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Sample records for blood cell labeling

  1. Blood cell labelling

    International Nuclear Information System (INIS)

    The labelling of blood cells in vitro for subsequent in vivo studies was one of the earliest applications of radioactive tracers in clinical medicine and laid the foundations for many important contributions to the advancement of knowledge of human blood cell pathophysiology. The characteristics required for satisfactory clinical studies, the mechanisms of cell labelling, the problems of radiation or chemical damage to the labelled cells and some examples of modern clinical applications are described and discussed. (Author)

  2. Recent developments in blood cell labeling research

    International Nuclear Information System (INIS)

    A number of recent developments in research on blood cell labeling techniques are presented. The discussion relates to three specific areas: (1) a new in vitro method for red blood cell labeling with /sup 99m/Tc; (2) a method for labeling leukocytes and platelets with /sup 99m/Tc; and (3) the use of monoclonal antibody technique for platelet labeling. The advantages and the pitfalls of these techniques are examined in the light of available mechanistic information. Problems that remain to be resolved are reviewed. An assessment is made of the progress as well as prospects in blood cell labeling methodology including that using the monoclonal antibody approach. 37 refs., 4 figs

  3. The antibody approach of labeling blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, S.C.

    1991-12-31

    Although the science of blood cell labeling using monoclonal antibodies directed against specific cellular antigens is still in its early stages, considerable progress has recently been accomplished in this area. The monoclonal antibody approach offers the promise of greater selectivity and enhanced convenience since specific cell types can be labeled in vivo, thus eliminating the need for complex and damaging cell separation procedures. This article focuses on these developments with primary emphasis on antibody labeling of platelets and leukocytes. The advantages and the shortcomings of the recently reported techniques are criticality assessed and evaluated.

  4. The antibody approach of labeling blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, S.C.

    1992-12-31

    Although the science of blood cell labeling using monoclonal antibodies directed against specific cellular antigens is still in its early stages, considerable progress has recently been accomplished in this area. The monoclonal antibody approach offers the promise of greater selectivity and enhanced convenience since specific cell types can be labeled in vivo, thus eliminating the need for complex and damaging cell separation procedures. This article focuses on these developments with primary emphasis on antibody labeling of platelets and leukocytes. The advantages and the shortcomings of the recently reported techniques are critically assessed and evaluated.

  5. The antibody approach of labeling blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, S.C.

    1991-01-01

    Although the science of blood cell labeling using monoclonal antibodies directed against specific cellular antigens is still in its early stages, considerable progress has recently been accomplished in this area. The monoclonal antibody approach offers the promise of greater selectivity and enhanced convenience since specific cell types can be labeled in vivo, thus eliminating the need for complex and damaging cell separation procedures. This article focuses on these developments with primary emphasis on antibody labeling of platelets and leukocytes. The advantages and the shortcomings of the recently reported techniques are criticality assessed and evaluated.

  6. The antibody approach of labeling blood cells

    International Nuclear Information System (INIS)

    Although the science of blood cell labeling using monoclonal antibodies directed against specific cellular antigens is still in its early stages, considerable progress has recently been accomplished in this area. The monoclonal antibody approach offers the promise of greater selectivity and enhanced convenience since specific cell types can be labeled in vivo, thus eliminating the need for complex and damaging cell separation procedures. This article focuses on these developments with primary emphasis on antibody labeling of platelets and leukocytes. The advantages and the shortcomings of the recently reported techniques are critically assessed and evaluated

  7. The antibody approach of labeling blood cells

    International Nuclear Information System (INIS)

    Although the science of blood cell labeling using monoclonal antibodies directed against specific cellular antigens is still in its early stages, considerable progress has recently been accomplished in this area. The monoclonal antibody approach offers the promise of greater selectivity and enhanced convenience since specific cell types can be labeled in vivo, thus eliminating the need for complex and damaging cell separation procedures. This article focuses on these developments with primary emphasis on antibody labeling of platelets and leukocytes. The advantages and the shortcomings of the recently reported techniques are criticality assessed and evaluated

  8. Current state of the art of blood cell labeling

    International Nuclear Information System (INIS)

    An update on some recent developments in the area of blood cell labeling is provided. Specific topics covered include red cell labeling with /sup 99m/Tc, platelet labeling using an antiplatelet monoclonal antibody, and the labeling of leukocytes with /sup 99m/Tc. Mechanistic information, where available, is discussed. A critical evaluation of current techniques, their pitfalls as well as advantages, and the problems that remain to be resolved, is presented. The promise shown by recent results using the antibody approach for cell labeling is emphasized. An assessment of the progress made in these areas is presented. 38 refs., 10 figs., 6 tabs

  9. Labelling of red blood cells with 99m pertechnetate

    International Nuclear Information System (INIS)

    This paper describes a method for labelling red blood cells with 99mTc in vitro, using electrolytically generated stannous ions as the reducing agent for 99mTc-pertechnetate. A labelling of 95% was found. A method for the in vivo labelling of red blood cells is also reported. This involves an injection of a stanno-DTPA-complex followed 20 minutes later by a 99mTc-pertechnetate solution scintillation camera images show more background activity when the in vivo method of labelling is used

  10. State of the science of blood cell labeling

    International Nuclear Information System (INIS)

    Blood cell labeling can be considered a science in as far as it is based on precise knowledge and can be readily reproduced. This benchmark criterion is applied to all current cell labeling modalities and their relative merits and deficiencies are discussed. Mechanisms are given where they are known as well as labeling yields, label stability, and cell functionality. The focus is on the methodology and its suitability to the clinical setting rather than on clinical applications per se. Clinical results are cited only as proof of efficacy of the various methods. The emphasis is on technetium as the cell label, although comparisons are made between technetium and indium, and all blood cells are covered. 52 refs., 6 figs., 7 tabs

  11. Clinical applications of indium-111-acetylacetone-labelled blood cells

    International Nuclear Information System (INIS)

    A method permitting red-cell labelling with 111In-acetylacetone was reported in 1974 for evaluating intestinal blood loss, the liver-spleen ratio and the red-cell volume. White blood cells can be tagged similarly. In white-cell labelling, simultaneous red-cell or platelet tagging is avoided. Several procedures (dextran separation and gradient centrifugations) have been combined, to develop a highly selective cell separation. In osteomyelitis it may not be as advantageous to use 67Ga-citrate, as in inflammatory soft tissue processes. The detection of inflammatory processes with labelled leukocytes could be of great importance for the scintigraphic diagnosis of osteomyelitidies. A group of 97 patients with suspected osteomyelitis have been examined using 111In-acetylacetone-labelled leukocytes (111In-AAL) immediately following positive routine skeletal scintigraphy. Images obtained 24 h post injection usually were the most satisfactory. In the followup group of 70 patients 21 true positives, 43 true negatives, 21 false negatives and 3 false positives were observed. These findings result in a specificity of 92%, sensitivity of 50% and accuracy of 70% with 111In-AAL for osteomyelitis. Preliminary investigations using 111In-acetylacetone-labelled thrombocytes (111In-AAT) were carried out to detect rejection of transplanted kidneys. The platelets were separated by means of additional special density gradient centrifugations but no dextran from 15-20 ml of autologous whole blood. Scans have been obtained 15 min, 2.5 h and 24 h post injection in an initial group of 10 patients. In acute rejection, a high transplant uptake has been detected, whereas patients without acute rejection showed no or only a minimum activity accumulation. Patients with chronic rejection have intermediate uptakes

  12. Some technetium complexes for labelling red blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Emery, M.F.

    1988-01-01

    A new approach to produce technetium labelled red blood cells, used routinely in diagnostic nuclear medicine, is reported. The enzyme Carbonic Anhydrase (CA), present in erythrocytes, is strongly inhibited by primary aromatic sulphonamides, which bind at the enzyme active site. Three types of ligand able to coordinate to technetium and suitable for modification to include a primary aromatic sulphonamide group were studied; bis(thiosemicarbazones), Schiff bases and some propylene amine oximes. The experimental conditions needed to label the ligands were determined. Both the thiosemicarbazone and propyleneamine oxime derivatives were labelled, but under no conditions attempted were the Schiff bases complexed by Technetium. The two major isozymes of Human Carbonic Anhydrase, HCA I and HCA II, were isolated from blood. The strength of binding of the free ligands SET, PN130 and PN135 with each of the isozymes was measured and expressed as the Dissociation Constant K{sub d}. The rate of uptake of the technetium complexes into washed RBCs and whole blood was measured and found to be much slower in whole blood. The biodistribution of both TcPN130 and TcPN135 in rats was determined and scintigraphic images for the TcPN130 complex were recorded. Attempts to synthesise the Tc-99 analogues on the milligram scale to allow chemical characterisation of these complexes were unsuccessful. (author).

  13. Some technetium complexes for labelling red blood cells

    International Nuclear Information System (INIS)

    A new approach to produce technetium labelled red blood cells, used routinely in diagnostic nuclear medicine, is reported. The enzyme Carbonic Anhydrase (CA), present in erythrocytes, is strongly inhibited by primary aromatic sulphonamides, which bind at the enzyme active site. Three types of ligand able to coordinate to technetium and suitable for modification to include a primary aromatic sulphonamide group were studied; bis(thiosemicarbazones), Schiff bases and some propylene amine oximes. The experimental conditions needed to label the ligands were determined. Both the thiosemicarbazone and propyleneamine oxime derivatives were labelled, but under no conditions attempted were the Schiff bases complexed by Technetium. The two major isozymes of Human Carbonic Anhydrase, HCA I and HCA II, were isolated from blood. The strength of binding of the free ligands SET, PN130 and PN135 with each of the isozymes was measured and expressed as the Dissociation Constant Kd. The rate of uptake of the technetium complexes into washed RBCs and whole blood was measured and found to be much slower in whole blood. The biodistribution of both TcPN130 and TcPN135 in rats was determined and scintigraphic images for the TcPN130 complex were recorded. Attempts to synthesise the Tc-99 analogues on the milligram scale to allow chemical characterisation of these complexes were unsuccessful. (author)

  14. False positive paediatric labelled white blood cell study

    International Nuclear Information System (INIS)

    Full text: An eight-month-old female presented for a technetium labelled white blood cell study (LWBC) to exclude an intra-abdominal abscess. Born premature, the child had surgery to repair a perforated bowel and had repeated presentations with diarrhoea, fevers, a tender right upper quadrant and a raised leucocyte count. Multiple imaging modalities failed to demonstrate recurrent bowel perforation, ischaemia or an intra-abdominal mass. A LWBC study was performed with whole body imaging at 1 and 5 hours post re-injection of the radiolabelled blood. No abnormal uptake was visualised in the abdomen but abnormal white cell accumulation was noted in the right hind foot and the length of the right lower leg. This activity appeared to lie along the course of the right tibia. Plain X-ray demonstrated no evidence of tibial osteomyelitis. Concern that the LWBC may be falsely negative in a patient on antibiotics, a gallium scan was immediately performed to re-examine the abdomen. The whole body gallium images demonstrated normal physiological uptake in the abdomen and no evidence of infection in the right leg. The patient had no clinical features to support right leg pathology. The abnormal LWBC localisation in the right lower leg/foot was therefore falsely positive. The most likely explanation is increased activation of the autologous LWBC by 'rough' handling during difficult venesection and re-injection through small veins and needles/cannulas. The slow flow through the veins draining the foot injection site would contribute to margination in these vessel walls. This is a potential cause for false positive LWBC studies- with significant implications for patient care. Copyright (2002) The Australian and New Zealand Society of Nuclear Medicine Inc

  15. Blood loss estimation in Schistosoma incognitum by the use of 51Cr labelled red cells

    International Nuclear Information System (INIS)

    C42 Red blood cells labelled with 51Cr were used to study the pathophysiology of S. incognitum infection. Blood volume, cell volume, faecal blood excretion and the half life of the red cells were determined. It was shown that in rabbits infected with the blood fluke, there was loss of blood, which may result in the development of anaemia in the infected animals. (author)

  16. Measurement of limb blood flow using technetium-labelled red blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Parkin, A; Robinson, P.J.; Wiggins, P.A.; Leveson, S.H.; Salter, M.C.P.; Matthews, I.F.; Ware, F.M.

    1986-05-01

    A method for measuring blood flow below the knee during reactive hyperaemia induced by 3 min of arterial occlusion has been developed. Subjects are positioned with lower limbs within the field of view of a gamma camera and pneumatic cuffs are placed below the knees to isolate the blood and induce a hyperaemic response. The remaining blood pool is labelled with /sup 99/Tcsup(m)-labelled red cells. Blood flows have been derived from the initial gradients of time-activity curves and from equilibrium blood sampling. The technique has been validated using a tissue-equivalent leg phantom and peristaltic pump. The method has been applied to a small group of patients with peripheral vascular disease and to normal controls. The mean value (+-SD) of limb perfusion for normal controls was found to be 16.4 +- 3.0 ml/100 ml/min and for patients with intermittent claudication was 5.1 +- 2.6 ml/100 ml/min. Flow measurements are found to correlate with clinical findings and with symptoms. Reproducibility (established by repeated measurements) is high. The method is well tolerated even by patients suffering from rest pain.

  17. The measurement of limb blood flow using technetium-labelled red blood cells

    International Nuclear Information System (INIS)

    A method for measuring blood flow below the knee during reactive hyperaemia induced by 3 min of arterial occlusion has been developed. Subjects are positioned with lower limbs within the field of view of a gamma camera and pneumatic cuffs are placed below the knees to isolate the blood and induce a hyperaemic response. The remaining blood pool is labelled with 99Tcsup(m)-labelled red cells. Blood flows have been derived from the initial gradients of time-activity curves and from equilibrium blood sampling. The technique has been validated using a tissue-equivalent leg phantom and peristaltic pump. The method has been applied to a small group of patients with peripheral vascular disease and to normal controls. The mean value (+-SD) of limb perfusion for normal controls was found to be 16.4+-3.0 ml/100 ml/min and for patients with intermittent claudication was 5.1+-2.6 ml/100 ml/min. Flow measurements are found to correlate with clinical findings and with symptoms. Reproducibility (established by repeated measurements) is high. The method is well tolerated even by patients suffering from rest pain. (author)

  18. Safety and radiation risks in the labelling of blood cells

    International Nuclear Information System (INIS)

    Risk in the management of radioactive material and biological exposition to infectious agents. Protocols and normative to observe GOOD RADIOPHARMACY Practices. Main infectious agents that may be transmitted during preparation of a blood cell radiopharmaceutical. Problems of contamination

  19. Tc-99m Labeled HMPAO white Blood Cell Scintigraphy in Pediatric Patients

    OpenAIRE

    Funda Aydın; Arzu Kın Cengiz; Fırat Güngör

    2012-01-01

    Objective: 99mTc labeled hexamethylpropylene amine oxime (HMPAO) white blood cell (WBC) scintigraphy is a frequently used option for acute infection, particularly in pediatric patients. This scintigraphy is applied to detect sites of infection/inflammation in patients with fever of unknown origin, to find and follow up osteomyelitis, and to detect suspicion of acute appendicitis. The aim of this retrospective study was to evaluate the value of 99mTc-HMPAO labeled WBC scintigraphy in pediatric...

  20. Use of indium-111-labeled white blood cells in the diagnosis of diabetic foot infections

    International Nuclear Information System (INIS)

    The diagnosis of bone infection in the patient with nonvirgin bone is a diagnostic dilemma. This is especially true in the diabetic patient with a soft tissue infection and an underlying osteoarthropathy. The authors present a retrospective study using the new scintigraphic technique of indium-111-labeled white blood cells as a method of attempting to solve this diagnostic dilemma

  1. In vitro preparation of radionuclides labeled blood cells: Status and requirements

    International Nuclear Information System (INIS)

    Labelled blood cells permit nuclear medicine imaging using their physiological behaviours. The radiolabeling must be performed in vitro because of the lack of specific markers and requires several highly technical stages of preparation. Labelled blood cells have not the medication drug status, so that the nuclear physician conducting the nuclear test is fully liable. In most cases, the physician delegates the technical responsibility to radio-pharmacists. Although the status of radiolabelled autologous cells is not legally defined and in the absence of a specific repository, it is essential that their preparation is subject to the requirements of the rules of French Good Manufacturing Practice published by Agence francaise de securite sanitaire des produits de sante (Afssaps). It would be desirable to harmonize the practices of radiolabeling cellular blood components by editing a repository. (authors)

  2. Hydrodynamic and label-free sorting of circulating tumor cells from whole blood

    Science.gov (United States)

    Geislinger, Thomas M.; Stamp, Melanie E. M.; Wixforth, Achim; Franke, Thomas

    2015-11-01

    We demonstrate continuous, passive, and label-free sorting of different in vitro cancer cell lines (MV3, MCF7, and HEPG2) as model systems for circulating tumor cells (CTCs) from undiluted whole blood employing the non-inertial lift effect as driving force. This purely viscous, repulsive cell-wall interaction is sensitive to cell size and deformability differences and yields highly efficient cell separation and high enrichment factors. We show that the performance of the device is robust over a large range of blood cell concentrations and flow rates as well as for the different cell lines. The collected samples usually contain more than 90% of the initially injected CTCs and exhibit average enrichment factors of more than 20 for sorting from whole blood samples.

  3. Optimization of [67Ga]-oxinate complex formation conditions for white blood cell labeling

    International Nuclear Information System (INIS)

    In this work, the effective factors on the preparation of 67Ga-oxinate complex for white blood cell labeling were determined. Gallium-67 was produced at AMIRS 30 MeV cyclotron via 68Zn(p,2n)67Ga reaction in the from of 67GaCl3, and was used for radiolabeling of oxinate complex at optimized conditions. A mixture of 67GaCl3 (3uL, 200uCi) and ethanolic oxine solution (lmg/ml, 100μl) was evaporated and reacted at 25degreeC for 1 h in the presence of NaOAc solution (pH. 5.5). ITLC was performed using a mixture of ammonium acetate and methanol solution (1:1) followed by recording the activity using radio thin layer chromatography scanner. The radiochemical purity of %95.18 at these conditions was obtained (specific activity of 1432 GBq/rnmol). Freshly prepared white blood cells were separated from human volunteers and used for labeling by the above mentioned complex at 37dgreeC. 67Ga- oxinate complex due to it's lipophilicity and suitable gamma rays is a suitable cell labeling agent and available for blood stem cell and microorganism studies.

  4. Label-free characterization of white blood cells by measuring 3D refractive index maps

    CERN Document Server

    Yoon, Jonghee; Park, HyunJoo; Choi, Chulhee; Jang, Seongsoo; Park, YongKeun

    2015-01-01

    The characterization of white blood cells (WBCs) is crucial for blood analyses and disease diagnoses. However, current standard techniques rely on cell labeling, a process which imposes significant limitations. Here we present three-dimensional (3D) optical measurements and the label-free characterization of mouse WBCs using optical diffraction tomography. 3D refractive index (RI) tomograms of individual WBCs are constructed from multiple two-dimensional quantitative phase images of samples illuminated at various angles of incidence. Measurements of the 3D RI tomogram of WBCs enable the separation of heterogeneous populations of WBCs using quantitative morphological and biochemical information. Time-lapse tomographic measurements also provide the 3D trajectory of micrometer-sized beads ingested by WBCs. These results demonstrate that optical diffraction tomography can be a useful and versatile tool for the study of WBCs.

  5. Multifocal peritoneal splenosis in Tc-99m-labeled heat-denatured red blood cell scintigraphy

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Min Ki; Hwang, Kyung Hoon; Choe, Won Sick [Gachon University Gil Medical Center, Incheon (Korea, Republic of)

    2006-06-15

    A 44-year-old man with a past medical history of splenectomy came to hospital because of epigastric pain abdominopelvic computed tomography(CT) showed a soft tissue mass and multifocal variable-sized nodules as well as finding suggestive of cholecystitis. Subsequently, he underwent Tc-99m-labeled heat- denatured red blood cell(RBC) scintigraphy to evaluate the mass and nodules. The scintigraphy confirmed multifocal peritoneal splenosis in the abdominopelvic cavity.

  6. Detection of gastritis by /sup 99m/Tc-labeled red-blood-cell scintigraphy

    International Nuclear Information System (INIS)

    Gastritis is a common condition, with a variety of causes, that is diagnosed most often by barium upper gastrointestinal tract series or endoscopy. The authors report a case in which gastritis without active bleeding was apparent in scintiscans obtained during the evaluation of GI bleeding using /sup 99m/Tc-labeled red blood cells (TcRBC). The scintigraphic findings that suggest gastritis are described

  7. Diagnosis of infection by preoperative scintigraphy with indium-labeled white blood cells

    International Nuclear Information System (INIS)

    Scintigraphy with indium-labeled white blood cells has been reported to be sensitive and specific in the diagnosis of low-grade sepsis of the musculoskeletal system. We reviewed the records of fifty patients who had suspected osteomyelitis or suspected infection about a total joint prosthesis and who underwent scintigraphy with technetium-99m methylene diphosphonate and scintigraphy with indium-111 oxine-labeled white blood cells before an open surgical procedure. Any patient who received preoperative antibiotics was not included in the study. For all of the patients, gram-stain examination of smears, evaluation of a culture of material from the operative site, and histological examination were done. The patients were divided into two groups. Group I was composed of twenty-four patients, each of whom had a prosthesis in place and complained of pain. Group II was composed of twenty-six patients for whom a diagnosis of chronic osteomyelitis had to be considered. With the indium scans alone, there was only one false-negative result (in Group II), but there were eighteen false-positive results (eight patients in Group II and ten patients in Group I). Although scintigraphy with indium-labeled white blood cells is quite sensitive, it is not specific in detecting chronic osteomyelitis; a negative scan should be considered highly suggestive that osteomyelitis is not present. Specificity can be increased by interpreting the indium scan in conjunction with the technetium scan

  8. Nifedipine effect on the labelling of blood cells and plasma proteins with Tc-99m

    International Nuclear Information System (INIS)

    The labeling of red blood cells (RBC) with Tc-99m depends on the presence of stannous ion (Sn) that helps this radionuclide's fixation on the hemoglobin molecule. Nifedipine is an agent capable to block a specific way where calcius (Ca) ion acrosses the cellular membrane and to bind itself on plasma proteins. The effect of nifedipine in the labeling of RBC and plasma proteins with Tc-99m was studied because of similarities between Ca and Sn ions. Blood with anticoagulant was treated with nifedipine concentration of 10-6M for 15 min at 370C. The labeling of RBC with Tc-99m was done incubating with Sn ion solution (3 uM) for different times. The % of radioactivity in RBC was determined. Samples of plasma were precipited with trichloroacetic acid and the % of radiocctivity in insoluble fraction was calculated. The same procedure was done using different nifedipine concentrations and the blood was incubated for 60 min with Sn ion. The determination of the % of Tc-99m labeled in RBC and plasma proteins showed that this drug does not have the capability to alter this incorporation because the results are similar to control. It is suggested that the Sn ions passage across RBC is not altered by nifedipine although this drug could bind to plasma protein, it does not modify the Tc-99m fixation on it. (author)

  9. Propanolol, Ciclosporine, Adryamicine, nifedipine and the in vitro labelling of red blood cells with 99mtechnetium

    International Nuclear Information System (INIS)

    Objectives: To evaluate the possible influence of Propanolol, Ciclosporine, Adryamicine and Nifedipine on the labelling in vitro of red blood cells. Materials And Methods: 20 ml of blood were withdrawn from 40 healthy volunteers that have not used drug seven days before of experiments. 2,0ml aliquots of each sample were incubated at 37 deg. C for 30 min with different concentrations of drugs to the two labelling method used. In the simple method 60ml of stannous chloride solution (10,2 m g/ml) were added and the samples centrifuged at 1000g for 5 min and plasma and blood cells were isolated. After that, 2,0ml of saline, 0,2 ml of EDTA (2,2%) and 7,4 MBq of 99mTc were also added. To the hypochlorite method the blood samples were incubated with SnCl2 (10,2m g/ml) for 5 min. After this period of time, 40ml of NaClO solution (1%) and all the reagents mentioned to simple method were added. The samples were centrifuged and labelling yield was calculated to both methods. Conclusions: The analysis of the results shows that using the two methods described there are no significant differences on the in vitro labelling of RBC with 99mTc at the used concentrations of all of these studied drugs. We can speculate that the interferences observed in vivo may be due the presence of active metabolites or interactions among different drugs

  10. Effects of chronic sucralose sweetener on the labeling of blood constituents with technetium-99m, morphology of red blood cells and the biodistribution of sodium pertechnetate in rats

    OpenAIRE

    Gabrielle de Souza Rocha; Marcia de Oliveira Pereira; Mônica Oliveira Benarroz; Jacques Natan Grinapel Frydman; Angélica Beatriz Garcia-Pinto; Mário José Pereira; Adenilson de Souza da Fonseca; Mario Bernardo-Filho

    2008-01-01

    This work evaluates effects of the sweetener with sucralose on the labeling of blood constituents with technetium-99m (99mTc), on the morphology of red blood cells (RBC) and on the biodistribution of sodium pertechnetate in Wistar rats. Animals were treated with sweetener for 8 days. Blood samples were withdrawn and the assay of labeling of blood constituents with 99mTc was performed. Blood cells (BC) and plasma (P) were isolated. Aliquots of BC and P were also precipitated, soluble and insol...

  11. Effect of an Arctium lappa (burdock) extract on the labeling of blood constituents with technetium-99m and on the morphology of the red blood cells

    OpenAIRE

    Rosane de Figueiredo Neves; Silvana Ramos Farias Moreno; Bernardo Machado Rebello; Luiz Querino de Araújo Caldas; Adenilson de Souza da Fonseca; Mario Bernardo-Filho; Aldo da Cunha Medeiros

    2007-01-01

    Arctium lappa (burdock) has been used to treat inflammatory processes. Blood constituents labeled with technetium-99m (99mTc) have been utilized in nuclear medicine. It was evaluated the influence of a burdock extract on the labeling of blood constituents with 99mTc and on the morphometry of red blood cells. Blood samples from Wistar rats were incubated with burdock extract and the radiolabeling procedure was carried out. Plasma and blood cells, soluble and insoluble fractions of plasma and b...

