this study is to investigate the biological effects of PHD2-shRNA after its transfection into H9C2 cells. pEGFP-N1 or eukaryotic shPHD2-EGFP plasmid was mixed with albumin-coated microbubbles and PEI to form complexes for transfection. After these were added into H9C2 cells, the cells were exposed to US with various sets of parameters. The cells were then harvested and analyzed for gene expression. UTMD/PEI was shown to be highly efficient in gene transfection. An US intensity of 1.5 W/cm2, a microbubble concentration of 300μl/ml, an exposure time of 45s, and a plasmid concentration of 15μg/ml were found to be optimal for transfection. UTMD/PEI-mediated PHD2-shRNA transfection in H9C2 cells significantly down regulated the expression of PHD2 and increased expression of HIF-1α and downstream angiogenesis factors VEGF, TGF-β and bFGF. UTMD/PEI, combined with albumin-coated microbubbles, warrants further investigation for therapeutic gene delivery.

  6. Young women with polycystic ovary syndrome have raised levels of circulating annexin V-positive platelet microparticles.

    Science.gov (United States)

    Willis, G R; Connolly, K; Ladell, K; Davies, T S; Guschina, I A; Ramji, D; Miners, K; Price, D A; Clayton, A; James, P E; Rees, D A

    2014-12-01

    Are circulating microparticles (MPs) altered in young women with polycystic ovary syndrome (PCOS)? Women with PCOS have elevated concentrations of circulating platelet-derived MPs, which exhibit increased annexin V binding and altered microRNA (miR) profiles compared with healthy volunteers. Some studies have shown that cardiovascular risk is increased in young women with PCOS but the mechanisms by which this occurs are uncertain. Circulating MPs are elevated in patients with cardiovascular disease but the characteristics of MPs in patients with PCOS are unclear. Case-control study comprising 17 women with PCOS (mean ± SD; age 31 ± 7 years, BMI 29 ± 6 kg/m(2)) and 18 healthy volunteers (age 31 ± 6 years, BMI 30 ± 6 kg/m(2)). The study was conducted in a University hospital. Nanoparticle tracking analysis (NTA) and flow cytometry (CD41 platelet, CD11b monocyte, CD144 endothelial) were used to determine MP size, concentration, cellular origin and annexin V positivity (reflecting phosphatidylserine exposure). Fatty acid analysis was performed by gas chromatography and MP miR expression profiles were compared by microarray. PCOS subjects showed increased MP concentrations compared with healthy volunteers (mean ± SD; 11.5 ± 5 × 10(12)/ml versus 10.0 ± 4 × 10(12)/ml, respectively; P = 0.03), which correlated with the homeostasis model of insulin resistance (r = 0.53, P = 0.03). This difference was predominantly seen in MPs whose size was in the small exosomal range (PV(+) MPs compared with healthy volunteers (84 ± 18 versus 74 ± 24%, respectively, P = 0.05) but the cellular origin of MPs, which were predominantly platelet-derived (PCOS: 99 ± 0.9%; controls: 99 ± 2.5%), did not differ. MP fatty acid concentration and composition was similar between groups but 16 miRs were differentially expressed (P < 0.05). Patients with PCOS were classified by the Rotterdam criteria, which describes a less severe metabolic phenotype than other definitions of the syndrome

  7. Apo and calcium-bound crystal structures of cytoskeletal protein alpha-14 giardin (annexin E1) from the intestinal protozoan parasite Giardia lamblia.

    Science.gov (United States)

    Pathuri, Puja; Nguyen, Emily Tam; Ozorowski, Gabriel; Svärd, Staffan G; Luecke, Hartmut

    2009-01-30

    Alpha-14 giardin (annexin E1), a member of the alpha giardin family of annexins, has been shown to localize to the flagella of the intestinal protozoan parasite Giardia lamblia. Alpha giardins show a common ancestry with the annexins, a family of proteins most of which bind to phospholipids and cellular membranes in a Ca(2+)-dependent manner and are implicated in numerous membrane-related processes including cytoskeletal rearrangements and membrane organization. It has been proposed that alpha-14 giardin may play a significant role during the cytoskeletal rearrangement during differentiation of Giardia. To gain a better understanding of alpha-14 giardin's mode of action and its biological role, we have determined the three-dimensional structure of alpha-14 giardin and its phospholipid-binding properties. Here, we report the apo crystal structure of alpha-14 giardin determined in two different crystal forms as well as the Ca(2+)-bound crystal structure of alpha-14 giardin, refined to 1.9, 1.6 and 1.65 A, respectively. Although the overall fold of alpha-14 giardin is similar to that of alpha-11 giardin, multiwavelength anomalous dispersion phasing was required to solve the alpha-14 giardin structure, indicating significant structural differences between these two members of the alpha giardin family. Unlike most annexin structures, which typically possess N-terminal domains, alpha-14 giardin is composed of only a core domain, followed by a C-terminal extension that may serve as a ligand for binding to cytoskeletal protein partners in Giardia. In the Ca(2+)-bound structure we detected five bound calcium ions, one of which is a novel, highly coordinated calcium-binding site not previously observed in annexin structures. This novel high-affinity calcium-binding site is composed of seven protein donor groups, a feature rarely observed in crystal structures. In addition, phospholipid-binding assays suggest that alpha-14 giardin exhibits calcium-dependent binding to

  8. The native structure of annexin A2 peptides in hydrophilic environment determines their anti-angiogenic effects.

    Science.gov (United States)

    Raddum, Aase M; Hollås, Hanne; Shumilin, Igor A; Henklein, Petra; Kretsinger, Robert; Fossen, Torgils; Vedeler, Anni

    2015-05-01

    The progression of aggressive cancer occurs via angiogenesis and metastasis makes these processes important targets for the development of anti-cancer agents. However, recent studies have raised the concern that selective inhibition of angiogenesis results in a switch towards increased tumour growth and metastasis. Since Annexin A2 (AnxA2) is involved in both angiogenesis and metastasis, it may serve as an ideal target for the simultaneous inhibition of both processes. Based on the discovery that domains I (D(I)) and IV (D(IV)) of AnxA2 are potent inhibitors of angiogenesis, we designed seven peptides derived from these domains based on AnxA2 crystal structures. The peptides were expressed as fusion peptides to increase their folding and solubility. Light scattering, far-UV circular dichroism and thermal transition analyses were employed to investigate their aggregation tendencies, α-helical propensity and stability, respectively. 2,2,2-trifluoroethanol (50%) increased the α-helical propensities of all peptides, indicating that they may favour a hydrophobic environment, but did not enhance their thermal stability. D(I)-P2 appears to be the most stable and folded peptide in a hydrophilic environment. The secondary structure of D(I)-P2 was confirmed by nuclear magnetic resonance spectra. The effect of the seven AnxA2 peptides on the formation and integrity of capillary-like networks was studied in a co-culture system mimicking many of the angiogenesis-related processes. Notably, D(I)-P2 inhibited significantly network formation in this system, indicating that the folded D(I)-P2 peptide interferes with vascular endothelial growth factor-dependent pro-angiogenic processes. Thus, this peptide has the potential of being developed further as an anti-angiogenic drug. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  9. Domains I and IV of annexin A2 affect the formation and integrity of in vitro capillary-like networks.

    Science.gov (United States)

    Raddum, Aase M; Evensen, Lasse; Hollås, Hanne; Grindheim, Ann Kari; Lorens, James B; Vedeler, Anni

    2013-01-01

    Annexin A2 (AnxA2) is a widely expressed multifunctional protein found in different cellular compartments. In spite of lacking a hydrophobic signal peptide, AnxA2 is found at the cell surface of endothelial cells, indicative of a role in angiogenesis. Increased extracellular levels of AnxA2 in tumours correlate with neoangiogenesis, metastasis and poor prognosis. We hypothesised that extracellular AnxA2 may contribute to angiogenesis by affecting endothelial cell-cell interactions and motility. To address this question, we studied the effect of heterotetrameric and monomeric forms of AnxA2, as well as its two soluble domains on the formation and maintenance of capillary-like structures by using an in vitro co-culture system consisting of endothelial and smooth muscle cells. In particular, addition of purified domains I and IV of AnxA2 potently inhibited the vascular endothelial growth factor (VEGF)-dependent formation of the capillary-like networks in a dose-dependent manner. In addition, these AnxA2 domains disrupted endothelial cell-cell contacts in preformed capillary-like networks, resulting in the internalisation of vascular endothelial (VE)-cadherin and the formation of VE-cadherin-containing filopodia-like structures between the endothelial cells, suggesting increased cell motility. Addition of monoclonal AnxA2 antibodies, in particular against Tyr23 phosphorylated AnxA2, also strongly inhibited network formation in the co-culture system. These results suggest that extracellular AnxA2, most likely in its Tyr phosphorylated form, plays a pivotal role in angiogenesis. The exogenously added AnxA2 domains most likely mediate their effects by competing with endogenous AnxA2 for extracellular factors necessary for the initiation and maintenance of angiogenesis, such as those involved in the formation/integrity of cell-cell contacts.

  10. The role of annexin A1 in expression of matrix metalloproteinase-9 and invasion of breast cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Hyereen [Department of Medicine, Graduate School, University of Ulsan, Pungnap-2 dong, Songpa-gu, Seoul (Korea, Republic of); Ko, Jesang [School of Life Sciences and Biotechnology, Korea University, Seoul 136-701 (Korea, Republic of); Jang, Sung-Wuk, E-mail: swjang@amc.seoul.kr [Department of Medicine, Graduate School, University of Ulsan, Pungnap-2 dong, Songpa-gu, Seoul (Korea, Republic of)

    2012-06-22

    Highlights: Black-Right-Pointing-Pointer We evaluated the effect of ANXA1 on promoting migration and invasion in MDA-MB-231 cells. Black-Right-Pointing-Pointer ANXA1 siRNA inhibits invasion and migration. Black-Right-Pointing-Pointer ANXA1 regulates MMP-9 expression and activity. Black-Right-Pointing-Pointer ANX-1 siRNA inhibits the activation of NF-{kappa}B in MDA-MB-231 cells. -- Abstract: Matrix metalloproteinase-9 (MMP-9) plays an important role in the invasion and metastasis of cancer cells. However, the regulatory mechanism of MMP-9 expression and its biological effects on breast cancer development remain obscure. In the current study, we examined the potential role of annexin A1 (ANXA1) in regulating migration and invasion in breast cancer cell lines. Both ANXA1 mRNA and protein are expressed in the highly invasive, hormone-insensitive human breast cancer cell lines MDA-MB-231 and SKBr3, but not in the hormone-responsive cell lines MCF-7 and T47D. Downregulation of ANXA1 expression with specific small interfering RNAs (ANXA1 siRNA) in MDA-MB-231 cells resulted in decreased cancer cell migration and invasion. Ablation of ANXA1 expression decreases the expression of MMP-9 at both the mRNA and protein levels and also reduces the proteolytic activity of MMP-9 in MDA-MB-231 cells. Moreover, silencing ANXA1 also decreases the transcriptional activity of MMP-9 by the suppression of nuclear factor kappa-B (NF-{kappa}B) activity. Collectively, these results indicate that ANXA1 functions as a positive regulator of MMP-9 expression and invasion of breast cancer cells through specific activation of the NF-{kappa}B signaling pathway.

  11. Expression profiles of Annexin A1, formylated peptide receptors and cyclooxigenase-2 in gastroesophageal inflammations and neoplasias.

    Science.gov (United States)

    Takaoka, Rodolfo T C; Sertório, Nathália D; Magalini, Lara P J; Dos Santos, Leandro M; Souza, Helena R; Iyomasa-Pilon, Melina M; Possebon, Lucas; Costa, Sara S; Girol, Ana P

    2018-02-01

    The anti-inflammatory protein Annexin-A1 (ANXA1) is associated to tumor invasion process and its actions can be mediated by formylated peptides receptors (FPRs). Therefore, we evaluated the expression and correlation of ANXA1, FPR and cyclooxygenase-2 (COX-2) enzyme in esophageal and stomach inflammations and neoplasias. The study of proteins was performed by immunohistochemistry in biopsies of esophagitis, Barrett's esophagus, squamous cell carcinoma and adenocarcinoma of the esophagus, as well as gastritis, stomach polypus and gastric adenocarcinoma. The intensity of the expressions was evaluated by densitometry. The immunohistochemical and densitometric analyzes showed specificity for the FPR1 receptor and modulation of the ANXA1, COX-2 and FPR1 expressions in the epithelial cells in the different studied conditions. Increased immunoreactivity of these proteins was observed in cases of inflammation and stomach polypus. Interestingly, moderate immunoreactivity for ANXA1 and FPR1 but increased immunolabeling for COX-2 were observed in Barrett́s esophagus and esophageal adenocarcinomas. Also, there was reduced expression of ANXA1 and FPR1 in esophageal carcinoma but COX-2 overexpression in this tumor. There was no expression of FPR2 but ANXA1 and FPR1 expressions were positively correlated in all clinical conditions studied. Positive correlation between ANXA1 and COX-2 were also observed in inflammation conditions while negative correlation between ANXA1 and COX-2 was observed in esophageal carcinoma. Our results demonstrate the unregulated expression of ANXA1 and COX-2 in precursor lesions of esophageal and stomach cancers, reinforcing their involvement in gastroesophageal carcinogenesis. In addition, the data show that the actions of ANXA1 in the inflammatory and neoplastic processes of the esophagus and stomach are specifically mediated by the FPR1 receptor. Copyright © 2017 Elsevier GmbH. All rights reserved.

  12. PET imaging of apoptosis with 64Cu-labeled streptavidin following pretargeting of phosphatidylserine with biotinylated annexin-V

    International Nuclear Information System (INIS)

    Cauchon, Nicole; Langlois, Rejean; Rousseau, Jacques A.; Tessier, Guillaume; Cadorette, Jules; Lecomte, Roger; Hunting, Darel J.; Lier, Johan E. van; Pavan, Roberto A.; Zeisler, Stefan K.

    2007-01-01

    In vivo detection of apoptosis is a diagnostic tool with potential clinical applications in cardiology and oncology. Radiolabeled annexin-V (anxV) is an ideal probe for in vivo apoptosis detection owing to its strong affinity for phosphatidylserine (PS), the molecular flag on the surface of apoptotic cells. Most clinical studies performed to visualize apoptosis have used 99m Tc-anxV; however, its poor distribution profile often compromises image quality. In this study, tumor apoptosis after therapy was visualized by positron emission tomography (PET) using 64 Cu-labeled streptavidin (SAv), following pre-targeting of apoptotic cells with biotinylated anxV. Apoptosis was induced in tumor-bearing mice by photodynamic therapy (PDT) using phthalocyanine dyes as photosensitizers, and red light. After PDT, mice were injected i.v. with biotinylated anxV, followed 2 h later by an avidin chase, and after another 2 h with 64 Cu-DOTA-biotin-SAv. PET images were subsequently recorded up to 13 h after PDT. PET images delineated apoptosis in treated tumors as early as 30 min after 64 Cu-DOTA-biotin-SAv administration, with tumor-to-background ratios reaching a maximum at 3 h post-injection, i.e., 7 h post-PDT. Omitting the administration of biotinylated anxV or the avidin chase failed to provide a clear PET image, confirming that all three steps are essential for adequate visualization of apoptosis. Furthermore, differences in action mechanisms between photosensitizers that target tumor cells directly or via initial vascular stasis were clearly recognized through differences in tracer uptake patterns detecting early or delayed apoptosis. This study demonstrates the efficacy of a three-step 64 Cu pretargeting procedure for PET imaging of apoptosis. Our data also confirm the usefulness of small animal PET to evaluate cancer treatment protocols. (orig.)

  13. Domains I and IV of annexin A2 affect the formation and integrity of in vitro capillary-like networks.

    Directory of Open Access Journals (Sweden)

    Aase M Raddum

    Full Text Available Annexin A2 (AnxA2 is a widely expressed multifunctional protein found in different cellular compartments. In spite of lacking a hydrophobic signal peptide, AnxA2 is found at the cell surface of endothelial cells, indicative of a role in angiogenesis. Increased extracellular levels of AnxA2 in tumours correlate with neoangiogenesis, metastasis and poor prognosis. We hypothesised that extracellular AnxA2 may contribute to angiogenesis by affecting endothelial cell-cell interactions and motility. To address this question, we studied the effect of heterotetrameric and monomeric forms of AnxA2, as well as its two soluble domains on the formation and maintenance of capillary-like structures by using an in vitro co-culture system consisting of endothelial and smooth muscle cells. In particular, addition of purified domains I and IV of AnxA2 potently inhibited the vascular endothelial growth factor (VEGF-dependent formation of the capillary-like networks in a dose-dependent manner. In addition, these AnxA2 domains disrupted endothelial cell-cell contacts in preformed capillary-like networks, resulting in the internalisation of vascular endothelial (VE-cadherin and the formation of VE-cadherin-containing filopodia-like structures between the endothelial cells, suggesting increased cell motility. Addition of monoclonal AnxA2 antibodies, in particular against Tyr23 phosphorylated AnxA2, also strongly inhibited network formation in the co-culture system. These results suggest that extracellular AnxA2, most likely in its Tyr phosphorylated form, plays a pivotal role in angiogenesis. The exogenously added AnxA2 domains most likely mediate their effects by competing with endogenous AnxA2 for extracellular factors necessary for the initiation and maintenance of angiogenesis, such as those involved in the formation/integrity of cell-cell contacts.

  14. Non-invasive controlled release from gold nanoparticle integrated photo-responsive liposomes through pulse laser induced microbubble cavitation.

    Science.gov (United States)

    Mathiyazhakan, Malathi; Yang, Yuanxiang; Liu, Yibo; Zhu, Caigang; Liu, Quan; Ohl, Claus-Dieter; Tam, Kam Chiu; Gao, Yu; Xu, Chenjie

    2015-02-01

    Drug-carriers, capable of releasing the drug at the target sites upon external stimuli, are attractive for theranostic applications. In recent years, photo-responsive nanoparticles (NPs) have received considerable attention because of their potentials in providing spatial, temporal, and dosage control over the drug release. However, most of the relevant technologies are still in the process of development and are unprocurable by the clinics. Here, we demonstrated facile fabrication of these photo-responsive NPs by loading hydrophilic gold NPs within thermo-responsive liposomes. Calcein was used as a model drug to evaluate the encapsulation efficiency and the release kinetic profile upon heat/light stimulation. Furthermore, we characterized their size, morphology, phase transition temperature and stability. Finally, we demonstrated that this photo-triggered release might be due to the membrane disruption caused by microbubble cavitation. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Pretest Score for Predicting Microbubble Contrast Agent Use in Stress Echocardiography: A Method to Increase Efficiency in the Echo Laboratory

    Directory of Open Access Journals (Sweden)

    Mathieu Bernier

    2009-01-01

    contrast. Logistic regression models were used to evaluate the association between individual characteristics and contrast use. An 11-point score was derived from the significant characteristics. Results. Variables associated with microbubble use were age, sex, smoking, presence of multiple risk factors, bodymass index (BMI, referral for dobutamine stress echocardiography, history of coronary artery disease, and abnormal baseline electrocardiogram. All variables except BMI were given a score of 1 if present and 0 if absent; BMI was given a score of 0 to 4 according to its value. An increased score was directly proportional to increased likelihood of contrast use. The score cutoff value to optimize sensitivity and specificity was 5. Conclusions. A pretest score can be computed from information available before imaging. It may facilitate contrast agent use through early identification of patients who are likely to benefit from improved endocardial border definition.

  16. High-Frequency Fiber-Optic Ultrasonic Sensor Using Air Micro-Bubble for Imaging of Seismic Physical Models

    Directory of Open Access Journals (Sweden)

    Tingting Gang

    2016-12-01

    Full Text Available A micro-fiber-optic Fabry-Perot interferometer (FPI is proposed and demonstrated experimentally for ultrasonic imaging of seismic physical models. The device consists of a micro-bubble followed by the end of a single-mode fiber (SMF. The micro-structure is formed by the discharging operation on a short segment of hollow-core fiber (HCF that is spliced to the SMF. This micro FPI is sensitive to ultrasonic waves (UWs, especially to the high-frequency (up to 10 MHz UW, thanks to its ultra-thin cavity wall and micro-diameter. A side-band filter technology is employed for the UW interrogation, and then the high signal-to-noise ratio (SNR UW signal is achieved. Eventually the sensor is used for lateral imaging of the physical model by scanning UW detection and two-dimensional signal reconstruction.

  17. Optical Microbubble Resonators with High Refractive Index Inner Coating for Bio-Sensing Applications: An Analytical Approach

    Directory of Open Access Journals (Sweden)

    Andrea Barucci

    2016-11-01

    Full Text Available The design of Whispering Gallery Mode Resonators (WGMRs used as an optical transducer for biosensing represents the first and crucial step towards the optimization of the final device performance in terms of sensitivity and Limit of Detection (LoD. Here, we propose an analytical method for the design of an optical microbubble resonator (OMBR-based biosensor. In order to enhance the OMBR sensing performance, we consider a polymeric layer of high refractive index as an inner coating for the OMBR. The effect of this layer and other optical/geometrical parameters on the mode field distribution, sensitivity and LoD of the OMBR is assessed and discussed, both for transverse electric (TE and transverse magnetic (TM polarization. The obtained results do provide physical insights for the development of OMBR-based biosensor.

  18. Effects of RNA interference combined with ultrasonic irradiation and SonoVue microbubbles on expression of STAT3 gene in keratinocytes of psoriatic lesions.

    Science.gov (United States)

    Ran, Li-Wei; Wang, Hao; Lan, Dong; Jia, Hong-Xia; Yu, Si-Si

    2017-04-01

    The most effective sequence of small interfering RNA (siRNA) silencing STAT3 of psoriatic keratinocytes (KCs) was screened out, and the effects of the most effective siRNA combined with ultrasonic irradiation and SonoVue microbubbles on the expression of STAT3 of KCs and the dose- and time-response were investigated. Three chemically-synthetic siRNAs targeting STAT3 carried by Lipofectamine 3000 were transfected into KCs, and the effects on STAT3 expression were detected, then the most effective siRNA was selected for the subsequent experiments. The negative controls of siRNA (siRNA-NC) labeled with Cy3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles were transfected into KCs, then the optimal parameters of ultrasonic irradiation were determined. The most effective siRNA carried by Li-pofectamine 3000 combined with ultrasonic irradiation at the optimal parameters and SonoVue microbubbles was transfected into KCs, and the dose- and time-response of RNA interference was determined. The effect of RNA interference by the most effective siRNA at the optimal time and dose carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (LUS group) was compared with that only carried by Li-pofectamine 3000 (L group). The results showed that siRNA-3 achieved the highest silencing efficacy. 0.5 W/cm2 and 30 s were selected as the parameters of ultrasonic irradiation. The siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles could effectively knock down the STAT3 expression at mRNA and protein levels in dose- and time-dependent manners determined at 100 nmol/L with maximum downregulation on mRNA at 48 h, and on protein at 72 h after transfection. The LUS group achieved the highest silencing efficacy. It was concluded that siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles could effectively knock down the STAT3

  19. Static response of coated microbubbles compressed between rigid plates: Simulations and asymptotic analysis including elastic and adhesive forces

    Science.gov (United States)

    Lytra, A.; Pelekasis, N.