  12. Effect of an Arctium lappa (burdock) extract on the labeling of blood constituents with technetium-99m and on the morphology of the red blood cells

    International Nuclear Information System (INIS)

    Arctium lappa (burdock) has been used to treat inflammatory processes. Blood constituents labeled with technetium-99m (99mTc) have been utilized in nuclear medicine. It was evaluated the influence of a burdock extract on the labeling of blood constituents with 99mTc and on the morphometry of red blood cells. Blood samples from Wistar rats were incubated with burdock extract and the radiolabeling procedure was carried out. Plasma and blood cells, soluble and insoluble fractions of plasma and blood cells were separated. The radioactivity in each fraction was counted and the percentages of radioactivity (%ATI) were determined. Morphology and morphometric (perimeter/area ratio) measurements of red blood cells (RBC) were performed. The incubation with burdock extract significantly (p99mTc obtained in this study. (author)

  13. Autoradiographic study on the incorporation of carbon-14 labeled formate and adenine into nucleic acid in blood-forming cells

    International Nuclear Information System (INIS)

    The incorporation of [14C]formate and [8-14C]adenine into nucleic acid in blood-forming cells was studied by the autoradiographic technique. The isotopic markers were injected subcutaneously into young rats weighting from 100 to 150 g three times every 24 hours and the animals were examined 3 hours after the last injection. In the case of [14C]formate injection, erythroblasts exhibited extremely strong labeling in contrast to weaker labeling of other blood-forming cells. In the case of [14C]adenine administration, on the other hand, immature cells of the granuclocytic series as well as immature reticulum cells (proliferating cells of reticular tissue) were much more heavily labeled than were other blood-forming cells, particularly the erythroblasts which revealed weak or no labeling. By digestion or extraction of DNA, RNA or both from cells with DNase, RNase or hot 10% perchloric acid treatment, respectively, it was confirmed that the observed heavy labeling of any type of cells with either [14C]formate or [14C]adenine was due chiefly to incorporation of the radioactive materials into nuclear DNA. The present results are discussed together with the findings of earlier studies on lymphoid cells which indicate that, in certain cell types, the patterns of [3H]deoxycytidine labeling differ considerably from the corresponding patterns of [3H]deoxycytidine labeling. The present and earlier findings provide evidence to substantiate that, among blood-forming cells, there are considerable variations in the labeling patterns of nuclear DNA depending on differences in the radioactive DNA precursors used as well as in the cell types. (author)

  14. Use of chromium-50 as a label for red blood cells in studies with pregnant women and premature infants

    International Nuclear Information System (INIS)

    A technique is described in which non-radioactive chromium-50 is used as a label for red blood cells in patients for whom radioactive labels are not permissible. The chromium-50, adsorbed on to donor blood in vitro, is infused in the circulatory system and measured, following collection, using neutron activation analyses and a high resolution germanium (lithium) diode gamma-ray spectrometer. The application of this technique to the measurement of blood cell survival time in pregnant women suspected of having haemolytic anaemia and to the measurement of intracranial bleeding in premature infants is described. (author)

  15. Clinical applications of cells labelling

    International Nuclear Information System (INIS)

    Blood cells labelled with radionuclides are reviewed and main applications are described. Red blood cell labelling by both random and specific principle. A table with most important clinical uses, 99mTc labelling of RBC are described pre tinning and in vivo reduction of Tc, in vitro labelling and administration of labelled RBC and in vivo modified technique. Labelled leucocytes with several 99mTc-complex radiopharmaceuticals by in vitro technique and specific monoclonal s for white cells(neutrofiles). Labelled platelets for clinical use and research by in vitro technique and in vivo labelling

  16. Tc-99m Labeled HMPAO white Blood Cell Scintigraphy in Pediatric Patients

    Directory of Open Access Journals (Sweden)

    Funda Aydın

    2012-04-01

    Full Text Available Objective: 99mTc labeled hexamethylpropylene amine oxime (HMPAO white blood cell (WBC scintigraphy is a frequently used option for acute infection, particularly in pediatric patients. This scintigraphy is applied to detect sites of infection/inflammation in patients with fever of unknown origin, to find and follow up osteomyelitis, and to detect suspicion of acute appendicitis. The aim of this retrospective study was to evaluate the value of 99mTc-HMPAO labeled WBC scintigraphy in pediatric patients. Material and Methods: The study was conducted between January 2006 and December 2008 and included 13 patients (5 boys, 8 girls; mean age 6.9±6.2 years. Those patients who had suspicion of bone infection (n=7, fever of unknown origin (n=3, and suspicion of acute appendicitis (n=3 were evaluated retrospectively. 99mTc-HMPAO labeled WBC scintigraphy imaging was performed to all patients. Diagnosis was done according to operation and pathological results or clinical follow-up. Results: 99mTc-HMPAO labeled WBC scintigraphy has been found to be true positive in 6 cases, true negative in 6 cases, and false negative in one patient who had fewer unknown origin. The false negative case has been found to have encephalitis with MRI. Conclusion: Leukocyte scintigraphy has been described as a useful diagnostic tool in the diagnosis of suspicion of bone infection, fever of unknown origin and suspicion of acute appendicitis. 99mTc-HMPAO labeled WBC scintigraphy is a rapid and very accurate method for detecting those pathologies. Our results showed that WBC scintigraphy might be reliably used for diagnosis of suspected bone infection and acute appendicitis, fever of unknown origin, and acute appendicitis, in pediatric patient population. (MIRT 2012;21:13-18

  17. Effect of an Arctium lappa (burdock) extract on the labeling of blood constituents with technetium-99m and on the morphology of the red blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Neves, Rosane de Figueiredo; Rebello, Bernardo Machado; Medeiros, Aldo da Cunha [Universidade Federal do Rio Grande do Norte, Natal, RN (Brazil). Programa de Pos-graduacao em Ciencias da Saude]. E-mail: nevesrosane@yahoo.com.br; Moreno, Silvana Ramos Farias; Fonseca, Adenilson de Souza da [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Inst. de Biologia Roberto Alcantara Gomes. Lab. de Radiofarmacia Experimental; Caldas, Luiz Querino de Araujo [Universidade Federal Fluminense, Niteroi, RJ (Brazil). Programa de Pos-Graduacao em Ciencias Medicas; Bernardo-Filho, Mario [Instituto Nacional do Cancer (INCa), Rio de Janeiro, RJ (Brazil). Coordenadoria de Pesquisa

    2007-09-15

    Arctium lappa (burdock) has been used to treat inflammatory processes. Blood constituents labeled with technetium-99m ({sup 99m}Tc) have been utilized in nuclear medicine. It was evaluated the influence of a burdock extract on the labeling of blood constituents with {sup 99m}Tc and on the morphometry of red blood cells. Blood samples from Wistar rats were incubated with burdock extract and the radiolabeling procedure was carried out. Plasma and blood cells, soluble and insoluble fractions of plasma and blood cells were separated. The radioactivity in each fraction was counted and the percentages of radioactivity (%ATI) were determined. Morphology and morphometric (perimeter/area ratio) measurements of red blood cells (RBC) were performed. The incubation with burdock extract significantly (p<0.05) altered the %ATI on the blood compartments and the perimeter/area ratio of RBC, as well as, induced modifications on the shape of RBC. Alterations on membrane could justify the decrease of labeling of blood cells with {sup 99m}Tc obtained in this study. (author)

  18. Label-free identification of white blood cell using optical diffraction tomography (Conference Presentation)

    Science.gov (United States)

    Yoon, Jonghee; Kim, Kyoohyun; Kim, Min-hyeok; Kang, Suk-Jo; Park, YongKeun

    2016-03-01

    White blood cells (WBC) have crucial roles in immune systems which defend the host against from disease conditions and harmful invaders. Various WBC subsets have been characterized and reported to be involved in many pathophysiologic conditions. It is crucial to isolate a specific WBC subset to study its pathophysiological roles in diseases. Identification methods for a specific WBC population are rely on invasive approaches, including Wright-Gimesa staining for observing cellular morphologies and fluorescence staining for specific protein markers. While these methods enable precise classification of WBC populations, they could disturb cellular viability or functions. In order to classify WBC populations in a non-invasive manner, we exploited optical diffraction tomography (ODT). ODT is a three-dimensional (3-D) quantitative phase imaging technique that measures 3-D refractive index (RI) distributions of individual WBCs. To test feasibility of label-free classification of WBC populations using ODT, we measured four subtypes of WBCs, including B cell, CD4 T cell, CD8 T cell, and natural killer (NK) cell. From measured 3-D RI tomograms of WBCs, we obtain quantitative structural and biochemical information and classify each WBC population using a machine learning algorithm.

  19. Detection of liver hemangiomas by Tc-99m-labeled red blood cells

    International Nuclear Information System (INIS)

    The differentiation of hemangiomas and liver cysts (5-15% in adult population) from single metastases, hepatocellular carcinomas, adenomas and focal nodular hyperplasia is important because of the current aggressive surgical approaches to the treatment of single malignant lesions. Very vascular lesions (focal nodular hyperplasia, some adenomas, some hepatocellular carcinomas, and vascular metastases) show radiological contrast in bolus in the first 30 seconds (the arterial dominant phase). During the delayed contrast enhancement phase (from 30 to 180 seconds), some hepatocellular carcinomas, metastases and occasional hemangiomas may be seen. The accumulation of tagged red blood cells in the hemangioma is much slower than of the contrast, and their activity persists for a much longer time (30 minutes - 2 hours). Data from literature show that hemangiomas larger than 2 cm are detected in 95 %% of cases. In our study we have labeled the red blood cells in vivo with Tc-99m-PYP or Tc-99m-DTPA. Planar scintigrams in several projections are made from 3 to 30 minutes, in matrix 256x8. In 11 of 42 cases one or two very vascular lesions, larger than 2 cm, were found. In four cases the increased activity accumulation was marked as suspect, and in others the increased activity accumulation was not detected. (author)

  20. Determination of the volume of circulating blood by means of in vivo labelled red blood cells with 99mTc pertechnetate and use of a Bulgarian kit

    International Nuclear Information System (INIS)

    A method was proposed for determination of the circulating blood volume (CBV) by means of in vivo labelled red blood cells, which was compared to the routine method with 51Cr-sodium chromate. To the patients concecutively was given 1 g of potassium perchlorate (for blocking of the organs, which actively absorbed the perchnetate ion) and 500 mkg of tin pyrophosphate (Bulgarian kit) with subsequent labelling of the red blood cells with 99mTc-pertechnate (1,8 - 3,7 MBq). The volume of the red blood cells, and hence also CBV, was measured with the use of a modified by the authors formula, in which correction for the individual effectiveness of the cell labelling was done. In comparison with the standard method for in vitro labelling of the red blood cells with 51Cr sodium chromate, the method proposed gave an insignificant difference of 4,16%, but when compared to the commercial tin pyrophosphate (of the firm Mallinckrot - Holland), the Bulgarian kit displayed equivalent qualities. It was concluded that the method has a high accuracy and was easy for execution, cause a low radiation burden of the patient and is suitable for application in nuclear cardiology and radionuclide angiography

  1. Assessment of the effect of phytic acid on the labeling of blood cells and plasma proteins with Technetium-99m

    International Nuclear Information System (INIS)

    Blood elements labeled with technetium-99m (99m Tc) have been used in various procedures in nuclear medicine. We have investigated if phytic acid (PHY) could alter the labeling of blood elements with 99m Tc. Blood was incubated with different concentrations of PHY. Stannous chloride and 99mTc, as sodium pertechnetate, were added. Blood was centrifuged and plasma (P) and blood cell (BC) were isolated. Samples of P and BC were also precipitated with trichloroacetic acid and centrifuged, and insoluble (IF) and soluble (SF) fractions were separated. The percentages of radioactivity (%ATI) in BC, IF-P and IF-BC were calculated. The %ATI decreased significantly (p 99m Tc with possible undesirable effects, it is relevant to verify the necessity to repeat the examination and to evaluate the increase of the radiation dose to the patient. (author)

  2. Relation between clinical mature and immature lymphocyte cells in human peripheral blood and their spatial label free scattering patterns

    Science.gov (United States)

    Zhang, Lu; Zhao, Xin; Zhang, Zhenxi; Zhao, Hong; Chen, Wei; Yuan, Li

    2016-07-01

    A single living cell's light scattering pattern (LSP) in the horizontal plane, which has been denoted as the cell's "2D fingerprint," may provide a powerful label-free detection tool in clinical applications. We have recently studied the LSP in spatial scattering planes, denoted as the cell's "3D fingerprint," for mature and immature lymphocyte cells in human peripheral blood. The effects of membrane size, morphology, and the existence of the nucleus on the spatial LSP are discussed. In order to distinguish clinical label-free mature and immature lymphocytes, the special features of the spatial LSP are studied by statistical method in both the spatial and frequency domains. Spatial LSP provides rich information on the cell's morphology and contents, which can distinguish mature from immature lymphocyte cells and hence ultimately it may be a useful label-free technique for clinical leukemia diagnosis.

  3. Effects of fenoprofen on the labeling of blood constituents with technetium-99m, the morphology of red blood cells and the plasmid

    Energy Technology Data Exchange (ETDEWEB)

    Pereira, Marcia de Oliveira; Rocha, Gabrielle de Souza [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil). Centro de Ciencias da Saude; Lombardi, Simone dos Santos; Santos-Filho, Sebastiao David; Fonseca, Adenilson de Souza da; Bernardo-Filho, Mario [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Instituto de Biologia Roberto Alcantara Gomes. Dept. de Biofisica e Biometria]. E-mail: adenilso@uerj.br; Pereira, Mario Jose [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Dept. de Fisiologia; Geller, Mauro [Centro Universitario Serra dos Orgaos, Teresopolis, RJ (Brazil). Centro de Ciencias da Saude

    2008-12-15

    The aim of this work was to evaluate the effect of fenoprofen on the labeling of blood constituents with technetium- 99m, on the morphology of red blood cells and on the plasmid DNA. Blood samples from Wistar rats were incubated with fenoprofen and the assay of labeling of blood constituents with technetium-99m ({sup 99m}Tc) was performed. Blood cells, plasma, soluble and insoluble fractions of blood cells and plasma were separated. The radioactivity in each fraction was counted and percentage of incorporated radioactivity (%ATI) was determined. Blood smears were prepared, fixed, stained and the qualitative and quantitative morphology of the red blood cells (RBC) was evaluated. Plasmid (pBSK) was incubated with fenoprofen with stannous chloride, and agarose gel electrophoresis procedure was carried out to evaluate genotoxic and the protection of this drug against stannous chloride effect on DNA. In conclusion, under the conditions used in this work, our data suggest that fenoprofen would not affect the fixation of the {sup 99m}Tc on the blood constituents, alter the RBC membrane and present genotoxic and redox effects. (author)

  4. Assessment of the effect of phytic acid on the labeling of blood cells and plasma proteins with Technetium-99m

    Energy Technology Data Exchange (ETDEWEB)

    Lima-Filho, Guilherme L.; Freitas, Rosimeire S.; Moreno, Silvana R.F.; Boasquevisque, Edson M.; Bernardo-Filho, Mario [Universidade do Estado, Rio de Janeiro, RJ (Brazil). Inst. de Biologia. Dept. de Biofisica e Biometria]. E-mail: gllf@hotmail.com; Lima, Glaydes M.T. [Pernambuco Univ., Recife, PE (Brazil). Hospital das Clinicas; Catanho, Maria T.J.A. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Biofisica e Radiobiologia

    2002-07-01

    Blood elements labeled with technetium-99m ({sup 99m} Tc) have been used in various procedures in nuclear medicine. We have investigated if phytic acid (PHY) could alter the labeling of blood elements with {sup 99m} Tc. Blood was incubated with different concentrations of PHY. Stannous chloride and {sup 99m}Tc, as sodium pertechnetate, were added. Blood was centrifuged and plasma (P) and blood cell (BC) were isolated. Samples of P and BC were also precipitated with trichloroacetic acid and centrifuged, and insoluble (IF) and soluble (SF) fractions were separated. The percentages of radioactivity (%ATI) in BC, IF-P and IF-BC were calculated. The %ATI decreased significantly (p < 0.05) in BC (95.08 {+-}1.94 to 80.68 {+-} 3.35), in IF-P (74.42 {+-}4.50 to 39.94{+-} 5.51) and in IF-BC (89.91{+-} 3.91 to 79.54 {+-} 5.42) in presence of PHY. These results suggest that the chelating property of PHY can modify the labeling of the BC, although other effects of PHY could be responsible. As PHY is found in many food and it could alter the labeling of blood elements with {sup 99m} Tc with possible undesirable effects, it is relevant to verify the necessity to repeat the examination and to evaluate the increase of the radiation dose to the patient. (author)

  5. Placental localization in abdominal pregnancy using technetium-99m-labeled red blood cells

    International Nuclear Information System (INIS)

    In a patient with third trimester abdominal pregnancy with fetal demise, technetium-99m-labeled erythrocytes (99mTc-RBCs) localized the placenta preoperatively, after nonvisualization by ultrasonography and arteriography. Extrauterine placental localization by blood-pool imaging may be useful when ultrasound fails

  6. In vivo/in vitro labeling of red blood cells with sup(99m)Tc and clinical applications

    International Nuclear Information System (INIS)

    A reliable and stabile in vivo/in vitro labeling technique of red blood cells (RBC) is described. The patients are injected 20% of the content of an unlabeled kit used for bone scintigraphy (TechneScan PYP, Byk-Mallinckrodt). 15 minutes later 3 ml blood are sampled in a heparinized syringe. The blood is incubated together with 30-40 mCi (1-1.5 GBq) sup(99m)Tc for 10 minutes in a water bath at 35-370C. After centrifugation at 500 g a dose of 15-25 mCi (0.6-1 GBq) sup(99m)Tc labeled RBC may be withdrawn in a volume of 1-1.5 ml. Mean labeling efficiency is 88%, without using the first eluat of a Tc-generator the yield is as high as 92%. Due to the small volume, the labeled RBC may be reinjected as bolus and first pass radionuclide angiocardiography can be performed. Using labeled RBC, scintigraphy of the intravasal space is possible up to 20 hours without deterioration in contrast or accumulation of radioactivity in the extravasal space or in other organs. Evaluation of heart function can be performed up to 10 hours. In addition, labeled RBC are useful in detecting unknown gastrointestinal bleeding. (orig.)

  7. Indium-111 labelled white blood cell scintigraphy in cranial and spinal septic lesions

    Energy Technology Data Exchange (ETDEWEB)

    Medina, M.; Lucano, A. [Div. of Neurosurgery, S. Croce e Carle Hospital, Cuneo (Italy); Viglietti, A.L.; Camuzzini, G. [Service of Nuclear Medicine, S. Croce e Carle Hospital, Cuneo (Italy); Gozzoli, L. [Service of Neuroradiology, S. Croce e Carle Hospital, Cuneo (Italy); Ravasi, L.; Lucignani, G. [INB-CNR, Univ. of Milan, H San Raffaele, Milan (Italy)

    2000-10-01

    Cranial and spinal infections are severe events that require timely diagnosis and treatment. Physical and neurological examination, laboratory tests and radiological imaging may be insufficient for assessing cranial and spinal septic lesions. This study aimed to evaluate the accuracy of indium-111 white blood cell (WBC) scan in assessing the presence of leucocytes in intracranial and spinal lesions, and in the diagnosis, management and follow-up of primary, post-traumatic and post-surgical infections. One hundred and twenty-four subjects were included in the study (48 with post-traumatic or post-surgical lesions, 73 with primary cerebral lesions, and 3 with spinal lesions). All patients underwent a diagnostic work-up including planar scans with {sup 111}In-labelled WBCs, at 4 and 24 h post tracer injection. All subjects underwent surgical treatment. Patients who did not recover from the infection as suggested by clinical evolution underwent further treatment (up to three times) and further WBC scans (up to four times). WBC scintigraphy correctly identified all the areas of leucocyte accumulation, as confirmed after surgery. WBC scintigraphy also correctly excluded the presence of leucocytes in all other lesions, as demonstrated at surgery. The results of this study confirm the accuracy of WBC scan for the assessment of patients with cranial and spinal lesions, in whom the demonstration of leucocyte accumulation can ease the diagnosis of infection, and indicate that the method is also accurate for the follow-up and management of neurosurgical patients. (orig.)

  8. Indium-111 labelled white blood cell scintigraphy in cranial and spinal septic lesions

    International Nuclear Information System (INIS)

    Cranial and spinal infections are severe events that require timely diagnosis and treatment. Physical and neurological examination, laboratory tests and radiological imaging may be insufficient for assessing cranial and spinal septic lesions. This study aimed to evaluate the accuracy of indium-111 white blood cell (WBC) scan in assessing the presence of leucocytes in intracranial and spinal lesions, and in the diagnosis, management and follow-up of primary, post-traumatic and post-surgical infections. One hundred and twenty-four subjects were included in the study (48 with post-traumatic or post-surgical lesions, 73 with primary cerebral lesions, and 3 with spinal lesions). All patients underwent a diagnostic work-up including planar scans with 111In-labelled WBCs, at 4 and 24 h post tracer injection. All subjects underwent surgical treatment. Patients who did not recover from the infection as suggested by clinical evolution underwent further treatment (up to three times) and further WBC scans (up to four times). WBC scintigraphy correctly identified all the areas of leucocyte accumulation, as confirmed after surgery. WBC scintigraphy also correctly excluded the presence of leucocytes in all other lesions, as demonstrated at surgery. The results of this study confirm the accuracy of WBC scan for the assessment of patients with cranial and spinal lesions, in whom the demonstration of leucocyte accumulation can ease the diagnosis of infection, and indicate that the method is also accurate for the follow-up and management of neurosurgical patients. (orig.)

  9. Effect of Peumus boldus on the labeling of red blood cells and plasma proteins with Technetium-99m

    International Nuclear Information System (INIS)

    Peumus boldus is used in popular medicine in Brazil. The influence of Peumus boldus on the labeling of red blood cells and plasma proteins with 99mTc was studied. Stannous chloride and 99mTc pertechnetate were incubated with blood and a tincture of Peumus boldus. Aliquots of plasma and blood cells were isolated from the mixture and treated with trichloroacetic acid (TCA). After separation, analysis of the soluble and insoluble fractions showed a rapid uptake of the radioactivity by blood cells in the presence of the drug, whereas there was a slight decrease in the amount of 99mTc radioactivity in the TCA-insoluble fraction of plasma

  10. Effect of Peumus boldus on the labeling of red blood cells and plasma proteins with Technetium-99m

    Energy Technology Data Exchange (ETDEWEB)

    Wancke Reiniger, Ingrid; Fonseca de Oliveira, Joelma; Caldeira-de-Araujo, Adriano [Departamento de Biofisica e Biometria, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ (Brazil); Bernardo-Filho, Mario [Instituto Nacional de Cancer, Centro de Pesquisa Basica, Rio de Janeiro, RJ (Brazil)

    1999-08-01

    Peumus boldus is used in popular medicine in Brazil. The influence of Peumus boldus on the labeling of red blood cells and plasma proteins with {sup 99m}Tc was studied. Stannous chloride and {sup 99m}Tc pertechnetate were incubated with blood and a tincture of Peumus boldus. Aliquots of plasma and blood cells were isolated from the mixture and treated with trichloroacetic acid (TCA). After separation, analysis of the soluble and insoluble fractions showed a rapid uptake of the radioactivity by blood cells in the presence of the drug, whereas there was a slight decrease in the amount of {sup 99m}Tc radioactivity in the TCA-insoluble fraction of plasma.

  11. Effect of Peumus boldus on the labeling of red blood cells and plasma proteins with technetium-99m.

    Science.gov (United States)

    Reiniger, I W; de Oliveira, J F; Caldeira-de-Araújo, A; Bernardo-Filho, M

    1999-08-01

    Peumus boldus is used in popular medicine in Brazil. The influence of Peumus boldus on the labeling of red blood cells and plasma proteins with 99mTc was studied. Stannous chloride and 99mTc pertechnetate were incubated with blood and a tincture of Peumus boldus. Aliquots of plasma and blood cells were isolated from the mixture and treated with trichloroacetic acid (TCA). After separation, analysis of the soluble and insoluble fractions showed a rapid uptake of the radioactivity by blood cells in the presence of the drug, whereas there was a slight decrease in the amount of 99mTc radioactivity in the TCA-insoluble fraction of plasma. PMID:10376326

  12. Influence of Momordica charantia L. on the red and white blood cells labeling with 99mTc

    International Nuclear Information System (INIS)

    Full text: Momordica charantia L. is popularly known in Brazil as bitter melon and it's commonly used to treat several diseases as cancer, diabetes and to heal skin injuries. Many papers have been published showing the potential radio pharmacological activity of this plant due to its linkage with 99mTc through some protein fractions of the extract. In this study, it was evaluated the influence of Momordica charantia L extract , labeling ( in vitro) of blood elements with sodium pertechnetate (Na 99mTcO4). In the labeling of red blood cells (in vitro), blood samples were obtained from Wistar rats and incubated with different concentrations of M. charantia, for control group was used NaCl 0.9% and added stannous chloride (SnCl2) and 99mTc. The plasma fractions (P) and the cells (C) were separated and, also, precipitated with trichloroacetic acid at 5%, obtaining the soluble (SF) and insoluble (IF) fractions. The radioactivity rate (%ATl) of each fraction was calculated. The same methodology was applied for white blood cells but these cells were separated in advance by centrifugation at 1800 rpm during 15 minutes. There weren't alterations in the labeling of red blood cells in the concentrations tested of the extract when compared with the rate of the control group neither in the insoluble fractions. However, on the white blood cells it was noticed an increase in 99mTc uptake in the presence of M. charantia extract. So its possible to conclude, based on previous results obtained by our group, that the M. charantia L. could be used to evaluate inflammatory processes. (author)

  13. Investigations into the use of radiolabeled monoclonal antibodies for selective cell labeling in whole blood: Progress report, January 1986-December 1986

    International Nuclear Information System (INIS)

    Techniques to effectively labeling blood cell specific monoclonal antibodies (MAbs), with In-111 using bifunctional chelating agents such as the cyclic or mixed anhydride of diethylenetriaminepentaacetic acid have been developed. Conditions for the optimal labeling of blood cells are reported. The specificity (Kd values) of In-111 labeled MAbs for the cell specific antigens are reported. In-111-MAb labeled canine platelets were used to image vascular thrombi. Under the best experimental conditions approximately 70% of the radiolabeled MAbs get incorporated into isolated blood cells. However, only about 35% of the added MAb-associated radioactivity would incorporate into the desired type of blood cells in whole blood. The chemical nature of the radioactivity that does not bind to the cells was investigated. 9 figs., 6 tabs

  14. Rapid and label-free separation of Burkitt's lymphoma cells from red blood cells by optically-induced electrokinetics.