    2018-03-01

    The static response of coated microbubbles is investigated with a novel approach employed for modeling contact between a microbubble and the cantilever of an atomic force microscope. Elastic tensions and moments are described via appropriate constitutive laws. The encapsulated gas is assumed to undergo isothermal variations. Due to the hydrophilic nature of the cantilever, an ultrathin aqueous film is formed, which transfers the force onto the shell. An interaction potential describes the local pressure applied on the shell. The problem is solved in axisymmetric form with the finite element method. The response is governed by the dimensionless bending, k^ b=kb/(χ R02 ), pressure, P^ A=(PAR0 )/χ , and interaction potential, W ^ =w0/χ . Hard polymeric shells have negligible resistance to gas compression, while for the softer lipid shells gas compressibility is comparable with shell elasticity. As the external force increases, numerical simulations reveal that the force versus deformation (f vs d) curve of polymeric shells exhibits a transition from the linear O(d) (Reissner) regime, marked by flattened shapes around the contact region, to a non-linear O(d1/2) (Pogorelov) regime dominated by shapes exhibiting crater formation due to buckling. When lipid shells are tested, buckling is bypassed as the external force increases and flattened shapes prevail in an initially linear f vs d curve. Transition to a curved upwards regime is observed as the force increases, where gas compression and area dilatation form the dominant balance providing a nonlinear regime with an O(d3) dependence. Asymptotic analysis recovers the above patterns and facilitates estimation of the shell mechanical properties.

  20. The partitioning of nanoparticles to endothelium or interstitium during ultrasound-microbubble-targeted delivery depends on peak-negative pressure

    Energy Technology Data Exchange (ETDEWEB)

    Hsiang, Y.-H.; Song, J.; Price, R. J., E-mail: rprice@virginia.edu [University of Virginia, Department of Biomedical Engineering (United States)

    2015-08-15

    Patients diagnosed with advanced peripheral arterial disease often face poor prognoses and have limited treatment options. For some patient populations, the therapeutic growth of collateral arteries (i.e. arteriogenesis) that bypass regions affected by vascular disease may become a viable treatment option. Our group and others are developing therapeutic approaches centered on the ability of ultrasound-activated microbubbles to permeabilize skeletal muscle capillaries and facilitate the targeted delivery of pro-arteriogenic growth factor-bearing nanoparticles. The development of such approaches would benefit significantly from a better understanding of how nanoparticle diameter and ultrasound peak-negative pressure affect both total nanoparticle delivery and the partitioning of nanoparticles to endothelial or interstitial compartments. Toward this goal, using Balb/C mice that had undergone unilateral femoral artery ligation, we intra-arterially co-injected nanoparticles (50 and 100 nm) with microbubbles, applied 1 MHz ultrasound to the gracilis adductor muscle at peak-negative pressures of 0.7, 0.55, 0.4, and 0.2 MPa, and analyzed nanoparticle delivery and distribution. As expected, total nanoparticle (50 and 100 nm) delivery increased with increasing peak-negative pressure, with 50 nm nanoparticles exhibiting greater tissue coverage than 100 nm nanoparticles. Of particular interest, increasing peak-negative pressure resulted in increased delivery to the interstitium for both nanoparticle sizes, but had little influence on nanoparticle delivery to the endothelium. Thus, we conclude that alterations to peak-negative pressure may be used to adjust the fraction of nanoparticles delivered to the interstitial compartment. This information will be useful when designing ultrasound protocols for delivering pro-arteriogenic nanoparticles to skeletal muscle.

  1. Changes in tumor vascularity precede microbubble contrast accumulation deficit in the process of dedifferentiation of hepatocellular carcinoma

    International Nuclear Information System (INIS)

    Maruyama, Hitoshi; Takahashi, Masanori; Ishibashi, Hiroyuki; Okabe, Shinichiro; Yoshikawa, Masaharu; Yokosuka, Osamu

    2010-01-01

    Purpose: To elucidate the changes in tumor vascularity and microbubble accumulation on contrast-enhanced sonograms, in relation to the dedifferentiation of hepatocellular carcinoma (HCC). Materials and methods: This prospective study enrolled 10 patients with histologically proven HCC (14.4-39.0 mm, 26.1 ± 7.4) showing nodule-in-nodule appearance upon contrast-enhanced computed tomography. Contrast-enhanced ultrasound was performed by harmonic imaging under a low mechanical index (0.22-0.25) during the vascular phase (agent injection to 1 min) and late phase (15 min) following the injection of Sonazoid TM (0.0075 ml/kg). Contrast enhancement in the inner and outer nodules was assessed in comparison with that in adjacent liver parenchyma as hyper-, iso-, or hypo-enhanced. Results: Vascular-phase enhancement of all 10 inner nodules was hyper-enhanced, and that of outer nodules was hyper-enhanced in 3, iso-enhanced in 2, and hypo-enhanced in 5. Late-phase enhancement of inner nodules was hypo-enhanced in 8 and iso-enhanced in 2. Furthermore, late-phase enhancement of outer nodules was iso-enhanced in the 7 lesions that showed iso- or hypo-enhancement in the vascular phase, and hypo-enhanced in the 3 with hyper-enhancement in the vascular phase. Late-phase hypo-enhancement was significantly more frequent in the nodules showing early-phase hyper-enhancement (11/13) than in the nodules showing early-phase iso- or hypo-enhancement (0/7) in both the inner and outer nodules. Conclusion: Dedifferentiation of HCC may be accompanied by changes in tumor vascularity prior to a reduction in microbubble accumulation. Observation of the vascular phase may be more useful than late-phase imaging for the early recognition of HCC dedifferentiation when using contrast-enhanced ultrasound with Sonazoid.

  2. The partitioning of nanoparticles to endothelium or interstitium during ultrasound-microbubble-targeted delivery depends on peak-negative pressure

    International Nuclear Information System (INIS)

    Hsiang, Y.-H.; Song, J.; Price, R. J.

    2015-01-01

    Patients diagnosed with advanced peripheral arterial disease often face poor prognoses and have limited treatment options. For some patient populations, the therapeutic growth of collateral arteries (i.e. arteriogenesis) that bypass regions affected by vascular disease may become a viable treatment option. Our group and others are developing therapeutic approaches centered on the ability of ultrasound-activated microbubbles to permeabilize skeletal muscle capillaries and facilitate the targeted delivery of pro-arteriogenic growth factor-bearing nanoparticles. The development of such approaches would benefit significantly from a better understanding of how nanoparticle diameter and ultrasound peak-negative pressure affect both total nanoparticle delivery and the partitioning of nanoparticles to endothelial or interstitial compartments. Toward this goal, using Balb/C mice that had undergone unilateral femoral artery ligation, we intra-arterially co-injected nanoparticles (50 and 100 nm) with microbubbles, applied 1 MHz ultrasound to the gracilis adductor muscle at peak-negative pressures of 0.7, 0.55, 0.4, and 0.2 MPa, and analyzed nanoparticle delivery and distribution. As expected, total nanoparticle (50 and 100 nm) delivery increased with increasing peak-negative pressure, with 50 nm nanoparticles exhibiting greater tissue coverage than 100 nm nanoparticles. Of particular interest, increasing peak-negative pressure resulted in increased delivery to the interstitium for both nanoparticle sizes, but had little influence on nanoparticle delivery to the endothelium. Thus, we conclude that alterations to peak-negative pressure may be used to adjust the fraction of nanoparticles delivered to the interstitial compartment. This information will be useful when designing ultrasound protocols for delivering pro-arteriogenic nanoparticles to skeletal muscle

  3. 99mTc-annexin V and 111In-antimyosin antibody uptake in experimental myocardial infarction in rats

    International Nuclear Information System (INIS)

    Sarda-Mantel, Laure; Rouzet, Francois; Martet, Genevieve; Raguin, Olivier; Vrigneaud, Jean-Marc; Guludec, Dominique Le; Michel, Jean-Baptiste; Louedec, Liliane; Vanderheyden, Jean-Luc; Hervatin, Florence; Khaw, Ban An

    2006-01-01

    99m Tc-annexin V (ANX) allows scintigraphic detection of apoptotic cells via specific binding to exposed phosphatidylserine. In myocardial infarction, apoptosis of myocytes is variable and depends especially on the presence or absence of coronary reperfusion. In this study, ANX uptake in non-reperfused experimental myocardial infarcts was compared with uptake of a marker of myocyte necrosis ( 111 In-antimyosin antibodies, AM) and an immunohistochemical marker of apoptosis (Apostain). The left anterior coronary artery was ligated in 47 Wistar rats, which were then injected with ANX (n=20), AM (n=21) or both (n=6). Myocardial uptake of ANX and AM was determined at 2 h (n=14), 4 h (n=14) and 24 h (n=19) after coronary ligation (CL), by quantitative autoradiography with (n=23) or without (n=24) gamma imaging. Heart-to-lung ratios (HLRs) and infarct-to-remote myocardium activity ratios (INRs) were calculated on the scintigrams and autoradiograms respectively. Cardiac sections were stained with haematoxylin-eosin and Apostain. The above studies were repeated in 12 normal rats. All rats with CL showed increased ANX and AM uptake in cardiac areas on scintigrams 24 h after CL, with HLRs higher than in controls: 3.1±0.6 versus 1.5±0.3 (p=0.001) for ANX and 1.99±0.44 versus 1.01±0.05 (p<0.0005) for AM. Autoradiography showed intense ANX and AM uptake in infarcts, with comparable topography and INRs at 2 h, 4 h and 24 h after CL (4.6±0.9 versus 5.0±1.8 at 24 h), while Apostain staining was very low (0.06±0.06% of cells). In this model of persistent CL, we observed increased ANX uptake in injured myocardium, comparable in intensity, topography and kinetics to that of AM. There was only minimal Apostain staining in the same areas. (orig.)

  4. Expression of Annexin-A1 and Galectin-1 Anti-Inflammatory Proteins and mRNA in Chronic Gastritis and Gastric Cancer

    Directory of Open Access Journals (Sweden)

    Yvana Cristina Jorge

    2013-01-01

    Full Text Available Objective. The anti-inflammatory proteins annexin-A1 and galectin-1 have been associated with tumor progression. This scenario prompted us to investigate the relationship between the gene and protein expression of annexin-A1 (ANXA1/AnxA1 and galectin-1 (LGALS1/Gal-1 in an inflammatory gastric lesion as chronic gastritis (CG and gastric adenocarcinoma (GA and its association with H. pylori infection. Methods. We analyzed 40 samples of CG, 20 of GA, and 10 of normal mucosa (C by the quantitative real-time PCR (qPCR technique and the immunohistochemistry assay. Results. High ANXA1 mRNA expression levels were observed in 90% (36/40 of CG cases (mean relative quantification RQ = 4.26 ± 2.03 and in 80% (16/20 of GA cases (mean RQ = 4.38 ± 4.77. However, LGALS1 mRNA levels were high (mean RQ = 2.44 ± 3.26 in 60% (12/20 of the GA cases, while low expression was found in CG (mean RQ = 0.43±3.13; P<0.01. Normal mucosa showed modest immunoreactivity in stroma but not in epithelium, while stroma and epithelium displayed an intense immunostaining in CG and GA for both proteins. Conclusion. These results have provided evidence that galectin-1 and mainly annexin-A1 are overexpressed in both gastritis and gastric cancer, suggesting a strong association of these proteins with chronic gastric inflammation and carcinogenesis.

  5. Monitoring Cell Death in Regorafenib-Treated Experimental Colon Carcinomas Using Annexin-Based Optical Fluorescence Imaging Validated by Perfusion MRI.

    Directory of Open Access Journals (Sweden)

    Philipp M Kazmierczak

    Full Text Available To investigate annexin-based optical fluorescence imaging (OI for monitoring regorafenib-induced early cell death in experimental colon carcinomas in rats, validated by perfusion MRI and multiparametric immunohistochemistry.Subcutaneous human colon carcinomas (HT-29 in athymic rats (n = 16 were imaged before and after a one-week therapy with regorafenib (n = 8 or placebo (n = 8 using annexin-based OI and perfusion MRI at 3 Tesla. Optical signal-to-noise ratio (SNR and MRI tumor perfusion parameters (plasma flow PF, mL/100mL/min; plasma volume PV, % were assessed. On day 7, tumors underwent immunohistochemical analysis for tumor cell apoptosis (TUNEL, proliferation (Ki-67, and microvascular density (CD31.Apoptosis-targeted OI demonstrated a tumor-specific probe accumulation with a significant increase of tumor SNR under therapy (mean Δ +7.78±2.95, control: -0.80±2.48, p = 0.021. MRI detected a significant reduction of tumor perfusion in the therapy group (mean ΔPF -8.17±2.32 mL/100 mL/min, control -0.11±3.36 mL/100 mL/min, p = 0.036. Immunohistochemistry showed significantly more apoptosis (TUNEL; 11392±1486 vs. 2921±334, p = 0.001, significantly less proliferation (Ki-67; 1754±184 vs. 2883±323, p = 0.012, and significantly lower microvascular density (CD31; 107±10 vs. 182±22, p = 0.006 in the therapy group.Annexin-based OI allowed for the non-invasive monitoring of regorafenib-induced early cell death in experimental colon carcinomas, validated by perfusion MRI and multiparametric immunohistochemistry.

  6. Influence of ultrasound power on acoustic streaming and micro-bubbles formations in a low frequency sono-reactor: mathematical and 3D computational simulation.

    Science.gov (United States)

    Sajjadi, Baharak; Raman, Abdul Aziz Abdul; Ibrahim, Shaliza

    2015-05-01

    This paper aims at investigating the influence of ultrasound power amplitude on liquid behaviour in a low-frequency (24 kHz) sono-reactor. Three types of analysis were employed: (i) mechanical analysis of micro-bubbles formation and their activities/characteristics using mathematical modelling. (ii) Numerical analysis of acoustic streaming, fluid flow pattern, volume fraction of micro-bubbles and turbulence using 3D CFD simulation. (iii) Practical analysis of fluid flow pattern and acoustic streaming under ultrasound irradiation using Particle Image Velocimetry (PIV). In mathematical modelling, a lone micro bubble generated under power ultrasound irradiation was mechanistically analysed. Its characteristics were illustrated as a function of bubble radius, internal temperature and pressure (hot spot conditions) and oscillation (pulsation) velocity. The results showed that ultrasound power significantly affected the conditions of hotspots and bubbles oscillation velocity. From the CFD results, it was observed that the total volume of the micro-bubbles increased by about 4.95% with each 100 W-increase in power amplitude. Furthermore, velocity of acoustic streaming increased from 29 to 119 cm/s as power increased, which was in good agreement with the PIV analysis. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Nanoparticles Formed by Acoustic Destruction of Microbubbles and Their Utilization for Imaging and Effects on Therapy by High Intensity Focused Ultrasound.

    Science.gov (United States)

    Blum, Nicholas T; Yildirim, Adem; Chattaraj, Rajarshi; Goodwin, Andrew P

    2017-01-01

    This work reports that when PEG-lipid-shelled microbubbles with fluorocarbon interior (C 4 F 10 , C 5 F 12 , or C 6 F 14 ) are subjected to ultrasound pulses, they produce metastable, fluid-filled nanoparticles that can be re-imaged upon administration of HIFU. The nanoparticles produced by destruction of the microbubbles (MBNPs) are of 150 nm average diameter and can be re-imaged for up to an hour after creation for C 4 F 10 , and for at least one day for C 5 F 12 . The active species were found to be fluid (gas or liquid) filled nanoparticles rather than lipid debris. The acoustic droplet vaporization threshold of the nanoparticles was found to vary with the vapor pressure of the encapsulated fluorocarbon, and integrated image brightness was found to increase dramatically when the temperature was raised above the normal boiling point of the fluorocarbon. Finally, the vaporization threshold decreases in serum as compared to buffer, and administration of HIFU to the nanoparticles caused breast cancer cells to completely detach from their culture substrate. This work demonstrates a new functionality of microbubbles that could serve as a platform technology for ultrasound-based theranostics.

  8. Expression of annexin-A1 and galectin-1 anti-inflammatory proteins and mRNA in chronic gastritis and gastric cancer.

    Science.gov (United States)

    Jorge, Yvana Cristina; Mataruco, Mayra Mioto; Araújo, Leandro Pires; Rossi, Ana Flávia Teixeira; de Oliveira, Juliana Garcia; Valsechi, Marina Curado; Caetano, Alaor; Miyazaki, Kenji; Fazzio, Célia Sebastiana de Jesus; Thomé, Jorge Alberto; Rahal, Paula; Oliani, Sonia Maria; Silva, Ana Elizabete

    2013-01-01

    The anti-inflammatory proteins annexin-A1 and galectin-1 have been associated with tumor progression. This scenario prompted us to investigate the relationship between the gene and protein expression of annexin-A1 (ANXA1/AnxA1) and galectin-1 (LGALS1/Gal-1) in an inflammatory gastric lesion as chronic gastritis (CG) and gastric adenocarcinoma (GA) and its association with H. pylori infection. We analyzed 40 samples of CG, 20 of GA, and 10 of normal mucosa (C) by the quantitative real-time PCR (qPCR) technique and the immunohistochemistry assay. High ANXA1 mRNA expression levels were observed in 90% (36/40) of CG cases (mean relative quantification RQ = 4.26  ±  2.03) and in 80% (16/20) of GA cases (mean RQ = 4.38  ±  4.77). However, LGALS1 mRNA levels were high (mean RQ = 2.44  ±  3.26) in 60% (12/20) of the GA cases, while low expression was found in CG (mean RQ = 0.43 ± 3.13; P gastritis and gastric cancer, suggesting a strong association of these proteins with chronic gastric inflammation and carcinogenesis.

  9. Proteomic Identification of Annexins, Calcium-Dependent Membrane Binding Proteins That Mediate Osmotic Stress and Abscisic Acid Signal Transduction in Arabidopsis

    Science.gov (United States)

    Lee, Sumin; Lee, Eun Jung; Yang, Eun Ju; Lee, Ji Eun; Park, Ae Ran; Song, Won Hyun; Park, Ohkmae K.

    2004-01-01

    Comparative proteomic analysis of the Arabidopsis thaliana root microsomal fraction was performed to identify novel components of salt stress signaling. Among the salt-responsive microsomal proteins, two spots that increased upon salt treatment on a two-dimensional gel were identified as the same protein, designated annexin 1 (AnnAt1). Annexins comprise a multigene family of Ca2+-dependent membrane binding proteins and have been extensively studied in animal cells. AnnAt1 is strongly expressed in root but rarely in flower tissue. In this study, the results suggest that salt stress induces translocation from the cytosol to the membrane and potential turnover of existing protein. This process is blocked by EGTA treatment, implying that AnnAt1 functions in stress response are tightly associated with Ca2+. T-DNA insertion mutants of annAt1 and a different isoform, annAt4, displayed hypersensitivity to osmotic stress and abscisic acid (ABA) during germination and early seedling growth. The results collectively suggest that AnnAt1 and AnnAt4 play important roles in osmotic stress and ABA signaling in a Ca2+-dependent manner. PMID:15161963

  10. Design, synthesis and SAR exploration of tri-substituted 1,2,4-triazoles as inhibitors of the annexin A2–S100A10 protein interaction

    Science.gov (United States)

    Reddy, Tummala R.K.; Li, Chan; Guo, Xiaoxia; Fischer, Peter M.; Dekker, Lodewijk V.

    2014-01-01

    Recent target validation studies have shown that inhibition of the protein interaction between annexin A2 and the S100A10 protein may have potential therapeutic benefits in cancer. Virtual screening identified certain 3,4,5-trisubstituted 4H-1,2,4-triazoles as moderately potent inhibitors of this interaction. A series of analogues were synthesized based on the 1,2,4-triazole scaffold and were evaluated for inhibition of the annexin A2–S100A10 protein interaction in competitive binding assays. 2-[(5-{[(4,6-Dimethylpyrimidin-2-yl)sulfanyl]methyl}-4-(furan-2-ylmethyl)-4H-1,2,4-triazol-3-yl)sulfanyl]-N-[4-(propan-2-yl)phenyl]acetamide (36) showed improved potency and was shown to disrupt the native complex between annexin A2 and S100A10. PMID:25172147

  11. Interaction between S100A8/A9 and annexin A6 is involved in the calcium-induced cell surface exposition of S100A8/A9.

    Science.gov (United States)

    Bode, Günther; Lüken, Aloys; Kerkhoff, Claus; Roth, Johannes; Ludwig, Stephan; Nacken, Wolfgang

    2008-11-14

    The calcium binding S100A8/A9 complex (MRP8/14; calgranulin) is considered as an important proinflammatory mediator in acute and chronic inflammation and has recently gained attention as a molecular marker up-regulated in various human cancers. Here, we report that S100A8/A9 is expressed in breast cancer cell lines and is up-regulated by interleukin-1beta and tumor necrosis factor-alpha in SKBR3 and MCF-7 cells. We identified the phospholipid-binding protein annexin A6 as a potential S100A8/A9 binding protein by affinity chromatography. This finding was verified by Southwestern overlay experiments and by coimmunoprecipitation with the S100A8/A9-specific monoclonal antibody 27E10. Immunocytochemical experiments demonstrated that S100A8/A9 and annexin A6 colocalize in SKBR3 breast cancer cells predominantly in membranous structures. Upon calcium influx both S100A8/A9 and annexin A6 are exposed on the cell surface of SKBR3 cells. Subcellular fractionation studies suggested that after A23187 stimulation membrane association of S100A8/A9 is not enhanced. However, both S100A8/A9 and annexin A6 are exposed on the cell surface of SKBR3 cells upon calcium influx. Experiments with artificial liposomes indicated that S100A8/A9 is able to associate with membranes independently of both annexin A6 and independently of calcium. Finally, cell surface expression of S100A8/A9 could not be observed in A23187-treated A431 and HaCaT cells. Both cell lines are known to be devoid of annexin A6. Repression of annexin A6 expression by small interfering RNA in SKBR3 cells abolishes the cell surface exposition of S100A8/A9 upon calcium influx, suggesting that annexin A6 contributes to the calcium-dependent cell surface exposition of the membrane associated-S100A8/A9 complex.

  12. Thrombolytic therapy with rt-PA and transcranial color Doppler ultrasound (TCCS) combined with microbubbles for embolic thrombus.

    Science.gov (United States)

    Ren, Xinping; Wang, Yong; Wang, Yi; Chen, Hong; Chen, Li; Liu, Yi; Xue, Chengshi

    2015-11-01

    To investigate the efficacy of transcranial color Doppler ultrasound (TCCS) combined with microbubbles (MBs) and rt-PA (recombinant tissue plasminogen activator) for thrombolysis in vivo. An arterial embolization model was established in Sprague-Dawley rats by retrograde placement of autologous thrombi into the common carotid artery followed by ligation of external carotid artery and temporary occlusion of the carotid artery and internal carotid artery by clamping. The animals were divided into 5 groups with 6 rats in each group: ① untreated control group, ② rt-PA group, ③ TCCS+MBs group, ④ TCCS+MBs+rt-PA group, ⑤ TCCS+MBs+1/2 rt-PA+group. Thrombolytic effects including recanalization rate and recanalization grade were evaluated with grayscale and color Doppler ultrasound post treatment. Examination of histological change in the involved vessel was performed at the end of the experiments. Thrombolysis in terms of recanalization rate and recanalization grade was observed in the treated groups in a time-dependent manner, except the TCCS+MBs group. Addition of rt-PA remarkably enhanced the therapeutic lysis in TCCS+MBs group. However, no difference of therapeutic effects existed between TCCS+MBs+rt-PA group and TCCS+MBs+1/2 rt-PA group. TCCS+MBs combined with rt-PA is a relatively effective approach for ischemic arterial thrombosis with an additive or synergistic effect. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Borneol Attenuates Ultrasound-Targeted Microbubble Destruction-Induced Blood–Brain Barrier Opening in Focal Cerebral Ischemia

    Directory of Open Access Journals (Sweden)

    Xiao-guang Zhang

    2017-12-01

    Full Text Available Ultrasound-targeted microbubble destruction (UTMD and the herb medicine borneol can both facilitate the delivery of therapeutic agents to diseased brain regions and serve as promising adjuvant neuroprotective therapies. Our preliminary experiments showed that UTMD could exacerbate ischemic blood–brain barrier (BBB opening, while borneol can protect the BBB. In this study, we tested the hypothesis that the combination of UTMD and borneol could attenuate UTMD-induced injury to the BBB under ischemic stroke conditions. Male albino mice were subjected to 60-min middle cerebral artery occlusion (MCAO with reperfusion. Borneol and UTMD was given to mice 3 days before and 24 h after MCAO induction. BBB permeability, brain water contents, ultrastructural changes of the BBB and histopathological alterations were evaluated. Our data demonstrated that UTMD aggravated the leakage of Evans blue dye, ultrastructural alterations of cerebral microvasculature, brain edema, and even induced cerebral hemorrhage in ischemic stroke mice. Pretreatment with borneol significantly attenuated the above detrimental effects of UTMD on the BBB. This study indicates that under ischemic stroke conditions, the BBB becomes vulnerable to UTMD intervention, and the combination of borneol can help to maintain the integrity of the BBB.

  14. Ultrasound Improves the Delivery and Therapeutic Effect of Nanoparticle-Stabilized Microbubbles in Breast Cancer Xenografts.

    Science.gov (United States)

    Snipstad, Sofie; Berg, Sigrid; Mørch, Ýrr; Bjørkøy, Astrid; Sulheim, Einar; Hansen, Rune; Grimstad, Ingeborg; van Wamel, Annemieke; Maaland, Astri F; Torp, Sverre H; Davies, Catharina de Lange

    2017-11-01

    Compared with conventional chemotherapy, encapsulation of drugs in nanoparticles can improve efficacy and reduce toxicity. However, delivery of nanoparticles is often insufficient and heterogeneous because of various biological barriers and uneven tumor perfusion. We investigated a unique multifunctional drug delivery system consisting of microbubbles stabilized by polymeric nanoparticles (NPMBs), enabling ultrasound-mediated drug delivery. The aim was to examine mechanisms of ultrasound-mediated delivery and to determine if increased tumor uptake had a therapeutic benefit. Cellular uptake and toxicity, circulation and biodistribution were characterized. After intravenous injection of NPMBs into mice, tumors were treated with ultrasound of various pressures and pulse lengths, and distribution of nanoparticles was imaged on tumor sections. No effects of low pressures were observed, whereas complete bubble destruction at higher pressures improved tumor uptake 2.3 times, without tissue damage. An enhanced therapeutic effect was illustrated in a promising proof-of-concept study, in which all tumors exhibited regression into complete remission. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  15. Bleomycin delivery into cancer cells in vitro with ultrasound and SonoVue® or BR14® microbubbles.

    Science.gov (United States)

    Lamanauskas, N; Novell, A; Escoffre, J-M; Venslauskas, M; Satkauskas, S; Bouakaz, A

    2013-05-01

    Cell exposure to ultrasound (US) in the presence of contrast agent microbubbles (MBs) can result in cell sonoporation that can be exploited for drug or gene delivery. Anticancer drug bleomycin (BLM), used in sonoporation, can effectively eliminate tumor cells in vitro and in vivo. Nevertheless, sonoporation mechanism is not known, thus different US parameters and MB types are used. Recently, we proposed that efficiency of cell sonoporation can be related to the efficiency of MB sonodestruction. We analyzed human tumor cells viability in response to BLM, US and MB treatment. Human glioblastoma astrocytoma (U-87 MG) or colon cancer (HCT-116) cells were exposed to US in the presence of BLM and either SonoVue® or BR14® MBs. MB sonodestruction was evaluated according to US signal attenuation. Both HCT-116 and U-87 MG cell viability following US exposure decreased up to 30%. Decrease in cell viability followed similar tendency as MB sonodestruction, which suggests direct relationship between MB sonodestruction and BLM intracellular delivery. Sonoporation is a feasible method to deliver BLM in to several types of human cancer cell lines. Efficiency of cell sonoporation correlated well with MB sonodestruction, providing a possibility to optimize US parameters by measuring MB sonodestruction.

  16. Intensity-Based Assessment of Microbubble-Enhanced Ultrasonography: Phase-Related Diagnostic Ability for Cellular Differentiation of Hepatocellular Carcinoma.

    Science.gov (United States)

    Kondo, Takayuki; Maruyama, Hitoshi; Kiyono, Soichiro; Sekimoto, Tadashi; Shimada, Taro; Takahashi, Masanori; Ogasawara, Sadahisa; Suzuki, Eiichiro; Ooka, Yoshihiko; Tawada, Akinobu; Chiba, Tetsuhiro; Kanai, Fumihiko; Yokosuka, Osamu

    2015-12-01

    This prospective study aimed to elucidate the effect of phase-related quantitative parameters of contrast-enhanced ultrasound (CEUS) with perflubutane microbubble agent to assess the cellular differentiation of hepatocellular carcinoma (HCC). Intensity was analyzed in 94 lesions (19.4 ± 4.9 mm, 86 patients), 47 well-differentiated HCCs (wHCCs) and 47 moderately-differentiated HCCs (mHCCs): I(e) (early phase) = I(te) (tumor) - I(le) (liver), I(p) (post-vascular phase) = I(tp) (tumor) - I(lp) (liver), I(ep) = I(e) - I(p). The area under the receiver operating characteristic curve with the best cutoff value (I(e), 13.2, I(p), -4.5, I(ep), 21.3) for discriminating between wHCC and mHCC was 0.6922 for Ie, 0.7680 for Ip and 0.7925 for Iep, which indicated a significantly greater ability to differentiate between wHCC and mHCC compared with visual/qualitative assessment (early phase, 0.6170, p = 0.04; post-vascular phase, 0.6702, p = 0.01; both phases, 0.7021, p = 0.04). In conclusion, I(ep) was found to have the highest diagnostic ability, suggesting it is a promising parameter for the cellular differentiation of HCCs with CEUS. Copyright © 2015 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  17. Microbubbles combined with ultrasound therapy in ischemic stroke: A systematic review of in-vivo preclinical studies.

    Directory of Open Access Journals (Sweden)

    Laurent Auboire

    Full Text Available Microbubbles (MBs combined with ultrasound sonothrombolysis (STL appears to be an alternative therapeutic strategy for acute ischemic stroke (IS, but clinical results remain controversial.The aim of this systematic review is to identify the parameters tested; to assess evidence on the safety and efficacy on preclinical data on STL; and to assess the validity and publication bias.Pubmed® and Web of ScienceTM databases were systematically searched from January 1995 to April 2017 in French and English. We included studies evaluating STL on animal stroke model. This systematic review was conducted in accordance with the PRISMA guidelines. Data were extracted following a pre-defined schedule by two of the authors. The CAMARADES criteria were used for quality assessment. A narrative synthesis was conducted.Sixteen studies met the inclusion criteria. The result showed that ultrasound parameters and types of MBs were heterogeneous among studies. Numerous positive outcomes on efficacy were found, but only four studies demonstrated superiority of STL versus recombinant tissue-type plasminogen activator on clinical criteria. Data available on safety are limited.Quality assessment of the studies reviewed revealed a number of biases.Further in vivo studies are needed to demonstrate a better efficacy and safety of STL compared to currently approved therapeutic options.http://syrf.org.uk/protocols/.

  18. Hypoxia due to shunts in pig lung treated with O2 and fluorocarbon-derived intravascular microbubbles.

    Science.gov (United States)

    Tyssebotn, Ingvald M; Lundgren, Claes E G; Olszowka, Albert J; Bergoe, Guri W

    2010-04-01

    Earlier work has shown that experimental conditions calling for improved tissue oxygenation could be assisted by i.v. infusion of a dodecafluoropentane emulsion (DDFPe) forming oxygen-transporting microbubbles. The present work investigated the effect of DDFPe on hypoxia due to experimental shunts in the pig lung. Nineteen O(2) breathing, anesthetized pigs had glass beads administered into the trachea so as to significantly depress arterial oxygen tension (PaO(2)). PaO(2) was recorded for up to 12 hrs while 0.1 ml/kg DDFPe was administered 1-3 times. The animals were divided into two groups based on arterial oxygen saturation (SaO(2)) after shunt induction, combined with oxygen breathing: the "SaO(2) >90% group" (n=6) and the "SaO(2) e.g. (1(st) infusion) from a PvO(2) of 41.4+/-2.3 to 49.9+/-4.2 mmHg (Pbreathing may be beneficial in severe right-to-left shunting in humans.

  19. Measurement of sliding velocity and induction time of a single micro-bubble under an inclined collector surface

    Energy Technology Data Exchange (ETDEWEB)

    Najafi, A.S.; Xu, Z.; Masliyah, J. [Alberta Univ., Edmonton, AB (Canada). Dept. of Chemical and Materials Engineering

    2008-12-15

    Flotation is a major process by which bitumens are recovered from oil sands slurries, and air bubble-bitumen attachment is necessary for the effective separation of aerated bitumen. This study investigated the interactions between a gas bubble and a solid, flat surface. Various physical and chemical conditions were studied in order to determine their impact on the sliding velocity and induction times of the micro-bubble as it slid underneath an inclined solid surface. The bubble's trajectory was monitored by a high-speed video imaging system. Tests were conducted using de-aerated process water in order to study the effect of dissolved gases on bubble sliding velocity. Simulated process water was also used to study the role of the natural surfactants contained in recycle process water. The study showed that terminal and sliding velocities of the bubbles were functions of temperature. Sliding velocities increased with increases in liquid temperature. Increases in liquid temperature also reduced the induction time used to quantify bubble-solid attachment. Induction times were also reduced with increases in surface hydrophobicity. Induction times measured for CO{sub 2} bubbles were shorter than those observed with air, oxygen, and hydrogen bubbles. It was concluded that the use of natural surface active agents in process water reduced bubble terminal rising velocity and increased the induction times of bubble-solid attachment. 36 refs., 2 tabs., 15 figs.

  20. Synthesis and evaluation of a {sup 18}F-labelled recombinant annexin-V derivative, for identification and quantification of apoptotic cells with PET

    Energy Technology Data Exchange (ETDEWEB)

    Zijlstra, S. E-mail: szijlstra@hdz-nrw.de; Gunawan, J.; Burchert, W

    2003-02-01

    In this report, we describe the synthesis of 4-[{sup 18}F]-fluorobenzoyl-annexin V (4-[{sup 18}F]FBA). In a four-step procedure, 4-[{sup 18}F]FBA was synthesised with a microcomputer controlled, automated module within 90 min. The radiochemical yield was in the range of 15-20% (corrected for decay) with a specific activity of more than 35 GBq/{mu}mol. The specific binding was confirmed by studies of 4-[{sup 18}F]FBA with phosphatidylserine-containing liposomes. The biological activity of 4-[{sup 18}F]FBA was verified by measuring its binding to Jurkat T-cell lymphoblasts after induction of apoptosis as compared to control cells without apoptosis. 4-[{sup 18}F]FBA seems to be a suited tracer to measure apoptotic activity in vivo.

  1. Role of lipoxygenases and the lipoxin A(4)/annexin 1 receptor in ischemia-reperfusion-induced gastric mucosal damage in rats.

    Science.gov (United States)

    Peskar, Brigitta M; Ehrlich, Karlheinz; Schuligoi, Rufina; Peskar, Bernhard A

    2009-01-01

    Rat gastric mucosal damage was induced by ischemia-reperfusion. The 5-lipoxygenase inhibitors MK886 and A63162, the 12-lipoxygenase inhibitor baicalein, the 15-lipoxygenase inhibitor PD146176 and the lipoxin (LX) A(4)/annexin 1 antagonist Boc1 increased mucosal damage in a dose-dependent manner. Low doses of these compounds, which have no effects on mucosal integrity, cause severe damage when combined with low doses of indomethacin, celecoxib or dexamethasone. 16,16-Dimethylprostaglandin (PG) E(2) and LXA(4) can replace each other in preventing mucosal injury induced by either cyclooxygenase or lipoxygenase inhibitors. The results suggest that not only cyclooxygenases, but also lipoxygenases have a role in limiting gastric mucosal damage during ischemia-reperfusion. Copyright 2009 S. Karger AG, Basel.

  2. Precision Medicine in Assisted Conception: A Multicenter Observational Treatment Cohort Study of the Annexin A5 M2 Haplotype as a Biomarker for Antithrombotic Treatment to Improve Pregnancy Outcome

    Directory of Open Access Journals (Sweden)

    Simon Fishel

    2016-08-01

    Interpretation: Pragmatic ANXA5 M5 screening and treatment with LMWH in couples undergoing IVF is associated with similar outcome to couples with more favorable prognostic factors. The difference in live birth outcome for treated male only carrier couples may be consistent with an additional maternal thrombophilic factor that may adversely affect pregnancy, although other mechanisms are possible. This study suggests that LMWH treatment should be started prior to clinical pregnancy.

  3. Ultrasound-guided delivery of siRNA and a chemotherapeutic drug by using microbubble complexes: In vitro and in vivo evaluations in a prostate cancer model

    International Nuclear Information System (INIS)

    Bae, Yun Jung; Yoon, Young Il; Lee, Hak Jong; Yoon, Tae Jong

    2016-01-01

    To evaluate the effectiveness of ultrasound and microbubble-liposome complex (MLC)-mediated delivery of siRNA and doxorubicin into prostate cancer cells and its therapeutic capabilities both in vitro and in vivo. Microbubble-liposome complexes conjugated with anti-human epidermal growth factor receptor type 2 (Her2) antibodies were developed to target human prostate cancer cell lines PC-3 and LNCaP. Intracellular delivery of MLC was observed by confocal microscopy. We loaded MLC with survivin-targeted small interfering RNA (siRNA) and doxorubicin, and delivered it into prostate cancer cells. The release of these agents was facilitated by ultrasound application. Cell viability was analyzed by MTT assay after the delivery of siRNA and doxorubicin. Survivin-targeted siRNA loaded MLC was delivered into the xenograft mouse tumor model. Western blotting was performed to quantify the expression of survivin in vivo. Confocal microscopy demonstrated substantial intracellular uptake of MLCs in LNCaP, which expresses higher levels of Her2 than PC-3. The viability of LNCaP cells was significantly reduced after the delivery of MLCs loaded with siRNA and doxorubicin (85.0 ± 2.9%), which was further potentiated by application of ultrasound (55.0 ± 3.5%, p = 0.009). Survivin expression was suppressed in vivo in LNCaP tumor xenograft model following the ultrasound and MLC-guided delivery of siRNA (77.4 ± 4.90% to 36.7 ± 1.34%, p = 0.027). Microbubble-liposome complex can effectively target prostate cancer cells, enabling intracellular delivery of the treatment agents with the use of ultrasound. Ultrasound and MLC-mediated delivery of survivin-targeted siRNA and doxorubicin can induce prostate cell apoptosis and block survivin expression in vitro and in vivo

  4. Ultrasound-Guided Delivery of siRNA and a Chemotherapeutic Drug by Using Microbubble Complexes: In Vitro and In Vivo Evaluations in a Prostate Cancer Model

    Energy Technology Data Exchange (ETDEWEB)

    Bae, Yun Jung [Department of Radiology, Seoul National University Bundang Hospital, Seongnam 13620 (Korea, Republic of); Department of Radiology, Seoul National University College of Medicine, Seoul 03080 (Korea, Republic of); Yoon, Young Il [Department of Radiology, Seoul National University Bundang Hospital, Seongnam 13620 (Korea, Republic of); Department of Radiology, Seoul National University College of Medicine, Seoul 03080 (Korea, Republic of); Program in Nano Science and Technology, Department of Transdisciplinary Studies, Seoul National University Graduate School of Convergence Science and Technology, Suwon 16229 (Korea, Republic of); Yoon, Tae-Jong [Department of Applied Bioscience, College of Life Science, CHA University, Pocheon 11160 (Korea, Republic of); College of Pharmacy, Ajou University, Suwon 16499 (Korea, Republic of); Lee, Hak Jong [Department of Radiology, Seoul National University Bundang Hospital, Seongnam 13620 (Korea, Republic of); Department of Radiology, Seoul National University College of Medicine, Seoul 03080 (Korea, Republic of); Program in Nano Science and Technology, Department of Transdisciplinary Studies, Seoul National University Graduate School of Convergence Science and Technology, Suwon 16229 (Korea, Republic of)

    2016-11-01

    To evaluate the effectiveness of ultrasound and microbubble-liposome complex (MLC)-mediated delivery of siRNA and doxorubicin into prostate cancer cells and its therapeutic capabilities both in vitro and in vivo. Microbubble-liposome complexes conjugated with anti-human epidermal growth factor receptor type 2 (Her2) antibodies were developed to target human prostate cancer cell lines PC-3 and LNCaP. Intracellular delivery of MLC was observed by confocal microscopy. We loaded MLC with survivin-targeted small interfering RNA (siRNA) and doxorubicin, and delivered it into prostate cancer cells. The release of these agents was facilitated by ultrasound application. Cell viability was analyzed by MTT assay after the delivery of siRNA and doxorubicin. Survivin-targeted siRNA loaded MLC was delivered into the xenograft mouse tumor model. Western blotting was performed to quantify the expression of survivin in vivo. Confocal microscopy demonstrated substantial intracellular uptake of MLCs in LNCaP, which expresses higher levels of Her2 than PC-3. The viability of LNCaP cells was significantly reduced after the delivery of MLCs loaded with siRNA and doxorubicin (85.0 ± 2.9%), which was further potentiated by application of ultrasound (55.0 ± 3.5%, p = 0.009). Survivin expression was suppressed in vivo in LNCaP tumor xenograft model following the ultrasound and MLC-guided delivery of siRNA (77.4 ± 4.90% to 36.7 ± 1.34%, p = 0.027). Microbubble-liposome complex can effectively target prostate cancer cells, enabling intracellular delivery of the treatment agents with the use of ultrasound. Ultrasound and MLC-mediated delivery of survivin-targeted siRNA and doxorubicin can induce prostate cell apoptosis and block survivin expression in vitro and in vivo.

  5. Ultrasound-Guided Delivery of siRNA and a Chemotherapeutic Drug by Using Microbubble Complexes: In Vitro and In Vivo Evaluations in a Prostate Cancer Model

    International Nuclear Information System (INIS)

    Bae, Yun Jung; Yoon, Young Il; Yoon, Tae-Jong; Lee, Hak Jong

    2016-01-01

    To evaluate the effectiveness of ultrasound and microbubble-liposome complex (MLC)-mediated delivery of siRNA and doxorubicin into prostate cancer cells and its therapeutic capabilities both in vitro and in vivo. Microbubble-liposome complexes conjugated with anti-human epidermal growth factor receptor type 2 (Her2) antibodies were developed to target human prostate cancer cell lines PC-3 and LNCaP. Intracellular delivery of MLC was observed by confocal microscopy. We loaded MLC with survivin-targeted small interfering RNA (siRNA) and doxorubicin, and delivered it into prostate cancer cells. The release of these agents was facilitated by ultrasound application. Cell viability was analyzed by MTT assay after the delivery of siRNA and doxorubicin. Survivin-targeted siRNA loaded MLC was delivered into the xenograft mouse tumor model. Western blotting was performed to quantify the expression of survivin in vivo. Confocal microscopy demonstrated substantial intracellular uptake of MLCs in LNCaP, which expresses higher levels of Her2 than PC-3. The viability of LNCaP cells was significantly reduced after the delivery of MLCs loaded with siRNA and doxorubicin (85.0 ± 2.9%), which was further potentiated by application of ultrasound (55.0 ± 3.5%, p = 0.009). Survivin expression was suppressed in vivo in LNCaP tumor xenograft model following the ultrasound and MLC-guided delivery of siRNA (77.4 ± 4.90% to 36.7 ± 1.34%, p = 0.027). Microbubble-liposome complex can effectively target prostate cancer cells, enabling intracellular delivery of the treatment agents with the use of ultrasound. Ultrasound and MLC-mediated delivery of survivin-targeted siRNA and doxorubicin can induce prostate cell apoptosis and block survivin expression in vitro and in vivo

  6. Ultrasound-guided delivery of siRNA and a chemotherapeutic drug by using microbubble complexes: In vitro and in vivo evaluations in a prostate cancer model

    Energy Technology Data Exchange (ETDEWEB)

    Bae, Yun Jung; Yoon, Young Il; Lee, Hak Jong [Dept. of Radiology, Seoul National University Bundang Hospital, Seongnam (Korea, Republic of); Yoon, Tae Jong [Dept. of Applied Bioscience, College of Life Science, CHA University, Pocheon (Korea, Republic of)

    2016-07-15

    To evaluate the effectiveness of ultrasound and microbubble-liposome complex (MLC)-mediated delivery of siRNA and doxorubicin into prostate cancer cells and its therapeutic capabilities both in vitro and in vivo. Microbubble-liposome complexes conjugated with anti-human epidermal growth factor receptor type 2 (Her2) antibodies were developed to target human prostate cancer cell lines PC-3 and LNCaP. Intracellular delivery of MLC was observed by confocal microscopy. We loaded MLC with survivin-targeted small interfering RNA (siRNA) and doxorubicin, and delivered it into prostate cancer cells. The release of these agents was facilitated by ultrasound application. Cell viability was analyzed by MTT assay after the delivery of siRNA and doxorubicin. Survivin-targeted siRNA loaded MLC was delivered into the xenograft mouse tumor model. Western blotting was performed to quantify the expression of survivin in vivo. Confocal microscopy demonstrated substantial intracellular uptake of MLCs in LNCaP, which expresses higher levels of Her2 than PC-3. The viability of LNCaP cells was significantly reduced after the delivery of MLCs loaded with siRNA and doxorubicin (85.0 ± 2.9%), which was further potentiated by application of ultrasound (55.0 ± 3.5%, p = 0.009). Survivin expression was suppressed in vivo in LNCaP tumor xenograft model following the ultrasound and MLC-guided delivery of siRNA (77.4 ± 4.90% to 36.7 ± 1.34%, p = 0.027). Microbubble-liposome complex can effectively target prostate cancer cells, enabling intracellular delivery of the treatment agents with the use of ultrasound. Ultrasound and MLC-mediated delivery of survivin-targeted siRNA and doxorubicin can induce prostate cell apoptosis and block survivin expression in vitro and in vivo.

  7. Ultrasound-targeted stromal cell-derived factor-1-loaded microbubble destruction promotes mesenchymal stem cell homing to kidneys in diabetic nephropathy rats

    Directory of Open Access Journals (Sweden)

    Wu S

    2014-12-01

    Full Text Available Shengzheng Wu,1 Lu Li,1 Gong Wang,1 Weiwei Shen,2 Yali Xu,1 Zheng Liu,1 Zhongxiong Zhuo,1 Hongmei Xia,1 Yunhua Gao,1 Kaibin Tan1 1Department of Ultrasound, 2Department of Orthopedics, Xinqiao Hospital, Third Military Medical University, Chongqing, People’s Republic of China Abstract: Mesenchymal stem cell (MSC therapy has been considered a promising strategy to cure diabetic nephropathy (DN. However, insufficient MSCs can settle in injured kidneys, which constitute one of the major barriers to the effective implementation of MSC therapy. Stromal cell-derived factor-1 (SDF-1 plays a vital role in MSC migration and involves activation, mobilization, homing, and retention, which are presumably related to the poor homing in DN therapy. Ultrasound-targeted microbubble destruction has become one of the most promising strategies for the targeted delivery of drugs and genes. To improve MSC homing to DN kidneys, we present a strategy to increase SDF-1 via ultrasound-targeted microbubble destruction. In this study, we developed SDF-1-loaded microbubbles (MBSDF-1 via covalent conjugation. The characterization and bioactivity of MBSDF-1 were assessed in vitro. Target release in the targeted kidneys was triggered with diagnostic ultrasound in combination with MBSDF-1. The related bioeffects were also elucidated. Early DN was induced in rats with streptozotocin. Green fluorescent protein-labeled MSCs were transplanted intravenously following the target release of SDF-1 in the kidneys of normal and DN rats. The homing efficacy was assessed by detecting the implanted exogenous MSCs at 24 hours. The in vitro results showed an impressive SDF-1 loading efficacy of 79% and a loading content of 15.8 µg/mL. MBSDF-1 remained bioactive as a chemoattractant. In the in vivo study, SDF-1 was successfully released in the targeted kidneys. The homing efficacy of MSCs to DN kidneys after the target release of SDF-1 was remarkably ameliorated at 24 hours compared with

  8. Preparation of Metallochelating Microbubbles and Study on Their Site-Specific Interaction with rGFP-HisTag as a Model Protein

    Czech Academy of Sciences Publication Activity Database

    Lukáč, R.; Kauerová, Z.; Mašek, J.; Bartheldyová, E.; Kulich, P.; Koudelka, Š.; Korvasová, Z.; Plocková, J.; Papoušek, František; Kolář, František; Schmidt, R.; Turánek, J.