    Directory of Open Access Journals (Sweden)

    Wenfeng Liang

    Full Text Available Early stage detection of lymphoma cells is invaluable for providing reliable prognosis to patients. However, the purity of lymphoma cells in extracted samples from human patients' marrow is typically low. To address this issue, we report here our work on using optically-induced dielectrophoresis (ODEP force to rapidly purify Raji cells' (a type of Burkitt's lymphoma cell sample from red blood cells (RBCs with a label-free process. This method utilizes dynamically moving virtual electrodes to induce negative ODEP force of varying magnitudes on the Raji cells and RBCs in an optically-induced electrokinetics (OEK chip. Polarization models for the two types of cells that reflect their discriminate electrical properties were established. Then, the cells' differential velocities caused by a specific ODEP force field were obtained by a finite element simulation model, thereby established the theoretical basis that the two types of cells could be separated using an ODEP force field. To ensure that the ODEP force dominated the separation process, a comparison of the ODEP force with other significant electrokinetics forces was conducted using numerical results. Furthermore, the performance of the ODEP-based approach for separating Raji cells from RBCs was experimentally investigated. The results showed that these two types of cells, with different concentration ratios, could be separated rapidly using externally-applied electrical field at a driven frequency of 50 kHz at 20 Vpp. In addition, we have found that in order to facilitate ODEP-based cell separation, Raji cells' adhesion to the OEK chip's substrate should be minimized. This paper also presents our experimental results of finding the appropriate bovine serum albumin concentration in an isotonic solution to reduce cell adhesion, while maintaining suitable medium conductivity for electrokinetics-based cell separation. In short, we have demonstrated that OEK technology could be a promising tool for

  15. Effect of an extract of Artemisia vulgaris L. (Mugwort) on the in vitro labeling of red blood cells and plasma proteins with technetium-99m

    International Nuclear Information System (INIS)

    The aim of this work was to evaluate the effect of an extract of the Artemisia vulgaris L. (mugwort) on the labeling of blood constituents with technetium-99m (99mTc). Blood samples from Wistar rats were incubated with a mugwort extract and the radiolabeling of blood constituents was carried out. Plasma and blood cells were separated by centrifugation. Aliquots of plasma and blood cells were also precipitated with trichloroacetic acid and centrifuged to isolate soluble and insoluble fractions of plasma and blood cells. Radioactivity in each fraction was counted and the percentages of radioactivity (%ATI) was calculated. Mugwort extract decreased significantly (p<0.05) the %ATI on the blood compartments and on the blood cells proteins (insoluble fraction). The analysis of the results indicates that the extract could have substances that could interfere on the transport of stannous through the erythrocyte membrane altering the labeling of blood cells with 99mTc. (author)

  16. Localization of lower gastrointestinal hemorrhage. Experience with red blood cells labeled in vitro with technetium Tc 99m

    International Nuclear Information System (INIS)

    Seventy-six patients clinically suspected of having lower gastrointestinal bleeding were studied by scintigraphy utilizing red blood cells labeled in vitro with technetium Tc 99m. Sixteen patients required emergency surgery; bleeding was accurately localized in 15 (94%). One patient (6%) had a normal scan. A 20-month mean follow-up of the 16 patients showed no recurrent bleeding. Of 60 patients not requiring emergency surgery, bleeding was localized in 11, but the bleeding ceased. Forty-nine of the 60 patients had normal scans and had no further hemorrhaging during hospitalization. A 21-month mean follow-up of 38 of the 49 patients showed no further bleeding episodes or surgical procedures in 29 patients; however, eight patients required surgical procedures, including seven for gastrointestinal malignancies. Scanning of red blood cells labeled in vitro with 99mTc is accurate and efficacious in localization of bleeding sites that require emergency surgery for lower gastrointestinal hemorrhage

  17. Histochemical evidence for the differential surface labeling, uptake, and intracellular transport of a colloidal gold-labeled insulin complex by normal human blood cells

    International Nuclear Information System (INIS)

    A colloidal gold-labeled insulin-bovine serum albumin (GIA) reagent has been developed for the ultrastructural visualization of insulin binding sites on the cell surface and for tracing the pathway of intracellular insulin translocation. When applied to normal human blood cells, it was demonstrated by both visual inspection and quantitative analysis that the extent of surface labeling, as well as the rate and degree of internalization of the insulin complex, was directly related to cell type. Further, the pathway of insulin (GIA) transport via round vesicles and by tubulo-vesicles and saccules and its subsequent fate in the hemic cells was also related to cell variety. Monocytes followed by neutrophils bound the greatest amount of labeled insulin. The majority of lymphocytes bound and internalized little GIA, however, between 5-10% of the lymphocytes were found to bind considerable quantities of GIA. Erythrocytes rarely bound the labeled insulin complex, while platelets were noted to sequester large quantities of the GIA within their extracellular canalicular system. GIA uptake by the various types of leukocytic cells appeared to occur primarily by micropinocytosis and by the direct opening of cytoplasmic tubulo-vesicles and saccules onto the cell surface in regions directly underlying surface-bound GIA. Control procedures, viz., competitive inhibition of GIA labeling using an excess of unlabeled insulin in the incubation medium, preincubation of the GIA reagent with an antibody directed toward porcine insulin, and the incorporation of 125I-insulin into the GIA reagent, indicated the specificity and selectivity of the GIA histochemical procedure for the localization of insulin binding sites

  18. Effects of chronic sucralose sweetener on the labeling of blood constituents with technetium-99m, morphology of red blood cells and the biodistribution of sodium pertechnetate in rats

    Energy Technology Data Exchange (ETDEWEB)

    Rocha, Gabrielle de Souza; Pereira, Marcia de Oliveira; Frydman, Jacques Natan Grinapel [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil). Centro de Ciencias da Saude; Benarroz, Monica de Oliveira; Garcia-Pinto, Angelica Beatriz; Fonseca, Adenilson de Souza da; Bernardo-Filho, Mario [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Instituto de Biologia Roberto Alcantara Gomes. Dept. de Biofisica e Biometria]. E-mail: adenilso@uerj.br; Pereira, Mario Jose [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Dept. de Fisiologia

    2008-12-15

    This work evaluates effects of the sweetener with sucralose on the labeling of blood constituents with technetium- 99m ({sup 99m}Tc), on the morphology of red blood cells (RBC) and on the biodistribution of sodium pertechnetate in Wistar rats. Animals were treated with sweetener for 8 days. Blood samples were withdrawn and the assay of labeling of blood constituents with {sup 99m}Tc was performed. Blood cells (BC) and plasma (P) were isolated. Aliquots of BC and P were also precipitated, soluble and insoluble fractions separated. The radioactivity in each fraction was counted and percentage of incorporated radioactivity (%ATI) determined. Blood smears were prepared, fixed, stained and the qualitative and quantitative morphology of the RBC was evaluated under optical microscopy. In biodistribution experiments, sodium pertechnetate was administrated, organs and tissues isolated, radioactivity was counted and percentage of incorporated radioactivity per gram (%ATI/g) determined. The data showed no significant alterations in %ATI, morphology of RBC and in %ATI/g in the studied organs. (author)

  19. Effects of chronic sucralose sweetener on the labeling of blood constituents with technetium-99m, morphology of red blood cells and the biodistribution of sodium pertechnetate in rats

    International Nuclear Information System (INIS)

    This work evaluates effects of the sweetener with sucralose on the labeling of blood constituents with technetium- 99m (99mTc), on the morphology of red blood cells (RBC) and on the biodistribution of sodium pertechnetate in Wistar rats. Animals were treated with sweetener for 8 days. Blood samples were withdrawn and the assay of labeling of blood constituents with 99mTc was performed. Blood cells (BC) and plasma (P) were isolated. Aliquots of BC and P were also precipitated, soluble and insoluble fractions separated. The radioactivity in each fraction was counted and percentage of incorporated radioactivity (%ATI) determined. Blood smears were prepared, fixed, stained and the qualitative and quantitative morphology of the RBC was evaluated under optical microscopy. In biodistribution experiments, sodium pertechnetate was administrated, organs and tissues isolated, radioactivity was counted and percentage of incorporated radioactivity per gram (%ATI/g) determined. The data showed no significant alterations in %ATI, morphology of RBC and in %ATI/g in the studied organs. (author)

  20. Tc-99m Labeled Red Blood Cell by Ultra Tag RBC Kit in Patients Suspected of Gastrointestinal Bleeding

    International Nuclear Information System (INIS)

    Twenty patients suspected of gastrointestinal bleeding who underwent Tc-99m labeled red blood cell (RBC) by ultraTag RBC kit at Division of Nuclear Medicine, Bumrungrad Hospital between January 2000 and December 2002 were studied. The histories of patients together with either endoscopic results or angiographic findings or pathological reports were used as gold standards. Two by Two decision matrix was used for data analysis and the sensitivity together with specificity were calculated. The results show that the sensitivity and specificity of Tc-99m labeled RBC by ultraTag RBC kit are 87.5% and 91.7%, respectively. We conclude that Tc-99m labeled RBC by ultraTag RBC kit gives high percentages of sensitivity and specificity. Moreover, the image quality is improved because of the absence of free Tc-99m pertechnetate uptake in the stomach in all patients

  1. Recurrent gastrointestinal bleeding diagnosed by delayed scintigraphy with Tc-99m-labeled red blood cells.

    Science.gov (United States)

    Nwakanma, Lois; Meyerrose, Gary; Kennedy, Shalyn; Rakvit, Ariwan; Bohannon, Todd; Silva, Micheal

    2003-08-01

    A 56-year-old woman presented with bright-red blood from the rectum. Esophagogastroduodenoscopy revealed mild gastritis. Colonoscopy demonstrated diverticulosis without active bleeding, and in vitro tagged red blood cell scintigraphy was unremarkable. There was no further evidence of bleeding and the patient was discharged home. The patient returned with recurrent bright-red blood from the rectum. Although delayed scintigraphic images seldom demonstrate the site of bleeding, delayed images at 12 hours demonstrated active bleeding near the hepatic flexure in this patient. This was confirmed with selective mesenteric angiography, and was treated with coil embolization of the tertiary branches of the right middle colic artery. PMID:12897664

  2. Morphologic alterations on red blood cells labeled with technetium-99m: the effect of Mentha crispa L. (hortela) extract

    International Nuclear Information System (INIS)

    The use of natural products, as medicinal plants, is very frequent in the world. Mentha crispa L. (M. crispa) is utilized in herbal medicine. Blood elements labeled with technetium-99m (99mTc) are used in nuclear medicine procedures and this labeling process may be altered by drugs. We have investigated the possibility of M. crispa extract being capable to alter the labeling of blood elements with 99mTc. Blood was incubated with M. crispa extract in various concentrations (6.25, 12.5, 25, 50 and 100%). Stannous chloride solution and Tc-99m, as sodium pertechnetate, were added. Blood was centrifuged and plasma (P) and blood cells (BC) were isolated. Samples of P and BC were also precipitated, centrifuged and insoluble (IF) and soluble (SF) separated. The percentage of radioactivity (%ATI) in BC, IF-P and IF-BC was calculated. Histological evaluations of the red blood cells (RBC) were performed with blood samples treated with various concentrations of M. Crispa L. and the morphology of the RBC was observed under optical microscope. Important morphological alterations expressed by mean of the perimeter/area of the RBC treated with M. crispa: 6.25% (0.67 ± 0.02), 12.5% (0.77 ± 0.03), 25% (0.73 ± 0.04), 50% (0.76 ± 0.04), 100% (0.69 ± 0.08) and the control cells (0.67 ± 0.05). The %ATI decreased: (i) on BC from 97.3 ± 1.92 to 60.0 ± 2.44; (ii) on IF-P from 74.8 ± 3.78 to 9.99 ± 3.61; (iii) on IF-BC from 88.6 ± 5.41 to 58.4 ± 11.55. The perimeter/area of the RBC showed significant differences (P>0.01) when compared 6.25% and 12.5%, and when compared 6.25% and 50% of M. Crispa L. extract. These findings could also justify the decrease of the labeling of BC with 99mTc in presence of M. Crispa extract

  3. On Orbit Immuno-Based, Label-Free, White Blood Cell Counting System with MicroElectroMechanical Sensor (MEMS) Technology (OILWBCS-MEMS) Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Aurora Flight Sciences Corporation and partner, Draper Laboratory, propose to develop an on-orbit immuno-based label-free white blood cell counting system using...

  4. On Orbit Immuno-Based, Label-Free, White Blood Cell Counting System with MicroElectroMechanical Sensor (MEMS) Technology (OILWBCS-MEMS) Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Aurora Flight Sciences Corporation and our partner, Draper Laboratory, propose to develop an on orbit immuno-based, label-free, white blood cell counting system for...

  5. Label-Free Detection of Rare Cell in Human Blood Using Gold Nano Slit Surface Plasmon Resonance

    Directory of Open Access Journals (Sweden)

    Mansoureh Z. Mousavi

    2015-03-01

    Full Text Available Label-free detection of rare cells in biological samples is an important and highly demanded task for clinical applications and various fields of research, such as detection of circulating tumor cells for cancer therapy and stem cells studies. Surface Plasmon Resonance (SPR as a label-free method is a promising technology for detection of rare cells for diagnosis or research applications. Short detection depth of SPR (400 nm provides a sensitive method with minimum interference of non-targets in the biological samples. In this work, we developed a novel microfluidic chip integrated with gold nanoslit SPR platform for highly efficient immunomagnetic capturing and detection of rare cells in human blood. Our method offers simple yet efficient detection of target cells with high purity. The approach for detection consists of two steps. Target cells are firs captured on functionalized magnetic nanoparticles (MNPs with specific antibody I. The suspension containing the captured cells (MNPs-cells is then introduced into a microfluidic chip integrated with a gold nanoslit film. MNPs-cells bind with the second specific antibody immobilized on the surface of the gold nanoslit and are therefore captured on the sensor active area. The cell binding on the gold nanoslit was monitored by the wavelength shift of the SPR spectrum generated by the gold nanoslits.

  6. Failure to label red blood cells adequately in daily practice using an in vivo method: methodological and clinical considerations

    International Nuclear Information System (INIS)

    This study was conducted to evaluate the frequency and possible causes of poor red blood cell (RBC) labelling when performing equilibrium gated blood pool (GBP) radionuclide angiography at rest with an in vivo method. The influence of the mode of administration on tagging efficiency was studied. The patients were subclassified into four groups according to the way both molecules involved in the tagging procedure were administered. When poor image quality was found, the labelling efficiency was quantified and the frequency of failed tagging in each group was calculated. A significant association was found between poor labelling and the use of a Teflon catheter or butterfly needle for the injection of the stannous agent. In another 737 patients a strict administration protocol was applied to analyse the frequency of poor tagging and its possible causes. Suboptimal image quality was present in 88 patients. Quantitatively confirmed poor tagging was present in 36 of the 88; the remaining 52 patients showed borderline normal labelling. Drug interference was studied by comparing the medications used by the 36 patients showing poor binding with those used by 44 control patients. A significant relationship was found between the use of heparin or chemotherapy and the tagging. A significant correlation was found between advanced age and poor labelling efficiency. Finally, in 36 patients with poor labelling, a second GBP test was performed. This allowed us to evaluate the accuracy of the obtained ejection fraction value when a suboptimal image set is used, and to assess the feasibility of using the new kit in daily practice. (orig.)

  7. The appearance of Paget's disease on a 99mTc-HMPAO labelled white blood cell scan

    International Nuclear Information System (INIS)

    Full text: A 73 year old female presented with pyrexia of unknown origin despite the use of antibiotics. She recently had a colonoscopic stent inserted to resolve a large bowel obstruction secondary to stage IV sigmoid colon cancer. The procedure complicated by a perforation of the sigmoid colon and the need for Hartmann's procedure. Urine and blood cultures v both clear however the white cell count was elevated. X rays of the chest and abdomen and CT of the abdomen and pe showed no sites of infection. A 99mTc-HMPAO labelled white blood cell scan was performed to localise the site of infection. The whole body and SPECT images showed patchy uptake consistent with the known liver metastases. Reduced bone marrow uptake was seen in the left hemipelvis and a patchy appearance in the right hemipelvis. The appearance of osteomyelitis could not be confirmed and a bone scan was performed. The blood pool images demonstrated hyperaemia in the region of the upper sacrum and superior iliac crests. The delayed images confirmed the presence of Paget's disease involving the right and left iliac crests and the sacrum. CT confirmed these findings. This case demonstrates that conditions such as Paget's disease, which displaces the marrow, may cause reduced uptake on a labelled white cell scan. The cause may be confirmed with other techniques, such as bone scans and x ray.

  8. Splenic sequestration of Tc-99m labeled heat treated red blood cells. [Feasibility of use for spleen imaging

    Energy Technology Data Exchange (ETDEWEB)

    Atkins, H L; Goldman, A G; Fairchild, R G; Oster, Z H; Som, P; Richards, P; Meinken, G E; Srivastava, S C

    1979-01-01

    The rate of blood clearance and spleen uptake as well as the total spleen uptake of heat damaged red blood cells labeled with Tc-99m was determined in eight patients, six of whom had chronic lymphatic leukemia, one had polycythemia vera and one had eosinophilia of unknown origin. Spleen uptake at 2 hrs was 72.0 +- 18.5%. Approximately 82.6% of the initial radioactivity was cleared from the blood by 2 hrs with a rapid T 1/2 component of 6.3 +- 4.7 min. The T 1/2 of splenic uptake was 8.3 +- 4.6 min with a plateauing of splenic radioactivity by 30 min. The preliminary results indicate that the method of preparation is reliable but the usefulness of the method for evaluating spleen function remains to be determined.

  9. Paramagnetic Gd(3+) labeled red blood cells for magnetic resonance angiography.

    Science.gov (United States)

    Aryal, Santosh; Stigliano, Cinzia; Key, Jaehong; Ramirez, Maricela; Anderson, Jeff; Karmonik, Christof; Fung, Steve; Decuzzi, Paolo

    2016-08-01

    Despite significant advances in contrast enhanced-magnetic resonance angiography, the lack of truly blood-pool agents with long circulating property is limiting the clinical impact of this imaging technique. The terminal half-life for blood elimination of most small molecular weight gadolinium (Gd) based extracellular fluid agents is about 1.5 h when administered intravenously to subjects with normal renal function. The small size of these extracellular fluid agents does not prevent them from extravasating, especially from damaged vessels which are generally hyperpermeable. Therefore, the development of novel, clinically relevant blood pool contrast agents is critically needed to improve outcomes in the prevention, detection, and treatment of vascular diseases. We have demonstrated the fusion strategies in which the Gd-liposome without any stealth property radically fuses with red blood cells (RBCs) forming MR glowing Gd-RBC with the order of magnitude enhancements in circulation half-life (t1/2 = 50 h) and r1 relaxivity (r1 = 19.0 mM(-1) s(-1)) of Gd. The in vivo contrast enhancement of Gd-RBC was studied by using 3T clinical MR scanner for extended period of time, which clearly visualized the abdominal aorta. In summary, the vascular delivery of blood pool agents may benefit from carriage by RBCs because it naturally stays within the vascular lumen. PMID:27192419

  10. Detection of acute gastrointestinal bleeding by means of technetium-99m in vivo labelled red blood cells

    International Nuclear Information System (INIS)

    Prognosis of gastrointestinal (GI) bleeding depends on the timely and accurate detection of the source of bleeding and sequential surgical or endoscopy therapy. Scintigraphy with red blood cells (RBCs) in vivo labelled by means of technetium-99m hastened detection of source of GI bleeding and improved management of the particular disease. Gastrointestinal endoscopy is the method of choice for the diagnostics of bleeding from upper tract and large bowel. For diagnostics of bleeding from the small bowel we can use scintigraphy with in vivo labelled autological red blood cells if pushenteroscopy, intra-operative enteroscopy or angiography are not available. 31 patients (13 men, 18 women, aged 20-91, mean 56 years) underwent this investigation from 1998 till 2001 at the Department of Nuclear Medicine. All patients had melaena or enterorrhagia associated with acute anaemia. Gastroscopy, colonoscopy, enteroclysis or X-ray angiography did not detect the source of bleeding. Twenty-one patients had positive scintigraphy with in vivo labelled RBCs - 9 patients were already positive on dynamic scintigraphy, and 12 patients were positive on static images. Scintigraphy with in vivo labelled RBCs was negative in 10 patients. GI bleeding stopped spontaneously in these 10 patients with negative scintigraphy. These patients did not undergo intra-operative enteroscopy or surgery. The final diagnosis of the 21 patients with positive scintigraphy was determined in 16 patients by push-enteroscopy (6 patients), intra-operative enteroscopy (6 patients) or by surgery (4 patients). Of these 16 patients the correct place of bleeding was determined by scintigraphy with labelled RBCs in 11 (69%) patients. Final diagnoses of our 16 patients with positive scintigraphy with autological labelled RBCs were: bleeding small bowel arteriovenous malformation (6 patients), uraemic enteritis with bleeding erosions in ileum and jejunum (2 patients), Osler-Rendu- Weber disease (1 patient), pseudocyst of

  11. 99mTc-besilesomab (Scintimun registered) in peripheral osteomyelitis: comparison with 99mTc-labelled white blood cells

    International Nuclear Information System (INIS)

    The diagnosis of osteomyelitis is a challenge for diagnostic imaging. Nuclear medicine procedures including white blood cell imaging have been successfully used for the identification of bone infections. This multinational, phase III clinical study in 22 European centres was undertaken to compare anti-granulocyte imaging using the murine IgG antibody besilesomab (Scintimun registered) with 99mTc-labelled white blood cells in patients with peripheral osteomyelitis. A total of 119 patients with suspected osteomyelitis of the peripheral skeleton received 99mTc-besilesomab and 99mTc-hexamethylpropyleneamine oxime (HMPAO)-labelled white blood cells (WBCs) in random order 2-4 days apart. Planar images were acquired at 4 and 24 h after injection. All scintigraphic images were interpreted in an off-site blinded read by three experienced physicians specialized in nuclear medicine, followed by a fourth blinded reader for adjudication. In addition, clinical follow-up information was collected and a final diagnosis was provided by the investigators and an independent truth panel. Safety data including levels of human anti-mouse antibodies (HAMA) and vital signs were recorded. The agreement in diagnosis across all three readers between Scintimun registered and 99mTc-HMPAO-labelled WBCs was 0.83 (lower limit of the 95% confidence interval 0.8). Using the final diagnosis of the local investigator as a reference, Scintimun registered had higher sensitivity than 99mTc-HMPAO-labelled WBCs (74.8 vs 59.0%) at slightly lower specificity (71.8 vs 79.5%, respectively). All parameters related to patient safety (laboratory data, vital signs) did not provide evidence of an elevated risk associated with the use of Scintimun registered except for two cases of transient hypotension. HAMA were detected in 16 of 116 patients after scan (13.8%). Scintimun registered imaging is accurate, efficacious and safe in the diagnosis of peripheral bone infections and provides comparable information to 99mTc-HMPAO-labelled

  12. Effect of an Arctium lappa (burdock extract on the labeling of blood constituents with technetium-99m and on the morphology of the red blood cells

    Directory of Open Access Journals (Sweden)

    Rosane de Figueiredo Neves

    2007-09-01

    Full Text Available Arctium lappa (burdock has been used to treat inflammatory processes. Blood constituents labeled with technetium-99m (99mTc have been utilized in nuclear medicine. It was evaluated the influence of a burdock extract on the labeling of blood constituents with 99mTc and on the morphometry of red blood cells. Blood samples from Wistar rats were incubated with burdock extract and the radiolabeling procedure was carried out. Plasma and blood cells, soluble and insoluble fractions of plasma and blood cells were separated. The radioactivity in each fraction was counted and the percentages of radioactivity (%ATI were determined. Morphology and morphometric (perimeter/area ratio measurements of red blood cells (RBC were performed. The incubation with burdock extract significantly (pArctium lappa (bardana tem sido utilizada na medicina popular para o tratamento de processos inflamatórios. Constituintes sangüíneos marcados com tecnécio-99m (99mTc são utilizados na medicina nuclear para obtenção de imagens. Neste trabalho foi avaliada a influência de um extrato de bardana na marcação de constituintes sangüíneos com 99mTc e na morfologia de hemácias. Amostras de sangue de ratos Wistar foram incubadas com extrato de bardana e o processo de radiomarcação de constituintes sangüíneos foi realizado. Plasma e células sangüíneas, frações solúvel e insolúvel do plasma e das células sangüíneas foram separadas, a radioatividade em cada fração foi contada e as porcentagens de radioatividade (%ATI foram determinadas. A morfologia e a relação perímetro/área das hemácias foram avaliadas. A incubação de sangue com o extrato de bardana alterou significativamente (p<0.05 a %ATI a distribuição de radioatividade nos compartimentos plasmático e celular. A relação perímetro/área de hemácias, bem como a forma das hemácias também sofreram alterações Modificações na membrana poderiam justificar a diminuição da marcação das c

  13. Demonstration of hematobilia using technetium-99m labeled red blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Lee, S.M.; Lee, R.G.; Clouse, M.E.; Hill, T.C.

    1986-01-01

    A 75-year-old woman, who presented with obstructive jaundice, was shown by percutaneous transhepatic cholangiography to have a markedly dilated biliary system and stones within the common bile duct. The stones were removed percutaneously using the transduodenal approach, and an internal drainage catheter was placed. Following the procedure, the patient experienced gastrointestinal bleeding manifested by melanotic stools. Blood-tinged bile was withdrawn from the biliary drainage catheter, leading to the suspicion that the bleeding might be originating from the biliary tract. A Tc-99m red blood cell (Tc-99m RBC) scan was performed to try to designate the biliary tract as the site of bleeding, and to determine if there were any other bleeding sites present. The study demonstrated bleeding from the biliary tract, which was confirmed by angiography and endoscopy. The technique for the detection of gastrointestinal bleeding using Tc-99m RBCs is well described. This case suggests that when doing studies to localize occult bleeding, the liver should be included in the field-of-view to exclude bleeding from the liver.