    2011-01-01

    Roč. 27, č. 8 (2011), s. 4829-4837 ISSN 0743-7463 R&D Projects: GA AV ČR KAN200520703 Grant - others:GA ČR(CZ) GAP304/10/1951; GA AV ČR(CZ) KAN200100801 Program:GA; KA Institutional research plan: CEZ:AV0Z50110509 Keywords : microbubble * ultrasound imaging * metallochelating bond * rGFP * liposome * contrast echocardiography * static light scattering * flow * cytometry * confocal Subject RIV: FA - Cardiovascular Diseases incl. Cardiotharic Surgery Impact factor: 4.186, year: 2011

  9. Transient disruption of vascular barriers using focused ultrasound and microbubbles for targeted drug delivery in the brain

    Science.gov (United States)

    Aryal, Muna

    The physiology of the vasculature in the central nervous system (CNS) which includes the blood-brain-barrier (BBB) and other factors, prevents the transport of most anticancer agents to the brain and restricts delivery to infiltrating brain tumors. The heterogeneous vascular permeability in tumor vessels (blood-tumor barrier; BTB), along with several other factors, creates additional hurdles for drug treatment of brain tumors. Different methods have been used to bypass the BBB/BTB, but they have their own limitations such as being invasive, non-targeted or requiring the formulation of new drugs. Magnetic Resonance Imaging guided Focused Ultrasound (MRIgFUS), when combined with circulating microbubbles, is an emerging noninvasive method to temporarily permeabilize the BBB and BTB. The purpose of this thesis was to use this alternative approach to deliver chemotherapeutic agents through the BBB/BTB for brain tumor treatment in a rodent model to overcome the hinderances encountered in prior approaches tested for drug delivery in the CNS. The results presented in thesis demonstrate that MRIgFUS can be used to achieve consistent and reproducible BBB/BTB disruption in rats. It enabled us to achieve clinically-relevant concentrations of doxorubicin (~ 4.8+/-0.5 microg/g) delivered to the brain with the sonication parameters (0.69 MHz; 0.55 MPa; 10 ms bursts; 1 Hz PRF; 60 s duration), microbubble concentration (Definity, 10 microl/kg), and liposomoal doxorubicin (Lipo-DOX) dose (5.67 mg/kg) used. The resulting doxorubicin concentration was reduced by 32% when the agent was injected 10 minute after the last sonication. Three weekly sessions of FUS and Lipo-DOX appeared to be safe in the rat brain, despite some minor tissue damage. Importantly, the severe neurotoxicity seen in earlier works using other approaches does not appear to occur with delivery via FUS-BBB disruption. The resuls from three weekly treatments of FUS and Lipo-DOX in a rat glioma model are highly

  10. Effectiveness of a Layer-by-Layer Microbubbles-Based Delivery System for Applying Minoxidil to Enhance Hair Growth.

    Science.gov (United States)

    Liao, Ai-Ho; Lu, Ying-Jui; Lin, Yi-Chun; Chen, Hang-Kang; Sytwu, Huey-Kang; Wang, Chih-Hung

    2016-01-01

    Minoxidil (Mx) is a conventional drug for treating androgenetic alopecia, preventing hair loss, and promoting hair growth. The solubility of Mx has been improved using chemical enhancement methods to increase its skin permeability over the long term. This study created a new ultrasound (US) contrast agent-albumin-shelled microbubbles (MBs) that absorb chitosan oligosaccharide lactate (COL) and Mx-and combined it with sonication by US energy in the water phase to enhance hair growth while shortening the treatment period. COL and Mx grafted with MBs (mean diameter of 1480 nm) were synthesized into self-assembled complexes of COL-MBs and Mx-COL-MBs that had mean diameters of 4150 and 4500 nm, respectively. The US was applied at 3 W/cm(2) for 1 min, and combined with Mx-COL-MBs containing 0.3% Mx. The diffusion of Mx through the dialysis membrane from Mx-COL-MB during US (US+Mx-COL-MB) was more rapid at pH 4 than at pH 7.4, which is favorable given that the environment of the scalp is mildly acidic (pH=4.5-5.5). In Franz diffusion experiments performed in vitro, the release rates at 18 hours in the US+Mx-COL-MBs and US+MBs+Mx groups resulted in 2.3 and 1.7 times the penetration and deposition, respectively, of Mx relative to the group with Mx alone. During 21 days treatment in animal experiments, the growth rates at days 10 and 14 in the US+Mx-COL-MBs group increased by 22.6% and 64.7%, respectively, and there were clear significant differences (phair shafts and the size of hair follicles without causing skin damage.

  11. Effect of acoustic parameters on the cavitation behavior of SonoVue microbubbles induced by pulsed ultrasound.

    Science.gov (United States)

    Lin, Yutong; Lin, Lizhou; Cheng, Mouwen; Jin, Lifang; Du, Lianfang; Han, Tao; Xu, Lin; Yu, Alfred C H; Qin, Peng

    2017-03-01

    SonoVue microbubbles could serve as artificial nuclei for ultrasound-triggered stable and inertial cavitation, resulting in beneficial biological effects for future therapeutic applications. To optimize and control the use of the cavitation of SonoVue bubbles in therapy while ensuring safety, it is important to comprehensively understand the relationship between the acoustic parameters and the cavitation behavior of the SonoVue bubbles. An agarose-gel tissue phantom was fabricated to hold the SonoVue bubble suspension. 1-MHz transmitting transducer calibrated by a hydrophone was used to trigger the cavitation of SonoVue bubbles under different ultrasonic parameters (i.e., peak rarefactional pressure (PRP), pulse repetition frequency (PRF), and pulse duration (PD)). Another 7.5-MHz focused transducer was employed to passively receive acoustic signals from the exposed bubbles. The ultraharmonics and broadband intensities in the acoustic emission spectra were measured to quantify the extent of stable and inertial cavitation of SonoVue bubbles, respectively. We found that the onset of both stable and inertial cavitation exhibited a strong dependence on the PRP and PD and a relatively weak dependence on the PRF. Approximate 0.25MPa PRP with more than 20μs PD was considered to be necessary for ultraharmonics emission of SonoVue bubbles, and obvious broadband signals started to appear when the PRP exceeded 0.40MPa. Moreover, the doses of stable and inertial cavitation varied with the PRP. The stable cavitation dose initially increased with increasing PRP, and then decreased rapidly after 0.5MPa. By contrast, the inertial cavitation dose continuously increased with increasing PRP. Finally, the doses of both stable and inertial cavitation were positively correlated with PRF and PD. These results could provide instructive information for optimizing future therapeutic applications of SonoVue bubbles. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Bioeffects of albumin-encapsulated microbubbles and real-time myocardial contrast echocardiography in an experimental canine model

    Directory of Open Access Journals (Sweden)

    P.M.M. Dourado

    2006-06-01

    Full Text Available Myocardial contrast echocardiography has been used for assessing myocardial perfusion. Some concerns regarding its safety still remain, mainly regarding the induction of microvascular alterations. We sought to determine the bioeffects of microbubbles and real-time myocardial contrast echocardiography (RTMCE in a closed-chest canine model. Eighteen mongrel dogs were randomly assigned to two groups. Nine were submitted to continuous intravenous infusion of perfluorocarbon-exposed sonicated dextrose albumin (PESDA plus continuous imaging using power pulse inversion RTMCE for 180 min, associated with manually deflagrated high-mechanical index impulses. The control group consisted of 3 dogs submitted to continuous imaging using RTMCE without PESDA, 3 dogs received PESDA alone, and 3 dogs were sham-operated. Hemodynamics and cardiac rhythm were monitored continuously. Histological analysis was performed on cardiac and pulmonary tissues. No hemodynamic changes or cardiac arrhythmias were observed in any group. Normal left ventricular ejection fraction and myocardial perfusion were maintained throughout the protocol. Frequency of mild and focal microhemorrhage areas in myocardial and pulmonary tissue was similar in PESDA plus RTMCE and control groups. The percentages of positive microscopical fields in the myocardium were 0.4 and 0.7% (P = NS in the PESDA plus RTMCE and control groups, respectively, and in the lungs they were 2.1 and 1.1%, respectively (P = NS. In this canine model, myocardial perfusion imaging obtained with PESDA and RTMCE was safe, with no alteration in cardiac rhythm or left ventricular function. Mild and focal myocardial and pulmonary microhemorrhages were observed in both groups, and may be attributed to surgical tissue manipulation.

  13. Ultrasound Molecular Imaging of Atherosclerosis for Early Diagnosis and Therapeutic Evaluation through Leucocyte-like Multiple Targeted Microbubbles.

    Science.gov (United States)

    Yan, Fei; Sun, Yu; Mao, Yang; Wu, Meiying; Deng, Zhiting; Li, Shuai; Liu, Xin; Xue, Li; Zheng, Hairong

    2018-01-01

    Cardiovascular diseases resulting from atherosclerosis have become a serious threat to human health. It is well-known that an ongoing inflammatory response is involved during atherosclerosis progression that ultimately results in the accumulation of lipids and formation of plaques. Monitoring the pathological changes during the inflammatory response will be of great significance for early diagnosis and therapeutic evaluation of atherosclerosis. Targeted contrast-enhanced ultrasonography has been shown to be a promising noninvasive imaging technique for evaluating the degree of atherosclerosis and may potentially be translated to clinical imaging in the future. However, inadequate cell adhesion of targeted microbubbles (MBs) in large arterial vessels still remains a great challenge. Methods: By mimicking the leucocytes that are recruited to the vessel wall during the initiation of atherosclerosis through selectin-dependent arrest and cell adhesion molecule-mediated firm cell adhesion, we developed VCAM-1/ICAM-1/P-selectin-targeted MB VIS by integrating VCAM-1 and ICAM-1 antibodies and synthetic polymeric sialyl Lewis X (sLe x ) onto the MB surface. Results: The resulting MB VIS had a high affinity to inflammatory bEnd.3 cells in both static and dynamic flow conditions. Significantly enhanced ultrasound imaging signals were achieved by MB VIS in detecting the atherosclerosis progress when compared with the single- or dual-targeted MBs. Taking advantage of the artificial MB VIS , less ultrasound imaging signals were found in the atorvastatin-treated, but not placebo-treated, ApoE-deficient mice with atherosclerosis, revealing a potential therapeutic efficacy of atorvastatin for early stage atherosclerosis. This was further confirmed by histologic staining examination. Conclusions: Our study provides a promising ultrasound molecular imaging probe for early-stage diagnosis and therapeutic evaluation of atherosclerosis.

  14. Homing of endogenous bone marrow mesenchymal stem cells to rat infarcted myocardium via ultrasound-mediated recombinant SDF-1α adenovirus in microbubbles.

    Science.gov (United States)

    Su, Gaofeng; Liu, Liyun; Yang, Lingjie; Mu, Yuming; Guan, Lina

    2018-01-02

    Stem cells can promote myocardial regeneration and accelerate the formation of new blood vessels. As such, transplanted stem cells represent a promising treatment modality for acute myocardial infarction (AMI). Stem cells spontaneously home to the infarcted myocardium using chemotaxis, in which the stromal cell-derived factor (SDF-1α) has been shown to be one of the most important chemokines. However, spontaneously secreted SDF-1α is short-lived, and therefore does not meet the needs of tissue repair. In this study, adenoviruses carrying SDF-1α genes were loaded on microbubble carriers and the adenoviruses were released into AMI rats by ultrasound targeted microbubble destruction. The possibility of in vivo self-transplantation of bone marrow mesenchymal stem cells (BMSCs) induced by overexpression of SDF-1α in the infarcted myocardium was explored by detecting the number of BMSCs homing from the peripheral blood to the myocardial infarcts. The concentration of SDF-1α in peripheral blood was significantly higher after transfection, and the number of BMSCs was significantly higher in the peripheral blood and infarcted area. Further analyses indicated that the number of homing BMSCs increased with increased SDF-1α expression. In conclusion, our results suggest that ultrasound mediated transduction of exogenous SDF-1α genes into myocardial infarcted AMI rats can effectively promote the homing of endogenous BMSCs into the heart. Moreover, the number of homing stem cells was controlled by the level of SDF-1α expression.

  15. Combined ultrasound-targeted microbubble destruction and polyethylenimine-mediated plasmid DNA delivery to the rat retina: enhanced efficiency and accelerated expression.

    Science.gov (United States)

    Li, Hongli; Qian, Jin; Yao, Chunfang; Wan, Caifeng; Li, Fenghua

    2016-04-01

    Gene therapy has potential in the treatment of refractory retinal diseases. It is important to develop an effective delivery system in the retina. The present study aimed to investigate the efficacy and safety of ultrasound (US)-targeted microbubble destruction (UTMD)-mediated polyethylenimine (PEI) to the rat retina. Gene transfer was examined by injecting PEI/plasmid DNA (pDNA) with or without microbubbles (MBs) into the subretinal space of rats that were then exposed to US. We investigated enhanced green fluorescent protein (eGFP) expression on flat fundus oculi and performed quantitative analysis. Hematoxylin and eosin staining was used to observe tissue damage. UTMD significantly enhanced PEI/pDNA transfection efficiency safely by increasing both the transgene expression per cell and the percentage of transfected cells of the retina. PEI/pDNA combined with UTMD significantly increased the number of DNA gene copies and the mRNA level in the retinal pigment epithelium (RPE) and neural retina, respectively, compared to PEI/pDNA alone. The present study demonstrates that enhanced and accelerated pDNA expression can be achieved in the retina/RPE cells in vivo by UTMD physical techniques combined with a PEI chemical vector. Our study provides useful information for further in vivo retinal gene therapy work. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  16. Synergistic effects of ultrasound-targeted microbubble destruction and TAT peptide on gene transfection: an experimental study in vitro and in vivo.

    Science.gov (United States)

    Zhou, Zhiyi; Zhang, Ping; Ren, Jianli; Ran, Haitao; Zheng, Yuanyi; Li, Pan; Zhang, Qunxia; Zhang, Maohui; Wang, Zhigang

    2013-09-28

    Cell-permeable peptides (CPPs) and ultrasound-targeted microbubble destruction (UTMD) have tremendous potential for gene delivery. However, their applications are limited due to nonspecificity of CPPs and low transfection efficiency of UTMD. Here, we developed a 'smart' gene delivery system by encapsulating TAT peptide (TATp) and hepatocyte growth factor (HGF) gene within lipid microbubbles, in which TATp was protected from being enzymatically cleaved and HGF gene was protected from degradation. This new strategy had synergistic effects of UTMD and TATp on gene transfection. We investigated the efficacy and safety of HGF gene transfection mediated by the combination of UTMD and TATp in vitro and in vivo. The results from MTT assay and flow cytometry analyses indicated that the combination of UTMD and TATp could enhance HGF gene expression in HUVECs without any significant side effect on cell viability. In rat myocardial infarction models, we demonstrated that the protein and mRNA expressions of HGF in myocardium caused by the combination of UTMD and TATp were the highest. Histopathological findings demonstrated that the combination of UTMD and TATp enhanced myocardial microvasculature and ameliorated myocardial fibrosis. In conclusion, the combination of UTMD and TATp might be a safe and efficient technique for gene delivery. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Ultrasonic Analysis of Peptide- and Antibody-Targeted Microbubble Contrast Agents for Molecular Imaging of αvβ3-Expressing Cells

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    Paul A. Dayton

    2004-04-01

    Full Text Available The goal of targeted ultrasound contrast agents is to significantly and selectively enhance the detection of a targeted vascular site. In this manuscript, three distinct contrast agents targeted to the αvβ3 integrin are examined. The αvβ3 integrin has been shown to be highly expressed on metastatic tumors and endothelial cells during neovascularization, and its expression has been shown to correlate with tumor grade. Specific adhesion of these contrast agents to αvβ3-expressing cell monolayers is demonstrated in vitro, and compared with that of nontargeted agents. Acoustic studies illustrate a backscatter amplitude increase from monolayers exposed to the targeted contrast agents of up to 13-fold (22 dB relative to enhancement due to control bubbles. A linear dependence between the echo amplitude and bubble concentration was observed for bound agents. The decorrelation of the echo from adherent targeted agents is observed over successive pulses as a function of acoustic pressure and bubble density. Frequency–domain analysis demonstrates that adherent targeted bubbles exhibit high-amplitude narrowband echo components, in contrast to the primarily wideband response from free microbubbles. Results suggest that adherent targeted contrast agents are differentiable from free-floating microbubbles, that targeted contrast agents provide higher sensitivity in the detection of angiogenesis, and that conventional ultrasound imaging techniques such as signal subtraction or decorrelation detection can be used to detect integrin-expressing vasculature with sufficient signal-to-noise.

  18. Inflammation and cancer: role of annexin A1 and FPR2/ALX in proliferation and metastasis in human laryngeal squamous cell carcinoma.

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    Thaís Santana Gastardelo

    Full Text Available The anti-inflammatory protein annexin A1 (ANXA1 has been associated with cancer progression and metastasis, suggesting its role in regulating tumor cell proliferation. We investigated the mechanism of ANXA1 interaction with formylated peptide receptor 2 (FPR2/ALX in control, peritumoral and tumor larynx tissue samples from 20 patients, to quantitate the neutrophils and mast cells, and to evaluate the protein expression and co-localization of ANXA1/FPR2 in these inflammatory cells and laryngeal squamous cells by immunocytochemistry. In addition, we performed in vitro experiments to further investigate the functional role of ANXA1/FPR2 in the proliferation and metastasis of Hep-2 cells, a cell line from larynx epidermoid carcinoma, after treatment with ANXA1(2-26 (annexin A1 N-terminal-derived peptide, Boc2 (antagonist of FPR and/or dexamethasone. Under these treatments, the level of Hep-2 cell proliferation, pro-inflammatory cytokines, ANXA1/FPR2 co-localization, and the prostaglandin signalling were analyzed using ELISA, immunocytochemistry and real-time PCR. An influx of neutrophils and degranulated mast cells was detected in tumor samples. In these inflammatory cells of peritumoral and tumor samples, ANXA1/FPR2 expression was markedly exacerbated, however, in laryngeal carcinoma cells, this expression was down-regulated. ANXA1(2-26 treatment reduced the proliferation of the Hep-2 cells, an effect that was blocked by Boc2, and up-regulated ANXA1/FPR2 expression. ANXA1(2-26 treatment also reduced the levels of pro-inflammatory cytokines and affected the expression of metalloproteinases and EP receptors, which are involved in the prostaglandin signalling. Overall, this study identified potential roles for the molecular mechanism of the ANXA1/FPR2 interaction in laryngeal cancer, including its relationship with the prostaglandin pathway, providing promising starting points for future research. ANXA1 may contribute to the regulation of tumor growth

  19. Contrast-Enhanced Ultrasound with VEGFR2-Targeted Microbubbles for Monitoring Regorafenib Therapy Effects in Experimental Colorectal Adenocarcinomas in Rats with DCE-MRI and Immunohistochemical Validation.

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    Ralf Stefan Eschbach

    Full Text Available To investigate contrast-enhanced ultrasound (CEUS with VEGFR2-targeted microbubbles for monitoring therapy effects of regorafenib on experimental colon carcinomas in rats with correlation to dynamic contrast-enhanced MRI (DCE-MRI and immunohistochemistry.Human colorectal adenocarcinoma xenografts (HT-29 were implanted subcutaneously in n = 21 (n = 11 therapy group; n = 10 control group female athymic nude rats (Hsd: RH-Foxn1rnu. Animals were imaged at baseline and after a one-week daily treatment with regorafenib or a placebo (10 mg/kg bodyweight, using CEUS with VEGFR2-targeted microbubbles and DCE-MRI. In CEUS tumor perfusion was assessed during an early vascular phase (wash-in area under the curve = WiAUC and VEGFR2-specific binding during a late molecular phase (signal intensity after 8 (SI8min and 10 minutes (SI10min, using a conventional 15L8 linear transducer (transmit frequency 7 MHz, dynamic range 80 dB, depth 25 mm. In DCE-MRI functional parameters plasma flow (PF and plasma volume (PV were quantified. For validation purposes, CEUS parameters were correlated with DCE-MRI parameters and immunohistochemical VEGFR2, CD31, Ki-67 and TUNEL stainings.CEUS perfusion parameter WiAUC decreased significantly (116,989 ± 77,048 a.u. to 30,076 ± 27,095a.u.; p = 0.005 under therapy with no significant changes (133,932 ± 65,960 a.u. to 84,316 ± 74,144 a.u.; p = 0.093 in the control group. In the therapy group, the amount of bound microbubbles in the late phase was significantly lower in the therapy than in the control group on day 7 (SI8min: 283 ± 191 vs. 802 ± 460 a.u.; p = 0.006; SI10min: 226 ± 149 vs. 645 ± 461 a.u.; p = 0.009. PF and PV decreased significantly (PF: 147 ± 58 mL/100 mL/min to 71 ± 15 mL/100 mL/min; p = 0.003; PV: 13 ± 3% to 9 ± 4%; p = 0.040 in the therapy group. Immunohistochemistry revealed significantly fewer VEGFR2 (7.2 ± 1.8 vs. 17.8 ± 4.6; p < 0.001, CD31 (8.1 ± 3.0 vs. 20.8 ± 5.7; p < 0.001 and Ki-67 (318.7

  20. Proteomic Identification of an Upregulated Isoform of Annexin A3 in the Spinal Cords of Rats in a Neuropathic Pain Model

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    Wangyuan Zou

    2017-09-01

    Full Text Available Neuropathic pain (NP is induced by nerve damage or a disturbance in the peripheral or central nervous systems. Nerve damage causes the activation of sensitizing mechanisms in the peripheral and central nervous systems, which induces transcriptional and post-transcriptional alterations in sensory nerves. However, the underlying mechanisms of NP remain elusive. In the study, Two-dimensional gel electrophoresis (2DGE-based comparative proteomics identified 38 differential gel spots, and 15 differentially expressed proteins (DEPs between the sham and the chronic constriction injury (CCI-induced neuropathic pain rats. Of them, Annexin A3 (ANXA3 was significantly increased after CCI with Western blot analysis and immunofluorescence imaging. A lentivirus delivering ANXA3 shRNA (LV-shANXA3 was administered intrathecally to determine the analgesic effects of ANXA3 on allodynia and hyperalgesia in a CCI-induced neuropathic pain model in rats. Further study showed that LV-shANXA3 reversed the upregulation of ANXA3, alleviated CCI-induced mechanical allodynia and thermal hyperalgesia. The study indicated that ANXA3 may play an important role in neuropathic pain.

  1. Comparative Label-Free Mass Spectrometric Analysis of Mildly versus Severely Affected mdx Mouse Skeletal Muscles Identifies Annexin, Lamin, and Vimentin as Universal Dystrophic Markers

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    Ashling Holland

    2015-06-01

    Full Text Available The primary deficiency in the membrane cytoskeletal protein dystrophin results in complex changes in dystrophic muscles. In order to compare the degree of secondary alterations in differently affected subtypes of skeletal muscles, we have conducted a global analysis of proteome-wide changes in various dystrophin-deficient muscles. In contrast to the highly degenerative mdx diaphragm muscle, which showed considerable alterations in 35 distinct proteins, the spectrum of mildly to moderately dystrophic skeletal muscles, including interosseus, flexor digitorum brevis, soleus, and extensor digitorum longus muscle, exhibited a smaller number of changed proteins. Compensatory mechanisms and/or cellular variances may be responsible for differing secondary changes in individual mdx muscles. Label-free mass spectrometry established altered expression levels for diaphragm proteins associated with contraction, energy metabolism, the cytoskeleton, the extracellular matrix and the cellular stress response. Comparative immunoblotting verified the differences in the degree of secondary changes in dystrophin-deficient muscles and showed that the up-regulation of molecular chaperones, the compensatory increase in proteins of the intermediate filaments, the fibrosis-related increase in collagen levels and the pathophysiological decrease in calcium binding proteins is more pronounced in mdx diaphragm as compared to the less severely affected mdx leg muscles. Annexin, lamin, and vimentin were identified as universal dystrophic markers.

  2. Comparative Label-Free Mass Spectrometric Analysis of Mildly versus Severely Affected mdx Mouse Skeletal Muscles Identifies Annexin, Lamin, and Vimentin as Universal Dystrophic Markers.

    Science.gov (United States)

    Holland, Ashling; Henry, Michael; Meleady, Paula; Winkler, Claudia K; Krautwald, Mirjam; Brinkmeier, Heinrich; Ohlendieck, Kay

    2015-06-19

    The primary deficiency in the membrane cytoskeletal protein dystrophin results in complex changes in dystrophic muscles. In order to compare the degree of secondary alterations in differently affected subtypes of skeletal muscles, we have conducted a global analysis of proteome-wide changes in various dystrophin-deficient muscles. In contrast to the highly degenerative mdx diaphragm muscle, which showed considerable alterations in 35 distinct proteins, the spectrum of mildly to moderately dystrophic skeletal muscles, including interosseus, flexor digitorum brevis, soleus, and extensor digitorum longus muscle, exhibited a smaller number of changed proteins. Compensatory mechanisms and/or cellular variances may be responsible for differing secondary changes in individual mdx muscles. Label-free mass spectrometry established altered expression levels for diaphragm proteins associated with contraction, energy metabolism, the cytoskeleton, the extracellular matrix and the cellular stress response. Comparative immunoblotting verified the differences in the degree of secondary changes in dystrophin-deficient muscles and showed that the up-regulation of molecular chaperones, the compensatory increase in proteins of the intermediate filaments, the fibrosis-related increase in collagen levels and the pathophysiological decrease in calcium binding proteins is more pronounced in mdx diaphragm as compared to the less severely affected mdx leg muscles. Annexin, lamin, and vimentin were identified as universal dystrophic markers.

  3. Influence of DNA-Microbubble Coupling on Contrast Ultrasound-Mediated Gene Transfection in Muscle and Liver.

    Science.gov (United States)

    Xie, Aris; Wu, Melinda D; Cigarroa, Gabriella; Belcik, J Todd; Ammi, Azzdine; Moccetti, Federico; Lindner, Jonathan R

    2016-08-01

    Contrast ultrasound-mediated gene delivery (CUMGD) is a promising approach for enhancing gene therapy that relies on microbubble (MB) cavitation to augment complementary deoxyribonucleic acid (cDNA) transfection. The aims of this study were to determine optimal conditions for charge-coupling cDNA to MBs and to evaluate the advantages of surface loading for gene transfection in muscle and liver. Charge coupling of fluorescently labeled cDNA to either neutral MBs (MBN) or cationic MBs (MB+) in low- to high-ionic conditions (0.3%-1.8% NaCl) was assessed by flow cytometry. MB aggregation from cDNA coupling was determined by electrozone sensing. Tissue transfection of luciferase in murine hindlimb skeletal muscle and liver was made by CUMGD with MBN or MB+ combined with subsaturated, saturated, or supersaturated cDNA concentrations (2.5, 50, and 200 μg/10(8) MBs). Charge-coupling of cDNA was detected for MB+ but not MBN. Coupling occurred over almost the entire range of ionic conditions, with a peak at 1.2% NaCl, although electrostatic interference occurred at >1.5% NaCl. DNA-mediated aggregation of MB+ was observed at ≤0.6% NaCl but did not reduce the ability to produce inertial cavitation. Transfection with CUMGD in muscle and liver was low for both MBs at subsaturation concentrations. In muscle, higher cDNA concentrations produced a 10-fold higher degree of transfection with MB+, which was approximately fivefold higher (P transfection with MBN equal to that of MB+. Efficient charge-coupling of cDNA to MB+ but not MBN occurs over a relatively wide range of ionic conditions without aggregation. Transfection with CUMGD is much more efficient with charge-coupling of cDNA to MBs and is not affected by supersaturation except in the liver, which is specialized for macromolecular and cDNA uptake. Copyright © 2016 American Society of Echocardiography. Published by Elsevier Inc. All rights reserved.

  4. Cell Lysis and Detoxification of Cyanotoxins Using a Novel Combination of Microbubble Generation and Plasma Microreactor Technology for Ozonation

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    Jagroop Pandhal

    2018-04-01

    Full Text Available There has been a steady rise in the incidences of algal blooms globally, and worryingly, there is increasing evidence that changes in the global climate are leading to a shift toward cyanobacterial blooms. Many cyanobacterial genera are harmful, producing several potent toxins, including microcystins, for which there are over 90 described analogues. There are a wide range of negative effects associated with these toxins including gastroenteritis, cytotoxicity, hepatotoxicity and neurotoxicity. Although a variety of oxidation based treatment methods have been described, ozonation and advanced oxidation are acknowledged as most effective as they readily oxidise microcystins to non-toxic degradation products. However, most ozonation technologies have challenges for scale up including high costs and sub-optimum efficiencies, hence, a low cost and scalable ozonation technology is needed. Here we designed a low temperature plasma dielectric barrier discharge (DBD reactor with an incorporated fluidic oscillator for microbubble delivery of ozone. Both technologies have the potential to drastically reduce the costs of ozonation at scale. Mass spectrometry analysis revealed very rapid (<2 min destruction of two pure microcystins (MC-LR and MC-RR, together with removal of by-products even at low flow rate 1 L min−1 where bubble size was 0.56–0.6 mm and the ozone concentration within the liquid was 20 ppm. Toxicity levels were calculated through protein phosphatase inhibition assays and indicated loss of toxicity as well as confirming the by-products were also non-toxic. Finally, treatment of whole Microcystis aeruginosa cells showed that even at these very low ozone levels, cells can be killed and toxins (MC-LR and Desmethyl MC-LR removed. Little change was observed in the first 20 min of treatment followed by rapid increase in extracellular toxins, indicating cell lysis, with most significant release at the higher 3 L min−1 flow rate compared to 1 L

  5. Epigallocatechin gallate induces an up-regulation of LDL receptor accompanied by a reduction of PCSK9 via the annexin A2-independent pathway in HepG2 cells.

    Science.gov (United States)

    Kitamura, Kohei; Okada, Yudai; Okada, Kenji; Kawaguchi, Yuya; Nagaoka, Satoshi

    2017-08-01

    In animal studies, epigallocatechin gallate (EGCG), the dominant catechin in green tea, has been shown to improve cholesterol metabolism. However, the molecular mechanisms of EGCG underlying these functions have not been fully understood. In this study, we aimed to clarify the molecular mechanisms of the effect of EGCG on cholesterol metabolism mainly in HepG2 cells. We found that EGCG induced a reduction of the extracellular proprotein convertase subtilisin/kexin 9 (PCSK9) level accompanied by an up-regulation of the LDL receptor (LDLR) in HepG2 cells. The EGCG-induced up-regulation of LDLR occurred via the extracellular signal-regulated kinase (ERK) signaling pathway. Moreover, we showed that EGCG induced a significant early reduction of the extracellular PCSK9 protein level. However, there were no significant changes in the PCSK9 mRNA and the intracellular PCSK9 protein levels induced by EGCG. Annexin A2 knockdown affected the basal LDLR expression and did not affect the EGCG-induced reduction of the extracellular PCSK9 protein level or the up-regulation of LDLR. Annexin A2 possesses an essential function for the basal LDLR expression in HepG2 cells. But, EGCG induces the suppression of PCSK9 accompanied by an up-regulation of LDLR in an annexin A2-independent manner. EGCG attenuates the statin-induced an increase in PCSK9 level. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Targeted gene delivery in tumor xenografts by the combination of ultrasound-targeted microbubble destruction and polyethylenimine to inhibit survivin gene expression and induce apoptosis

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    Qiu Ri-Xiang

    2010-11-01

    Full Text Available Abstract Background Noninvasive and tissue-specific technologies of gene transfection would be valuable in clinical gene therapy. This present study was designed to determine whether it could enhance gene transfection in vivo by the combination of ultrasound-targeted microbubble destruction (UTMD with polyethylenimine (PEI in tumor xenografts, and illuminate the effects of gene silencing and apoptosis induction with short hairpin RNA (shRNA interference therapy targeting human survivin by this novel technique. Methods Two different expression vectors (pCMV-LUC and pSIREN were incubated with PEI to prepare cationic complexes (PEI/DNA and confirmed by the gel retardation assay. Human cervical carcinoma (Hela tumors were planted subcutaneously in both flanks of nude mice. Tumor-bearing mice were administered by tail vein with PBS, plasmid, plasmid and SonoVue microbubble, PEI/DNA and SonoVue microbubble. One tumor was exposed to ultrasound irradiation, while the other served as control. The feasibility of targeted delivery and tissue specificity facilitated by UTMD and PEI were investigated. Moreover, immunohistochemistry analyses about gene silencing and apoptosis induction were detected. Results Electrophoresis experiment revealed that PEI could condense DNA efficiently. The application of UTMD significantly increases the tissue transfection. Both expression vectors showed that gene expressions were present in all sections of tumors that received ultrasound exposure but not in control tumors. More importantly, the increases in transgene expression were related to UTMD with the presence of PEI significantly. Silencing of the survivin gene could induce apoptosis effectively by downregulating survivin and bcl-2 expression, also cause up-regulation of bax and caspase-3 expression. Conclusions This noninvasive, novel combination of UTMD with PEI could enhance targeted gene delivery and gene expression in tumor xenografts at intravenous administration

  7. {sup 99m}Tc-annexin V and {sup 111}In-antimyosin antibody uptake in experimental myocardial infarction in rats

    Energy Technology Data Exchange (ETDEWEB)

    Sarda-Mantel, Laure; Rouzet, Francois; Martet, Genevieve; Raguin, Olivier; Vrigneaud, Jean-Marc; Guludec, Dominique Le [Bichat Hospital AP-HP, EA 3512, Nuclear Medicine Department, Paris (France); Michel, Jean-Baptiste; Louedec, Liliane [INSERM U460, UFR Bichat, Paris (France); Vanderheyden, Jean-Luc [Theseus Imaging Corporation, Boston, MA (United States); Hervatin, Florence [Bichat Hospital AP-HP, EA 3512, Nuclear Medicine Department, Paris (France); CGA/SHFS, Orsay (France); Khaw, Ban An [Bouve College of Pharmacy and Health Sciences, Center for Drug Targeting and Analysis, Boston, MA (United States)

    2006-03-15

    {sup 99m}Tc-annexin V (ANX) allows scintigraphic detection of apoptotic cells via specific binding to exposed phosphatidylserine. In myocardial infarction, apoptosis of myocytes is variable and depends especially on the presence or absence of coronary reperfusion. In this study, ANX uptake in non-reperfused experimental myocardial infarcts was compared with uptake of a marker of myocyte necrosis ({sup 111}In-antimyosin antibodies, AM) and an immunohistochemical marker of apoptosis (Apostain). The left anterior coronary artery was ligated in 47 Wistar rats, which were then injected with ANX (n=20), AM (n=21) or both (n=6). Myocardial uptake of ANX and AM was determined at 2 h (n=14), 4 h (n=14) and 24 h (n=19) after coronary ligation (CL), by quantitative autoradiography with (n=23) or without (n=24) gamma imaging. Heart-to-lung ratios (HLRs) and infarct-to-remote myocardium activity ratios (INRs) were calculated on the scintigrams and autoradiograms respectively. Cardiac sections were stained with haematoxylin-eosin and Apostain. The above studies were repeated in 12 normal rats. All rats with CL showed increased ANX and AM uptake in cardiac areas on scintigrams 24 h after CL, with HLRs higher than in controls: 3.1{+-}0.6 versus 1.5{+-}0.3 (p=0.001) for ANX and 1.99{+-}0.44 versus 1.01{+-}0.05 (p<0.0005) for AM. Autoradiography showed intense ANX and AM uptake in infarcts, with comparable topography and INRs at 2 h, 4 h and 24 h after CL (4.6{+-}0.9 versus 5.0{+-}1.8 at 24 h), while Apostain staining was very low (0.06{+-}0.06% of cells). In this model of persistent CL, we observed increased ANX uptake in injured myocardium, comparable in intensity, topography and kinetics to that of AM. There was only minimal Apostain staining in the same areas. (orig.)

  8. Myocardial uptake of {sup 99m}Tc-annexin-V and {sup 111}In-antimyosin-antibodies after ischemia-reperfusion in rats

    Energy Technology Data Exchange (ETDEWEB)

    Sarda-Mantel, Laure [Universite Denis Diderot-Paris 7, UMR S773, Paris (France); AP-HP, Groupe Hospitalier Bichat-Beaujon, Service de Medecine Nucleaire, Paris (France); INSERM, U773, Paris (France); Hopital Bichat, Service de Medecine Nucleaire, Paris (France); Hervatin, Florence [Universite Denis Diderot-Paris 7, UMR S773, Paris (France); CEA, DSV/DRM/SHFJ, Orsay (France); Michel, Jean-Baptiste; Louedec, Liliane [INSERM, U698, Paris (France); Martet, Genevieve [Universite Denis Diderot-Paris 7, UMR S773, Paris (France); INSERM, U773, Paris (France); Rouzet, Francois; Lebtahi, Rachida; Merlet, Pascal; Le Guludec, Dominique [Universite Denis Diderot-Paris 7, UMR S773, Paris (France); AP-HP, Groupe Hospitalier Bichat-Beaujon, Service de Medecine Nucleaire, Paris (France); INSERM, U773, Paris (France); Khaw, Ban-An [Bouve College of Pharmacy and Health Sciences, Center for Drug Targeting and Analysis, Boston, MA (United States)

    2008-01-15

    Phosphatidylserin exposure on cell surfaces occurs early during apoptosis and is detected in vivo by using {sup 99m}Tc-annexin-V (ANX). Cardiomyocyte membrane disruption is detected in vivo by using {sup 111}In-antimyosin-antibodies (AM). We aimed to determine if ANX and AM allow evaluation of the time-course of these two distinct cell death events after myocardial ischemia-reperfusion. Coronary tying (20 min) followed by reperfusion (IR) was performed in 31 rats. Twelve of the rats were injected with ANX, 11 with AM, and eight with both tracers. Myocardial uptake of tracers was studied 1-2 h, 4 h, or 24 h after IR by scintigraphy (ANX, n = 14) and autoradiography (all cases), and compared to histology and Apostain staining. Scintigraphy was positive in all rats 2 h after IR and in three of five rats at 24 h. On autoradiography, ANX activity was intense in myocardial lesions as early as 1 h post-IR, whereas AM activity was mild at 2 h then increased at 4 h post-IR. ANX and AM uptakes evolved from mid-myocardium to endocardial and epicardial regions from 2 h to 24 h post-IR. Apostain staining was significant in myocardial lesions (p < 10{sup 6} compared to six sham-operated rats). On histology, myocardial lesion was characterized by interstitial oedema, myocytes necrosis, and dramatic thinning at 24 h. These data suggest that ANX and AM allow temporal and regional evaluations of PS exposure and membrane disruption, respectively, during myocytes death after 20-min myocardial ischemia followed by reperfusion. Also, (i) apoptosis starts very early in injured myocardium, (ii) myocyte necrosis occurs later (3-4 h post-reperfusion), and (iii) most dead cells are removed from mid-myocardium between 6 h and 24 h after reperfusion. (orig.)

  9. Anti-inflammatory effects of Tat-Annexin protein on ovalbumin-induced airway inflammation in a mouse model of asthma

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Sun Hwa; Kim, Dae Won; Kim, Hye Ri; Woo, Su Jung; Kim, So Mi; Jo, Hyo Sang [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of); Jeon, Seong Gyu [Department of Life Science, Pohang University of Science and Technology, Pohang 790-784 (Korea, Republic of); Cho, Sung-Woo [Department of Biochemistry and Molecular Biology, University of Ulsan, College of Medicine, Seoul 138-736 (Korea, Republic of); Park, Jong Hoon [Department of Biological Science, Sookmyung Women' s University, Seoul 140-742 (Korea, Republic of); Won, Moo Ho [Department of Neurobiology, School of Medicine, Kangwon National University, Chuncheon 200-701 (Korea, Republic of); Park, Jinseu [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of); Eum, Won Sik, E-mail: wseum@hallym.ac.kr [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of); Choi, Soo Young, E-mail: sychoi@hallym.ac.kr [Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702 (Korea, Republic of)

    2012-01-20

    Highlights: Black-Right-Pointing-Pointer We construct a cell permeable Tat-ANX1 fusion protein. Black-Right-Pointing-Pointer We examined the protective effects of Tat-ANX1 protein on OVA-induced asthma in animal models. Black-Right-Pointing-Pointer Transduced Tat-ANX1 protein protects from the OVA-induced production of cytokines and eosinophils in BAL fluid. Black-Right-Pointing-Pointer Tat-ANX1 protein markedly reduced OVA-induced MAPK in lung tissues. Black-Right-Pointing-Pointer Tat-ANX1 protein could be useful as a therapeutic agent for lung disorders including asthma. -- Abstract: Chronic airway inflammation is a key feature of bronchial asthma. Annexin-1 (ANX1) is an anti-inflammatory protein that is an important modulator and plays a key role in inflammation. Although the precise action of ANX1 remains unclear, it has emerged as a potential drug target for inflammatory diseases such as asthma. To examine the protective effects of ANX1 protein on ovalbumin (OVA)-induced asthma in animal models, we used a cell-permeable Tat-ANX1 protein. Mice sensitized and challenged with OVA antigen had an increased amount of cytokines and eosinophils in their bronchoalveolar lavage (BAL) fluid. However, administration of Tat-ANX1 protein before OVA challenge significantly decreased the levels of cytokines (interleukin (IL)-4, IL-5, and IL-13) and BAL fluid in lung tissues. Furthermore, OVA significantly increased the activation of mitogen-activated protein kinase (MAPK) in lung tissues, whereas Tat-ANX1 protein markedly reduced phosphorylation of MAPKs such as extracellular signal-regulated protein kinase, p38, and stress-activated protein kinase/c-Jun N-terminal kinase. These results suggest that transduced Tat-ANX1 protein may be a potential protein therapeutic agent for the treatment of lung disorders including asthma.

  10. Reciprocal regulation of annexin A2 and EGFR with Her-2 in Her-2 negative and herceptin-resistant breast cancer.

    Directory of Open Access Journals (Sweden)

    Praveenkumar K Shetty

    Full Text Available Alternative survival pathways are commonly seen to be upregulated upon inhibition of receptor tyrosine kinases (RTK, including Her-2. It is established that treatment with Herceptin leads to selective overexpression and activation of epidermal growth factor receptor (EGFR and Src which further contributes to oncogenesis in Herceptin resistant and triple negative breast cancer (TNBC patients. Here, we show a co-regulated upregulation in the expression of Annexin A2 (AnxA2, a known substrate of Src and one of the regulators of EGFR receptor endocytosis, in Herceptin resistant and Her-2 negative breast cancer. Immunohistochemical expression analysis revealed a reciprocal regulation between Her-2 and AnxA2 in breast cancer clinical samples as well as in cell lines as confirmed by protein and RNA analysis. The siRNA and Herceptin mediated downregulation/inhibition of Her-2 in Her-2 amplified cells induced AnxA2 expression and membrane translocation. In this study we report a possible involvement of AnxA2 in maintaining constitutively activated EGFR downstream signaling intermediates and hence in cell proliferation, migration and viability. This effect was consistent in Herceptin resistant JIMT-1 cells as well as in Her-2 negative breast cancer. The siRNA mediated AnxA2 downregulation leads to increased apoptosis, decreased cell viability and migration. Our studies further indicate the role of AnxA2 in EGFR-Src membrane bound signaling complex and ligand induced activation of downstream signaling pathways. Targeting this AnxA2 dependent positive regulation of EGFR signaling cascade may be of therapeutic value in Her-2 negative breast cancer.

  11. Biological studies in animal models using [99mTc](CO)3 recombinant annexin V as diagnostic agent of apoptotic processes

    International Nuclear Information System (INIS)

    Teran, Mariella Adriana; Martinez, Elena; Reyes, Ana L.; Paolino, Andrea; Vital, Marcelo; Esperon, Patricia; Pacheco, Jose P.; Savio, Eduardo

    2011-01-01

    Introduction: There are many diseases associated with variations in the expression of apoptosis such as organ rejection after transplantation, myocardial ischemia or infarct and neurodegenerative diseases. For this reason, the early visualization of this process is relevant to set fast and effective therapeutic strategies. Methods: The precursor was prepared according to the procedure reported by R. Alberto, R. Schibli, P. Schubiger, U. Abram, and T. Kaden [Reactions with the technetium and rhenium carbonyl complexes (NEt 4 )[MX 3 (CO) 3 ]. Synthesis and structure of Tc(CN-But) 3 (CO) 3 ](NO 3 ) and (Net 4 )[Tc 2 (μ-SCH 2 CH 2 OH) 3 (CO) 3 ], Polyhedron 1996;15: 1079-89]. Recombinant annexin V was incubated with [ 99m Tc](H 2 O)3(CO) 3 + solution, previously neutralized with buffer. Biodistribution studies were performed in 8-week-old female Wistar rats. Animals were housed and treated in compliance with institutional guidelines related to animal experimentation. Work protocol was previously approved by the Animal Ethics Committee of the university. Two groups of rats were defined. One was used as control and the other group was previously injected with 150 mg/kg ip of cyclophosphamide to induce apoptosis. Results: The synthesis of carbonyl precursor achieved yields higher than 90%, and the radiolabeled protein was obtained with 92% of radiochemical purity and high stability in vitro. An important uptake in apoptotic tissues was confirmed by biodistributions, scintigraphic images and histological studies. Conclusions: Biodistribution studies revealed hepatobiliary elimination, high stability in vivo and important uptake in the reticuloendothelial system. In the pathologic model, higher uptake values correspond to the liver, spleen, lungs and femur. Histological studies confirmed the development of apoptosis at 8 and 24 h postinduction in the spleen and lymphocyte bulks in the peribronchial area. Scintigraphic images confirmed high uptake both the spleen and the

  12. Annexin A1 in blood mononuclear cells from patients with coronary artery disease: Its association with inflammatory status and glucocorticoid sensitivity.