  14. Effect of a peel passion fruit flour (Passiflora edulis f. flavicarpa) extract on the labeling of blood constituents with technetium-99m and on the morphology of red blood cells

    International Nuclear Information System (INIS)

    Passiflora edulis f. flavicarpa (maracuja) is a fruit consumed in Brazil and worldwide. Blood constituents labeled with technetium-99m (99mTc) are used in nuclear medicine. The effect of P. flavicarpa extract on the radiolabeling of blood constituents and on red blood cells morphology was evaluated. Blood samples from Wistar rats was incubated with P. flavicarpa extract. After that, the labeling of blood constituents with 99mTc was carried out. Samples of plasma and blood cells were precipitated with trichloroacetic acid to isolate the soluble and insoluble fractions of plasma and blood cells. The radioactivity in each fractions was counted and the percentage of radioactivity was determined. Blood smears were also prepared to morphological evaluation and perimeter/area ratio determination. P. flavicarpa extract altered (p99mTc on plasma proteins and the perimeter/area ratio of red blood cells. Substances present in P. flavicarpa extract could affect the labeling of blood constituents with 99mTc acting in specific targets as membrane of red blood cells. (author)

  15. Effect of a peel passion fruit flour (Passiflora edulis f. flavicarpa) extract on the labeling of blood constituents with technetium-99m and on the morphology of red blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Rebello, Bernardo Machado; Moreno, Silvana Ramos Farias; Ribeiro, Camila Godinho; Neves, Rosane de Figueiredo; Fonseca, Adenilson de Souza da; Bernardo-Filho, Mario; Medeiros, Aldo da Cunha [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil). Programa de Pos-graduacao em Ciencias da Saude]. E-mail: rebellobm@uol.com.br; Caldas, Luis Querino de Araujo [Universidade Federal Fluminense (UFF), Niteroi, RJ (Brazil). Programa de Pos-Graduacao em Ciencias Medicas

    2007-09-15

    Passiflora edulis f. flavicarpa (maracuja) is a fruit consumed in Brazil and worldwide. Blood constituents labeled with technetium-99m (99mTc) are used in nuclear medicine. The effect of P. flavicarpa extract on the radiolabeling of blood constituents and on red blood cells morphology was evaluated. Blood samples from Wistar rats was incubated with P. flavicarpa extract. After that, the labeling of blood constituents with 99mTc was carried out. Samples of plasma and blood cells were precipitated with trichloroacetic acid to isolate the soluble and insoluble fractions of plasma and blood cells. The radioactivity in each fractions was counted and the percentage of radioactivity was determined. Blood smears were also prepared to morphological evaluation and perimeter/area ratio determination. P. flavicarpa extract altered (p<0.05) the fixation of {sup 99m}Tc on plasma proteins and the perimeter/area ratio of red blood cells. Substances present in P. flavicarpa extract could affect the labeling of blood constituents with {sup 99m}Tc acting in specific targets as membrane of red blood cells. (author)

  16. Effects of chronic sucralose sweetener on the labeling of blood constituents with technetium-99m, morphology of red blood cells and the biodistribution of sodium pertechnetate in rats

    Directory of Open Access Journals (Sweden)

    Gabrielle de Souza Rocha

    2008-12-01

    Full Text Available This work evaluates effects of the sweetener with sucralose on the labeling of blood constituents with technetium-99m (99mTc, on the morphology of red blood cells (RBC and on the biodistribution of sodium pertechnetate in Wistar rats. Animals were treated with sweetener for 8 days. Blood samples were withdrawn and the assay of labeling of blood constituents with 99mTc was performed. Blood cells (BC and plasma (P were isolated. Aliquots of BC and P were also precipitated, soluble and insoluble fractions separated. The radioactivity in each fraction was counted and percentage of incorporated radioactivity (%ATI determined. Blood smears were prepared, fixed, stained and the qualitative and quantitative morphology of the RBC was evaluated under optical microscopy. In biodistribution experiments, sodium pertechnetate was administrated, organs and tissues isolated, radioactivity was counted and percentage of incorporated radioactivity per gram (%ATI/g determined. The data showed no significant alterations in %ATI, morphology of RBC and in %ATI/g in the studied organs.Neste estudo foram avaliados efeitos do adoçante com sucralose na marcação de constituintes sangüíneos com 99mTc, na morfologia de hemácias e na biodistribuição do pertecnetato de sódio em ratos Wistar. Animais foram tratados com adoçante durante 8 dias. Amostras de sangue foram retiradas e a marcação de constituintes sangüíneos com 99mTc foi realizada. Células sangüíneas (CS e plasma (P foram isolados. Alíquotas de CS e P foram precipitadas, frações insolúvel e solúvel foram separadas. A radioatividade em cada fração foi contada e o percentual de radioatividade incorporada (%ATI, determinado. Distensões sangüíneas foram preparadas, fixadas, coradas e análise morfológica, qualitativa e quantitativa, de hemácias foi avaliada sob microscopia óptica. Nos experimentos de biodistribuição, pertecnetato de sódio foi administrado, órgãos e tecidos isolados, a

  17. In-111-labeled white blood cell uptake in noninfected closed fracture in humans: prospective study

    International Nuclear Information System (INIS)

    Since indium-111 white blood cell (In-111 WBC) scintigraphy is often used to evaluate for osteomyelitis in bone fractures, it is important to know if noninfected fractures have In-111 WBC uptake. Twenty-seven noninfected closed fracture sites in 19 patients were prospectively evaluated with technetium-99m methylene diphosphonate bone scintigraphy and In-111 WBC scintigraphy. In-111 WBC uptake was present in 41% of the 27 sites. In the 11 positive sites, the In-111 WBC uptake was 1+ (definite but minimal) in 55%, 2+ (moderate) in 36%, and 3+ (marked) in 9%. The visual intensity of the radioactive uptake on In-111 WBC scintigrams relative to that on bone scintigrams was less in 82%, equal in 9%, and greater in 9%. The visual size of the area of uptake on In-111 WBC scintigrams and bone scintigrams was smaller in 36%, equal in 55%, and greater in 9%. Factors that may help distinction of In-111 WBC uptake due to fracture alone from infection associated with fracture are discussed

  18. Hyperemic peripheral red marrow in a patient with sickle cell anemia demonstrated on Tc-99m labeled red blood cell venography

    International Nuclear Information System (INIS)

    A 25-year-old gravid woman, homozygous for sickle cell anemia, with a history of recent deep venous thrombosis, was examined using Tc-99m labeled red blood cell venography for recurrent thrombosis. Although negative for thrombus, the study presented an unusual incidental finding: the patient's peripheral bone marrow was hyperemic in a distribution consistent with peripheral red bone marrow expansion. Such a pattern has not been documented before using this technique. This report supports other literature that has demonstrated hyperemia of peripheral red bone marrow in other hemolytic anemias. This finding may ultimately define an additional role of scintigraphy in assessing the pathophysiologic status of the sickle cell patient

  19. In vitro studies using Tc-99m-labelled human serum albumin millimicrospheres for assessing the contribution to phagocytosis of mononuclear blood cells

    International Nuclear Information System (INIS)

    Tc-99m-labelled HSA millimicrospheres were incubated with whole blood for various periods of time, in order to analyse the rate of fixation of HSA millimicrospheres to monocytes, and their interaction. It was not possible with the technique applied to differentiate between a genuine process of phagocytosis and a conceivable mere fixation of particles to the cells. (MBC)

  20. Labelling of T cell subsets under field conditions in tropical countries. Adaptation of the immuno-alkaline phosphatase staining method for blood smears

    DEFF Research Database (Denmark)

    Lisse, I M; Whittle, H; Aaby, P;

    1990-01-01

    Immuno-alkaline phosphatase (AP) staining for T cell subsets (CD4 and CD8) of smears from fingerprick blood functioned well under tropical climatic conditions when smears were stored frozen with silica gel before being labelled. Unlabelled smears were stored for up to 12 months and could be...

  1. Clinical evaluation of technetium-99m labeled red blood cell scan in the detection of gastrointestinal hemorrhage

    International Nuclear Information System (INIS)

    A three years' experience at Chigasaki Tokushukai Medical Center with Technetium-99m labeled red blood cell scintiscans (Tc-99m-RBC Scan) performed on 68 cases of suspected lower gastrointestinal (LGI) bleeding was reviewed in retrospect. Of the 36 cases performed within 24 hours from the clinical detection of active LGI hemorrhage on emergency basis, 27 cases were positive (75 % positivity). Of the total 45 positive cases, 21 (47 %) became positive one or more hours post injection, confirming the intermittent nature of the LGI hemorrhage. All studies were completed without untoward effects and confirmed the noninvasive nature of the study, as compared to the other invasive modalities such as endoscopy and angiography. Detection and localization of the site of LGI bleeding directed the precise modality to be chosen subsequently and greatly facilitated in performing the invasive procedure with selectivity further improving its sensitivity. Tc-99m-RBC scan is a completely noninvasive, simple and sensitive procedure, which may be used initially in routine to detect and localize LGI bleeding. (author)

  2. Detection of acute osteomyelitis with indium-111 labeled white blood cells in a patient with sickle cell disease

    International Nuclear Information System (INIS)

    A young patient with sickle cell disease (SCD) and multiple hospitalizations for crisis was admitted because of suspected osteomyelitis. Initial laboratory work, radiographs, and bone images were not contributory. An In-111 white blood cell (WBC) study demonstrated two areas of increased radionuclide uptake consistent with osteomyelitis. One of these had associated soft tissue infection. No other areas of active osteomyelitis were visualized, in spite of the presence of several additional infection sites. Imaging with In-111 WBC is probably not justified for routine diagnosis of acute osteomyelitis in areas free of previous disease, where conventional bone images are highly efficient. In-111 WBC imaging, however, may be helpful in detecting osteomyelitis in selected patients with SCD in whom Tc-99m bone images and radiographs are usually abnormal and difficult to interpret due to previous bone infarcts. Localization of the infection focus is very important in choosing the aspiration site for bacteriologic studies. A negative study, however, should be interpreted cautiously

  3. Bleeding rates necessary for detecting acute gastrointestinal bleeding with technetium-99m-labeled red blood cells in an experimental model

    International Nuclear Information System (INIS)

    Proponents of [/sup 99m/Tc]sulfur colloid for GI bleeding studies argue that, although labeled red blood cells are useful for intermittent bleeding, they are not capable of detecting low bleeding rates. Studies of dogs with experimental GI bleeding have indicated bleeding rates of 0.05 ml/min can be detected with [/sup 99m/Tc]sulfur colloid. Since similar data in the dog model were unavailable for /sup 99m/Tc-labeled red blood cells, we undertook this study. To simulate lower GI bleeding, catheters were inserted into the bowel lumen. Each dog's blood was labeled with /sup 99m/Tc using an in vitro technique. Venous blood was then withdrawn and re-infused into the lumen of the bowel using a Harvard pump. Fourteen dogs were studied, ten receiving a bleeding rate from 4.6-0.02 ml/min in the descending colon and four with proximal jejunal bleeds of 0.20-0.02 ml/min. Bleeding rates of 4.6-0.2 ml/min were detected within 10 min in the colon and bleeding rates as low as 0.04 ml/min were seen by 55 min. Slower bleeding rates were not detected. Similar findings were noted for proximal jejunal bleeds. Based on the time of appearance, a minimum volume of approximately 2-3 ml labeled blood was necessary to detect bleeding. We conclude that /sup 99m/Tc-labeled RBCs are sensitive for low bleeding rates in the dog model. The rates are comparable to those described for [/sup 99m/Tc]sulfur colloid in this experimental setting. The time of appearance of activity is related to the bleeding rate

  4. Hyperemic peripheral red marrow in a patient with sickle cell anemia demonstrated on Tc-99m labeled red blood cell venography

    Energy Technology Data Exchange (ETDEWEB)

    Heiden, R.A.; Locko, R.C.; Stent, T.R. (Columbia Univ. College of Physicians and Surgeons, New York, NY (USA))

    1991-03-01

    A 25-year-old gravid woman, homozygous for sickle cell anemia, with a history of recent deep venous thrombosis, was examined using Tc-99m labeled red blood cell venography for recurrent thrombosis. Although negative for thrombus, the study presented an unusual incidental finding: the patient's peripheral bone marrow was hyperemic in a distribution consistent with peripheral red bone marrow expansion. Such a pattern has not been documented before using this technique. This report supports other literature that has demonstrated hyperemia of peripheral red bone marrow in other hemolytic anemias. This finding may ultimately define an additional role of scintigraphy in assessing the pathophysiologic status of the sickle cell patient.

  5. Effect of the extract of Ricinus communis L. on the osmotic fragility, labeling of red blood cells with Technetium-99m and morphology of the cells

    Directory of Open Access Journals (Sweden)

    Kristiana Cerqueira Mousinho

    2008-12-01

    Full Text Available The aim of this study was to evaluate the influence of the proteic extract of R. communis on the cell physiology by the osmotic fragility, labeling of the blood elements with the 99mTc and cell morphology. To evaluate the osmotic fragility, the blood samples of the Wistar rats were incubated with the concentrations of R. communis and with the solutions of NaCl (0.4; 0.7; 0.9%. In the labeling of the blood elements procedure, the rat blood was treated with a solution of Tc-99m and TCA at 5%, determining the rate of radioactivity (%ATI in the plasma (P and in the red blood cells (RBC. The soluble and insoluble fractions of the plasma were also evaluated. The cells morphology submitted to the extract was evaluated by the optical microscopy (x40. The results indicated that the rate of the hemolysis increased in the presence of 0.125 mg/mL of the extract. There was a decay of 49.69% in the rate of ATI in the insoluble fraction of the cells, with the morphological alterations in the red blood cells. These results suggested that the extract changed the capability of binding of the red blood cells due to the stannous ion oxidation, modifying the cells structure.Produtos naturais são usados freqüentemente por muitas pessoas no tratamento do câncer. O Ricinus communis L é uma Euforbiaceae que apresenta propriedades laxativas, purgativas e antitumorais. O objetivo deste trabalho é estudar a influência da fração protéica do extrato hidroalcoólico de R. communis L. na fisiologia celular através da fragilidade osmótica, da marcação de elementos sanguíneo com 99mTc e da morfologia celular. Para avaliar a fragilidade osmótica, amostras de sangue de ratos Wistar foram incubadas com concentrações de R. communis e com soluções de NaCl (0,4; 0,7; 0,9%. No procedimento de marcação de elementos sanguíneos, as amostras de sangue foram tratadas com solução de Tc-99m e TCA à 5%, determinando o percentual de radioatividade (%ATI no plasma (P e

  6. Transcriptomic Profile of Whole Blood Cells from Elderly Subjects Fed Probiotic Bacteria Lactobacillus rhamnosus GG ATCC 53103 (LGG) in a Phase I Open Label Study

    OpenAIRE

    Solano-Aguilar, Gloria; Molokin, Aleksey; Botelho, Christine; Fiorino, Anne-Maria; Vinyard, Bryan; Li, Robert; Chen, Celine; Urban, Joseph; Dawson, Harry; Andreyeva, Irina; Haverkamp, Miriam; Hibberd, Patricia L.

    2016-01-01

    We examined gene expression of whole blood cells (WBC) from 11 healthy elderly volunteers participating on a Phase I open label study before and after oral treatment with Lactobacillus rhamnosus GG-ATCC 53103 (LGG)) using RNA-sequencing (RNA-Seq). Elderly patients (65–80 yrs) completed a clinical assessment for health status and had blood drawn for cellular RNA extraction at study admission (Baseline), after 28 days of daily LGG treatment (Day 28) and at the end of the study (Day 56) after LG...

  7. Integrating Cell Phone Imaging with Magnetic Levitation (i-LEV) for Label-Free Blood Analysis at the Point-of-Living.

    Science.gov (United States)

    Baday, Murat; Calamak, Semih; Durmus, Naside Gozde; Davis, Ronald W; Steinmetz, Lars M; Demirci, Utkan

    2016-03-01

    There is an emerging need for portable, robust, inexpensive, and easy-to-use disease diagnosis and prognosis monitoring platforms to share health information at the point-of-living, including clinical and home settings. Recent advances in digital health technologies have improved early diagnosis, drug treatment, and personalized medicine. Smartphones with high-resolution cameras and high data processing power enable intriguing biomedical applications when integrated with diagnostic devices. Further, these devices have immense potential to contribute to public health in resource-limited settings where there is a particular need for portable, rapid, label-free, easy-to-use, and affordable biomedical devices to diagnose and continuously monitor patients for precision medicine, especially those suffering from rare diseases, such as sickle cell anemia, thalassemia, and chronic fatigue syndrome. Here, a magnetic levitation-based diagnosis system is presented in which different cell types (i.e., white and red blood cells) are levitated in a magnetic gradient and separated due to their unique densities. Moreover, an easy-to-use, smartphone incorporated levitation system for cell analysis is introduced. Using our portable imaging magnetic levitation (i-LEV) system, it is shown that white and red blood cells can be identified and cell numbers can be quantified without using any labels. In addition, cells levitated in i-LEV can be distinguished at single-cell resolution, potentially enabling diagnosis and monitoring, as well as clinical and research applications. PMID:26523938

  8. Assessment of the effect of Mentha crispa L. (hortela) extract on the labeling of red blood cells and plasma proteins with technetium-99m

    Energy Technology Data Exchange (ETDEWEB)

    Santos-Filho, Sebastiao D. [UNIFOA - Centro Universitario de Volta Redonda, RJ (Brazil). Dept. de Biofisica; Dire, Glaucio L.; Lima, Elaine [Universidade do Estado, Rio de Janeiro, RJ (Brazil). Inst. de Biologia. Dept. de Biofisica e Biometria; Pereira, Mario [Universidade do Estado, Rio de Janeiro, RJ (Brazil). Inst. de Biologia. Dept. de Anatomia; Bernardo-Filho, Mario [Universidade do Estado, Rio de Janeiro, RJ (Brazil). Inst. de Biologia. Dept. de Biofisica e Biometria]|[Instituto Nacional do Cancer, Rio de Janeiro, RJ (Brazil). Centro de Pesquisa Basica

    2002-07-01

    We have investigated the possibility of M. Crispa L. extract being capable to alter the labeling of blood elements with 99mTc. Blood was incubated with M. Crispa L. extract. Stannous chloride solution and Tc-99m, as sodium pertechnetate, were added. Blood was centrifuged and plasma (P) and blood cells (BC) were isolated. Samples of P and BC were also precipitated, centrifuged and insoluble (IF) and soluble (SF) separated. The percentage of radioactivity (% ATI) in BC, IF-P and IF-BC was calculated. Histological evaluations were performed and the morphology of the red blood cells was observed under optical microscopy showing important morphological alterations on the shape of the RBC treated with 6.25% M. Crispa L. extract. The % ATI decreased: on BC from 97.3 {+-} 1.92 to 60.0 {+-} 2.44; on IF-P from 74.8 {+-} 3.78 to 9.99 {+-} 3.61; on IF-BC from 88.6 {+-} 5.41 to 58.4 {+-} 11.55. The substances of the M. Crispa L. extract could increase the valence of these stannous (+2) ions to stannic (+4) and this fact would decrease the % ATI on blood elements and indicates the possible presence of oxidant agents in the M. Crispa L. extract. (author)

  9. Assessment of the effect of Mentha crispa L. (hortela) extract on the labeling of red blood cells and plasma proteins with technetium-99m

    International Nuclear Information System (INIS)

    We have investigated the possibility of M. Crispa L. extract being capable to alter the labeling of blood elements with 99mTc. Blood was incubated with M. Crispa L. extract. Stannous chloride solution and Tc-99m, as sodium pertechnetate, were added. Blood was centrifuged and plasma (P) and blood cells (BC) were isolated. Samples of P and BC were also precipitated, centrifuged and insoluble (IF) and soluble (SF) separated. The percentage of radioactivity (% ATI) in BC, IF-P and IF-BC was calculated. Histological evaluations were performed and the morphology of the red blood cells was observed under optical microscopy showing important morphological alterations on the shape of the RBC treated with 6.25% M. Crispa L. extract. The % ATI decreased: on BC from 97.3 ± 1.92 to 60.0 ± 2.44; on IF-P from 74.8 ± 3.78 to 9.99 ± 3.61; on IF-BC from 88.6 ± 5.41 to 58.4 ± 11.55. The substances of the M. Crispa L. extract could increase the valence of these stannous (+2) ions to stannic (+4) and this fact would decrease the % ATI on blood elements and indicates the possible presence of oxidant agents in the M. Crispa L. extract. (author)

  10. Labelling by 3H-N-ethylmaleimide of diamide-oxidized thiol groups in sheep red blood cell (SRBC) membranes

    International Nuclear Information System (INIS)

    Exposure of SRBC to the thiol oxidant diamide activates K:Cl cotransport, reversed upon metabolic restoration of cellular glutathione suggesting redox control of the K:Cl cotransporter, as well as by subsequent exposure to dithiothreitol (DTT). The thiols crucial for activation may be either on the transporter or on a membrane or cytoplasmic regulator. To test this hypothesis, the authors attempted to label with 3H-N-ethylmaleimide (3H-NEM) the thiols protected by diamide oxidation and reduced subsequently by DTT. SRBC were first treated with a diamide concentration activating K:Cl cotransport, followed by a second exposure to unlabeled (cold) NEM to block any non-oxidized thiol, and then hemolyzed to obtain white ghosts. The ghosts were again treated with cold NEM and after reduction by DTT exposed to 3H-NEM with and without cold NEM. Saturation labelling by 3H-NEM of diamide protected groups occurred in the range of CTT concentrations inactivating the diamide-stimulated K:Cl cotransport. Saturation labelling with 3H-NEM occurred at about 25μM NEM suggesting a Ki of less than 10μM NEM. The number of diamide protected thiols was about 5-10,000/cell membrane. At 100μM 3H-NEM, SRBC not treated with diamide possess at least 100,000 thiols cell and this number is likely to rise by tenfold at higher NEM concentrations. Thus, diamide protected about 1/1,000 of the membrane thiols in both genetically low and high K SRBC, assayed under conditions where K:Cl cotransport is activated in intact cells. Therefore, at least some of the thiols crucial for potential regulation of K:Cl cotransport reside within the plasma membrane

  11. Microfluidic bead-based multienzyme-nanoparticle amplification for detection of circulating tumor cells in the blood using quantum dots labels

    International Nuclear Information System (INIS)

    Graphical abstract: A microfluidic beads-based nucleic acid sensor for sensitive detection of circulating tumor cells (CTCs) in the blood using multienzyme-nanoparticle amplification and quantum dots labels was developed. The chip-based CTCs analysis could detect reverse transcription-polymerase chain reaction (RT-PCR) products of tumor cell as low as 1 tumor cell (e.g. CEA expressing cell) in 1 mL blood sample. This microfluidic beads-based nucleic acid sensor is a promising platform for disease-related nucleic acid molecules at the lowest level at their earliest incidence. -- Highlights: •Combination of microfluidic bead-based platform and enzyme–probe–AuNPs is proposed. •The developed nucleic acid sensor could respond to 5 fM of tumor associated DNA. •Microfluidic platform and multienzyme-labeled AuNPs greatly enhanced sensitivity. •The developed nucleic acid sensor could respond to RT-PCR products of tumor cell as low as 1 tumor cell in 1 mL blood sample. •We report a sensitive nucleic acid sensor for detection of circulating tumor cells. -- Abstract: This study reports the development of a microfluidic bead-based nucleic acid sensor for sensitive detection of circulating tumor cells in blood samples using multienzyme-nanoparticle amplification and quantum dot labels. In this method, the microbeads functionalized with the capture probes and modified electron rich proteins were arrayed within a microfluidic channel as sensing elements, and the gold nanoparticles (AuNPs) functionalized with the horseradish peroxidases (HRP) and DNA probes were used as labels. Hence, two signal amplification approaches are integrated for enhancing the detection sensitivity of circulating tumor cells. First, the large surface area of Au nanoparticle carrier allows several binding events of HRP on each nanosphere. Second, enhanced mass transport capability inherent from microfluidics leads to higher capture efficiency of targets because continuous flow within micro

  12. Microfluidic bead-based multienzyme-nanoparticle amplification for detection of circulating tumor cells in the blood using quantum dots labels

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, He, E-mail: mzhang_he@126.com; Fu, Xin; Hu, Jiayi; Zhu, Zhenjun

    2013-05-24

    Graphical abstract: A microfluidic beads-based nucleic acid sensor for sensitive detection of circulating tumor cells (CTCs) in the blood using multienzyme-nanoparticle amplification and quantum dots labels was developed. The chip-based CTCs analysis could detect reverse transcription-polymerase chain reaction (RT-PCR) products of tumor cell as low as 1 tumor cell (e.g. CEA expressing cell) in 1 mL blood sample. This microfluidic beads-based nucleic acid sensor is a promising platform for disease-related nucleic acid molecules at the lowest level at their earliest incidence. -- Highlights: •Combination of microfluidic bead-based platform and enzyme–probe–AuNPs is proposed. •The developed nucleic acid sensor could respond to 5 fM of tumor associated DNA. •Microfluidic platform and multienzyme-labeled AuNPs greatly enhanced sensitivity. •The developed nucleic acid sensor could respond to RT-PCR products of tumor cell as low as 1 tumor cell in 1 mL blood sample. •We report a sensitive nucleic acid sensor for detection of circulating tumor cells. -- Abstract: This study reports the development of a microfluidic bead-based nucleic acid sensor for sensitive detection of circulating tumor cells in blood samples using multienzyme-nanoparticle amplification and quantum dot labels. In this method, the microbeads functionalized with the capture probes and modified electron rich proteins were arrayed within a microfluidic channel as sensing elements, and the gold nanoparticles (AuNPs) functionalized with the horseradish peroxidases (HRP) and DNA probes were used as labels. Hence, two signal amplification approaches are integrated for enhancing the detection sensitivity of circulating tumor cells. First, the large surface area of Au nanoparticle carrier allows several binding events of HRP on each nanosphere. Second, enhanced mass transport capability inherent from microfluidics leads to higher capture efficiency of targets because continuous flow within micro

  13. {sup 99m}Tc-besilesomab (Scintimun {sup registered}) in peripheral osteomyelitis: comparison with {sup 99m}Tc-labelled white blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Richter, Wolf S. [Pharmtrace Klinische Entwicklung GmbH, Berlin (Germany); University Clinics Magdeburg (Germany). Clinics for Radiology and Nuclear Medicine; Ivancevic, Velimir [Nuclear Medicini, Celle (Germany); Meller, Johannes [University Medicine, Goettingen (Germany). Dept. of Nuclear Medicine; Lang, Otto [UH Kralovske, Prague (Czech Republic). Dept. of Nclear Medicine; Le Guludec, Dominique [CHU Bichat-Claude Bernard, Paris (France). Service de Medecine Nucleaire; Szilvazi, Istvan [Orszagos Gyogyintezeti Koezpont, Budapest (Hungary). Dept. of Nuclear Medicine; Amthauer, Holger [University Clinics Magdeburg (Germany). Clinics for Radiology and Nuclear Medicine; Chossat, Florence; Dahmane, Amel [IBA/CIS Bio International, Gif sur Yvette (France); Schwenke, Carsten [SCOSSIS, Berlin (Germany); Signore, Alberto [University of Rome, Faculty of Medicine, Nuclear Medicine Unit, Rome (Italy)

    2011-05-15

    The diagnosis of osteomyelitis is a challenge for diagnostic imaging. Nuclear medicine procedures including white blood cell imaging have been successfully used for the identification of bone infections. This multinational, phase III clinical study in 22 European centres was undertaken to compare anti-granulocyte imaging using the murine IgG antibody besilesomab (Scintimun {sup registered}) with {sup 99m}Tc-labelled white blood cells in patients with peripheral osteomyelitis. A total of 119 patients with suspected osteomyelitis of the peripheral skeleton received {sup 99m}Tc-besilesomab and {sup 99m}Tc-hexamethylpropyleneamine oxime (HMPAO)-labelled white blood cells (WBCs) in random order 2-4 days apart. Planar images were acquired at 4 and 24 h after injection. All scintigraphic images were interpreted in an off-site blinded read by three experienced physicians specialized in nuclear medicine, followed by a fourth blinded reader for adjudication. In addition, clinical follow-up information was collected and a final diagnosis was provided by the investigators and an independent truth panel. Safety data including levels of human anti-mouse antibodies (HAMA) and vital signs were recorded. The agreement in diagnosis across all three readers between Scintimun {sup registered} and {sup 99m}Tc-HMPAO-labelled WBCs was 0.83 (lower limit of the 95% confidence interval 0.8). Using the final diagnosis of the local investigator as a reference, Scintimun {sup registered} had higher sensitivity than {sup 99m}Tc-HMPAO-labelled WBCs (74.8 vs 59.0%) at slightly lower specificity (71.8 vs 79.5%, respectively). All parameters related to patient safety (laboratory data, vital signs) did not provide evidence of an elevated risk associated with the use of Scintimun {sup registered} except for two cases of transient hypotension. HAMA were detected in 16 of 116 patients after scan (13.8%). Scintimun {sup registered} imaging is accurate, efficacious and safe in the diagnosis of peripheral

  14. Conference on radionuclide labelled cellular blood elements

    International Nuclear Information System (INIS)

    The South African Medical Research Council presented this conference on radionuclide labelled cellular blood elements with application in atherosclerosis and thrombosis. The conference was held in Bloemfontein from 3-6 February 1986. This work only consists of the abstracts of the seminars that were delivered on the conference. The radioisotopes that occur most of the time in the abstracts include Indium 111, Indium 114, Chromium 51, Iodine 125, Iodine 131 and Carbon 14. Especially Indium 111 seems to be the method of choice for all labelling

  15. Separation of uncompromised whole blood mixtures for single source STR profiling using fluorescently-labeled human leukocyte antigen (HLA) probes and fluorescence activated cell sorting (FACS).