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    Ida Bergström

    Full Text Available Annexin A1 (AnxA1 is a key player in resolution of inflammation and a mediator of glucocorticoid actions. In atherosclerotic tissue, increased expression of AnxA1 has been associated with protective plaque-stabilizing effects. Here, we investigated the expression of AnxA1 in peripheral blood mononuclear cells (PBMCs from patients with coronary artery disease (CAD. Blood was collected from 57 patients with stable CAD (SCAD and 41 healthy controls. We also included a minor group (n = 10 with acute coronary syndrome (ACS. AnxA1 mRNA was measured in PBMCs. Expression of AnxA1 protein (total and surface-bound and glucocorticoid receptors (GR were detected in PBMC subsets by flow cytometry. Also, salivary cortisol, interleukin(IL-6 and IL-10 in plasma, and LPS-induced cytokine secretion from PBMCs, with or without dexamethasone, were assessed. AnxA1 mRNA was found to be slightly increased in PBMCs from SCAD patients compared with controls. However, protein expression of AnxA1 or GRs in PBMC subsets did not differ between SCAD patients and controls, despite SCAD patients showing a more proinflammatory cytokine profile ex vivo. Only surface expression of AnxA1 on monocytes correlated with dexamethasone-mediated suppression of cytokines. In ACS patients, a marked activation of AnxA1 was seen involving both gene expression and translocation of protein to cell surface probably reflecting a rapid glucocorticoid action modulating the acute inflammatory response in ACS. To conclude, surface expression of AnxA1 on monocytes may reflect the degree of glucocorticoid sensitivity. Speculatively, "normal" surface expression of AnxA1 indicates that anti-inflammatory capacity is impaired in SCAD patients.

  13. Annexin A1 in blood mononuclear cells from patients with coronary artery disease: Its association with inflammatory status and glucocorticoid sensitivity.

    Science.gov (United States)

    Bergström, Ida; Lundberg, Anna K; Jönsson, Simon; Särndahl, Eva; Ernerudh, Jan; Jonasson, Lena

    2017-01-01

    Annexin A1 (AnxA1) is a key player in resolution of inflammation and a mediator of glucocorticoid actions. In atherosclerotic tissue, increased expression of AnxA1 has been associated with protective plaque-stabilizing effects. Here, we investigated the expression of AnxA1 in peripheral blood mononuclear cells (PBMCs) from patients with coronary artery disease (CAD). Blood was collected from 57 patients with stable CAD (SCAD) and 41 healthy controls. We also included a minor group (n = 10) with acute coronary syndrome (ACS). AnxA1 mRNA was measured in PBMCs. Expression of AnxA1 protein (total and surface-bound) and glucocorticoid receptors (GR) were detected in PBMC subsets by flow cytometry. Also, salivary cortisol, interleukin(IL)-6 and IL-10 in plasma, and LPS-induced cytokine secretion from PBMCs, with or without dexamethasone, were assessed. AnxA1 mRNA was found to be slightly increased in PBMCs from SCAD patients compared with controls. However, protein expression of AnxA1 or GRs in PBMC subsets did not differ between SCAD patients and controls, despite SCAD patients showing a more proinflammatory cytokine profile ex vivo. Only surface expression of AnxA1 on monocytes correlated with dexamethasone-mediated suppression of cytokines. In ACS patients, a marked activation of AnxA1 was seen involving both gene expression and translocation of protein to cell surface probably reflecting a rapid glucocorticoid action modulating the acute inflammatory response in ACS. To conclude, surface expression of AnxA1 on monocytes may reflect the degree of glucocorticoid sensitivity. Speculatively, "normal" surface expression of AnxA1 indicates that anti-inflammatory capacity is impaired in SCAD patients.

  14. Proteomic Characterization of Annexin l (ANX1 and Heat Shock Protein 27 (HSP27 as Biomarkers for Invasive Hepatocellular Carcinoma Cells.

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    Ruo-Chiau Wang

    Full Text Available To search for reliable biomarkers and drug targets for management of hepatocellular carcinoma (HCC, we performed a global proteomic analysis of a pair of HCC cell lines with distinct differentiation statuses using 2-DE coupled with MALDI-TOF MS. In total, 106 and 55 proteins were successfully identified from the total cell lysate and the cytosolic, nuclear and membrane fractions in well-differentiated (HepG2 and poorly differentiated (SK-Hep-1 HCC clonal variants, respectively. Among these proteins, nine spots corresponding to proteins differentially expressed between HCC cell types were selected and confirmed by immunofluorescence staining and western blotting. Notably, Annexin 1 (ANX1, ANX-2, vimentin and stress-associated proteins, such as GRP78, HSP75, HSC-70, protein disulfide isomerase (PDI, and heat shock protein-27 (HSP27, were exclusively up-regulated in SK-Hep-1 cells. Elevated levels of ANX-4 and antioxidant/metabolic enzymes, such as MnSOD, peroxiredoxin, NADP-dependent isocitrate dehydrogenase, α-enolase and UDP-glucose dehydrogenase, were observed in HepG2 cells. We functionally demonstrated that ANX1 and HSP27 were abundantly overexpressed only in highly invasive types of HCC cells, such as Mahlavu and SK-Hep-1. Knockdown of ANX1 or HSP27 in HCC cells resulted in a severe reduction in cell migration. The in-vitro observations of ANX1 and HSP27 expressions in HCC sample was demonstrated by immunohistochemical stains performed on HCC tissue microarrays. Poorly differentiated HCC tended to have stronger ANX1 and HSP27 expressions than well-differentiated or moderately differentiated HCC. Collectively, our findings suggest that ANX1 and HSP27 are two novel biomarkers for predicting invasive HCC phenotypes and could serve as potential treatment targets.

  15. Comparison of the synergistic effect of lipid nanobubbles and SonoVue microbubbles for high intensity focused ultrasound thermal ablation of tumors

    Directory of Open Access Journals (Sweden)

    Yuanzhi Yao

    2016-02-01

    Full Text Available Microbubbles (MBs are considered as an important enhancer for high intensity focused ultrasound (HIFU treatment of benign or malignant tumors. Recently, different sizes of gas-filled bubbles have been investigated to improve the therapeutic efficiency of HIFU thermal ablation and reduce side effects associated with ultrasound power and irradiation time. However, nanobubbles (NBs as an ultrasound contrast agent for synergistic therapy of HIFU thermal ablation remain controversial due to their small nano-size in diameter. In this study, phospholipid-shell and gas-core NBs with a narrow size range of 500–600 nm were developed. The synergistic effect of NBs for HIFU thermal ablation was carefully studied both in excised bovine livers and in breast tumor models of rabbits, and made a critical comparison with that of commercial SonoVue microbubbles (SonoVue MBs. In addition, the pathological changes of the targeted area in tumor tissue after HIFU ablation were further investigated. Phosphate buffer saline (PBS was used as the control. Under the same HIFU parameters, the quantitative echo intensity of B-mode ultrasound image and the volume of coagulative necrosis in lipid NBs groups were significantly higher and larger than that in PBS groups, but could not be demonstrated a difference to that in SonoVue MBs groups both ex vivo and in vivo. These results showed that the synergistic effect of lipid NBs for HIFU thermal ablation were similar with that of SonoVue MBs, and further indicate that lipid NBs could potentially become an enhancer for HIFU thermal ablation of tumors.

  16. Comparison of the synergistic effect of lipid nanobubbles and SonoVue microbubbles for high intensity focused ultrasound thermal ablation of tumors.

    Science.gov (United States)

    Yao, Yuanzhi; Yang, Ke; Cao, Yang; Zhou, Xuan; Xu, Jinshun; Liu, Jianxin; Wang, Qi; Wang, Zhigang; Wang, Dong

    2016-01-01

    Microbubbles (MBs) are considered as an important enhancer for high intensity focused ultrasound (HIFU) treatment of benign or malignant tumors. Recently, different sizes of gas-filled bubbles have been investigated to improve the therapeutic efficiency of HIFU thermal ablation and reduce side effects associated with ultrasound power and irradiation time. However, nanobubbles (NBs) as an ultrasound contrast agent for synergistic therapy of HIFU thermal ablation remain controversial due to their small nano-size in diameter. In this study, phospholipid-shell and gas-core NBs with a narrow size range of 500-600 nm were developed. The synergistic effect of NBs for HIFU thermal ablation was carefully studied both in excised bovine livers and in breast tumor models of rabbits, and made a critical comparison with that of commercial SonoVue microbubbles (SonoVue MBs). In addition, the pathological changes of the targeted area in tumor tissue after HIFU ablation were further investigated. Phosphate buffer saline (PBS) was used as the control. Under the same HIFU parameters, the quantitative echo intensity of B-mode ultrasound image and the volume of coagulative necrosis in lipid NBs groups were significantly higher and larger than that in PBS groups, but could not be demonstrated a difference to that in SonoVue MBs groups both ex vivo and in vivo. These results showed that the synergistic effect of lipid NBs for HIFU thermal ablation were similar with that of SonoVue MBs, and further indicate that lipid NBs could potentially become an enhancer for HIFU thermal ablation of tumors.

  17. Enhanced effect of nuclear localization signal peptide during ultrasound‑targeted microbubble destruction‑mediated gene transfection.

    Science.gov (United States)

    Cao, Sheng; Zhou, Qing; Chen, Jin-Ling; Jiang, Nan; Wang, Yi-Jia; Deng, Qing; Hu, Bo; Guo, Rui-Qiang

    2017-07-01

    Ultrasound‑targeted microbubble destruction (UTMD) can promote the entry of plasmid DNA (pDNA) into the cell cytoplasm, by increasing the permeability of the cell membrane. But the transfection efficiency remains low due to inability of the pDNA to enter the nucleus. Various methods have been explored to improve the UTMD transfection efficiency, but with little success. In cells, the classic nuclear localization signal (cNLS) peptide is an amino acid sequence that signals proteins that are due for nuclear transport. The present study aimed to investigate whether binding of a cNLS peptide to the pDNA may improve the transfection efficiency of UTMD. Four experimental groups were analyzed: Control group (UTMD + pDNA), group with cNLS (UTMD + pDNA + cNLS), group with mutated NLS (mNLS; UTMD + pDNA + mNLS), and group with cNLS and the nuclear import blocker, wheat germ agglutinin (WGA; UTMD + pDNA + cNLS + WGA). The NLS was labeled by fluorescein isothiocyanate, whereas pDNA was labeled with Cy3. Different molar ratios were tested for the NLS and pDNA combination in order to achieve optimal binding of the two molecules. Human umbilical vein endothelial cells were then transfected using the optimum ultrasonic irradiation parameters and NLS/pDNA molar ratio. At 6 h post‑transfection, the rates of Cy3‑labeled pDNA inside the cells and their nuclei were detected by flow cytometry and laser confocal microscopy, and the cellular vs. nuclear uptake of pDNA was calculated. In order to further evaluate the effect of NLS on UTMD‑mediated gene transfection, the transfection efficiency and relative expression levels of mRNA and protein were detected at 48 h post‑transfection. The results demonstrated that the optimal molar ratio of NLS with pDNA was 104:1. The rates of pDNA successful entry into the cell and nucleus were significantly higher in the cNLS group compared with the control group. The transfection efficiency, and relative expression levels of mRNA and protein

  18. 32 CFR 352a.5 - Relationships.

    Science.gov (United States)

    2010-07-01

    ... 32 National Defense 2 2010-07-01 2010-07-01 false Relationships. 352a.5 Section 352a.5 National Defense Department of Defense (Continued) OFFICE OF THE SECRETARY OF DEFENSE (CONTINUED) ORGANIZATIONAL CHARTERS DEFENSE FINANCE AND ACCOUNTING SERVICE (DFAS) § 352a.5 Relationships. (a) In the performance of...

  19. MIEN1, a novel interactor of Annexin A2, promotes tumor cell migration by enhancing AnxA2 cell surface expression.

    Science.gov (United States)

    Kpetemey, Marilyne; Dasgupta, Subhamoy; Rajendiran, Smrithi; Das, Susobhan; Gibbs, Lee D; Shetty, Praveenkumar; Gryczynski, Zygmunt; Vishwanatha, Jamboor K

    2015-08-15

    Migration and invasion enhancer 1 (MIEN1) is a novel gene found to be abundantly expressed in breast tumor tissues and functions as a critical regulator of tumor cell migration and invasion to promote systemic metastases. Previous studies have identified post-translational modifications by isoprenylation at the C-terminal tail of MIEN1 to favor its translocation to the inner leaflet of plasma membrane and its function as a membrane-bound adapter molecule. However, the exact molecular events at the membrane interface activating the MIEN1-driven tumor cell motility are vaguely understood. MIEN1 was first studied using in-silico analysis on available RNA sequencing data of human breast tissues and its expression was ascertained in breast cells. We performed several assays including co-immunoprecipitation, wound healing, western blotting and immunofluorescence to decipher the molecular events involved in MIEN1-mediated tumor cell migration. Clinically, MIEN1 is predominantly overexpressed in Her-2 and luminal B subtypes of breast tumors, and its increased expression correlates with poor disease free survival. Molecular studies identified a phosphorylation-dependent activation signal in the immunoreceptor tyrosine based activation motif (ITAM) of MIEN1 and the phosphorylation-deficient MIEN1-mutants (Y39F/50 F) to regulate filopodia generation, migration and invasion. We found that ITAM-phosphorylation of MIEN1 is significantly impaired in isoprenylation-deficient MIEN1 mutants indicating that prenylation of MIEN1 and membrane association is required for cross-phosphorylation of tyrosine residues. Furthermore, we identified MIEN1 as a novel interactor of Annexin A2 (AnxA2), a Ca(2+) -dependent phospholipid binding protein, which serves as an extracellular proteolytic center regulating plasmin generation. Fluorescence resonance energy transfer (FRET) confirmed that MIEN1 physically interacts with AnxA2 and functional studies revealed that they mutually cooperate to

  20. Biological studies in animal models using [{sup 99m}Tc](CO){sub 3} recombinant annexin V as diagnostic agent of apoptotic processes

    Energy Technology Data Exchange (ETDEWEB)

    Teran, Mariella Adriana, E-mail: mteran@fq.edu.u [Catedra de Radioquimica, Departamento Estrella Campos, Facultad de Quimica, Universidad de la Republica, P.O. 11800, Montevideo (Uruguay); Martinez, Elena; Reyes, Ana L.; Paolino, Andrea [Catedra de Radioquimica, Departamento Estrella Campos, Facultad de Quimica, Universidad de la Republica, P.O. 11800, Montevideo (Uruguay); Vital, Marcelo; Esperon, Patricia [Catedra de Biologia Molecular, Departamento de Bioquimica Clinica, Facultad de Quimica, Universidad de la Republica, Montevideo (Uruguay); Pacheco, Jose P. [Instituto de Patobiologia, Facultad de Veterinaria, Universidad de la Republica, Montevideo (Uruguay); Savio, Eduardo [Catedra de Radioquimica, Departamento Estrella Campos, Facultad de Quimica, Universidad de la Republica, P.O. 11800, Montevideo (Uruguay)

    2011-02-15

    Introduction: There are many diseases associated with variations in the expression of apoptosis such as organ rejection after transplantation, myocardial ischemia or infarct and neurodegenerative diseases. For this reason, the early visualization of this process is relevant to set fast and effective therapeutic strategies. Methods: The precursor was prepared according to the procedure reported by R. Alberto, R. Schibli, P. Schubiger, U. Abram, and T. Kaden [Reactions with the technetium and rhenium carbonyl complexes (NEt{sub 4})[MX{sub 3}(CO){sub 3}]. Synthesis and structure of Tc(CN-But){sub 3}(CO){sub 3}](NO{sub 3}) and (Net{sub 4})[Tc{sub 2}({mu}-SCH{sub 2}CH{sub 2}OH){sub 3}(CO){sub 3}], Polyhedron 1996;15: 1079-89]. Recombinant annexin V was incubated with [{sup 99m}Tc](H{sub 2}O)3(CO){sub 3}{sup +} solution, previously neutralized with buffer. Biodistribution studies were performed in 8-week-old female Wistar rats. Animals were housed and treated in compliance with institutional guidelines related to animal experimentation. Work protocol was previously approved by the Animal Ethics Committee of the university. Two groups of rats were defined. One was used as control and the other group was previously injected with 150 mg/kg ip of cyclophosphamide to induce apoptosis. Results: The synthesis of carbonyl precursor achieved yields higher than 90%, and the radiolabeled protein was obtained with 92% of radiochemical purity and high stability in vitro. An important uptake in apoptotic tissues was confirmed by biodistributions, scintigraphic images and histological studies. Conclusions: Biodistribution studies revealed hepatobiliary elimination, high stability in vivo and important uptake in the reticuloendothelial system. In the pathologic model, higher uptake values correspond to the liver, spleen, lungs and femur. Histological studies confirmed the development of apoptosis at 8 and 24 h postinduction in the spleen and lymphocyte bulks in the peribronchial area

  1. Anti-allergic cromones inhibit histamine and eicosanoid release from activated human and murine mast cells by releasing Annexin A1.

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    Samia Yazid

    Full Text Available Although the 'cromones' (di-sodium cromoglycate and sodium nedocromil are used to treat allergy and asthma, their 'mast cell stabilising' mechanism of pharmacological action has never been convincingly explained. Here, we investigate the hypothesis that these drugs act by stimulating the release of the anti-inflammatory protein Annexin-A1 (Anx-A1 from mast cells.We used biochemical and immuno-neutralisation techniques to investigate the mechanism by which cromones suppress histamine and eicosanoid release from cord-derived human mast cells (CDMCs or murine bone marrow-derived mast cells (BMDMCs from wild type and Anx-A1 null mice.CDMCs activated by IgE-FcRε1 crosslinking, released histamine and prostaglandin (PG D2, which were inhibited (30-65% by 5 min pre-treatment with cromoglycate (10 nM or nedocromil (10 nM, as well as dexamethasone (2 nM and human recombinant Anx-A1 (1-10 nM. In CDMCs cromones potentiated (2-5 fold protein kinase C (PKC phosphorylation and Anx-A1 phosphorylation and secretion (3-5 fold. Incubation of CDMCs with a neutralising anti-Anx-A1 monoclonal antibody reversed the cromone inhibitory effect. Nedocromil (10 nM also inhibited (40-60% the release of mediators from murine bone marrow derived-mast cells from wild type mice activated by compound 48/80 and IgE-FcRε1 cross-linking, but were inactive in such cells when these were prepared from Anx-A1 null mice or when the neutralising anti-Anx-A1 antibody was present.We conclude that stimulation of phosphorylation and secretion of Anx-A1 is an important component of inhibitory cromone actions on mast cells, which could explain their acute pharmacological actions in allergy. These findings also highlight a new pathway for reducing mediator release from these cells.

  2. Sodium butyrate induces growth inhibition and apoptosis in human prostate cancer DU145 cells by up-regulation of the expression of annexin A1.

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    Dawei Mu

    Full Text Available BACKGROUND: Sodium butyrate, a histone deacetylase inhibitor, has emerged as a promising anticancer drug for multiple cancers. Recent studies have indicated that sodium butyrate could inhibit the progression of prostate cancer; however, the exact mechanism is still unclear. The aim of this study was to investigate the mechanism of sodium butyrate action in prostate cancer DU145 cells. METHODS: The inhibitory effects of NaB on cell growth were detected by the 3-(4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrrazolium bromide assay. Cell apoptosis was determined by flow cytometric analysis of DU145 cells stained with annexin V and PI. Hoechst 33258 and fluorescence microscopes were used to observe the nuclear morphology of DU145 cells after treatment with NaB. ANXA1 knockdown cells were established through transfection with ANXA1 siRNA. ANXA1 mRNA levels were measured by qRT-PCR. Bcl-2, Bax, ANXA1, ERK1/2 and pERK1/2 were detected by western blot. RESULTS: NaB significantly inhibited the growth and induction apoptosis of DU145 and PC3 cells in a dose-dependent manner. Expression of the anti-apoptosis gene Bcl-xl and Bcl-2 in DU145 cells are decreased and expression of the pro-apoptosis gene Bax and Bak increased after NaB treatment. Further studies have demonstrated that NaB up-regulated the expression of ANXA1 and that the tumor inhibition action of NaB was reduced markedly through knockdown of the ANXA1 gene in DU145 cells. Moreover, the siANXA1 cells showed that cell proliferation increased and cell apoptosis was induced by the inactivation of extracellular regulated kinase (ERK. CONCLUSION: Our results support a significant correlation between NaB functions and ANXA1 expression in prostate cancer, and pave the way for further studying the molecular mechanism of NaB actions in cancers.

  3. Dexamethasone-induced and estradiol-induced CREB activation and annexin 1 expression in CCRF-CEM lymphoblastic cells: evidence for the involvement of cAMP and p38 MAPK

    Directory of Open Access Journals (Sweden)

    M. Castro-caldas

    2003-01-01

    Full Text Available Aims: Annexin 1 (ANXA1, a member of the annexin family of calcium-binding and phospholipid-binding proteins, is a key mediator of the anti-inflammatory actions of steroid hormones. We have previously demonstrated that, in the human lymphoblastic CCRF-CEM cell line, both the synthetic glucocorticoid hormone, dexamethasone (Dex, and the estrogen hormone, 17β-estradiol (E2β, induce the synthesis of ANXA1, by a mechanism independent of the activation of their nuclear receptors. Recently, it was reported that the gene coding for ANXA1 contains a cAMP-responsive element (CRE. In this work, we investigated whether Dex and E2β were able to induce the activation of CRE binding proteins (CREB in the CCRF-CEM cells. Moreover, we studied the intracellular signalling pathways involved in CREB activation and ANXA1 synthesis in response to Dex and E2β; namely, the role of cAMP and the p38 mitogen-activated protein kinase (MAPK.

  4. Ultrasound-Targeted Microbubble Destruction Improves the Migration and Homing of Mesenchymal Stem Cells after Myocardial Infarction by Upregulating SDF-1/CXCR4: A Pilot Study

    Directory of Open Access Journals (Sweden)

    Lu Li

    2015-01-01

    Full Text Available Mesenchymal stem cell (MSC therapy shows considerable promise for the treatment of myocardial infarction (MI. However, the inefficient migration and homing of MSCs after systemic infusion have limited their therapeutic applications. Ultrasound-targeted microbubble destruction (UTMD has proven to be promising to improve the homing of MSCs to the ischemic myocardium, but the concrete mechanism remains unclear. We hypothesize that UTMD promotes MSC homing by upregulating SDF-1/CXCR4, and this study was aimed at exploring this potential mechanism. We analyzed SDF-1/CXCR4 expression after UTMD treatment in vitro and in vivo and counted the number of homing MSCs in MI areas. The in vitro results demonstrated that UTMD not only led to elevated secretion of SDF-1 but also resulted in an increased proportion of MSCs that expressed surface CXCR4. The in vivo findings show an increase in the number of homing MSCs and higher expression of SDF-1/CXCR4 in the UTMD combined with MSCs infusion group compared to other groups. In conclusion, UTMD can increase SDF-1 expression in the ischemic myocardium and upregulate the expression of surface CXCR4 on MSCs, which provides a molecular mechanism for the homing of MSCs assisted by UTMD via SDF-1/CXCR4 axis.

  5. Enhancing effects of SonoVue, a microbubble sonographic contrast agent, on high-intensity focused ultrasound ablation in rabbit livers in vivo.

    Science.gov (United States)

    Luo, Wen; Zhou, Xiaodong; Ren, Xiaolong; Zheng, Minjuan; Zhang, Jun; He, Guangbin

    2007-04-01

    The purpose of this study was to explore the enhancing biological effects of SonoVue (Bracco SpA, Milan, Italy), a sulfur hexafluoride sonographic contrast agent, on high-intensity focused ultrasound (HIFU) ablation in vivo. Forty-five rabbits were randomly divided into 3 groups and underwent laparotomy. Animals in group 1 were given injections of 0.2 mL of SonoVue intravenously; animals in group 2 were given physiologic saline; and those in group 3 were not given injections as control. The exposure time was set at 2 seconds with the acoustic power at 600 W. After HIFU ablations, volumes of coagulated regions were measured. Liver tissues were examined under light microscopy with hematoxylin-eosin staining and under transmission electron microscopy. Coagulated volumes in group 1 (mean +/- SD, 2.41 +/- 0.44 cm(3)) were larger than those in group 2 (0.80 +/- 0.13 cm(3)) and group 3 (0.83 +/- 0.16 cm(3)) (P SonoVue can substantially enhance the ablation effects of HIFU, suggesting that microbubble contrast agents may be useful for improving HIFU efficiency.