    Science.gov (United States)

    Dean, Lee; Kwon, Ye Jin; Philpott, M Katherine; Stanciu, Cristina E; Seashols-Williams, Sarah J; Dawson Cruz, Tracey; Sturgill, Jamie; Ehrhardt, Christopher J

    2015-07-01

    Analysis of biological mixtures is a significant problem for forensic laboratories, particularly when the mixture contains only one cell type. Contributions from multiple individuals to biologic evidence can complicate DNA profile interpretation and often lead to a reduction in the probative value of DNA evidence or worse, its total loss. To address this, we have utilized an analytical technique that exploits the intrinsic immunological variation among individuals to physically separate cells from different sources in a mixture prior to DNA profiling. Specifically, we applied a fluorescently labeled antibody probe to selectively bind to one contributor in a mixture through allele-specific interactions with human leukocyte antigen (HLA) proteins that are expressed on the surfaces of most nucleated cells. Once the contributor's cells were bound to the probe, they were isolated from the mixture using fluorescence activated cell sorting (FACS)-a high throughput technique for separating cell populations based on their optical properties-and then subjected to STR analysis. We tested this approach on two-person and four-person whole blood mixtures where one contributor possessed an HLA allele (A*02) that was not shared by other contributors to the mixture. Results showed that hybridization of the mixture with a fluorescently-labeled antibody probe complimentary to the A*02 allele's protein product created a cell population with a distinct optical profile that could be easily differentiated from other cells in the mixture. After sorting the cells with FACS, genetic analysis showed that the STR profile of this cell population was consistent with that of the contributor who possessed the A*02 allele. Minor peaks from the A*02 negative contributor(s) were observed but could be easily distinguished from the profile generated from A*02 positive cells. Overall, this indicates that HLA antibody probes coupled to FACS may be an effective approach for generating STR profiles of

  16. Evaluation of ethinylestradiol effect on labelling red blood cells with Tc-99m; Avaliacao do efeito do etinilestradiol sobre a marcacao de hemacias com tecnecio-99m

    Energy Technology Data Exchange (ETDEWEB)

    Braga, A.C.S.; Oliveira, J.F.; Santos, J.S.; Oliveira, M.B.N.; Gutfilen, B.; Bernardo-Filho, Mario [Universidade do Estado, Rio de Janeiro, RJ (Brazil). Inst. de Biologia Roberto Alcantara Gomes. Dept. de Biofisica e Biometria

    1999-11-01

    Significant alterations on the radiopharmaceutical distribution in humans are caused by drug interactions. The labeling red blood cells with technetium-99m is a daily routine procedure in nuclear medicine. Here, we investigated if the ethinylestradiol, an oral contraceptive, could alter the labeling of RBC with Tc-99m. Samples of blood with acid citrate dextrose were incubated with ethynilestradiol. Then, different concentrations of Sn C L{sub 2} were added and, after that, Tc-99m was added. Samples were centrifuged and plasma (P) and cells (C) were separated. the results showed that the drug studied decreased the uptake of radioactivity (%ATI) in the C to the reducing agent in the concentration of 1.2 (from 92.3 to 78.0) and increased in 12.0 (18.8 to 36.0) and in 24.0 (22.8 to 32.0){mu}/ml of Sn C L{sub 2}. The obtained results can be explained by the fact that this drug could alter the membrane permeability to the transport of stannous and/or pertechnetate ions. (author) 18 refs., 2 tabs.

  17. Microfluidic bead-based multienzyme-nanoparticle amplification for detection of circulating tumor cells in the blood using quantum dots labels.

    Science.gov (United States)

    Zhang, He; Fu, Xin; Hu, Jiayi; Zhu, Zhenjun

    2013-05-24

    This study reports the development of a microfluidic bead-based nucleic acid sensor for sensitive detection of circulating tumor cells in blood samples using multienzyme-nanoparticle amplification and quantum dot labels. In this method, the microbeads functionalized with the capture probes and modified electron rich proteins were arrayed within a microfluidic channel as sensing elements, and the gold nanoparticles (AuNPs) functionalized with the horseradish peroxidases (HRP) and DNA probes were used as labels. Hence, two signal amplification approaches are integrated for enhancing the detection sensitivity of circulating tumor cells. First, the large surface area of Au nanoparticle carrier allows several binding events of HRP on each nanosphere. Second, enhanced mass transport capability inherent from microfluidics leads to higher capture efficiency of targets because continuous flow within micro-channel delivers fresh analyte solution to the reaction site which maintains a high concentration gradient differential to enhance mass transport. Based on the dual signal amplification strategy, the developed microfluidic bead-based nucleic acid sensor could discriminate as low as 5 fM (signal-to-noise (S/N)3) of synthesized carcinoembryonic antigen (CEA) gene fragments and showed a 1000-fold increase in detection limit compared to the off-chip test. In addition, using spiked colorectal cancer cell lines (HT29) in the blood as a model system, the detection limit of this chip-based approach was found to be as low as 1 HT29 in 1 mL blood sample. This microfluidic bead-based nucleic acid sensor is a promising platform for disease-related nucleic acid molecules at the lowest level at their earliest incidence. PMID:23663673

  18. Effects of Momordica charantia on osmotic fragility and label red blood cells and plasmatic protein with 99m-Tc in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Magnata, Simey S.L.P. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Energia Nuclear]. E-mail: sfmagnata@terra.com.br; Correia, Marilia B.L.; Brandao, Jose Odinilson C.; Souza, Grace M.L.; Catanho, Maria Teresa J.A. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Biofisica e Radiobiologia; Terra, Daniele A.; Amorim, Lucia F. [Rio Grande do Norte Univ., Natal, RN (Brazil). Dept. de Fisiologia

    2005-07-01

    The use of natural products in the treatment physiopathology awaken the interest in the inquiry of the action mechanisms. The Momordica charantia, Melao de Sao Caetano, is used in the Caribbean and Orient for the diseases as stomatitis, cancer and diabetes. This work aims to verify the effect of the Momordica charantia's aqueous extract leaves on osmotic fragility and on labeling red blood cells (RBC) and plasmatic proteins with {sup 99m}Tc in vitro. To evaluate the osmotic fragility, samples of heparinized blood (500 mL) was incubed for 1 hour with brut extract (500 mL) in different concentrations (0; 10; 50 and 100% v/v); after centrifugation, the RCB were submitted the incubation (1 hour) with a gradient of NaCl (0;0,1;0,25;0,4;0,7 and 0.9%), the OD of supernatant was determined. With regards to label red blood cells and plasmatic proteins with {sup 99m}Tc in vitro was carried out by incubating of anticoagulant whole blood (500 mL) for 1 hour with brut extract (500 mL) in different concentrations (0; 10; 50 and 100% v/v). A stannous chloride solution of 1,2 {mu}g/mL was added the incubation for 60 minutes. After this the {sup 99m}Tc (3,7 MBq) was added and the incubation was continued for another 10 minutes. Those were centrifuged, precipitated with trichloroacetic acid 5% and mensured in a counter. The results shows that with regard to osmotic fragility, only the extract in the concentration of 100% provoked hemolysis. The Momordica charantia's extract is an agent who modify the fixation of {sup 99m}Tc in red blood cells. The results show with regard to osmotic fragility, only the extract in the quantity 100% provoked hemolysis. It is concluded that the Momordica charantia's extract is an agent who unchains the cellular fragility and {sup 99m}Tc fixation, showing a reduction effect. (author)

  19. Effects of Momordica charantia on osmotic fragility and label red blood cells and plasmatic protein with 99m-Tc in vitro

    International Nuclear Information System (INIS)

    The use of natural products in the treatment physiopathology awaken the interest in the inquiry of the action mechanisms. The Momordica charantia, Melao de Sao Caetano, is used in the Caribbean and Orient for the diseases as stomatitis, cancer and diabetes. This work aims to verify the effect of the Momordica charantia's aqueous extract leaves on osmotic fragility and on labeling red blood cells (RBC) and plasmatic proteins with 99mTc in vitro. To evaluate the osmotic fragility, samples of heparinized blood (500 mL) was incubed for 1 hour with brut extract (500 mL) in different concentrations (0; 10; 50 and 100% v/v); after centrifugation, the RCB were submitted the incubation (1 hour) with a gradient of NaCl (0;0,1;0,25;0,4;0,7 and 0.9%), the OD of supernatant was determined. With regards to label red blood cells and plasmatic proteins with 99mTc in vitro was carried out by incubating of anticoagulant whole blood (500 mL) for 1 hour with brut extract (500 mL) in different concentrations (0; 10; 50 and 100% v/v). A stannous chloride solution of 1,2 μg/mL was added the incubation for 60 minutes. After this the 99mTc (3,7 MBq) was added and the incubation was continued for another 10 minutes. Those were centrifuged, precipitated with trichloroacetic acid 5% and mensured in a counter. The results shows that with regard to osmotic fragility, only the extract in the concentration of 100% provoked hemolysis. The Momordica charantia's extract is an agent who modify the fixation of 99mTc in red blood cells. The results show with regard to osmotic fragility, only the extract in the quantity 100% provoked hemolysis. It is concluded that the Momordica charantia's extract is an agent who unchains the cellular fragility and 99mTc fixation, showing a reduction effect. (author)

  20. Extensive hemangiomatosis diagnosed by scintigraphy with 99mTc-labeled red blood cells in a patient with lower gastrointestinal bleeding

    International Nuclear Information System (INIS)

    Full text: Introduction: The gastrointestinal bleeding may be caused by vascular tumors and other lesions like inflammatory disorders, intestinal obstruction or vascular malformation. The Klippel-Trenaunay syndrome and blue rubber bleb nevus syndrome are hemangiomatosis diseases that may involve the gastrointestinal tract and cause recurrent hemorrhage. The signs and symptoms usually appear at childhood. Case report: male patient, 31 years old, presenting three days of gastrointestinal bleeding and an hemorrhage shock (Hb=3,9). Previous reports of small volume bleeding since childhood and schistossomosis. Dilated veins, hemorrhoid and port wine stain lesions were detected at physical examination in perineal region, penis and scrotum. Inferior limbs were symmetric at inspection. The upper endoscopy showed esophageal varices with no signs of active bleeding. The scintigraphy with 99mTc-labeled red blood cells showed active hemorrhage at recto-sigmoid topography during the first hour of study. Extensive and heterogeneous uptake was seen in gluteus, posterior right thigh and scrotum at the second and fifth hours of study. Then the hypothesis of vascular tumor was considered. The magnetic resonance (MR) of pelvis demonstrated extensive hemangiomatosis at the regions described by the scintigraphy. The clinical and imaging findings suggested the diagnosis of Klippel-Trenaunay syndrome. Discussion: The Klippel-Trenaunay syndrome is a rare disease characterized by congenital vascular and lymphatic malformations (port wine stain lesions, congenital varices) and bone growth and soft tissue disorder. Dilated veins may involve abdominal and pelvic structures, with rectal bleeding and haematuria occurring on average of 20%. The clinical investigation must approach the type, the extent and the severity of the malformation, since the morbidity and the mortality depends on the visceral involvement. The Doppler ultrasound, scanometry of lower extremities, MR, angiography and

  1. Extensive hemangiomatosis diagnosed by scintigraphy with 99mTc-labeled red blood cells in a patient with lower gastrointestinal bleeding

    Energy Technology Data Exchange (ETDEWEB)

    Souza, D.S.F.; Ichiki, W.A.; Borges, A.C.; Coura Filho, G.B.; Vecchia, J.F.; Sapienza, M.T.; Ono, C.R.; Watanabe, T.; Costa, P.L.A.; Hironaka, F.; Cerri, G.G.; Buchpiguel, C.A. [Universidade de Sao Paulo (FM/USP), SP (Brazil). Inst. de Radiologia. Servico de Medicina Nuclear

    2008-07-01

    Full text: Introduction: The gastrointestinal bleeding may be caused by vascular tumors and other lesions like inflammatory disorders, intestinal obstruction or vascular malformation. The Klippel-Trenaunay syndrome and blue rubber bleb nevus syndrome are hemangiomatosis diseases that may involve the gastrointestinal tract and cause recurrent hemorrhage. The signs and symptoms usually appear at childhood. Case report: male patient, 31 years old, presenting three days of gastrointestinal bleeding and an hemorrhage shock (Hb=3,9). Previous reports of small volume bleeding since childhood and schistossomosis. Dilated veins, hemorrhoid and port wine stain lesions were detected at physical examination in perineal region, penis and scrotum. Inferior limbs were symmetric at inspection. The upper endoscopy showed esophageal varices with no signs of active bleeding. The scintigraphy with {sup 99m}Tc-labeled red blood cells showed active hemorrhage at recto-sigmoid topography during the first hour of study. Extensive and heterogeneous uptake was seen in gluteus, posterior right thigh and scrotum at the second and fifth hours of study. Then the hypothesis of vascular tumor was considered. The magnetic resonance (MR) of pelvis demonstrated extensive hemangiomatosis at the regions described by the scintigraphy. The clinical and imaging findings suggested the diagnosis of Klippel-Trenaunay syndrome. Discussion: The Klippel-Trenaunay syndrome is a rare disease characterized by congenital vascular and lymphatic malformations (port wine stain lesions, congenital varices) and bone growth and soft tissue disorder. Dilated veins may involve abdominal and pelvic structures, with rectal bleeding and haematuria occurring on average of 20%. The clinical investigation must approach the type, the extent and the severity of the malformation, since the morbidity and the mortality depends on the visceral involvement. The Doppler ultrasound, scanometry of lower extremities, MR, angiography and

  2. Effect of a peel passion fruit flour (Passiflora edulis f. flavicarpa extract on the labeling of blood constituents with technetium-99m and on the morphology of red blood cells

    Directory of Open Access Journals (Sweden)

    Bernardo Machado Rebello

    2007-09-01

    Full Text Available Passiflora edulis f. flavicarpa (maracuja is a fruit consumed in Brazil and worldwide. Blood constituents labeled with technetium-99m (99mTc are used in nuclear medicine. The effect of P. flavicarpa extract on the radiolabeling of blood constituents and on red blood cells morphology was evaluated. Blood samples from Wistar rats was incubated with P. flavicarpa extract. After that, the labeling of blood constituents with 99mTc was carried out. Samples of plasma and blood cells were precipitated with trichloroacetic acid to isolate the soluble and insoluble fractions of plasma and blood cells. The radioactivity in each fractions was counted and the percentage of radioactivity was determined. Blood smears were also prepared to morphological evaluation and perimeter/area ratio determination. P. flavicarpa extract altered (pPassiflora edulis f. flavicarpa (maracujá é um fruto consumido no Brasil e no mundo. O efeito de um extrato de farinha da casca de maracujá na marcação dos constituintes sangüíneos com tecnécio-99m e na morfologia de hemácias foi avaliado. Amostras de sangue de ratos Wistar, foram incubadas com extrato de P. flavicarpa. Em seguida, o procedimento de marcação de constituintes sangüíneos com Tc-99m foi realizado. Amostras de plasma e células sangüíneas foram separadas e alíquotas destas frações foram precipitadas com ácido tricloroacético para isolamento das frações solúvel e insolúvel do plasma e das células sangüíneas. A radiatividade em cada fração foi contada a porcentagem de radioatividade (%ATI foi calculada. Distensões sangüíneas foram também preparadas para avaliação morfológica e da relação perímetro/área de hemácias. O extrato de P. flavicarpa alterou a fixação do 99mTc nas proteínas plasmáticas e a relação perímetro/área das hemácias. Substâncias presentes no extrato de P. flavicarpa poderiam afetar a marcação de constituintes sangüíneos com 99mTc atuando em alvos

  3. Technetium-99m-labelled red blood cell imaging in the diagnosis of hepatic haemangiomas: the role of SPECT/CT with a hybrid camera

    International Nuclear Information System (INIS)

    Delayed liver single-photon emission computed tomography (SPECT) after 99mTc red blood cell (RBC) labelling is helpful in detecting hepatic haemangiomas; however, diagnosis can be difficult when lesions are situated adjacent to structures like the inferior vena cava, the heart or hepatic vessels, where blood activity persists. The aims of this study were to evaluate the usefulness of RBC SPECT and transmission computed tomography (RBC SPECT/CT) performed simultaneously with a hybrid imaging system for correct characterisation of hepatic lesions in patients with suspected haemangioma, and to assess the additional value of fused images compared with SPECT alone. Twelve patients with 24 liver lesions were studied. The acquisitions of both anatomical (CT) and functional (SPECT) data were performed during a single session. SPECT images were first interpreted alone and then re-evaluated after adding the transmission anatomical maps. Image fusion was successful in all patients, with perfect correspondence between SPECT and CT data, allowing the precise anatomical localisation of sites of increased blood pool activity. SPECT/CT had a significant impact on results in four patients (33.3%) with four lesions defined as indeterminate on SPECT images, accurately characterising the hot spot foci located near vascular structures. In conclusion, RBC SPECT/CT imaging using this hybrid SPECT/CT system is feasible and useful in the identification or exclusion of suspected hepatic haemangiomas located near regions with high vascular activity. (orig.)

  4. Technetium-99m-labelled red blood cell imaging in the diagnosis of hepatic haemangiomas: the role of SPECT/CT with a hybrid camera

    Energy Technology Data Exchange (ETDEWEB)

    Schillaci, Orazio; Danieli, Roberta; Manni, Carlo; Capoccetti, Francesca; Simonetti, Giovanni [Department of Biopathology and Diagnostic Imaging, University ' ' Tor Vergata' ' , Rome (Italy)

    2004-07-01

    Delayed liver single-photon emission computed tomography (SPECT) after {sup 99m}Tc red blood cell (RBC) labelling is helpful in detecting hepatic haemangiomas; however, diagnosis can be difficult when lesions are situated adjacent to structures like the inferior vena cava, the heart or hepatic vessels, where blood activity persists. The aims of this study were to evaluate the usefulness of RBC SPECT and transmission computed tomography (RBC SPECT/CT) performed simultaneously with a hybrid imaging system for correct characterisation of hepatic lesions in patients with suspected haemangioma, and to assess the additional value of fused images compared with SPECT alone. Twelve patients with 24 liver lesions were studied. The acquisitions of both anatomical (CT) and functional (SPECT) data were performed during a single session. SPECT images were first interpreted alone and then re-evaluated after adding the transmission anatomical maps. Image fusion was successful in all patients, with perfect correspondence between SPECT and CT data, allowing the precise anatomical localisation of sites of increased blood pool activity. SPECT/CT had a significant impact on results in four patients (33.3%) with four lesions defined as indeterminate on SPECT images, accurately characterising the hot spot foci located near vascular structures. In conclusion, RBC SPECT/CT imaging using this hybrid SPECT/CT system is feasible and useful in the identification or exclusion of suspected hepatic haemangiomas located near regions with high vascular activity. (orig.)

  5. Effect of an extract of Artemisia vulgaris L. (Mugwort on the in vitro labeling of red blood cells and plasma proteins with technetium-99m

    Directory of Open Access Journals (Sweden)

    Danielle Amorim Terra

    2007-09-01

    Full Text Available The aim of this work was to evaluate the effect of an extract of the Artemisia vulgaris L. (mugwort on the labeling of blood constituents with technetium-99m (99mTc. Blood samples from Wistar rats were incubated with a mugwort extract and the radiolabeling of blood constituents was carried out. Plasma and blood cells were separated by centrifugation. Aliquots of plasma and blood cells were also precipitated with trichloroacetic acid and centrifuged to isolate soluble and insoluble fractions of plasma and blood cells. Radioactivity in each fraction was counted and the percentages of radioactivity (%ATI was calculated. Mugwort extract decreased significantly (pO objetivo desse trabalho foi avaliar o efeito da Artemisia vulgaris L.(artemisa na marcação dos constituintes sangüíneos com tecnécio-99m (99mTc. Amostras de sangue obtidas de ratos Wistar foram incubadas com um extrato de artemisa e o processo de radiomarcação dos constituintes sangüíneos foi realizado. Plasma e células sangüíneas foram isoladas por centrifugação. Alíquotas de plasma e células sangüíneas foram também precipitadas com ácido tricloroacético para isolamento de frações solúvel e insolúvel. A radiatividade em cada fração foi contada e as porcentagens de radioatividade (%ATI foram calculadas. O extrato de artemisa diminuiu significantemente (p<0,05 a %ATI nas células sanguíneas e nas proteínas celulares. A análise dos resultados indicou que o extrato de artemisa apresentaria substâncias que interferir no transporte de íons estanoso e/ou pertecnetato através da membrana do eritrócito alterando a marcação das células sangúineas com 99mTc.

  6. The diagnostic value of angioscintigraphy with sup(99m)Tc-labelled red blood cells for detection of deep vein thrombosis

    International Nuclear Information System (INIS)

    Angioscintigraphy with in vivo sup(99m)Tc-labelled red blood cells is a technically simple, non-invasive method which allows simultaneous bilateral visualization of the pelvis, deep femoral and crural veins up to 24 hours after the labelling. The radiation dose is significantly lower than with X-ray phlebography and thus angioscintigraphy may be used repeatedly and if urgent during pregnancy. In a 'blind' study involving the results from three hospitals, angioscintigraphy correctly classified about 80% of patients with clinical signs of deep vein thrombosis (D.V.T.) when a modern gamma camera was used; inferior results were obtained with an older type of gamma camera. The results also showed that a normal angioscintigram with very high probability excludes the presence of D.V.T. but an abnormal scintigram is not specific for D.V.T. Thus the rather unpleasant conventional phlebography can be omitted in patients with normal angioscintigraphy and angioscintigraphy should be used as first choice. (U.K.)

  7. The role of 99mTc-labelled stannous colloid white blood cells scanning in the evaluation of and differentiation between cellulitis and osteomyelitis

    International Nuclear Information System (INIS)

    Full text: Sequential 99mTc-MDP and 67Ga-citrate scanning have been extensively used in the evaluation of infection. 99mTc-labelled stannous colloid white blood cell (Tc-WBC) is not widely reported. A prospective study was undertaken in 92 patients to assess Tc-WBC and Tc-MDP scanning in this clinical application. Labelling efficiency (LE) was calculated with a mean ± SD of 94 ± 4.8 %. Patients (6) who were treated with gentamycin had an LE ± SD of 70 ± 9%. Twenty six patients treated as osteomyelitis had congruent abnormalities on Tc-WBC and Tc-MDP scans. Twenty one patients treated as cellulitis had abnormalities demonstrated on Tc-WBC but not on Tc-MDP scans. Tc-WBC appeared to define areas of inflammation of bone and soft-tissue in the hands or feet more distinctly than Tc-MDP. Fourteen patients had progress Tc-WBC scans 4-6 weeks following antibiotic treatment. Changes in scan appearances seemed to mirror the clinical course of the patients. Tc-WBC and Tc-MDP scans were useful complementary studies in the diagnosis, evaluation and assessment of treatment in patients with cellulitis and osteomyelitis

  8. On-Orbit, Immuno-Based, Label-Free White Blood Cell Counting System with Microelectromechanical Sensor Technology (OILWBCS-MEMS)

    Science.gov (United States)

    Edmonds, Jessica

    2015-01-01

    Aurora Flight Sciences, in partnership with Draper Laboratory, has developed a miniaturized system to count white blood cells in microgravity environments. The system uses MEMS technology to simultaneously count total white blood cells, the five white blood cell differential subgroups, and various lymphocyte subtypes. The OILWBCS-MEMS detection technology works by immobilizing an array of white blood cell-specific antibodies on small, gold-coated membranes. When blood flows across the membranes, specific cells' surface protein antigens bind to their corresponding antibodies. This binding can be measured and correlated to cell counts. In Phase I, the partners demonstrated surface chemistry sensitivity and specificity for total white blood cells and two lymphocyte subtypes. In Phase II, a functional prototype demonstrated end-to-end operation. This rugged, miniaturized device requires minimal blood sample preparation and will be useful for both space flight and terrestrial applications.

  9. Localization of the acute lower gastrointestinal hemorrhage in vivo-in vitro labeling of red blood cells with sup(99m)Tc

    International Nuclear Information System (INIS)

    For the detection and localization of acute lower gastrointestinal hemorrhage in vivo-in vitro labeling of red blood cells with sup(99m)Tc and sup(99m)Tc sulfur colloid has been sugested. The procedure for labeling RBC with sup(99m)Tc consisted in injecting IV 1 mg of ClSn; 20 minutes after injection of tin 10 cc of blood were withdrawn in a syringe containing 20 mCi of sup(99m)Tc; this was incubated for 10 minutes and then injected IV. Scintigraphy of the abdominal cavity was done in supine position and performed with a large field gamma camera with a parallel hole-low energy colimator. Computer adquisition of images was started 5 minutes after RBC injection and made at the rate of one enery 5 minutes for 45 minutes. 14 patients were studied divided in: a) control: 6 patients. b) with active gastrointestinal hemorrhage: 4 patients had positive scintigraphy. The hemorrhage was documented with superior mesenteric arteriography, endoscopy and/or necropsy. The sensitivity was 100%. In 4 out of 14 patients scintigraphy with sup(99m)Tc RBC compared with simultaneous sup(99m)Tc sulfur colloid demonstrated that all patients with positive sup(99m)Tc RBC had also positive sup(99m)Tc sulfur colloid scintigraphy. c) without active gastrointestinal hemorrhage: all of them had negative scintigraphy (specificity 100%). Abdominal scintigraphy with sup(99m)Tc RBC or sulfur colloid are both sensitive for detection and localization of lower gastrointestinal bleeding and the negative study suggests the absence of active hemorrhage. It is suggested that the sup(99m)Tc sulfur colloid scintigraphy should be the initial procedure to study these patients and abdominal arteriography should be performed only in patients with positive abdominal scintigraphy. (M.E.L.)

  10. Immunogold labels: cell-surface markers in atomic force microscopy

    NARCIS (Netherlands)

    Putman, Constant A.J.; Grooth, de Bart G.; Hansma, Paul K.; Hulst, van Niek F.; Greve, Jan

    1993-01-01

    The feasibility of using immunogold labels as cell-surface markers in atomic force microscopy is shown in this paper. The atomic force microscope (AFM) was used to image the surface of immunogold-labeled human lymphocytes. The lymphocytes were isolated from whole blood and labeled by an indirect imm

  11. Tc-99m HMPAO labelled white blood cell scintigraphy in the assessment of the extent and severity of active ulcerative colitis

    International Nuclear Information System (INIS)

    Full text: Aim: In this study it was aimed to assess the role of Tc-99m HMPAO labelled white blood cell (WBC) scintigraphy on different imaging times in the assessment of disease extent and severity in active ulcerative colitis (UC). Materials and Methods: Twenty-one consecutive patients (10 women, 11 men; mean age: 42.4 12 years) with active UC were prospectively studied. Clinical evaluation, laboratory analysis, endoscopy-biopsy, and scintigraphy were performed within three days before any treatment. Scintigraphy was performed 1, 2, and 4 h after 111-185 MBq Tc-99m HMPAO labelled WBC injection. Large bowel was evaluated in 4 segments (recto-sigmoid, descending, transverse and ascending colon) during the endoscopy and scintigraphy. Clinical activity score (CAS), endoscopic activity score (EAS), and scintigraphic activity score (SAS) at 1 hr, 2 hr, and 4 hr were calculated for each patient. Results: Scintigraphy showed 80%, 83%, and 80% agreement in the number of pathological segment with comparing endoscopy, at 1 hr, 2 hr and 4 hr imaging, respectively. Sensitivity, specificity, and accuracy values of Tc-99m HMPAO labelled WBC scintigraphy were calculated as follow, respectively: 1 hr imaging: 86%, 73%, 83%; 2 hr imaging: 89 %, 74 %, 86 %; 4 hr imaging: 90%, 58%, 83% in the detecting of active inflammatory segments using by endoscopy as gold standard. Statistically significant correlations were found between EAS and SAS for 2 hr and 4 hr imaging (r=0.64 and r=0.66, respectively). There were also significant correlations between CAS and SAS for 2 hr and 4 hr imaging (r=0.59 and r=0.62, respectively). Conclusion: Tc-99m HMPAO labelled WBC scintigraphy at 2 hr well correlated with endoscopy in the assessment of the extent of UC. Because of the higher correlation values of 4 hr SAS than 2 hr SAS with CAS and EAS, our results indicated that late scintigraphic imaging (4 h) might be useful in the evaluation of the severity of UC. (author)

  12. Contribution to the study of the red blood cells labelled with chromium-51 and technetium-99 m

    International Nuclear Information System (INIS)

    Although the bindings of Cr-51 and Tc-99 m were both in the β chain of hemoglobin molecule, the results obtained after previous incubations of the RBC with chromium and technetium, and the determinations of the efficiency of the labeling of RBC showed that the points of fixing of chromium and technetium with β chain of hemoglobin were probably different. The observations through the optic microscope allowed the verification that, at the concentration of 100 mg/ml of Cr-50, there were morphologic in the RBC. These modifications were not found after the other treatments. The comparison between scintigraphy obtained with Tc-99 m or Cr-51 RBC suggested that the technique which employs Tc-99 m can be more adequate than the one with Cr-51. (author)

  13. Diagnosis of deep vein thrombosis of the lower limbs with scintigraphy of red blood cells labelled with 99m technetium

    International Nuclear Information System (INIS)

    The clinical diagnosis of leg deep vein thrombosis (DVT) is notoriously unreliable. It must be supplemented by objective techniques which all have drawbacks. 99mTc-RBC venography also has its limitations, yet it is a simple, safe, and useful test for diagnosing DVT of the lower limb. When done carefully, it is a rewarding procedure with good sensitivity and specificity for the condition both in the calf and ilio-femoral regions. Blood pool venography is readily accessible to all nuclear medicine department for the diagnosis of thrombophlebitis and also the follow-up of treated patients

  14. Improving the diagnosis of acute appendicitis in children with atypical clinical findings using the technetium-99m hexamethylpropylene amine oxime-labelled white-blood-cell abdomen scan

    International Nuclear Information System (INIS)

    Heading AbstractBackground. Diagnosing acute appendicitis in children with equivocal signs and symptoms may be difficult. The usual approach is hospital observation and frequent re-examination. However, many surgeons are reluctant to delay surgery because of the risk of perforation and a negative laparotomy.Objective. To assess and compare the value of the technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO)-labelled white-blood-cell (WBC) abdomen scan in the diagnosis of acute appendicitis in children with atypical clinical presentation.Patients and methods. Fifty children with acute right lower quadrant abdominal pain and possible acute appendicitis, but atypical findings were included. After IV injection of 99mTc-HMPAO-labelled WBCs, serial anterior abdomen scans were obtained using a gamma camera.Results. Thirty-three children underwent surgery, while 17 children were managed conservatively and were followed up for at least 1 month. Four children had false-positive results and one child had a false-negative scan result. The overall sensitivity, specificity, accuracy, positive predictive value and negative predictive value of the scan to diagnose acute appendicitis in children with atypical findings was 96.7, 80.0, 90.0, 87.8 and 94.1%, respectively.Conclusions. The 99mTc-HMPAO WBC abdomen scan is a potential tool for diagnosing acute appendicitis in children with atypical clinical findings. The high sensitivity and negative predictive value allows early discharge from the emergency department to avoid costly observation in hospital and potentially unnecessary surgery in those patients with negative scans. (orig.)

  15. Transcriptomic Profile of Whole Blood Cells from Elderly Subjects Fed Probiotic Bacteria Lactobacillus rhamnosus GG ATCC 53103 (LGG in a Phase I Open Label Study.

    Directory of Open Access Journals (Sweden)

    Gloria Solano-Aguilar

    Full Text Available We examined gene expression of whole blood cells (WBC from 11 healthy elderly volunteers participating on a Phase I open label study before and after oral treatment with Lactobacillus rhamnosus GG-ATCC 53103 (LGG using RNA-sequencing (RNA-Seq. Elderly patients (65-80 yrs completed a clinical assessment for health status and had blood drawn for cellular RNA extraction at study admission (Baseline, after 28 days of daily LGG treatment (Day 28 and at the end of the study (Day 56 after LGG treatment had been suspended for 28 days. Treatment compliance was verified by measuring LGG-DNA copy levels detected in host fecal samples. Normalized gene expression levels in WBC RNA were analyzed using a paired design built within three analysis platforms (edgeR, DESeq2 and TSPM commonly used for gene count data analysis. From the 25,990 transcripts detected, 95 differentially expressed genes (DEGs were detected in common by all analysis platforms with a nominal significant difference in gene expression at Day 28 following LGG treatment (FDR<0.1; 77 decreased and 18 increased. With a more stringent significance threshold (FDR<0.05, only two genes (FCER2 and LY86, were down-regulated more than 1.5 fold and met the criteria for differential expression across two analysis platforms. The remaining 93 genes were only detected at this threshold level with DESeq2 platform. Data analysis for biological interpretation of DEGs with an absolute fold change of 1.5 revealed down-regulation of overlapping genes involved with Cellular movement, Cell to cell signaling interactions, Immune cell trafficking and Inflammatory response. These data provide evidence for LGG-induced transcriptional modulation in healthy elderly volunteers because pre-treatment transcription levels were restored at 28 days after LGG treatment was stopped. To gain insight into the signaling pathways affected in response to LGG treatment, DEG were mapped using biological pathways and genomic data mining

  16. Drug interaction with radiopharmaceuticals: effect on the labeling of red blood cells with technetium-99m and on the bioavailability of radiopharmaceuticals

    OpenAIRE

    Gomes Maria Luisa; Oliveira Marcia B. Nunes de; Bernardo-Filho Mario

    2002-01-01

    The evidence that natural and synthetic drugs can affect radiolabeling or bioavailability of radiopharmaceuticals in setting of nuclear medicine clinic is already known. However, this drug interaction with radiopharmaceuticals (DIR) is not completely understood. Several authors have described the effect of drugs on the labeling of blood elements with technetium-99m (99mTc) and on the biodistribution of radiopharmaceuticals. When the DIR is known, if desirable or undesirable, the natural conse...

  17. Intraindividual comparison of {sup 99m}Tc-labelled anti-SSEA-1 antigranulocyte antibody and {sup 99m}Tc-HMPAO labelled white blood cells for the imaging of infection

    Energy Technology Data Exchange (ETDEWEB)

    Gratz, S.; Behr, T.; Herrmann, A.; Becker, W. [Goettingen Univ. (Germany). Abt. fuer Nuklearmedizin; Dresing, K.; Stuermer, K.M. [Department of Trauma, Plastic and Reconstructive Surgery, University of Goettingen (Germany); Tarditi, L.; Franceschini, R. [SORIN Biomedica, Saluggia (Italy); Rhodes, B. [RHOMED, Albuquerque, New Mexiko (United States)

    1998-04-01

    The aim of this study was to compare the diagnostic accuracy of the {sup 99m}Tc-anti-SSEA-1 Mab with that of {sup 99m}Tc-HMPAO labelled white blood cells (WBCs). To this end, 17 patients with 23 proven infectious foci were examined with 555 MBq {sup 99m}Tc-anti-SSEA-1 MAb and with 370 MBq {sup 99m}Tc-HMPAO labelled autologous leucocytes within a period of 7 days. All the infections were confirmed by culturd of 7 days. Whole-body images and planar spot views with the antibody were performed at 1-h, 4-h and 24-h post injection; the biodistribution of the antibody was quantified, absorbed radiation doses were calculated and the diagnostic results were compared with the {sup 99m}Tc-HMPAO WBC images. Human anti-mouse antibody (HAMA) evaluation was performed in all patients before and 3 months after antibody injection. Blood was drawn at different times after {sup 99m}Tc-anti-SSEA-1 MAb injection to determine the amount of granulocyte-associated radioactivity and to calculate recovery. {sup 99m}Tc-anti-SSEA-1 MAb scintigraphy detected all 23 lesions, while 21 were detected with {sup 99m}Tc-HMPAO WBC scan. In this small group of patients, the sensitivity and specificity of {sup 99m}Tc-anti-SSEA-1 MAb scintigraphy were 95% and 96% respectively, as compared with 91% and 82% respectively for {sup 99m}Tc-HMPAO WBC scan. An increasing uptake of the injected activity in the lesion at different time points was indicative of high affinity and of specific PMN binding.There was no HAMA formation. In four of five patients investigated, a transient mild leukopenia was found at 15 min p.i. There was increased uptake of the antibody in liver and spleen and normal uptake in kidneys and bone marrow.The estimated radiation doses for the whole body and the red bone marrow were 1.1 x 10{sup -2} cGy/37 MBq and 5.3 x 10{sup -2} cGy/37 MBq, respectively. The activity associated to the PMNs in vivo was 33.5%, 30.6%, 21.3% and 9% at 5, 15, 30 and 45 min. post-injection, respectively

  18. Drug interaction with radiopharmaceuticals: effect on the labeling of red blood cells with Technetium-99m and on the bioavailability of radiopharmaceuticals

    Energy Technology Data Exchange (ETDEWEB)

    Gomes, Maria Luisa; Oliveira, Marcia B. Nunes de [Universidade do Estado, Rio de Janeiro, RJ (Brazil). Inst. de Biologia. Dept. de Biofisica e Biometria; Bernardo-Filho, Mario [Universidade do Estado, Rio de Janeiro, RJ (Brazil). Inst. de Biologia. Dept. de Biofisica e Biometria; Instituto Nacional do Cancer, Rio de Janeiro, RJ (Brazil). Coordenadoria de Pesquisa

    2002-09-01

    The evidence that natural and synthetic drugs can affect radiolabeling or bioavailability of radiopharmaceuticals in setting of nuclear medicine clinic is already known. However, this drug interaction with radiopharmaceuticals (DIR) is not completely understood. Several authors have described the effect of drugs on the labeling of blood elements with Technetium-99m (99mTc) and on the biodistribution of radiopharmaceuticals. When the DIR is known, if desirable or undesirable, the natural consequence is a correct diagnosis. However, when it is unknown, it is undesirable and the consequences are the possibility of misdiagnosis and/or the repetition of the examination with and increase of radiation dose to the patient. The possible explanation to the appearance of DIR are radiopharmaceutical modification, alternation of the labeling efficiency of the radiopharmaceutical, modification of the target, modification of no target and/or the alteration of the binding of the radiopharmaceutical on the blood proteins. The effect of drugs on the labeling of blood elements with 99 mTc might be explained by a direct inhibition (chelating action) of the stannous and pertechnetate ions, damage induced in the plasma membrane, competition of the cited ions for the same binding sites, possible generation of reactive oxygen species that could oxidize the stannous ion and/or (v) direct oxidation of the stannous ion. In conclusion, the development of biological models to study the D IR is highly relevant. (author)

  19. Drug interaction with radiopharmaceuticals: effect on the labeling of red blood cells with Technetium-99m and on the bioavailability of radiopharmaceuticals

    International Nuclear Information System (INIS)

    The evidence that natural and synthetic drugs can affect radiolabeling or bioavailability of radiopharmaceuticals in setting of nuclear medicine clinic is already known. However, this drug interaction with radiopharmaceuticals (DIR) is not completely understood. Several authors have described the effect of drugs on the labeling of blood elements with Technetium-99m (99mTc) and on the biodistribution of radiopharmaceuticals. When the DIR is known, if desirable or undesirable, the natural consequence is a correct diagnosis. However, when it is unknown, it is undesirable and the consequences are the possibility of misdiagnosis and/or the repetition of the examination with and increase of radiation dose to the patient. The possible explanation to the appearance of DIR are radiopharmaceutical modification, alternation of the labeling efficiency of the radiopharmaceutical, modification of the target, modification of no target and/or the alteration of the binding of the radiopharmaceutical on the blood proteins. The effect of drugs on the labeling of blood elements with 99 mTc might be explained by a direct inhibition (chelating action) of the stannous and pertechnetate ions, damage induced in the plasma membrane, competition of the cited ions for the same binding sites, possible generation of reactive oxygen species that could oxidize the stannous ion and/or (v) direct oxidation of the stannous ion. In conclusion, the development of biological models to study the D IR is highly relevant. (author)

  20. Acute toxicity of phyto medicine Mulher Ativa and antioxidant properties on the labeling of blood cells and plasmatic proteins with 99mTc in vitro

    International Nuclear Information System (INIS)

    Full text: Medicinal plants originate natural products that are biologically active and widely employed as an alternative source in health care. Mulher Ativa is a phyto medicine used in several gynecological pathologies composed of eight medicinal plants which exhibits estrogen properties in the reproductive tract. The objective of this work was determining the acute toxicity studies investigated of Mulher Ativa (Ma) were performed in mice and antioxidant properties on the labeling of blood cells and plasmatic protein with 99mTc in vitro. For these studies, mice were divided in two groups, containing 05 animals each. The treated group received Ma in doses of 10, 100, 200, 300, 600, 1000, 2000, 3000 mg/kg of animal weight. Mice were carefully observed 0.5, 1, 2, 3, 4, 24, 48, and 72h after the treatment to assess possible clinical or toxicological symptoms. The second experiment was realized incubating heparin with blood carried out the experiments. Different concentrations of Mulher Ativa were chosen (200; 100; 50; 25; 12,5 mg/mL). A stannous chloride solution was also added and incubation was kept for 60 minutes. After this, 99mTc was added and the incubation was continued for 10 minutes. The mixture was centrifuged, precipitated with thichloroacetic acid 5% and soluble (SF) and insoluble fractions (IF) were separated. The radioactivities in the groups P, BC, IF-P, SF-P, IF- BC, SF-BC were determined in counter. The analysis of radioactivity in the samples of P and BC isolated from samples of whole blood treated with Mulher Ativa showed decrease significant (*p99mTc in the TCA-insoluble fraction of plasma. It is also concluded that presents antioxidant properties. As part of this pharmacological study, the acute toxicity of Ma in mice was first investigated. In these doses, the median lethal dose LD50 was determined to be higher than highest dose tested i.e 2.0 gkg -1 b.w. From this data, the estimated LD50 was 2060.1 mg/kg. The product was classified as slightly

  1. The value of 99mTc-HMPAO labelled white blood cell scintigraphy in acute appendicitis patients with an equivocal clinical presentation

    International Nuclear Information System (INIS)

    Various imaging studies can be performed in the evaluation of patients with a clinical presentation equivocal for acute appendicitis. One of these studies is technetium-99m hexamethylpropylene amine oxime (HMPAO) labelled white blood cell (WBC) scintigraphy. The aim of this study was to evaluate the accuracy and clinical value of 99mTc-HMPAO WBC scintigraphy in the aforementioned group of patients. Forty-one patients who had acute right lower quadrant abdominal pain with a clinical presentation equivocal for acute appendicitis were included in the study. The anterior abdomen and pelvis were imaged up to 4 h after the injection of 125-300 MBq 99mTc-HMPAO WBCs and the results were immediately reported to the surgeon before a decision was taken on whether to perform laparotomy. Diagnostic accuracy was established by the intra-operative findings and the histopathology in operated patients. In non-operated patients, absence of abdominal symptoms 1 month after scintigraphy and/or identification of another cause of abdominal pain was used to rule out acute appendicitis. There were 16 patients with positive scintigraphy and 81% of these patients were positive within 2 h post injection. There were no false-positive or false-negative results. We operated on 17 (41.4%) patients, and only one patient (5.9%) underwent unnecessary laparotomy. We conclude that 99mTc-HMPAO WBC scintigraphy is a rapid, highly accurate method for the exclusion of acute appendicitis and that its use can lower the unnecessarily high laparotomy rate in patients with an equivocal clinical presentation. (orig.)

  2. Immunogold labels: cell-surface markers in atomic force microscopy

    OpenAIRE

    Putman, Constant A.J.; Grooth, de, B.G.; Hansma, Paul K.; Hulst, van der, R.W.M.; Greve, Jan

    1993-01-01

    The feasibility of using immunogold labels as cell-surface markers in atomic force microscopy is shown in this paper. The atomic force microscope (AFM) was used to image the surface of immunogold-labeled human lymphocytes. The lymphocytes were isolated from whole blood and labeled by an indirect immunolabeling method using the monoclonal antibody anti-CD3 and a secondary antibody (Goat-anti-Mouse) linked to 30 nm colloidal gold particles. Some of the samples were enhanced by silver deposition...

  3. Donating Peripheral Blood Stem Cells

    Science.gov (United States)

    ... this page Print this page Donating peripheral blood stem cells Peripheral blood stem cell (PBSC) donation is a nonsurgical procedure to collect ... Donating bone marrow Donor experiences videos Peripheral blood stem cell (PBSC) donation is one of two methods of ...

  4. Red blood cells, sickle cell (image)

    Science.gov (United States)

    Sickle cell anemia is an inherited blood disease in which the red blood cells produce abnormal pigment (hemoglobin). ... abnormal hemoglobin causes deformity of the red blood cells into crescent or sickle-shapes, as seen in this photomicrograph.

  5. The effect of drugs on the labeling of blood elements with technetium-99m.

    Science.gov (United States)

    Braga, A C; Oliveira, M B; Feliciano, G D; Reiniger, I W; Oliveira, J F; Silva, C R; Bernardo-Filho, M

    2000-07-01

    The influence of drugs on the labeling of red blood cells and plasma proteins with 99mTc has been reported. Any drug, which alters the labeling of the tracer, could be expected to modify the disposition of the radiopharmaceuticals. Red blood cells (RBC) labeled with technetium-99m (99mTc) are used for several evaluations in nuclear medicine. We have evaluated the effect of Thuya occidentalis, Peumus boldus and Nicotiana tabacum (tobacco) extracts on the labeling of RBC and plasma and cellular proteins with 99mTc. Blood was incubated with the drugs. Stannous chloride (SnCl2) solutions and 99mTc were added. Plasma (P) and blood cells (BC) were separated. The percentage of radioactivity (%ATI) bound to P and BC was determined. The %ATI on the plasma and cellular proteins was also evaluated by precipitation of P and BC samples with trichloroacetic acid (TCA) and isolation of soluble (SF) and insoluble (IF) fractions. The analysis of the results shows that there is a decrease in %ATI (from 97.64 to 75.89 percent) in BC with Thuya occidentalis extract. The labeling of RBC and plasma proteins can be decreased in presence of tobacco. This can be due either a direct or indirect effect (reactive oxygen species) of tobacco. The analysis of radioactivity in samples of P and BC isolated from samples of whole blood treated with Peumus boldus showed a rapid uptake of the radioactivity by blood cells in the presence of the Peumus boldus, whereas there was a slight decrease in the amount of 99mTc radioactivity in the TCA-insoluble fraction of plasma. This study shows that extracts of some medicinal plants can affect the radiolabeling of red blood cells with 99mTc using an in vitro technique. PMID:10903389

  6. Inverse correlation between cerebral blood flow measured by continuous arterial spin-labeling (CASL) MRI and neurocognitive function in children with sickle cell anemia (SCA)

    OpenAIRE

    Strouse, John J.; Cox, Christiane S.; Melhem, Elias R.; Lu, Hanzhang; Kraut, Michael A.; Razumovsky, Alexander; Yohay, Kaleb; van Zijl, Peter C.; Casella, James F.

    2006-01-01

    Overt stroke, clinically “silent” cerebral infarct, and neurocognitive impairment are frequent complications of sickle cell anemia (SCA). Current imaging techniques have limited sensitivity and specificity to identify children at risk for neurocognitive impairment. We prospectively evaluated 24 children with SCA with a neurologic exam, complete blood count, transcranial Doppler ultrasound (TCD), measurement of intelligence quotient (IQ), and magnetic resonance imaging (MRI) with measurement o...

  7. Diagnosing osteomyelitis in the diabetic foot: a pilot study to examine the sensitivity and specificity of Tc(99m) white blood cell-labelled single photon emission computed tomography/computed tomography.

    Science.gov (United States)

    Przybylski, Mallory M; Holloway, Samantha; Vyce, Steven D; Obando, Antonio

    2016-06-01

    Diabetic foot ulceration poses a significant threat of osteomyelitis (OM) and subsequent amputation. The diagnosis of OM via imaging studies is difficult as radiographic findings do not present immediately and advanced imaging studies may be contraindicated or unavailable. A novel diagnostic tool has been developed which synthesises technetium-99 white blood cell-labelled single-photon emission computed tomography and computed tomography (Tc(99m) WBC labelled-SPECT/CT) imaging, effectively enhancing anatomic detail. The aim of this pilot study was to determine the validity and reliability of this novel imaging technique in patients with diabetic foot ulcers in a Veterans Affairs healthcare facility. A retrospective review was performed on consecutive patients who met the inclusion criteria (n = 14) and underwent Tc(99m) WBC-labelled SPECT/CT for suspected OM. Histopathologic analysis of bone specimen (when available) and International Working Group on the Diabetic Foot consensus criteria were used as a reference standard. The sensitivity and specificity of Tc(99m) WBC-labelled SPECT/CT were 87·50% [confidence interval (CI): 64·58-110·42%] and 71·43% (CI: 37·96-104·90%), respectively. Negative predictive value (NPV) and positive predictive value (PPV) were 83·33% (CI: 53·51-113·15%) and 77·78% (CI: 50·62-104·94%), respectively, with a likelihood ratio (LR) of 3·063 and an accuracy of 80%. These findings suggest Tc(99m) WBC-labelled SPECT/CT can be useful in imaging OM in patients with diabetic foot ulcers. PMID:24976368

  8. Photomodification of human immunocompetent blood cells

    International Nuclear Information System (INIS)

    In this paper, processes of photomodification of lymphoid cells in human blood, developing immediately after exposure to visible radiation and also in the late stages after irradiation, were investigated by methods of spontaneous and immune rosette formation and the blast transformation test, combined with treatment with the antioxidant alpha-tocopherol and the radioactive assessment of spontaneous and stimulated DNA synthesis by tritium-thymidine-labelled cells

  9. Drug interaction with radiopharmaceuticals: effect on the labeling of red blood cells with technetium-99m and on the bioavailability of radiopharmaceuticals

    Directory of Open Access Journals (Sweden)

    Maria Luisa Gomes

    2002-09-01

    Full Text Available The evidence that natural and synthetic drugs can affect radiolabeling or bioavailability of radiopharmaceuticals in setting of nuclear medicine clinic is already known. However, this drug interaction with radiopharmaceuticals (DIR is not completely understood. Several authors have described the effect of drugs on the labeling of blood elements with technetium-99m (99mTc and on the biodistribution of radiopharmaceuticals. When the DIR is known, if desirable or undesirable, the natural consequence is a correct diagnosis. However, when it is unknown, it is undesirable and the consequences are the possibility of misdiagnosis and/or the repetition of the examination with an increase of radiation dose to the patient. The possible explanation to the appearance of DIR are (a radiopharmaceutical modification, (b alteration of the labeling efficiency of the radiopharmaceutical, (c modification of the target, (d modification of no target and/or the (e alteration of the binding of the radiopharmaceutical on the blood proteins. The effect of drugs on the labeling of blood elements with 99mTc might be explained by (i a direct inhibition (chelating action of the stannous and pertechnetate ions, (ii damage induced in the plasma membrane, (iii competition of the cited ions for the same binding sites, (iv possible generation of reactive oxygen species that could oxidize the stannous ion and/or (v direct oxidation of the stannous ion. In conclusion, the development of biological models to study the DIR is highly relevant.A evidência de que drogas naturais ou sintéticas podem afetar a radiomarcação ou a biodisponibilidade de radiofármacos nos procedimentos de medicina nuclear já é bem conhecida. Entretanto, essa interação de droga com radiofármacos (IDR não está completamente compreendida. Vários autores têm descrito o efeito de drogas na marcação de elementos sanguíneos com tecnécio-99m (99mTce na biodistribuição de radiofármacos. Quando a

  10. Estimation of anti-D IgG in red blood cell eluates using the specific radioactivity of 125I-labeled IgG: effect of unlabeled, cytophilic IgG

    Energy Technology Data Exchange (ETDEWEB)

    Masouredis, S.P.; Mahan, L.C.; Sudora, E.J.; Langley, J.W.; Victoria, E.J.

    1981-01-01

    The specific radioactivity of conventionally prepared 125I IgG anti-D eluates is significantly less (from 1/5 to 1/20) than that of the 125I IgG fraction used to prepare the eluate. This discrepancy is due to the release of unlabeled, cytophilic IgG from normal red blood cells during eluate preparation and does not represent an underestimation of the eluate anti-D IgG content. Cytophilic IgG content of eluates plays an important role in reducing the nonimmunologic binding of labeled antibody IgG. The results justify the assumption used in numerous studies that the specific radioactivity of 125I IgG fractions can be used to provide a valid estimate of the anti-D IgG content of eluates.

  11. Stable isotope labeling of oligosaccharide cell surface antigens

    Energy Technology Data Exchange (ETDEWEB)

    Unkefer, C.J.; Silks, L.A. III; Martinez, R.A. [and others

    1998-12-31

    The overall goal of this Laboratory Directed Research and Development (LDRD) project was to develop new methods for synthesis of {sup 13}C-labeled oligosaccharides that are required for nuclear magnetic resonance (NMR) studies of their solution conformation. Oligosaccharides are components of the cell`s outer surface and are involved in important processes such as cell-cell recognition and adhesion. Recently, Danishefsky and coworkers at Slone-Kettering Cancer Center developed a method for the solid-phase chemical synthesis of oligosaccharides. The specific goal of this LDRD project was to prepare uniform {sup 13}C-labeled aldohexose precursors required for the solid-phase synthesis of the Lewis blood-group antigenic determinants. We report the synthesis of {sup 13}C-labeled D-glucal, D-galactal and Fucosyl precursors. We have been collaborating with the Danishefsky group on the synthesis of the Lewis oligosaccharides and the NMR analysis of their solution conformation.

  12. Assessment of the effect of Bacopa monnieri (L.) Wettst. extract on the labeling of blood elements with technetium-99m and on the morphology of red blood cells Avaliação do efeito do extrato de Bacopa monnieri (L.) Wettst. na marcação de elementos sanguíneos com tecnécio-99m e na morfologia de células vermelhas do sangue

    OpenAIRE

    Kakali De; Susmita Chandra; Mridula Misra

    2009-01-01

    Bacopa monnieri (L.) Wettst. (BM), a traditional Ayurvedic medicine, used for centuries as a memory enhancing, anti-inflammatory, antipyretic, sedative and antiepileptic agent. BM extract have been extensively investigated by several authors for their neuropharmacological effects. In nuclear medicine, red blood cells (RBC) labeled with technetium-99m (99mTc) have several clinical applications. However, data have demonstrated that synthetic or natural drugs could modify the labeling of RBC wit...

  13. Blinded retrospective comparison of MR imaging and Tc-99m-labeled red blood cell SPECT for definitive diagnosis of hepatic hemangiomas

    International Nuclear Information System (INIS)

    This paper reports how, to define the roles of MR imaging versus Tc-99m red blood cell (RBC) single photon emission CT (SPECT) in imaging hepatic hemangiomas, 32 patients with 63 suspected (US or CT) hemangiomas were imaged by each method. Seventeen patients had known cancer. T2-weighted multiecho axial SE images (2,000/50, 100 [repetition time msec/echo time msec] 0.5T) were obtained; flow, immediate static, and 90 minutes static Tc-99m RBC SPECT (axial, coronal, sagittal) was performed. The studies were retrospectively evaluated on a 5-point scale by blinded readers who knew the location of the reference lesion(s) and whether the patient had cancer. Fifty-eight lesions were hemangiomas, two FNH, two metastases, and one focal normal liver. In 36 hemangiomas, SPECT and MR imaging agreed at the highest confidence levels. In three, only SPECT characterized hemangioma at the highest levels. In nine, MR imaging correctly characterized lesions not identified with SPECT. In three, neither modality was correct. The greatest improvement in MR diagnosis was in those lesions smaller than 1.9 cm and near the dome of the liver

  14. Image acquisition and interpretation criteria for {sup 99m}Tc-HMPAO-labelled white blood cell scintigraphy: results of a multicentre study

    Energy Technology Data Exchange (ETDEWEB)

    Erba, Paola A. [University of Pisa Medical School (Italy). Regional Center of Nuclear Medicine; Glaudemans, Andor W.J.M.; Dierckx, Rudi A.J.O. [University Medical Center Groningen (Netherlands). Dept. of Nuclear Medicine and Molecular Imaging; Veltman, Niels C. [Jeroen Bosch Hospital, ' s-Hertogenbosch (Netherlands). Dept. of Nuclear Medicine; Sollini, Martina [Arcisprdale S. Maria Nuova - IRCCS, Reggio Emilia (Italy). Nuclear Medicine Unit; Pacilio, Marta; Galli, Filippo [Sapienza Univ., Rome (Italy). Nuclear Medicine Unit; Signore, Alberto [University Medical Center Groningen (Netherlands). Dept. of Nuclear Medicine and Molecular Imaging; Sapienza Univ., Rome (Italy). Nuclear Medicine Unit; Sapienza Univ., Rome (Italy). Ospedale S. Andrea Medicina Nucleare

    2014-04-15

    There is no consensus yet on the best protocol for planar image acquisition and interpretation of radiolabelled white blood cell (WBC) scintigraphy. This may account for differences in reported diagnostic accuracy amongst different centres. This was a multicentre retrospective study analysing 235 WBC scans divided into two groups. The first group of scans (105 patients) were acquired with a fixed-time acquisition protocol and the second group (130 patients) were acquired with a decay time-corrected acquisition protocol. Planar images were interpreted both qualitatively and semiquantitatively. Three blinded readers analysed the images. The most accurate imaging acquisition protocol comprised image acquisition at 3 - 4 h and at 20 - 24 h in time mode with acquisition times corrected for isotope decay. Using this protocol, visual analysis had high sensitivity and specificity in the diagnosis of infection. Semiquantitative analysis could be used in doubtful cases, with no cut-off for the percentage increase in radiolabelled WBC over time, as a criterion to define a positive scan. (orig.)