  6. Delivery of TFPI-2 using ultrasound with a microbubble agent (SonoVue) inhibits intimal hyperplasia after balloon injury in a rabbit carotid artery model.

    Science.gov (United States)

    Zhou, Jie; Wang, Yuxue; Xiong, Yufang; Wang, Hongxing; Feng, Youmei; Chen, Juan

    2010-11-01

    Here we report a new, simple and efficient method by using ultrasound and a microbubble agent (SonoVue) for delivering a gene to balloon-injured carotid arteries for restenosis prophylaxis. The tissue factor pathway inhibitor-2 (TFPI-2) has been shown to inhibit the postinjury intimae hyperplasia in atherosclerotic vessels. New Zealand white rabbits were divided into 4 groups with 14 in each, a treatment control for balloon injury, a gene vehicle control, a gene delivery of TFPI-2 without using ultrasound and a gene delivery of TFPI-2 using ultrasound. After four weeks, the injured artery neointimal proliferation was significantly lower in the TFPI-2 group with ultrasound than the control groups (p < 0.01) according to the measurement of the mean luminal diameters by B-mode ultrasonography. The ratio of intimal/media area and the stenosis rate in the gene delivery facilitated by ultrasound were significantly lower than those of the nonultrasound gene delivering method (p < 0.01). Copyright © 2010 World Federation for Ultrasound in Medicine & Biology. All rights reserved.

  7. Hypoxic Preconditioning Combined with Microbubble-Mediated Ultrasound Effect on MSCs Promote SDF-1/CXCR4 Expression and its Migration Ability: An In Vitro Study.

    Science.gov (United States)

    Li, Lu; Wu, Shengzheng; Li, Peijing; Zhuo, Lisha; Gao, Yunhua; Xu, Yali

    2015-12-01

    Our objective is to investigate the promoting effect of hypoxic preconditioning combined with microbubble (MB)-mediated ultrasound (US) on the SDF-1/CXCR4 expression and the migration ability of mesenchymal stem cells (MSCs). Based on the uniform design, the parameters of MB-mediated US, such as the total treatment time (T), acoustic intensity (Q), and the dosage of MBs, were optimized firstly. The results were assessed by regression analysis. Using the optimum irradiation parameters, the concentration of SDF-1 in the supernatant, the expression levels of membrane CXCR4, and the cell viability of hypoxic MSCs or normoxic MSCs were compared. The in vitro transwell migration assay was performed as well. The best combination of parameters for more SDF-1 secretion and less MSCs death was T = 30 s, A = 0.6 W/cm(2), and MB = 10(6)/ml. After 24 h of hypoxic preconditioning, the expression of SDF-1 and surface CXCR4 was increased in the hypoxic MSC group as compared to the normoxic MSC group (P SDF-1/CXCR4 with the optimum parameters (P SDF-1/CXCR4 expression and improve the migration ability in MSCs.

  8. Synthesis of Pt nanoparticles as catalysts of oxygen reduction with microbubble-assisted low-voltage and low-frequency solution plasma processing

    Science.gov (United States)

    Horiguchi, Genki; Chikaoka, Yu; Shiroishi, Hidenobu; Kosaka, Shinpei; Saito, Morihiro; Kameta, Naohiro; Matsuda, Naoki

    2018-04-01

    In the preparation of metallic nanoparticles by conventional solution plasma (SP) techniques, unstable plasma emission becomes an issue when the voltage and frequency of the waves applied between two electrodes placed in solution are lowered to avoid the boiling of the solution. In this study, we confirm that, in the presence of microbubbles, plasma is generated stably at low voltage (440 V) and low frequency (50-100 Hz) and small-size (≤10 nm) Pt nanoparticles (PtNPs) are synthesized in succession using a flow cell. The smallest PtNPs, ∼3.3 nm in diameter, are obtained using half-wave rectification, a tungsten wire anode, and a platinum wire cathode. The PtNPs are characterized by X-ray diffraction, X-ray photoelectron spectroscopy, transmission electron microscopy, and thermogravimeter-differential thermal analysis. The oxygen reduction reaction (ORR) is investigated in 0.1 M HClO4 solution on carbon-supported PtNPs using a rotating ring-disk electrode. The catalytic activities per initial electrochemical active surface area of the carbon-supported PtNPs synthesized employing the low-voltage, low-frequency (LVLF)-SP technique is higher than that of the commercially available 20 wt% Pt on Vulcan XC-72R. These results indicate that the LVLF-SP technique is a promising approach to producing carbon-supported PtNPs that catalyze ORR with low energy consumption.

  9. The diagnostic value of small bowel wall vascularity after sulfur hexafluoride-filled microbubble injection in patients with Crohn's disease. Correlation with the therapeutic effectiveness of specific anti-inflammatory treatment

    International Nuclear Information System (INIS)

    Quaia, Emilio; Migaleddu, Vincenzo; Baratella, Elisa; Pizzolato, Riccardo; Rossi, Alexia; Grotto, Maurizio; Cova, Maria Assunta

    2009-01-01

    Purpose: To assess the value of small bowel wall vascularity after microbubble contrast agent injection in evaluating the therapeutic effectiveness of specific anti-inflammatory treatment in patients with Crohn's disease. Materials and methods: Fifteen patients (7 male and 8 female; mean age ± SD, 40 years ± 6) with a biopsy-proven diagnosis of Crohn's disease - Crohn's disease activity index (CDAI) > 150 (n = 12 patients) or 5 mm) were included. In each patient the terminal loop was scanned by contrast-enhanced ultrasound (CEUS) after sulfur hexafluoride-filled microbubble injection before and after 6-month anti-inflammatory treatment. The vascularity of the terminal loop was quantified in gray-scale levels (0-255) by a manually drawn ROI encompassing the thickened bowel wall and it was correlated with CDAI. Result: Before the beginning of the specific treatment all patients revealed diffuse transparietal contrast enhancement after microbubble injection, except for 3 patients who revealed contrast enhancement limited to the submucosa. In 13 patients the slope of the first ascending tract and the area under the enhancement curve were significantly lower after anti-inflammatory treatment (P < 0.05; Wilcoxon test) with a significant correlation with the CDAI score (ρ = 0.85, P < 0.05). In 2 patients no significant vascularity changes were found even though a mild reduction of CDAI score was identified (from 200 to 150 gray-scale levels). Conclusion: CEUS is a useful method to assess the therapeutic effectiveness of specific medical anti-inflammatory treatment in patients with Crohn's disease.

  10. Biological evaluation of octahydropyrazin[2,1-a:5,4-a']diisoquinoline derivatives as potent anticancer agents.

    Science.gov (United States)

    Gornowicz, Agnieszka; Pawłowska, Natalia; Czajkowska, Anna; Czarnomysy, Robert; Bielawska, Anna; Bielawski, Krzysztof; Michalak, Olga; Staszewska-Krajewska, Olga; Kałuża, Zbigniew

    2017-06-01

    In this study, we evaluated the cytotoxic activity and antiproliferative potency of novel octahydropyrazin[2,1-a:5,4-a']diisoquinoline derivatives (1-7) in MCF-7 and MDA-MB-231 breast cancer cell lines. Annexin V binding assay and disruption of the mitochondrial potential were performed to determine apoptosis. The activity of caspases 3, 8, 9, and 10 was measured after 24 h of incubation with tested compounds to explain detailed molecular mechanism of induction of apoptosis. The results from experiments were compared with effects obtained after incubation in the presence of camptothecin and etoposide. Our study demonstrated that the most active compounds in both analyzed breast cancer cell lines were compounds 3 and 4. We also observed that all compounds induced apoptosis. We demonstrated the higher activity of caspases 3, 8, 9, and 10, which confirmed that induction of apoptosis is associated with external and internal cell death pathway. Our study revealed that the novel compounds in group of diisoquinoline derivatives are promising candidates in anticancer treatment by activation of both extrinsic and intrinsic apoptotic pathways.

  11. Molecular imaging probes spy on the body's inner workings: miniaturized microscopes, microbubbles, 7- and 15-T scanners, diffusion-tensor MRI, and other molecular-imaging technologies are pushing molecular imaging into the future.

    Science.gov (United States)

    Mertz, Leslie

    2013-01-01

    Molecular imaging is one of the hot-button areas within medical imaging. This technology employs imaging techniques in concert with molecular probes, or biomarkers, that together noninvasively spy on cellular function and molecular processes. In some cases, this technology may be able to detect the very earliest stages of diseases and eliminate them on the spot. This paper discusses how miniaturized microscopes, microbubbles, 7T and 15T scanners, diffusion-tensor MRI and other molecular imaging technologies are pushing molecular imaging into the future.

  12. Apolipoprotein A5 in health and disease

    Czech Academy of Sciences Publication Activity Database

    Hubáček, J. A.; Adámková, V.; Vrablík, M.; Kadlecová, Michaela; Zicha, Josef; Kuneš, Jaroslav; Piťha, J.; Suchánek, P.; Poledne, R.

    2009-01-01

    Roč. 58, Suppl.2 (2009), S101-S109 ISSN 0862-8408 R&D Projects: GA MŠk(CZ) 1M0510 Grant - others:IKEM(CZ) 00023001; GA MŠk(CZ) MEB060808; GA MZd(CZ) NR8895; GAMZd(CZ) NR9393 Institutional research plan: CEZ:AV0Z50110509 Keywords : apolipoprotein A5 * plasma triglycerides * myocardial infarction Subject RIV: FB - Endocrinology, Diabetology, Metabolism, Nutrition Impact factor: 1.430, year: 2009

  13. Safety of thrombolytic therapy with rt-PA and transcranial color Doppler ultrasound (TCCS) combined with microbubbles: a histopathologic study on rabbit brain tissues.

    Science.gov (United States)

    Ren, Xinping; Wang, Yong; Wang, Yi; Chen, Hong; Chen, Li; Liu, Yi; Xu, Chengshi

    2015-04-01

    To evaluate effect of thrombolytic therapy with rt-PA (recombinant tissue plasminogen activator) and transcranial color Doppler ultrasound (TCCS) combined with microbubbles on histology of brain tissue. New Zealand rabbits were subjected to TCCS based thrombolytic therapy, in 8 groups depending on dose of rt-PA, exposure duration of TCCS and presence of attenuation by skull bone window, 2 animals/group: (1) skull+1/2 rt-PA+TCCS+MBs, 10 min, (2) skull+rt-PA+TCCS+MBs, 10 min, (3) skull+1/2 rt-PA+TCCS+MBs, 20 min, (4) skull+rt-PA+TCCS+MBs, 20 min, (5) skull+1/2 rt-PA+TCCS+MBs, 30 min, (6) skull+rt-PA+TCCS+MBs, 30 min, (7) 1/2 rt-PA+TCCS+MBs, 10 min, (8) 1/2 rt-PA+TCCS+MBs, 20 min. The brain tissues were harvested after therapies and submitted for microscopic, electronic microscope and immunohistochemical examination. The histological changes were scored. TCCS caused exposure duration dependent brain tissue damage. With attenuation by bone window, TCCS based therapies for 10-20 min caused minimal tissue damage. However, significant tissue damage was observed upon TCCS for 30 min in presence of skull bone window, presenting as hemorrhage, misdistribution of organelles, demyelination of nerve fibers, and thinning of basement membrane in blood-brain barrier, which was milder than that after 20 min of exposure to TCCS in absence of bone window. Dose of rt-PA did not affect brain histology in all groups. Short treatment of brain tissue with TCCS through a bone window is relative safe. And skull bone window protected brain tissue from TCCS induced damage. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Ultrasound enhanced prehospital thrombolysis using microbubbles infusion in patients with acute ST elevation myocardial infarction: Rationale and design of the Sonolysis study

    Directory of Open Access Journals (Sweden)

    van Dijk Arie PJ

    2008-12-01

    Full Text Available Abstract Background - Experimental studies have shown that ultrasound contrast agents enhance the effectiveness of thrombolytic agents in the presence of ultrasound in vitro and in vivo. Recently, we have launched a clinical pilot study, called "Sonolysis", to study this effect in patients with ST-elevation myocardial infarction based on proximal lesions of the infarct-related artery. Methods/design - In our multicenter, randomized, placebo controlled clinical trial we will include patients between 18 and 80 years of age with their first ST-elevation myocardial infarction based on a proximal lesion of the infarct-related artery. After receiving a single bolus alteplase 50 mg IV (Actilyse® Boehringer Ingelheim GmbH, a loading dose of aspirin 500 mg, and heparin 5000 IU in the ambulance according to the prehospital thrombolysis protocol, patients, following oral informed consent, are randomized to undergo 15 minutes of pulsatile ultrasound with intravenous administration of ultrasound contrast agent or placebo without ultrasound. Afterwards coronary angiography and, if indicated, percutaneous coronary intervention will take place. A total of 60 patients will be enrolled in approximately 1 year. The primary endpoints are based on the coronary angiogram and consist of TIMI flow, corrected TIMI frame count, and myocardial blush grade. Follow-up includes 12-lead ECG, 2D-echocardiography, cardiac MRI, and enzyme markers to obtain our secondary endpoints, including the infarct size, wall motion abnormalities, and the global left ventricular function. Discussion - The Sonolysis study is the first multicenter, randomized, placebo controlled clinical trial investigating the therapeutic application of ultrasound and microbubbles in acute ST-elevation myocardial infarction patients. A positive finding may stimulate further research and technical innovations to implement the treatment in the ambulance and maybe obtain even more patency at an earlier stage

  15. Enhanced delivery of PEAL nanoparticles with ultrasound targeted microbubble destruction mediated siRNA transfection in human MCF-7/S and MCF-7/ADR cells in vitro

    Directory of Open Access Journals (Sweden)

    Teng Y

    2015-08-01

    Full Text Available Yanwei Teng,1,2,* Min Bai,3,* Ying Sun,2 Qi Wang,1,2 Fan Li,3 Jinfang Xing,3 Lianfang Du,3 Tao Gong,1 Yourong Duan2 1Key Laboratory of Drug Targeting and Novel Drug Delivery Systems, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu, Sichuan, People’s Republic of China; 2State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, People’s Republic of China; 3Department of Ultrasound, Shanghai First People’s Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, People’s Republic of China *These authors contributed equally to this work Abstract: The gene knockdown activity of small interfering RNA (siRNA has led to their use as potential therapeutics for a variety of diseases. However, successful gene therapy requires safe and efficient delivery systems. In this study, we choose mPEG-PLGA-PLL nanoparticles (PEAL NPs with ultrasound targeted microbubble destruction (UTMD to efficiently deliver siRNA into cells. An emulsification-solvent evaporation method was used to prepare siRNA-loaded PEAL NPs. The NPs possessed an average size of 132.6±10.3 nm (n=5, with a uniform spherical shape, and had an encapsulation efficiency (EE of more than 98%. As demonstrated by MTT assay, neither PEAL NPs nor siRNA-loaded PEAL NPs showed cytotoxicity even at high concentrations. The results of cellular uptake showed, with the assistance of UTMD, the siRNA-loaded PEAL NPs can be effectively internalized and can subsequently release siRNA in cells. Taken together, PEAL NPs with UTMD may be highly promising for siRNA delivery, making it possible to fully exploit the potential of siRNA-based therapeutics. Keywords: gene delivery, mPEG-PLGA-PLL, UTMD, emulsification-solvent evaporation method, orthogonal design

  16. The Effect of Docetaxel-Loaded Micro-Bubbles Combined with Low-Frequency Ultrasound in H22 Hepatocellular Carcinoma-Bearing Mice.

    Science.gov (United States)

    Ren, Shu-Ting; Shen, Shu; He, Xin-Ying; Liao, Yi-Ran; Sun, Peng-Fei; Wang, Bing; Zhao, Wen-Bao; Han, Shui-Ping; Wang, Yi-Li; Tian, Tian

    2016-02-01

    A novel lipid micro-bubble (MB) loaded with docetaxel (DOC-MB) was investigated in a previous study. However, its anti-tumor effects and mechanism of action in combination with low-frequency ultrasound (LFUS) in vivo are still unclear. DOC-MBs containing 5.0 mg of DOC were prepared by lyophilization with modification via ultrasonic emulsification. Then, the effects of DOC-MBs combined with LFUS on tumor growth, proliferating cell nuclear antigen (PCNA) expression and cell apoptosis, as well as local DOC delivery, were investigated in H22 hepatocellular carcinoma (HCC)-bearing mice. Compared with the previously prepared DOC-MBs (1.6 mg of DOC loaded), the encapsulation efficiency (81.2% ± 3.89%) and concentration ([7.94 ± 0.04] × 10(9) bubbles/mL) of the DOC-MBs containing 5.0 mg of DOC were higher, but the bubble size (1.368 ± 0.004 μm) was smaller. After treatment with the DOC-MBs and LFUS, the H22 HCC growth inhibition rate was significantly increased, PCNA expression in tumor tissue was significantly inhibited and local release of DOC was induced. In conclusion, new DOC-MBs containing 5.0 mg of DOC were successfully prepared with a high encapsulation efficiency and superior bubble size and concentration, and their combination with LFUS significantly enhanced the anti-tumor effect of DOC in H22 HCC-bearing mice by inhibiting tumor cell proliferation and increasing local drug delivery. Copyright © 2016 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  17. [Differential patterns of contrast enhancement in different focal liver lesions after injection of the microbubble US contrast agent SonoVue].

    Science.gov (United States)

    Quaia, Emilio; Degobbis, Francesca; Tona, Giuseppe; Mosconi, Elisa; Bertolotto, Michele; Pozzi Mucelli, Roberto

    2004-03-01

    To identify differential contrast enhancement patterns in different focal hepatic lesions after injection of the microbubble contrast agent SonoVue using high or low acoustic power imaging. Forty-seven focal hepatic lesions (1-8 cm) were detected in 45 patients at unenhanced gray-scale ultrasound (US) and evaluated by color Doppler (CD) US with spectral analysis of tumoral vessels. Lesions were subsequently evaluated by US contrast specific modes after IV bolus administration of 2,4-4,8 ml of SonoVue, by intermittent high acoustic power (18 patients) or by continous low acoustic power imaging (27 patients), during arterial, portal and late phase. Subjective evaluation of lesions appearance before and after SonoVue injection was performed. For final diagnosis multiphasic helical CT (21 patients) and/or fine needle US guided biopsy (24 patients) were considered as the reference procedures. Final diagnoses comprised 22 hepatocellular carcinomas (HCCs; 1,5-6 cm), 2 macroregenerative nodules (RNNs; 1-2 cm), 10 metastasis (2-3,5 cm), 10 hemangiomas (2-6 cm) and 3 focal nodular hyperplasias (FNHs; 1-3 cm). On CD evaluation HCCs revealed peripheral basket shaped (12/22) or intranodular (10/22) arterial pattern while, after SonoVue injection HCCs revealed diffuse contrast enhancement during arterial phase with contrast washout during portal and late phase. Metastases did not reveal flow signals on CD or contrast enhancement after SonoVue injection, except for 2 metastases which revealed peripheral and central vessels on CD and a diffuse contrast enhancement during arterial phase, appearing hypoechoic to the adjacent liver during portal and late phase. RNNs revealed dotted contrast-enhancement during portal and late phase with isoechoic appearance to the adjacent liver. Hemangiomas revealed some peripheral venous flows on CD and a peripheral nodular contrast enhancement during arterial phase with a centripetal fill-in during portal and late phase. FNHs revealed low

  18. Ultrasound-targeted microbubble destruction of calcium channel subunit α 1D siRNA inhibits breast cancer via G protein-coupled receptor 30.

    Science.gov (United States)

    Ji, Yanlei; Han, Zhen; Shao, Limei; Zhao, Yuehuan

    2016-10-01

    Estrogen has been closely associated with breast cancer. Several studies reported that Ca2+ signal and Ca2+ channels act in estrogen-modulated non-genomic pathway of breast cancer, however little was revealed on the function of L-type Ca2+ channels. The L-type Ca2+ channel subunit α 1D, named Cav1.3 was found in breast cancer cells. We aimed to investigate the expression and activity of Cav1.3 in human breast cancer, and reveal the effect of estrogen in regulating the expression of Cav1.3. The qRT-PCR and western blotting were employed to show that Cav1.3 was highly expressed in breast cancer tissues. E2 exposure rapidly upregulated the expression of Cav1.3 in dosage- and time-dependent manner, and promoted Ca2+ influx. The silencing of G protein-coupled estrogen receptor 30 (GPER1/GPR30) using siRNA transfection inhibited the upregulation of Cav1.3 and Ca2+ influx induced by E2. Moreover, the inhibition of Cav1.3 by siRNA transfection suppressed E2-induced second peak of Ca2+ signal, the expression of p-ERK1/2, and the cell proliferation. Ultrasound-targeted microbubble destruction (UTMD) of Cav1.3 siRNA was used in MCF-7 cells in vitro and in the tumor xenografts mice in vivo. The application of UTMD significantly suppressed the tumor growth and promoted the survival rate. In conclusion, E2 upregulated the expression of Cav1.3 for Ca2+ influx to promote the expression of p-ERK1/2 for cell proliferation. The study confirmed that the mechanism of E2 inducing the expression of Cav1.3 through a non-genomic pathway, and highlighted that UTMD of Cav1.3 siRNA is a powerful promising technology for breast cancer gene therapy.

  19. Ultrasound-targeted microbubble destruction-mediated Foxp3 knockdown may suppress the tumor growth of HCC mice by relieving immunosuppressive Tregs function.

    Science.gov (United States)

    Shi, Chunying; Zhang, Yu; Yang, Haichao; Dong, Tianxiu; Chen, Yaodong; Xu, Yutong; Yang, Xiuhua; Liu, Pengfei

    2018-01-01

    The aim of the present study was to investigate the effect of Forkhead family transcription factor P3 (Foxp3) knockdown on the function of cluster of differentiation (CD)4 + CD25 + regulatory T cell (Tregs) and the tumor growth of a hepatocellular carcinoma (HCC) mouse model. CD4 + CD25 + Tregs and CD4 + CD25 - T cells were sorted from peripheral blood mononuclear cells (PBMCs) of patients with HCC. Then, ultrasound-targeted microbubble destruction (UTMD)-mediated Foxp3-microRNA (miRNA) was transfected into Tregs. Subsequently, CD4 + CD25 - T cells were co-cultured with PBMC and Tregs without Foxp3-miRNA (Foxp3 + Tregs) or Tregs with Foxp3-miRNA (Foxp3 - Tregs) and the proliferation-inhibition ratio of CD4 + CD25 - T cells was detected using a Cell Counting Kit-8. Additionally, HCC mice were treated with UTMD-mediated Foxp3-shRNA, the tumor volume was calculated and the content of CD4 + and CD25 + T cells in the blood were detected using flow cytometry. The content of interferon-γ (IFN-γ), interleukin (IL)-2, IL-10, transforming growth factor-β (TGF-β) and vascular endothelial growth factor (VEGF) in cultural supernatant and serum were detected by ELISA analysis. Foxp3 - Tregs significantly reduced the inhibition effect of Foxp3 + Tregs on the proliferation of CD4 + CD25 - T cells (P<0.01). The content of IFN-γ and IL-2 significantly increased, while IL-10 and TGF-β significantly decreased in the co-cultured system of Foxp3 - Tregs compared with the co-cultured system of Foxp3 + Tregs (P<0.01). Following treatment with Foxp3-shRNA, the average tumor volume, ratio of Tregs/CD4 + T cells and level of IL-10, TGF-β and VEGF significantly decreased, however, the level of IFN-γ and IL-2 significantly increased compared with un-treated HCC mice (P<0.05). Foxp3 knockdown may suppress the tumor growth of HCC mice through relieving the immunosuppressive function of Tregs.