  15. Image acquisition and interpretation criteria for 99mTc-HMPAO-labelled white blood cell scintigraphy: results of a multicentre study

    International Nuclear Information System (INIS)

    There is no consensus yet on the best protocol for planar image acquisition and interpretation of radiolabelled white blood cell (WBC) scintigraphy. This may account for differences in reported diagnostic accuracy amongst different centres. This was a multicentre retrospective study analysing 235 WBC scans divided into two groups. The first group of scans (105 patients) were acquired with a fixed-time acquisition protocol and the second group (130 patients) were acquired with a decay time-corrected acquisition protocol. Planar images were interpreted both qualitatively and semiquantitatively. Three blinded readers analysed the images. The most accurate imaging acquisition protocol comprised image acquisition at 3 - 4 h and at 20 - 24 h in time mode with acquisition times corrected for isotope decay. Using this protocol, visual analysis had high sensitivity and specificity in the diagnosis of infection. Semiquantitative analysis could be used in doubtful cases, with no cut-off for the percentage increase in radiolabelled WBC over time, as a criterion to define a positive scan. (orig.)

  16. Leukemic cell kinetics in peripheral blood, 2

    International Nuclear Information System (INIS)

    The in vivo kinetics of autologous leukemic cells labeled in vitro with indium-111-oxine was studied in 10 patients with acute non-lymphocytic leukemia (ANLL), consisting of 7 patients with acute myeloblastic leukemia (AML), 2 with acute myelomonocytic leukemia (AMML) and 1 with acute monocytic leukemia (AMoL). Leukemic cell disappearance curves showed a single exponential line. The half tims of disappearance (T1/2) in AML was 18.6 +- 8.3 hours (mean +- s.d.), and was longer than that of normal neutrophils. In AMML and AMoL, T1/2 was 11.5 +- 1.4 hours, and tended to be shorter than that in AML (p < 0.1). Total blood leukemic cell pool (TBLCP) size correlated with blood leukemic cell count (LC) (Y = 1.11 + 2.01X, r = 0.95). The ratio of marginal (MLCP) to circulating leukemic cell pool (CLCP) size was 2.38 +- 0.99 in AML. There was no significant correlation between leukemic cell turnover rate (LCTR) and TBLCP size. As for organ distribution, labeled leukemic cells passed immediately through lungs, are then accumulated markedly in the spleen and liver in that order. Initial pulmonary radioactivity was observed in only one of the AMML patients. Only in AMoL, hepatic radioactivity 30 minutes after the injection surpassed splenic radioactivity. Accumulation of radioactivity in the bone marrow was observed in 6 out of 8 patients studied. Radioactivity of the leukemic cells isolated from the bone marrow in 4 patients was larger than that expected from mixing of peripheral blood leukemic cells, suggesting that a portion of blood leukemic cells returned to the bone marrow. (author)

  17. Guava extract (Psidium guajava) alters the labelling of blood constituents with technetium-99m

    Institute of Scientific and Technical Information of China (English)

    ABREU P.R.C.; ALMEIDA M.C.; BERNARDO R.M.; BERNARDO L.C.; BRITO L.C.; GARCIA E.A.C.; FONSECA A.S.; BERNARDO-FILHO M.

    2006-01-01

    Psidium guajava (guava) leaf is a phytotherapic used in folk medicine to treat gastrointestinal and respiratory disturbances and is used as anti-inflammatory medicine. In nuclear medicine, blood constituents (BC) are labelled with technetium-99m (99mTc) and used to image procedures. However, data have demonstrated that synthetic or natural drugs could modify the labelling of BC with 99mTc. The aim of this work was to evaluate the effects of aqueous extract of guava leaves on the labelling of BC with 99mTc. Blood samples of Wistar rats were incubated with different concentrations of guava extract and labelled with 99mTc after the percentage of incorporated radioactivity (%ATI) in BC was determined. The results suggest that aqueous guava extract could present antioxidant action and/or alters the membrane structures involved in ion transport into cells, thus decreasing the radiolabelling of BC with 99mTc. The data showed significant (P<0.05) alteration of ATI in BC from blood incubated with guava extract.

  18. Radiolabeled blood cells: radiation dosimetry and significance

    International Nuclear Information System (INIS)

    Over the past few years blood cells labeled with In-111 have become increasingly useful in clinical diagnosis and biomedical research. Indium-111 by the virtue of its physical characteristics and ability to bind to cell cytoplasmic components, provides an excellent cell tracer and thereby, allows investigators to monitor in vivo cell distribution by external imaging and help determine a course of regimen in treating life threatening diseases. Due to natural phenomena such as margination, blood pool, and reticuloendothelial cell activity, in the normal state, depending upon the cell type and the quality of cell preparations, 30%-50% of the administered radioactivity is immediately distributed in the liver, spleen and bone marrow. Over a period of time the radioactivity in these organs slightly increases and decays with a physical half-life of In-111. The resulting radiation dose to these organs ranges between 1-25 rads/mCi In-111 administered. The authors have developed a new In-111 labeling technique which preserves platelet ultrastructure and shown that human lymphocytes labeled with In-111 in mixed leukocytes preparations a) are only 0.003% of the total -body lymphocytes population and b) are killed. The consequence if any may be considered insignificant, particularly because 5.6% metaphases from normal men and 6.5% metaphases from normal women in the US have at least one chromosome aberration. Calculations have shown that the risk of fatal hematological malignancy, over a 30 year period, in recipients of 100 million lymphocytes labeled with 100 μCi In-111 is 1/million patients studied. This risk is less than 0.025% of the 1981 spontaneous cancer patient rate in the country. 32 references, 10 tables

  19. Approaches to radiolabelling blood cells: past, present and future

    International Nuclear Information System (INIS)

    The importance of cellular blood elements in health and disease can never be overemphasized. Associated with every organic illness there is an involvement of blood cells. Using radiolabelled blood cells, researchers have made fundamental contributions in the basic knowledge of cell kinetics and physiology. Further development in cell labelling techniques, in conjunction with the advancements in nuclear imaging have made it possible to use radiolabelled blood cells as a non-invasive means of diagnosing diseases. Useful as it may be, we have become increasingly aware of the current limitations in the cell labelling technique. The object of this article is to highlight the past and present approaches to the technique, emphasize the current problems and discuss future directions that might help to fetch solutions. (Auth.)

  20. Deep Learning in Label-free Cell Classification

    Science.gov (United States)

    Chen, Claire Lifan; Mahjoubfar, Ata; Tai, Li-Chia; Blaby, Ian K.; Huang, Allen; Niazi, Kayvan Reza; Jalali, Bahram

    2016-03-01

    Label-free cell analysis is essential to personalized genomics, cancer diagnostics, and drug development as it avoids adverse effects of staining reagents on cellular viability and cell signaling. However, currently available label-free cell assays mostly rely only on a single feature and lack sufficient differentiation. Also, the sample size analyzed by these assays is limited due to their low throughput. Here, we integrate feature extraction and deep learning with high-throughput quantitative imaging enabled by photonic time stretch, achieving record high accuracy in label-free cell classification. Our system captures quantitative optical phase and intensity images and extracts multiple biophysical features of individual cells. These biophysical measurements form a hyperdimensional feature space in which supervised learning is performed for cell classification. We compare various learning algorithms including artificial neural network, support vector machine, logistic regression, and a novel deep learning pipeline, which adopts global optimization of receiver operating characteristics. As a validation of the enhanced sensitivity and specificity of our system, we show classification of white blood T-cells against colon cancer cells, as well as lipid accumulating algal strains for biofuel production. This system opens up a new path to data-driven phenotypic diagnosis and better understanding of the heterogeneous gene expressions in cells.

  1. White Blood Cell Disorders

    Science.gov (United States)

    ... where they are needed, and then kill and digest the harmful organism or substance (see White blood ... Patel Hello Everyone! Hello to all of you readers! I know you will be seeing my biography, ...

  2. Fine structure of reactive cells in injured nervous tissue labeled with 3H-thymidine injected before injury

    International Nuclear Information System (INIS)

    To examine the fine structure of blood mononuclear cells in injured nervous tissue, mice were given repeated injections of 3H-thymidine. Under ether anesthesia the animals either were given a stab wound to the spinal cord or had their left hypoglossal nerve transected. Tissue sections were prepared for electron microscopic radioautography, and all labeled cells were photographed. About half the labeled cells in the injured spinal cords and almost all the labeled cells in the nuclei of the injured hypoglossal nerves had nuclei with dark staining peripheral heterochromatin, dark cytoplasm with long cisternae of granular endoplasmic reticulum, and other ultrastructural features characteristic of the cells usually identifed as microglia. The remaining labeled cells in the injured spinal cords were macrophages, fibroblasts, and vascular cells. Since uninjured nervous tissue has extremely few labeled cells, most of the labeled cells in this experiment should be derived from blood mononuclear cells

  3. Label-Free Biosensors for Cell Biology

    OpenAIRE

    Ye Fang

    2011-01-01

    Label-free biosensors for studying cell biology have finally come of age. Recent developments have advanced the biosensors from low throughput and high maintenance research tools to high throughput and low maintenance screening platforms. In parallel, the biosensors have evolved from an analytical tool solely for molecular interaction analysis to powerful platforms for studying cell biology at the whole cell level. This paper presents historical development, detection principles, and applicat...

  4. Dual labeling of neural crest cells and blood vessels within chicken embryos using Chick(GFP) neural tube grafting and carbocyanine dye DiI injection.

    Science.gov (United States)

    Delalande, Jean-Marie; Thapar, Nikhil; Burns, Alan J

    2015-01-01

    All developing organs need to be connected to both the nervous system (for sensory and motor control) as well as the vascular system (for gas exchange, fluid and nutrient supply). Consequently both the nervous and vascular systems develop alongside each other and share striking similarities in their branching architecture. Here we report embryonic manipulations that allow us to study the simultaneous development of neural crest-derived nervous tissue (in this case the enteric nervous system), and the vascular system. This is achieved by generating chicken chimeras via transplantation of discrete segments of the neural tube, and associated neural crest, combined with vascular DiI injection in the same embryo. Our method uses transgenic chick(GFP) embryos for intraspecies grafting, making the transplant technique more powerful than the classical quail-chick interspecies grafting protocol used with great effect since the 1970s. Chick(GFP)-chick intraspecies grafting facilitates imaging of transplanted cells and their projections in intact tissues, and eliminates any potential bias in cell development linked to species differences. This method takes full advantage of the ease of access of the avian embryo (compared with other vertebrate embryos) to study the co-development of the enteric nervous system and the vascular system. PMID:26065540

  5. Cigarette smoking increases white blood cell aggregation in whole blood.

    OpenAIRE

    Bridges, A B; Hill, A; Belch, J J

    1993-01-01

    We studied the effect of chronic cigarette smoking on white blood cell aggregation, increased aggregation predisposes to microvascular occlusion and damage. Current smokers had significantly increased white blood cell aggregation when compared with non smokers. The presence of chronically activated white blood cells in current smokers may be relevant in the pathogenesis of ischaemic vascular disease.

  6. Rare-cell enrichment by a rapid, label-free, ultrasonic isopycnic technique for medical diagnostics

    OpenAIRE

    Bourquin, Yannyk; Syed, Abeer; Reboud, Julien; Ranford-Cartwright, Lisa C.; Barrett, Michael P.; Cooper, Jonathan M.

    2014-01-01

    One significant challenge in medical diagnostics lies in the development of label-free methods to separate different cells within complex biological samples. Here we demonstrate a generic, low-power ultrasonic separation technique, able to enrich different cell types based upon their physical properties. For malaria, we differentiate between infected and non-infected red blood cells in a fingerprick-sized drop of blood. We are able to achieve an enrichment of circulating cells infected by the...

  7. Rare-Cell Enrichment by a Rapid, Label-Free, Ultrasonic Isopycnic Technique for Medical Diagnostics**

    OpenAIRE

    Bourquin, Yannyk; Syed, Abeer; Reboud, Julien; Ranford-Cartwright, Lisa C.; Barrett, Michael P.; Cooper, Jonathan M.

    2014-01-01

    One significant challenge in medical diagnostics lies in the development of label-free methods to separate different cells within complex biological samples. Here we demonstrate a generic, low-power ultrasonic separation technique, able to enrich different cell types based upon their physical properties. For malaria, we differentiate between infected and non-infected red blood cells in a fingerprick-sized drop of blood. We are able to achieve an enrichment of circulating cells infected by the...

  8. Cinnamomum zeylanicum extract on the radiolabelling of blood constituents and the morphometry of red blood cells: In vitro assay

    Energy Technology Data Exchange (ETDEWEB)

    Benarroz, M.O. [Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biofisica e Biometria, Universidade do Estado do Rio de Janeiro, Avenida 28 de Setembro, 87, 4o Andar, Vila Isabel, 20551-030 Rio de Janeiro, RJ (Brazil); Universidade Federal do Rio Grande do Norte, Programa de Pos-Graduacao em Ciencias da Saude, Avenida General Gustavo Cordeiro de Farias, s/n, 59010-180 Natal, RN (Brazil); Fonseca, A.S. [Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biofisica e Biometria, Universidade do Estado do Rio de Janeiro, Avenida 28 de Setembro, 87, 4o Andar, Vila Isabel, 20551-030 Rio de Janeiro, RJ (Brazil)], E-mail: adenilso@uerj.br; Rocha, G.S. [Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biofisica e Biometria, Universidade do Estado do Rio de Janeiro, Avenida 28 de Setembro, 87, 4o Andar, Vila Isabel, 20551-030 Rio de Janeiro, RJ (Brazil); Frydman, J.N.G. [Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biofisica e Biometria, Universidade do Estado do Rio de Janeiro, Avenida 28 de Setembro, 87, 4o Andar, Vila Isabel, 20551-030 Rio de Janeiro, RJ (Brazil); Universidade Federal do Rio Grande do Norte, Programa de Pos-Graduacao em Ciencias da Saude, Avenida General Gustavo Cordeiro de Farias, s/n, 59010-180 Natal, RN (Brazil); Rocha, V.C.; Pereira, M.O. [Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biofisica e Biometria, Universidade do Estado do Rio de Janeiro, Avenida 28 de Setembro, 87, 4o Andar, Vila Isabel, 20551-030 Rio de Janeiro, RJ (Brazil)] (and others)

    2008-02-15

    Effects of Cinnamomum zeylanicum (cinnamon) on the labelling of blood constituents with technetium-99 m({sup 99m}Tc) and on the morphology of red blood cells were studied. Blood samples from Wistar rats were incubated with cinnamon extract for 1hour or with 0.9% NaCl, as control. Labelling of blood constituents with {sup 99m}Tc was performed. Plasma (P) and blood cells (BC), soluble (SF-P and SF-BC) and insoluble (IF-P and IF-BC) fractions were separated. The radioactivity in each fraction was counted and the percentage of radioactivity incorporated (%ATI) was calculated. Blood smears were prepared, fixed, stained and the qualitative and quantitative morphological analysis of the red blood cells was evaluated. The data showed that the cinnamon extract decreased significantly (p<0.05) the %ATI on BC, IF-P and IF-BC. No modifications were verified on shape of red blood cells. Cinnamon extracts could alter the labelling of blood constituents with {sup 99m}Tc, and although our results were obtained with animals, precaution is suggested in interpretations of nuclear medicine examinations involving the labelling of blood constituents in patients who are using cinnamon.

  9. To what extent can diverse types of liver lesions mimic hemangiomas? A retrospective quantitative analysis of masses found to be positive in SPECT/CT with labeled blood cells – a preliminary report

    International Nuclear Information System (INIS)

    Although specificity of SPECT/CT examination using technetium-99m radiolabeled red blood cells (Tc-99m-RBC) for detection of liver hemangiomas is very high, it is still not perfect. It is possible to overlook a malignancy. Moreover, the difference in accumulation of RBCs between a hemangioma and uninvolved liver remains unknown. The aim of the study is to determine the quotients of accumulation of Tc-99m-RBC in hemangiomas and in normal liver parenchyma (HEM/liv), and to verify, whether the quotient could be potentially helpful in distinguishing hemangiomas from other RBC-accumulating liver masses. 34 liver lesions larger than 1.5 cm classified scintigraphically (qualitatively) in our Department as either typical or suspicious of hemangioma 1.5–4 years earlier were enrolled in this retrospective study. Their SPECT/CT images were acquired 1 hour after in vivo labeling of RBCs with Tc-99m. In reconstructed images, ellipsoidal regions of interest (ROIs) with diameters of about 1.5 cm were created in the assessed lesions (HEM) and in the uninvolved liver parenchyma (liv). The HEM/liv quotients were calculated for each mass. The results were compared with radiological data. 31 lesions were found to be clinically and radiologically typical for hemangiomas, their HEM/liv ratios were at least 1.6 (smaller masses) or 1.8 (larger masses). One lesion with HEM/liv ratio equal to 1.21 was classified as metastasis. Two lesions with HEM/liv 1.42 and 1.46 were classified as benign foci other than hemangioma. The quantitative analysis can be preliminarily proposed as a helpful tool in the assessment of possible liver hemangiomas

  10. Quantitative measurement of blood cells

    International Nuclear Information System (INIS)

    Full text: We are observing and measuring the varying development reaction stages of blood cells to different saline solutions. The imaging process is based on a common path interferometer which is realized with a spatial light modulator (SLM) in the Fourier plane after the microscope objective. With the SLM we can shift the phase of the transmitted light with respect to the phase of signal wave. This principle is used for the phase contrast microscopy method where we take four pictures of the same image with different phase shifts in order to calculate the complex field of the measured cell. This microscope technique obtains quantitative data about the blood cell's surface in different development stages, amplitude and phase differences inside the cell itself. (author)

  11. Red Cell Volume Can Be Accurately Determined in Sheep Using a Non-radioactive Biotin Label

    OpenAIRE

    Mock, Donald M.; Mock, Nell I.; Lankford, Gary L.; Burmeister, Leon F.; Strauss, Ronald G.; Widness, John A.

    2008-01-01

    The sheep has served as an informative animal model for investigation of human fetal and newborn erythropoiesis and red blood cell (RBC) kinetics. We previously validated the permanent label (14C)cyanate for measuring red cell volume (RCV) in sheep. Here we validate biotin labeling of RBCs as a nonradioactive method for measuring RCV in sheep with the anticipation that it can be applied in studies of human infants. The RCV was determined simultaneously using two techniques for quantitation of...

  12. Circulating blood cells function as a surveillance system for damaged tissue in Drosophila larvae

    OpenAIRE

    Babcock, Daniel T.; Brock, Amanda R.; Fish, Greg S.; Wang, Yan; Perrin, Laurent; Krasnow, Mark A.; Galko, Michael J.

    2008-01-01

    Insects have an open circulatory system in which the heart pumps blood (hemolymph) into the body cavity, where it directly bathes the internal organs and epidermis. The blood contains free and tissue-bound immune cells that function in the inflammatory response. Here, we use live imaging of transgenic Drosophila larvae with fluorescently labeled blood cells (hemocytes) to investigate the circulatory dynamics of larval blood cells and their response to tissue injury. We find that, under normal...

  13. Induction and identification of rabbit peripheral blood derived dendritic cells

    Science.gov (United States)

    Zhou, Jing; Yang, FuYuan; Chen, WenLi

    2012-03-01

    Purpose: To study a method of the induction of dendritic cells (DCs) from rabbit peripheral blood. Methods: Peripheral blood cells were removed from rabbit, filtered through nylon mesh. Peripheral blood mononuclear cells (PBMC) were isolated from the blood cells by Ficoll-Hypaque centrifugation (density of 1.077g/cm3).To obtain DCs, PBMC were cultured in RPMI1640 medium containing 10% fetal calf serum, 50U/mL penicillin and streptomycin, referred to subsequently as complete medium, at 37°C in 5% CO2 atmosphere for 4 hours. Nonadherent cells were aspirated, adherent cells were continued incubated in complete medium, supplemented with granulocyte/macrophage colony-stimulating factor (GM-CSF, 50ng/ml),and interleukin 4 (IL-4, 50ng/ml) for 9 days. Fluorescein labeled antibodies(anti-CD14, anti-HLA-DR, anti-CD86) were used to sign cells cultured for 3,6,9 days respectively, Then flow cytometry was performed. Results: Ratio of anti-HLA-DR and anti-CD86 labeled cells increased with induction time extension, in contrast with anti-CD14. Conclusion: Dendritic cells can be effectively induced by the method of this experiment, cell maturation status increased with induction time extension.

  14. Carboxyfluorescein Diacetate Succinimidyl Ester Fluorescent Dye for Cell Labeling

    Institute of Scientific and Technical Information of China (English)

    Xiao-Qi WANG; Xiu-Mei DUAN; Li-Hua LIU; Yan-Qiu FANG; Yan TAN

    2005-01-01

    Our objective was to study the properties of the carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and the methodology of cell labeling using CFDA-SE fluorescent dye. First, we analyzed the kinetics of CFDA-SE fluorescent dye intensity over time. Second, we determined the optimal concentration of CFDA-SE fluorescent dye for cell labeling. Third, we tested the toxicity of CFDA-SE fluorescent dye on labeled cells. Finally, we determined the optimal staining time of CFDA-SE fluorescent dye for cell labeling.The results show that the optimal concentration of CFDA-SE fluorescent dye for cell labeling varies according to different cell types. CFDA-SE fluorescent dye is non-toxic to cells as the cell death rate caused by CFDASE labeling is below 5%. The optimal cell labeling time was determined to be 8 min of incubation with CFDA-SE fluorescent dye. We concluded that the advantages of using CFDA-SE fluorescent dye for cell labeling are as follows: (1) the binding of CFDA-SE fluorescent dye to cells is stable; (2) CFDA-SE fluorescent dye is not toxic and does not modify the viability of labeled cells; and (3) CFDA-SE fluorescent dye is a suitable fluorochrome for cell labeling.

  15. Evaluation of copper-labeled bifunctional chelate-albumin conjugates for blood pool imaging

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, C.J.; Rocque, P.A.; Welch, M.J. (Washington Univ., St. Louis, MO (United States). Edward Mallinckrodt Inst. of Radiology); Weinheimer, C.J. (Washington Univ., St. Louis, MO (United States). School of Medicine)

    1993-05-01

    [sup 62]Cu is a generator-produced positron-emitting radionuclide with a half-life amenable to blood-pool imaging with PET. Three bifunctional chelates [cyclic anhydride of diethylenetriamine-pentaacetic acid (cDTPAA), 6-bromoacetamidobenzyl-1,4,8,11-tetraazacyclotetradecane-N,N',N'', N''' tetraacetic acid (BAT), and p-carboxyethylphenylglyoxal-bis-([sup 4])N-methyl-thiosemicarbazone (CE-DTS)] were conjugated to HSA and labeled with [sup 67]Cu. Blood clearance and biodistribution of these three [sup 67]Cu-labeled conjugates were determined in rats. Of the three [sup 67]Cu-labeled bifunctional chelate-HSA conjugates, [sup 67]Cu-benzyl-TETA-HSA remained in the blood pool the longest, achieving stable blood levels at times longer than 24 h post-injection. The [sup 67]Cu radioactivity cleared the blood within 60 min post-injection of [sup 67]Cu-DTS-HSA, and within 10 min after administration of [sup 67]Cu-DTPA-HSA, indicating the dissociation of Cu[sup 2+] from these conjugates. Copper-labeled DTS-HSA achieved stable blood concentrations for at least 30 min post-injection and was therefore evaluated as a vascular imaging agent. DTS-HSA and benzy-TETA-HSA were labeled with [sup 62]Cu and administered to a dog for blood-pool imaging using PET. Because of the high labeling efficiency, DTS-HSA can be labeled with [sup 62]Cu without purification, making it more practical than [sup 62]Cu-benzyl-TETA-HSA as a blood-pool imaging agent. Generator-produced [sup 62]Cu-DTS-HSA should be a viable alternative blood pool agent to cyclotron-produced C[sup 15]O for PET facilities without cyclotrons. (author).

  16. Influence on bionomics of endothelial progenitor cells labeling with magnetic nanoparticles

    International Nuclear Information System (INIS)

    Objective: To explore the influence of home synthesize magnetic iron oxide (called Fe2O3-PLL) labeling on peripheral blood endothelial progenitor cells (EPCs) bionomics to provide experimental foundation for MR imaging ex and in vivo. Methods: Fe2O3 was incubated with PLL for 2 hours to obtain a complex of Fe2O3-PLL. Rabbit peripheral blood mononuclear cells were isolated and EPCs were selected by adherence method. Fe2O3-PLL was used to label EPCs. Prussian blue stain and electron microscope was used for showing intracellular iron. MTF assay was assessed to evaluate the difference of growth curve between unlabeled and labeled with 25 mg/L Fe2O3-PLL. Flow cytometry was performed to analyze cell cycle, cell apoptosis and the expression of surface markers of labeled and unlabeled cells. Expressions of eNOS, KDR and vWF at mRNA levels among unlabeled and labeled EPCs were detected by real-time polymerase chain reaction. Calcium ion channel and membrane fluidity were observed and analyzed by laser confocal microscopy. Statistical analyses were used with ANOVA and t test. Results: Almost 100% cells were labeled by Fe2O3-PLL, iron-containing vesicles were intracytoplasma. There was no statistical difference in cells growth curve, cell life cycle [(93.74±3.52)%, (94.57±3.66)%] and cell apoptosis rate (12.89±1.81)%, (11.67±1.18)%) between labeling with Fe2O3-PLL at a concentration of 25 mg/L and unlabeled cells (t=0.283, P>0.05; t=0.977, P>0.05). There was also no statistical difference in relative amount of eNOS, KDR and vWF at mRNA levels and the expression of surface phenotypic markers (CD34, CD106, CD146 and KDR) between two groups (P>0.05). In addition, Labeling had little influence on calcium ion channel and didn't significantly alter cell membrane fluidity. Conclusions: The rabbit peripheral blood EPCs can be effective labeled with Fe2O3-PLL and without significant influence on cells bionomics at a low concentration of 25 mg/L. Almost every cell can be labeled

  17. Becoming a Blood Stem Cell Donor

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    Full Text Available ... Find out why Close Becoming a Blood Stem Cell Donor NCIcancertopics Subscribe Subscribed Unsubscribe 352 352 Loading... ... considered becoming a bone marrow or blood stem cell donor? Follow this true story of a former ...