  20. Tissue plasminogen activator induces microglial inflammation via a noncatalytic molecular mechanism involving activation of mitogen-activated protein kinases and Akt signaling pathways and AnnexinA2 and Galectin-1 receptors.

    Science.gov (United States)

    Pineda, David; Ampurdanés, Coral; Medina, Manel G; Serratosa, Joan; Tusell, Josep Maria; Saura, Josep; Planas, Anna M; Navarro, Pilar

    2012-04-01

    Inflammatory responses mediated by glial cells play a critical role in many pathological situations related to neurodegeneration such as Alzheimer's disease. Tissue plasminogen activator (tPA) is a serine protease which best-known function is fibrinolysis, but it is also involved in many other physiological and pathological events as microglial activation. Here, we found that tPA is required for Aβ-mediated microglial inflammatory response and tumor necrosis factor-α release. We further investigated the molecular mechanism responsible for tPA-mediated microglial activation. We found that tPA induces a catalytic-independent rapid and sustained activation of extracellular signal-regulated kinase (ERK)1/2, Jun N-terminal kinase (JNK), Akt, and p38 signaling pathways. Inhibition of ERK1/2 and JNK resulted in a strong inhibition of microglial activation, whereas Akt inhibition led to increased inflammatory response, suggesting specific functions for each signaling pathway in the regulation of microglial activation. Furthermore, we demonstrated that AnnexinA2 and Galectin-1 receptors are involved in tPA signaling and inflammatory response in glial cells. This study provides new evidences supporting that tPA plays a cytokine-like role in glial activation by triggering receptor-mediated intracellular signaling circuits and opens new therapeutic strategies for the treatment of neurological disorders in which neuroinflammation plays a pathogenic role. Copyright © 2011 Wiley-Liss, Inc.

  1. In Vivo Microbubble Cavitation Imaging

    NARCIS (Netherlands)

    Vignon, F.; Shi, W.; Liu, J.; Xie, F.; Gao, S.; Drvol, L.; Lof, J.; Everbach, C.; Porter, T.; Powers, J.

    2011-01-01

    Stroke is the second cause of death and leading cause of disabilityworldwide. Less than 5% of ischemic stroke patients receive the state-of-the art treatment of a thrombolytic drug tPA, and only about 10% of these gain additional benefit from it. Ultrasound (US)-inducedmicrobubble (MB) cavitation

  2. Annexin A2 in Proliferative Vitreoretinopathy

    Science.gov (United States)

    2017-10-01

    histologic analysis. The eye was enucleated, and a nick created in the cornea with a #11 blade prior to incubation in 4% paraformaldehyde. The cornea and lens...autofluorescence was quenched with NH4Cl, and sections then blocked with normal donkey serum. For A2 staining, incubation with primary and secondary antibodies...n=7-11 mice per group) representing Anxa2-/- or Anxa2+/+ mice at 2, 4, or 6 weeks and were scored using a standard algorithm . Inter-observer

  3. Structure of a C-terminal AHNAK peptide in a 1:2:2 complex with S100A10 and an acetylated N-terminal peptide of annexin A2

    International Nuclear Information System (INIS)

    Ozorowski, Gabriel; Milton, Saskia; Luecke, Hartmut

    2013-01-01

    Structure of a 20-amino-acid peptide of AHNAK bound asymmetrically to the AnxA2–S100A10A heterotetramer (1:2:2 symmetry) provides insights into the atomic level interactions that govern this membrane-repair scaffolding complex. AHNAK, a large 629 kDa protein, has been implicated in membrane repair, and the annexin A2–S100A10 heterotetramer [(p11) 2 (AnxA2) 2 )] has high affinity for several regions of its 1002-amino-acid C-terminal domain. (p11) 2 (AnxA2) 2 is often localized near the plasma membrane, and this C2-symmetric platform is proposed to be involved in the bridging of membrane vesicles and trafficking of proteins to the plasma membrane. All three proteins co-localize at the intracellular face of the plasma membrane in a Ca 2+ -dependent manner. The binding of AHNAK to (p11) 2 (AnxA2) 2 has been studied previously, and a minimal binding motif has been mapped to a 20-amino-acid peptide corresponding to residues 5654–5673 of the AHNAK C-terminal domain. Here, the 2.5 Å resolution crystal structure of this 20-amino-acid peptide of AHNAK bound to the AnxA2–S100A10 heterotetramer (1:2:2 symmetry) is presented, which confirms the asymmetric arrangement first described by Rezvanpour and coworkers and explains why the binding motif has high affinity for (p11) 2 (AnxA2) 2 . Binding of AHNAK to the surface of (p11) 2 (AnxA2) 2 is governed by several hydrophobic interactions between side chains of AHNAK and pockets on S100A10. The pockets are large enough to accommodate a variety of hydrophobic side chains, allowing the consensus sequence to be more general. Additionally, the various hydrogen bonds formed between the AHNAK peptide and (p11) 2 (AnxA2) 2 most often involve backbone atoms of AHNAK; as a result, the side chains, particularly those that point away from S100A10/AnxA2 towards the solvent, are largely interchangeable. While the structure-based consensus sequence allows interactions with various stretches of the AHNAK C-terminal domain, comparison

  4. Ac2-26 Mimetic Peptide of Annexin A1 Inhibits Local and Systemic Inflammatory Processes Induced by Bothrops moojeni Venom and the Lys-49 Phospholipase A2 in a Rat Model.

    Directory of Open Access Journals (Sweden)

    Bruna Stuqui

    Full Text Available Annexin A1 (AnxA1 is an endogenous glucocorticoid regulated protein that modulates anti-inflammatory process and its therapeutic potential has recently been recognized in a range of systemic inflammatory disorders. The effect of the N-terminal peptide Ac2-26 of AnxA1 on the toxic activities of Bothrops moojeni crude venom (CV and its myotoxin II (MjTX-II were evaluated using a peritonitis rat model. Peritonitis was induced by the intraperitoneal injection of either CV or MjTX-II, a Lys-49 phospholipase A2. Fifteen minutes after the injection, the rats were treated with either Ac2-26 or PBS. Four hours later, the CV and MjTX-II-induced peritonitis were characterized by neutrophilia (in the peritoneal exudate, blood and mesentery and increased number of mesenteric degranulated mast cells and macrophages. At 24 hours post-injection, the local inflammatory response was attenuated in the CV-induced peritonitis while the MjTX-II group exhibited neutrophilia (peritoneal exudates and blood. Ac2-26 treatment prevented the influx of neutrophils in MjTX-II-induced peritonitis and diminished the proportion of mesenteric degranulated mast cells and macrophages in CV-induced peritonitis. Additionally, CV and MjTX-II promoted increased levels of IL-1β and IL-6 in the peritoneal exudates which were significantly reduced after Ac2-26 treatment. At 4 and 24 hours, the endogenous expression of AnxA1 was upregulated in the mesenteric neutrophils (CV and MjTX-II groups and mast cells (CV group. In the kidneys, CV and MjTX-II administrations were associated with an increased number of macrophages and morphological alterations in the juxtamedullary nephrons in proximal and distal tubules. Ac2-26 promoted significant recovery of the juxtamedullary structures, decreased the number of macrophages and diminished the AnxA1 in epithelial cells from distal tubules and renal capsules. Our results show that Ac2-26 treatment significantly attenuates local and systemic

  5. 15 CFR 4a.5 - Duration of classification.

    Science.gov (United States)

    2010-01-01

    ... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Duration of classification. 4a.5 Section 4a.5 Commerce and Foreign Trade Office of the Secretary of Commerce CLASSIFICATION..., except as provided in § 1.6(d) of E.O. 12958. Under E.O. 12958, information may be exempted from...

  6. 49 CFR 178.33a-5 - Material.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 2 2010-10-01 2010-10-01 false Material. 178.33a-5 Section 178.33a-5 Transportation Other Regulations Relating to Transportation PIPELINE AND HAZARDOUS MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION HAZARDOUS MATERIALS REGULATIONS SPECIFICATIONS FOR PACKAGINGS...

  7. Profile and regulation of annexin II expression during early embryogenesis in cattle Perfil e regulação da expressão da anexina II durante a embriogênese em bovinos

    Directory of Open Access Journals (Sweden)

    L.F.S. Costa

    2007-12-01

    Full Text Available The presence of annexin II (Ann-II during the initial stages of bovine embryo development and the regulation of Ann-II expression by retinol and insulin-like growth factor I (IGF-I were studied. Bovine embryos at different stages of development were produced in vitro on Synthetic Oviductal Fluid (SOF medium (control group, SOF supplemented with retinol (retinol group; 0.1ng/ml, or IGF-I (IGF-I group; 10ng/ml. The embryos were processed for mRNA extraction, cDNA production and polymerase chain reaction (PCR using Ann-II-specific oligonucleotides. Ann-II was detected in all stages of early embryo development, except for the 16-cell stage. The blastocyst rates were significantly higher (PForam estudadas a presença de anexina II (Ann-II durante a fase inicial do desenvolvimento embrionário bovino e sua regulação pelo retinol e pelo fator de crescimento semelhante à insulina (IGF-I. Embriões bovinos em diferentes estádios de desenvolvimento foram produzidos in vitro em fluido sintético de oviduto (SOF sem suplementação (grupo-controle ou suplementado com retinol (grupo retinol; 0,1ng/ml medium ou IGF-I (grupo IGF-I; 10 ng/ml de meio. Os embriões foram processados para extração de mRNA, produção de cDNA e posterior análise por reação em cadeia da polimerase (PCR com oligonucleotídeos específicos para Ann-II. Em todos os estádios de desenvolvimento embrionário, Ann-II foi detectada, exceto no estádio de 16 células. Os índices de blastocisto foram significativamente maiores (P<0,05 no grupo suplementado com retinol (37,8%, 45/119 durante o cultivo in vitro de embriões (PIV que aqueles obtidos no grupo controle (20,5%, 24/117 ou no IGF-I (25,8%, 24/93. Análise semiquantitativa da expressão de Ann-II em embriões produzidos em meio suplementado com IGF-I ou retinol revelou uma menor expressão desse gene quando comparado com embriões cultivados somente em SOF (P<0,05. A expressão de Ann-II não foi diferente em embri

  8. A 5.5 MeV electrostatic accelerator

    International Nuclear Information System (INIS)

    Zhan Furu; Hu Suhua; Hu Chundong; Yu Zengliang; Wang Shaohu

    1998-05-01

    The author has introduced the principle and the basic structure of a 5.5 MeV electrostatic accelerator presented by Texas University of American, and the main parameters and their measurements are shown as well

  9. Entropy evolution in warm inflation from a 5 D vacuum

    International Nuclear Information System (INIS)

    Romero, J.; Bellini, M.

    2009-01-01

    Using some ideas of Modern Kaluza-Klein theory, we examine the evolution of entropy on a 4 D Friedmann-Robertson-Walker (FRW) brane from a 5 D vacuum state, which is defined on a 5 D background Riemann-flat metric. We found that entropy production is sufficiently important during inflation: S > 10 90 ., for all the initial values of temperature T 0 GU .

  10. Potential use of microbubbles (MBs) as contrast material in x-ray dark field (DF) imaging: How does the DF signal change with the characteristic parameters of the MBs?

    Science.gov (United States)

    Zhang, Ran; Qin, Bin; Ge, Yongshuai; Whiting, Bruce; Li, Ke; Villanueva, Flordeliza; Chen, Guang-Hong

    2016-03-01

    One of the most exciting aspects of the grating based x-ray differential phase contrast (DPC) acquisition method is the concurrent generation of the so-called dark field (DF) signal, along with the classical absorption signal and the novel DPC signal. The DF signal is associated with local distribution of small angle scatterers in an image object, while the absorption signal and DPC signal are often used to characterize the relatively uniform structure of the image object. Besides the endogenous image contrast, exogenous contrast media are often used in x-ray imaging to locally enhance the image signal. This paper proposes a potential contrast medium for DF signal enhancement: microbubbles (MBs). MBs have already been developed for clinical use in ultrasound imaging, and recent experimental studies have shown that MBs may also enhance the DF signal, although it remained unclear how the physical characteristics of the MBs quantitatively impact the DF signal. In this paper, a systematic study was performed to investigate the quantitative relationships between the DF signal and the following properties of MBs: size, concentration, shell thickness, size uniformity, and whether gold nanoparticles were attached. The experimental results demonstrated that, an increased MB size (about 4 microns) may generate a stronger DF signal for our DPC imaging system; additionally, a moderately increased shell thickness and the use of gold nanoparticles on the shell surface also resulted in further enhancement of the DF signal. These findings may provide critical information needed for using MBs as the contrast agent of x-ray DF imaging.

  11. Clinicopathological Spectrum of Ovarian Tumors: A 5‑Year ...

    African Journals Online (AJOL)

    Clinicopathological Spectrum of Ovarian Tumors: A 5‑Year Experience in a Tertiary Health Care Center. ... Materials and Methods: It was a retrospective observational study. The study was conducted in. Department of Pathology, B. J. Medical College Pune, India from July 2006 to June 2011. All the histopathology slides of ...

  12. Phacomatosis cesioflammea in a 5-week-old infant

    Directory of Open Access Journals (Sweden)

    Khushboo Gupta

    2016-01-01

    Full Text Available A 5-week-old male infant presented with extensive lesions of nevus flammeus and Mongolian spots affecting the face, trunk, buttocks and extremities. In addition, the child had ocular melanosis and a café-au-lait spot on the trunk. The case is being reported on account of its rarity.

  13. Apolipoprotein a5 and hypertriglyceridemia in prague hypertriglyceridemic rats

    Czech Academy of Sciences Publication Activity Database

    Kadlecová, Michaela; Hojná, Silvie; Bohuslavová, R.; Hubáček, J. A.; Zicha, Josef; Kuneš, Jaroslav

    2006-01-01

    Roč. 55, č. 4 (2006), s. 373-379 ISSN 0862-8408 R&D Projects: GA MŠk(CZ) 1M0510; GA ČR(CZ) GA305/03/0769 Institutional research plan: CEZ:AV0Z50110509 Keywords : metabolic syndrome * apolipoprotein A5 * rat Subject RIV: ED - Physiology Impact factor: 2.093, year: 2006

  14. Elevated zinc concentrations in a 5 months old infant

    DEFF Research Database (Denmark)

    Petersen, Eva Rabing Brix; Mortensen, Sven; Nybo, Mads

    2018-01-01

    Pre-analytical errors account for the majority of laboratory-associated errors. In a 5 months old infant hospitalised with lung dysfunction due to prematurity, a routine measurement of zinc revealed an unexpected elevated concentration of 20.2 µmol/L (reference interval 10.0 - 19.0 µmol/L) compar...

  15. Biosynthesis of beta-glucosidase by Aspergillus niger a-5

    Energy Technology Data Exchange (ETDEWEB)

    Atev, A.; Panayotov, C.; Bubareva, L.; Benadova, R.; Kolev, E.

    1984-01-01

    Aspergillus niger A-5 produced beta-glucosidase, exocellobihydrolase (C1 enzyme) and endo-1, 4-beta-glucanase (Cx enzyme) in a culture medium containing farm residues of plant origin: wheat straw, ground maize stalks, wheat bran, and micricell as substrates. Maize stalk and wheat bran were the best inducers of the cellulase complex. Intensive aeration stimulated growth and enzyme synthesis. The highest beta-glucosidase activity (54 units/mL) was observed after 96 h of cultivation.

  16. 75 FR 59067 - Airworthiness Directives; International Aero Engines AG V2500-A1, V2522-A5, V2524-A5, V2525-D5...

    Science.gov (United States)

    2010-09-27

    ...) of this AD. Definitions (h) For the purpose of this AD, an engine shop visit is the induction of an... Airworthiness Directives; International Aero Engines AG V2500-A1, V2522-A5, V2524-A5, V2525-D5, V2527-A5, V2527E-A5, V2527M-A5, V2528-D5, V2530-A5, and V2533-A5 Turbofan Engines AGENCY: Federal Aviation...

  17. Lysinuric protein intolerance in a 5-month-old girl

    Directory of Open Access Journals (Sweden)

    Viplav Narayan Deogaonkar

    2016-01-01

    Full Text Available Lysinuric protein intolerance (LPI, also known as cationic aminoaciduria, hyperdibasic aminoaciduria type 2, or familial protein intolerance, is an autosomal recessive defect of diamino acid transport. LPI is characterized by the inability of the body to digest and utilize certain amino acids, namely lysine, arginine, and ornithine. As a result, there is an increased excretion of these amino acids, which in turn affects the liver, the gastrointestinal tract, lungs, immune system, spleen, and organs producing blood. We report a 5-month-old girl born of third degree consanguineous marriage who presented with hepatosplenomegaly with sepsis and worsening jaundice due to LPI.

  18. Comparison of sulfur hexafluoride microbubble (SonoVue)-enhanced myocardial contrast echocardiography with gated single-photon emission computed tomography for detection of significant coronary artery disease: a large European multicenter study.

    Science.gov (United States)

    Senior, Roxy; Moreo, Antonella; Gaibazzi, Nicola; Agati, Luciano; Tiemann, Klaus; Shivalkar, Bharati; von Bardeleben, Stephan; Galiuto, Leonarda; Lardoux, Hervé; Trocino, Giuseppe; Carrió, Ignasi; Le Guludec, Dominique; Sambuceti, Gianmario; Becher, Harald; Colonna, Paolo; Ten Cate, Folkert; Bramucci, Ezio; Cohen, Ariel; Bezante, Gianpaolo; Aggeli, Costantina; Kasprzak, Jaroslaw D

    2013-10-08

    The purpose of this study was to compare sulfur hexafluoride microbubble (SonoVue)-enhanced myocardial contrast echocardiography (MCE) with single-photon emission computed tomography (SPECT) relative to coronary angiography (CA) for assessment of coronary artery disease (CAD). Small-scale studies have shown that myocardial perfusion assessed by SonoVue-enhanced MCE is a viable alternative to SPECT for CAD assessment. However, large multicenter studies are lacking. Patients referred for myocardial ischemia testing at 34 centers underwent rest/vasodilator SonoVue-enhanced flash-replenishment MCE, standard (99m)Tc-labeled electrocardiography-gated SPECT, and quantitative CA within 1 month. Myocardial ischemia assessments by 3 independent, blinded readers for MCE and 3 readers for SPECT were collapsed into 1 diagnosis per patient per technique and were compared to CA (reference standard) read by 1 independent blinded reader. Of 628 enrolled patients who received SonoVue (71% males; mean age: 64 years; >1 cardiovascular [CV] risk factor in 99% of patients) 516 patients underwent all 3 examinations, of whom 161 (31.2%) had ≥70% stenosis (131 had single-vessel disease [SVD]; 30 had multivessel disease), and 310 (60.1%) had ≥50% stenosis. Higher sensitivity was obtained with MCE than with SPECT (75.2% vs. 49.1%, respectively; p SonoVue-enhanced MCE demonstrated superior sensitivity but lower specificity for detection of CAD compared to SPECT in a population with a high incidence of CV risk factors and intermediate-high prevalence of CAD. (A phase III study to compare SonoVue® enhanced myocardial echocardiography [MCE] to single photon emission computerized tomography [ECG-GATED SPECT], at rest and at peak of low-dose Dipyridamole stress test, in the assessment of significant coronary artery disease [CAD] in patients with suspect or known CAD using Coronary Angiography as Gold Standard-SonoVue MCE vs SPECT; EUCTR2007-003492-39-GR). Copyright © 2013 American College of

  19. Project and characteristics of a 5MW experimental fast reactor

    International Nuclear Information System (INIS)

    Ishiguro, Y.; Nascimento, J.A. do.

    1986-05-01

    Characteristics of a 5 MW experimental fast reactor are reported. The reactor is designed with emphasis on fuel and materials irradiation and uses fuel assemblies of a standard structure. The reference core consist of 37 fuel assemblies, each of which contains 19 pins of metallic Pu/Zr fuel. With a core height of 17.6 cm the core volume is 11.4 liter and the central fast (E >=100 KeV) flux is 0.9 x 10 15 n/cm 2 sec. In addition to twelve control rod assemblies with a total reactivity worth of 5.5% Δk, 42 assemblies for reactivity compensation are placed in the two rings outside the core. Replacing these assemblies with driver, blanket, or refletor-shield assemblies, large reactivities can be added to make the central assembly position available for test irradiations and to assure high levels of burnup of driver assemblies. (Author) [pt

  20. Evolution of the thermopsin peptidase family (A5.

    Directory of Open Access Journals (Sweden)

    Neil D Rawlings

    Full Text Available Thermopsin is a peptidase from Sulfolobus acidocaldarius that is active at low pH and high temperature. From reversible inhibition with pepstatin, thermopsin is thought to be an aspartic peptidase. It is a member of the only family of peptidases to be restricted entirely to the archaea, namely peptidase family A5. Evolution within this family has been mapped, using a taxonomic tree based on the known classification of archaea. Homologues are found only in archaeans that are both hyperthermophiles and acidophiles, and this implies lateral transfer of genes between archaea, because species with homologues are not necessarily closely related. Despite the remarkable stability and activity in extreme conditions, no tertiary structure has been solved for any member of the family, and the catalytic mechanism is unknown. Putative catalytic residues have been predicted here by examination of aligned sequences.

  1. Glucose Regulates the Expression of the Apolipoprotein A5 Gene

    Energy Technology Data Exchange (ETDEWEB)

    Fruchart, Jamila; Nowak, Maxime; Helleboid-Chapman, Audrey; Jakel, Heidelinde; Moitrot, Emmanuelle; Rommens, Corinne; Pennacchio, Len A.; Fruchart-Najib, Jamila; Fruchart, Jean-Charles

    2008-04-07

    The apolipoprotein A5 gene (APOA5) is a key player in determining triglyceride concentrations in humans and mice. Since diabetes is often associated with hypertriglyceridemia, this study explores whether APOA5 gene expression is regulated by alteration in glucose homeostasis and the related pathways. D-glucose activates APOA5 gene expression in a time- and dose-dependent manner in hepatocytes, and the glycolytic pathway involved was determined using D-glucose analogs and metabolites. Together, transient transfections, electrophoretic mobility shift assays and chromatin immunoprecipitation assays show that this regulation occurs at the transcriptional level through an increase of USF1/2 binding to an E-box in the APOA5 promoter. We show that this phenomenon is not due to an increase of mRNA or protein expression levels of USF. Using protein phosphatases 1 and 2A inhibitor, we demonstrate that D-glucose regulates APOA5 gene via a dephosphorylation mechanism, thereby resulting in an enhanced USF1/2-promoter binding. Last, subsequent suppressions of USF1/2 and phosphatases mRNA through siRNA gene silencing abolished the regulation. We demonstrate that APOA5 gene is up regulated by D-glucose and USF through phosphatase activation. These findings may provide a new cross talk between glucose and lipid metabolism.

  2. Biosurfactants for microbubble preparation and application

    Science.gov (United States)

    Biosurfactants can be classified by their chemical composition and their origin. This review briefly describes the type of biosurfactants based on their origin. Some of the widely used biosurfactants are introduced. The current statues and future trends in the production of biosurfactants are discus...

  3. Ultrasound acoustic energy for microbubble manipulation

    Science.gov (United States)

    Bakhtiari-Nejad, Marjan; Elnahhas, Ahmed; Jung, Sunghwan; Shahab, Shima

    2017-04-01

    Many bio-medical applications entail the problems of spatially manipulating of bubbles by means of acoustic radiation. The examples are ultrasonic noninvasive-targeted drug delivery and therapeutic applications. This paper investigates the n