  18. Becoming a Blood Stem Cell Donor

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    Full Text Available ... total__ Find out why Close Becoming a Blood Stem Cell Donor NCIcancertopics Subscribe Subscribed Unsubscribe 361 361 Loading... ... Ever considered becoming a bone marrow or blood stem cell donor? Follow this true story of a former ...

  19. Becoming a Blood Stem Cell Donor

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    Full Text Available ... Find out why Close Becoming a Blood Stem Cell Donor NCIcancertopics Subscribe Subscribed Unsubscribe 361 361 Loading... ... considered becoming a bone marrow or blood stem cell donor? Follow this true story of a former ...

  20. Becoming a Blood Stem Cell Donor

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    Full Text Available ... on Jul 19, 2011 Ever considered becoming a bone marrow or blood stem cell donor? Follow this ... Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation (BMT) and peripheral blood stem cell ...

  1. Becoming a Blood Stem Cell Donor

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    Full Text Available ... total__ Find out why Close Becoming a Blood Stem Cell Donor NCIcancertopics Subscribe Subscribed Unsubscribe 350 350 Loading... ... Ever considered becoming a bone marrow or blood stem cell donor? Follow this true story of a former ...

  2. Becoming a Blood Stem Cell Donor

    Science.gov (United States)

    ... total__ Find out why Close Becoming a Blood Stem Cell Donor NCIcancertopics Subscribe Subscribed Unsubscribe 350 350 Loading... ... Ever considered becoming a bone marrow or blood stem cell donor? Follow this true story of a former ...

  3. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... on Jul 19, 2011 Ever considered becoming a bone marrow or blood stem cell donor? Follow this true ... Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation (BMT) and peripheral blood stem cell transplantation ( ...

  4. Becoming a Blood Stem Cell Donor

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    Full Text Available ... Find out why Close Becoming a Blood Stem Cell Donor NCIcancertopics Subscribe Subscribed Unsubscribe 351 351 Loading... ... considered becoming a bone marrow or blood stem cell donor? Follow this true story of a former ...

  5. Becoming a Blood Stem Cell Donor

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    Full Text Available ... total__ Find out why Close Becoming a Blood Stem Cell Donor NCIcancertopics Subscribe Subscribed Unsubscribe 351 351 Loading... ... Ever considered becoming a bone marrow or blood stem cell donor? Follow this true story of a former ...

  6. Becoming a Blood Stem Cell Donor

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    Full Text Available ... total__ Find out why Close Becoming a Blood Stem Cell Donor NCIcancertopics Subscribe Subscribed Unsubscribe 360 360 Loading... ... Ever considered becoming a bone marrow or blood stem cell donor? Follow this true story of a former ...

  7. 75 FR 73107 - Guidance for Industry and Food and Drug Administration Staff; Blood Lancet Labeling; Availability

    Science.gov (United States)

    2010-11-29

    ... HUMAN SERVICES Food and Drug Administration Guidance for Industry and Food and Drug Administration Staff... ``Guidance for Industry and Food and Drug Administration Staff; Blood Lancet Labeling.'' FDA is issuing this....regulations.gov . To receive ``Guidance for Industry and Food and Drug Administration Staff; Blood...

  8. Uptake of carnitine by red blood cells

    Energy Technology Data Exchange (ETDEWEB)

    Campa, M.; Borum, P.

    1986-05-01

    A significant amount of blood carnitine (70% of cord blood and 40% of blood from healthy adults) is partitioned into the red blood cell compartment of whole blood. Data indicate that the plasma compartment and the red blood cell compartment of whole blood represent different metabolic pools of carnitine. There are no data to indicate that red blood cells synthesize carnitine, but our understanding of the uptake of carnitine by red blood cells is negligible. Red blood cells were obtained from healthy adults, washed twice with normal saline, and used for uptake experiments. When the cells were incubated at 37/sup 0/C in the presence of /sup 14/C-carnitine, radioactivity was found both in the soluble cytosolic and membrane fractions of the cells following lysis. The uptake was dependent upon the time of incubation, temperature of incubation, and carnitine concentration in the incubation medium. Washed red blood cell membranes incubated with /sup 14/C-carnitine showed specific binding of radioactivity. These data are consistent with the hypothesis that red blood cells have an uptake mechanism for L-carnitine.

  9. Uptake of carnitine by red blood cells

    International Nuclear Information System (INIS)

    A significant amount of blood carnitine (70% of cord blood and 40% of blood from healthy adults) is partitioned into the red blood cell compartment of whole blood. Data indicate that the plasma compartment and the red blood cell compartment of whole blood represent different metabolic pools of carnitine. There are no data to indicate that red blood cells synthesize carnitine, but our understanding of the uptake of carnitine by red blood cells is negligible. Red blood cells were obtained from healthy adults, washed twice with normal saline, and used for uptake experiments. When the cells were incubated at 370C in the presence of 14C-carnitine, radioactivity was found both in the soluble cytosolic and membrane fractions of the cells following lysis. The uptake was dependent upon the time of incubation, temperature of incubation, and carnitine concentration in the incubation medium. Washed red blood cell membranes incubated with 14C-carnitine showed specific binding of radioactivity. These data are consistent with the hypothesis that red blood cells have an uptake mechanism for L-carnitine

  10. Effect of Ginkgo biloba on the labeling of blood elements with technetium-99m: in vitro study

    Directory of Open Access Journals (Sweden)

    Silvana Ramos Farias Moreno

    2002-01-01

    Full Text Available Ginkgo biloba is the phytoterapic most used in popular medicine in the treatment of cerebral senescence. Red blood cells (RBC labeled with technetium-99m (Tc-99m is used for several evaluations in nuclear medicine. This labeling depends on a reducing agent, usually the stannous ion. Any drug, which alters the labeling of the tracer, could be expected to modify the disposition of the radiopharmaceutical. We have evaluated the influence of the Ginkgo biloba extract on the labeling of RBC and plasma proteins with Tc-99m. Blood was withdrawn and incubated with Ginkgo biloba extract (0; 0.004; 0.04; 0.4; 4; 20 and 40 mg/ml. Stannous chloride (1.2 ml/ml was added and, then, Tc-99m was added. Plasma (P and blood cells (RBC were isolated, also precipitated with trichloroacetic acid and soluble (SF and insoluble fractions (IF separated. The analysis of the results shows that there is a decrease in the radioactivity (from 97.7 ± 0.7 to 49.5 ± 3.9% in RBC with the drug (4 mg/ml. In the labeling process of RBC with Tc-99m, the stannous and pertechnetate ions pass though the membrane, so, we suggest that the Ginkgo biloba effect can be explained by (i an inhibition of the transport of these ions, (ii damage in membrane, (iii competition with the cited ions for the same binding sites, or (iv possible generation of reactive oxygen species that could oxidize the stannous ion.

  11. The use of radiolabeled monoclonal antibodies for cell labeling in vivo

    International Nuclear Information System (INIS)

    The authors have evaluated the potential of in vivo cell surface labeling using radiolabeled monoclonal antibodies (MoAbs) directed against their surface antigens. Two MoAbs, a specific antibody (anti-Thy-1 OX7) and a nonspecific control antibody (anti-CEA) were coupled with DTPA, labeled with /sup 111/In and evaluated against rat thymocytes, marrow cells, and lymphoma cells (all known to be Thy-1 positive) both in vitro and in vivo. Enumeration of the cells which bound the radiolabeled MoAb was done by detecting the antibody on the cell surface with a Fl-F(ab')/sub 2/ goat anti-mouse IgG and analyzing fluorescence (F1) in a flow cytometer (FACS). The thymocytes, which could be labeled in whole blood, showed a labeling efficiency of 80-100%. The labeling, which could be inhibited by cold antibody, was stable up to 72 hours and did not interfere with either cell viability or functional integrity. Following IV injection of the MoAbs in normal rats, there was very good visualization of the bone marrow not seen with the control. Analysis of the marrow cells on the FACS showed that at two hours over 60% of the marrow cells were specifically labeled as against 2% for the control. Within 15 minutes of injecting /sup 111/In-OX7 into rats with lymphoma, 70% of the activity in blood was bound to circulating lymphoma cells. The ability to stably label, rapidly target, and image specific cell populations in vivo has wide ranging diagnostic and therapeutic implications

  12. 21 CFR 640.10 - Red Blood Cells.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Red Blood Cells. 640.10 Section 640.10 Food and... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Red Blood Cells § 640.10 Red Blood Cells. The proper name of this product shall be Red Blood Cells. The product is defined as red blood cells...

  13. Cinnamomum zeylanicum extract on the radiolabelling of blood constituents and the morphometry of red blood cells: In vitro assay

    International Nuclear Information System (INIS)

    Effects of Cinnamomum zeylanicum (cinnamon) on the labelling of blood constituents with technetium-99 m(99mTc) and on the morphology of red blood cells were studied. Blood samples from Wistar rats were incubated with cinnamon extract for 1hour or with 0.9% NaCl, as control. Labelling of blood constituents with 99mTc was performed. Plasma (P) and blood cells (BC), soluble (SF-P and SF-BC) and insoluble (IF-P and IF-BC) fractions were separated. The radioactivity in each fraction was counted and the percentage of radioactivity incorporated (%ATI) was calculated. Blood smears were prepared, fixed, stained and the qualitative and quantitative morphological analysis of the red blood cells was evaluated. The data showed that the cinnamon extract decreased significantly (p99mTc, and although our results were obtained with animals, precaution is suggested in interpretations of nuclear medicine examinations involving the labelling of blood constituents in patients who are using cinnamon

  14. Avoiding Anemia: Boost Your Red Blood Cells

    Science.gov (United States)

    ... link, please review our exit disclaimer . Subscribe Avoiding Anemia Boost Your Red Blood Cells If you’re ... and sluggish, you might have a condition called anemia. Anemia is a common blood disorder that many ...

  15. Becoming a Blood Stem Cell Donor

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    Full Text Available ... cell donation experience at the National Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation ( ... About Cord Blood Banking - Duration: 49:19. Children's Health 26,035 views 49:19 Scott: Donating Blood ...

  16. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... cell donation experience at the National Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation ( ... About Cord Blood Banking - Duration: 49:19. Children's Health 27,845 views 49:19 Scott: Donating Blood ...

  17. Human neutrophil kinetics: modeling of stable isotope labeling data supports short blood neutrophil half-lives.

    Science.gov (United States)

    Lahoz-Beneytez, Julio; Elemans, Marjet; Zhang, Yan; Ahmed, Raya; Salam, Arafa; Block, Michael; Niederalt, Christoph; Asquith, Becca; Macallan, Derek

    2016-06-30

    Human neutrophils have traditionally been thought to have a short half-life in blood; estimates vary from 4 to 18 hours. This dogma was recently challenged by stable isotope labeling studies with heavy water, which yielded estimates in excess of 3 days. To investigate this disparity, we generated new stable isotope labeling data in healthy adult subjects using both heavy water (n = 4) and deuterium-labeled glucose (n = 9), a compound with more rapid labeling kinetics. To interpret results, we developed a novel mechanistic model and applied it to previously published (n = 5) and newly generated data. We initially constrained the ratio of the blood neutrophil pool to the marrow precursor pool (ratio = 0.26; from published values). Analysis of heavy water data sets yielded turnover rates consistent with a short blood half-life, but parameters, particularly marrow transit time, were poorly defined. Analysis of glucose-labeling data yielded more precise estimates of half-life (0.79 ± 0.25 days; 19 hours) and marrow transit time (5.80 ± 0.42 days). Substitution of this marrow transit time in the heavy water analysis gave a better-defined blood half-life of 0.77 ± 0.14 days (18.5 hours), close to glucose-derived values. Allowing the ratio of blood neutrophils to mitotic neutrophil precursors (R) to vary yielded a best-fit value of 0.19. Reanalysis of the previously published model and data also revealed the origin of their long estimates for neutrophil half-life: an implicit assumption that R is very large, which is physiologically untenable. We conclude that stable isotope labeling in healthy humans is consistent with a blood neutrophil half-life of less than 1 day. PMID:27136946

  18. Life span and tissue distribution of 111indium-labeled blood platelets in hypomagnesemic lambs

    International Nuclear Information System (INIS)

    Circulating platelets may be activated by exposed triple-helical collagen in atherosclerotic lesions in Mg-deficient ruminants. Autologous platelets, labeled in vitro with 111In and determined to be active, were injected into 5 hypomagnesemic and 3 control lambs fed semipurified diets with 100 or 2,000 mg of Mg/kg of feed for 3 months. During the first 68 hours, 111In concentrations were 11 times higher in packed cells than in plasma. Packed-cell 111In increased 60% during the first 2 hours, probably due to initial tissue sequestration and later release of labeled platelets. Thereafter, platelet half-life span averaged 60 and 63 hours for hypomagnesemic and control lambs. After 68 hours, lambs were injected with native vascular collagen fibrils at 500 micrograms/kg of body weight to initiate reversible platelet aggregation. Within 1 minute, 83% of packed-cell 111In disappeared from circulation. Thirty minutes later, the lambs were euthanatized and necropsied and in the lungs, liver, and spleen, 111In averaged 24%, 19%, and 9%, respectively, of 111In injected 68 hours earlier. Organ deposits were not affected by Mg intake, but 111In in the lungs was somewhat lower in 2 lambs injected with inactivated collagen. Pathologic changes induced by reversible platelet aggregation were compatible with right ventricular failure complicated by pulmonary edema, similar to changes in hypomagnesemic lambs that died spontaneously. Platelets in blood exposed to vascular lesions in hypomagnesemic ruminants could be a major mortality risk factor in grass tetany disease

  19. Evaluation of the effect of an extract of sabugueiro (Sambucus australis) on the labeling of blood constituents with technetium-99m

    Energy Technology Data Exchange (ETDEWEB)

    Ribeiro, Camila Godinho; Rebello, Bernardo Machado; Neves, Rosane de Figueiredo; Santos-Filho, Sebastiao David; Fonseca, Adenilson de Souza da [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Inst. de Biologia Roberto Alcantara Gomes. Lab. de Radiofarmacia Experimental]. E-mail: cacagr@yahoo.com.br; Medeiros, Aldo da Cunha; Bernardo-Filho, Mario [Universidade Federal do Rio Grande do Norte, Natal, RN (Brazil). Centro de Ciencias da Saude. Programa de Pos-graduacao em Ciencias da Saude; Catanho, Maria Teresa Jansem de Almeida [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de Biofisica e Radiobiologia

    2007-09-15

    Sambucus australis (sabugueiro) has been used to treat inflammatory and rheumatologic disorders. Blood constituents labeled with technetium-99m (99mTc) have been used in nuclear medicine to obtain diagnostic images. The aim of this work was to evaluate the effect of a sabugueiro extract on the labeling of blood cells with 99mTc. Blood samples from Wistar rats were incubated with sabugueiro extract and the radiolabeling assay of blood constituents was carried out. After centrifugation, samples of plasma and blood cells were separated. Aliquots of plasma and blood cells were precipitated with trichloroacetic acid and centrifuged to isolate soluble and insoluble fractions. The radioactivity in each fraction was counted and the percentage of activity (%ATI) was determined. Incubation with sabugueiro extract altered significantly (p<0.05) the %ATI incorporated to the blood constituents. These results could be explained due the presence of chemical substances in the sabugueiro extract that present redox and/or chelating action altering the labeling of the blood constituents with 99mTc. (author)

  20. Metabolic labeling with (14C)-glucose of bloodstream and cell culture trypanosoma cruzi trypomastigotes:

    International Nuclear Information System (INIS)

    Trypomastigote forms of Trypanosoma cruzi from infected mouse blood and from cell culture were metabolically labeled by incubation with D-(14C)-glucose. Analysis by polyacrylamide gel electrophoresis of lysates from parasites of two strains (RA and CA1) showed a significantly different pattern. The difference was mainly quantitative when the blood and cell culture trypomastigotes of the RA strain were compared. Analysis of the culture medium by paper electrophoresis showed an anionic exometabolite only in the blood forms of both strains. (Author)

  1. When Blood Cells Bend: Understanding Sickle Cell Disease

    Science.gov (United States)

    ... please review our exit disclaimer . Subscribe When Blood Cells Bend Understanding Sickle Cell Disease For people who don’t suspect they ... Cells Bend Wise Choices Links Living with Sickle Cell Disease See a sickle cell disease expert regularly. ...

  2. Residual nanoparticle label immunosensor for wash-free C-reactive protein detection in blood.

    Science.gov (United States)

    Huttunen, Roope J; Näreoja, Tuomas; Mariani, Laura; Härmä, Harri

    2016-09-15

    Current diagnostic immunotechnologies are universally based on the measurement of the bound label-antibody fraction in direct binding or sandwich-assay type approaches with various detection techniques (e.g. enzyme-linked immunosorbent assay or ELISA) on solid stationary phase surface. Here an alternative reciprocal approach is presented based on the detection of the non-bound fraction of nanoparticle-labelled antibodies using microparticles as solid support. The advantage of detecting the non-bound fraction of the labelled antibody instead of the bound fraction is the high dynamics and the suggested increased flexibility in the selection of the detection mode. No actual washing steps are required as the bound and non-bound fractions of the detection nanoparticle label are separated using physical separation rather than consecutive washing repeats. The quantitative proof-of-concept set-up was demonstrated through blood-based detection of C-reactive protein (CRP). A blood sample containing CRP was diluted 1/50 and measured in 15-min resulting in a linear response at a range from 1 to 30μg/ml. The lowest limit of detection was below 0.03μg/ml and the assay coefficient of variation ranged from 0.3 to 9%. The nanoparticle-based residual label detection outperformed the corresponding molecular label method providing wider applicability with nearly an order of magnitude higher signal-to-background ratio for novel assay configurations in clinical diagnostics practices. PMID:27104585

  3. Sucralose sweetener in vivo effects on blood constituents radiolabeling, red blood cell morphology and radiopharmaceutical biodistribution in rats

    International Nuclear Information System (INIS)

    Effects of sucralose sweetener on blood constituents labelled with technetium-99m (99mTc) on red blood cell (RBC) morphology, sodium pertechnetate (Na99mTcO4) and diethylenetriaminepentaacetic acid labeled with 99mTc (99mTc-DTPA) biodistribution in rats were evaluated. Radiolabeling on blood constituents from Wistar rats was undertaken for determining the activity percentage (%ATI) on blood constituents. RBC morphology was also evaluated. Na99mTcO4 and 99mTc-DTPA biodistribution was used to determine %ATI/g in organs. There was no alteration on RBC blood constituents and morphology %ATI. Sucralose sweetener was capable of altering %ATI/g of the radiopharmaceuticals in different organs. These findings are associated to the sucralose sweetener in specific organs.

  4. Sucralose sweetener in vivo effects on blood constituents radiolabeling, red blood cell morphology and radiopharmaceutical biodistribution in rats

    Energy Technology Data Exchange (ETDEWEB)

    Rocha, G.S.; Pereira, M.O. [Universidade do Estado do Rio de Janeiro, Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biofisica e Biometria, Avenida 28 de Setembro, 87, Vila Isabel, 20551030 Rio de Janeiro (Brazil); Universidade Federal do Rio Grande do Norte, Programa de Pos-Graduacao em Ciencias da Saude, Avenida General Gustavo Cordeiro de Farias, s/n, 59010180 Natal, Rio Grande do Norte (Brazil); Benarroz, M.O.; Frydman, J.N.G.; Rocha, V.C. [Universidade do Estado do Rio de Janeiro, Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biofisica e Biometria, Avenida 28 de Setembro, 87, Vila Isabel, 20551030 Rio de Janeiro (Brazil); Pereira, M.J. [Universidade do Estado do Rio de Janeiro, Instituto de Biologia Roberto Alcantara Gomes, Departamento de Fisiologia, Avenida 28 de Setembro, 87, Vila Isabel, 20551030 Rio de Janeiro (Brazil); Fonseca, A.S., E-mail: adnfonseca@ig.com.b [Universidade do Estado do Rio de Janeiro, Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biofisica e Biometria, Avenida 28 de Setembro, 87, Vila Isabel, 20551030 Rio de Janeiro (Brazil); Universidade Federal do Estado do Rio de Janeiro, Instituto Biomedico, Departamento de Ciencias Fisiologicas, Rua Frei Caneca, 94, Rio de Janeiro 20211040 (Brazil); Medeiros, A.C. [Universidade Federal do Rio Grande do Norte, Programa de Pos-Graduacao em Ciencias da Saude, Avenida General Gustavo Cordeiro de Farias, s/n, 59010180 Natal, Rio Grande do Norte (Brazil); Bernardo-Filho, M. [Universidade do Estado do Rio de Janeiro, Instituto de Biologia Roberto Alcantara Gomes, Departamento de Biofisica e Biometria, Avenida 28 de Setembro, 87, Vila Isabel, 20551030 Rio de Janeiro (Brazil); Instituto Nacional do Cancer, Coordenadoria de Pesquisa Basica, Praca Cruz Vermelha, 23, 20230130 Rio de Janeiro (Brazil)

    2011-01-15

    Effects of sucralose sweetener on blood constituents labelled with technetium-99m ({sup 99m}Tc) on red blood cell (RBC) morphology, sodium pertechnetate (Na{sup 99m}TcO{sub 4}) and diethylenetriaminepentaacetic acid labeled with {sup 99m}Tc ({sup 99m}Tc-DTPA) biodistribution in rats were evaluated. Radiolabeling on blood constituents from Wistar rats was undertaken for determining the activity percentage (%ATI) on blood constituents. RBC morphology was also evaluated. Na{sup 99m}TcO{sub 4} and {sup 99m}Tc-DTPA biodistribution was used to determine %ATI/g in organs. There was no alteration on RBC blood constituents and morphology %ATI. Sucralose sweetener was capable of altering %ATI/g of the radiopharmaceuticals in different organs. These findings are associated to the sucralose sweetener in specific organs.

  5. Instant magnetic labeling of tumor cells by ultrasound in vitro

    Science.gov (United States)

    Mo, Runyang; Yang, Jian; Wu, Ed X.; Lin, Shuyu

    2011-09-01

    Magnetic labeling of living cells creates opportunities for numerous biomedical applications. Here we describe an instantly cell magnetic labeling method based on ultrasound. We present a detailed study on the ultrasound performance of a simple and efficient labeling protocol for H-22 cells in vitro. High frequency focus ultrasound was investigated as an alternative method to achieve instant cell labeling with the magnetic particles without the need for adjunct agents or initiating cell cultures. Mean diameter of 168 nm dextran-T40 coated superparamagnetic iron oxide (SPIO) nanoparticles were prepared by means of classical coprecipitation in solution in our laboratory. H-22 tumor cells suspended in phosphate-buffered saline (PBS, pH=7.2) were exposed to ultrasound at 1.37 MHz for up to 120 s in the presence of SPIOs. The cellular uptake of iron oxide nanoparticles was detected by prussion blue staining. The viability of cells was determined by a trypan blue exclusion test. At 2 W power and 60 s ultrasound exposure in presence of 410 μg/ml SPIOs, H-22 cell labeling efficiency reached 69.4±6.3% and the labeled cells exhibited an iron content of 10.38±2.43 pg per cell. Furthermore, 95.2±3.2% cells remained viable. The results indicated that the ultrasound protocol could be potentially applied to label cells with large-sized magnetic particles. We also calculated the shear stress at the 2 W power and 1.37 MHz used in experiments. The results showed that the shear stress threshold for ultrasonically induced H-22 cell reparable sonoporation was 697 Pa. These findings provide a quantitative guidance in designing ultrasound protocols for cell labeling.

  6. Synthesis of 5-nitro-2-[N-3-(4-azidophenyl)-propylamino]-benzoic acid: Photoaffinity labeling of human red blood cell ghosts with a 5-nitro-2-(3-phenylpropylamino)-benzoic acid analog

    International Nuclear Information System (INIS)

    A photoaffinity analog of the potent epithelial chloride channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoic acid has been synthesized and characterized. In the dark, this reagent, 5-nitro-2-[N-3-(4-azidophenyl)-propylamino]-benzoic acid, and the parent compound reversibly inhibited chloride efflux in human red blood cell ghosts. Irradiation of ghost membranes with 350 microM arylazide analog reduced the rate of chloride efflux to 33% of the control value. The photoinactivation process was not reversed by exhaustive washing of ghost membranes. Covalent incorporation of the photoaffinity reagent was supported by difference ultraviolet spectroscopy, which indicated the attachment of the substituted 2-amino-5-nitrobenzoic acid chromophore to ghost membranes. The novel photolabeling agent described here should be a useful structural probe for chloride channels in erythrocyte membranes and epithelial cells

  7. Effects of Cinnamomum zeylanicum treatment on radiolabeling of blood constituents and morphology of red blood cells in Wistar rats

    International Nuclear Information System (INIS)

    The aim of this study was to evaluate the effect of in vivo treatment with an aqueous cinnamon extract on the labeling of blood constituents with 99mTc and on the morphology of red blood cells from Wistar rats. Animals were treated with cinnamon extract at different doses and for different periods of time. As controls, animals treated with 0.9% NaCl. Labeling of blood constituents with 99mTc was performed. Plasma, blood cells and insoluble fractions were isolated. Radioactivity in each fraction was counted and the percentage of radioactivity (%ATI) was calculated. Also, blood smears were prepared to morphological analysis of red blood cells from. Data showed that in vivo cinnamon extract did not significantly (p>0.05) modify the %ATI of blood constituents and morphology of red blood cells. The results suggest that in vivo aqueous cinnamon could not affect the membrane structures involved in transport of ions or the oxidation state of stannous and pertechnetate ions. (author)

  8. Effects of Cinnamomum zeylanicum treatment on radiolabeling of blood constituents and morphology of red blood cells in Wistar rats

    Energy Technology Data Exchange (ETDEWEB)

    Benarroz, Monica Oliveira; Fonseca, Adenilson de Souza da; Bernardo-Filho, Mario [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Dept. de Biofisica e Biometria]. E-mail: adenilso@uerj.br; Rocha, Gabrielle de Souza; Pereira, Marcia Oliveira [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil); Geller, Mauro [Centro Universitario Serra dos Orgaos, Teresopolis, RJ (Brazil). Centro de Ciencias da Saude; Presta, Giuseppe Antonio [Universidade Federal do Estado do Rio de Janeiro (UNIRIO), Rio de Janeiro, RJ (Brazil). Inst. Biomedico. Dept. de Fisiologia Humana

    2008-12-15

    The aim of this study was to evaluate the effect of in vivo treatment with an aqueous cinnamon extract on the labeling of blood constituents with {sup 99m}Tc and on the morphology of red blood cells from Wistar rats. Animals were treated with cinnamon extract at different doses and for different periods of time. As controls, animals treated with 0.9% NaCl. Labeling of blood constituents with {sup 99}mTc was performed. Plasma, blood cells and insoluble fractions were isolated. Radioactivity in each fraction was counted and the percentage of radioactivity (%ATI) was calculated. Also, blood smears were prepared to morphological analysis of red blood cells from. Data showed that in vivo cinnamon extract did not significantly (p>0.05) modify the %ATI of blood constituents and morphology of red blood cells. The results suggest that in vivo aqueous cinnamon could not affect the membrane structures involved in transport of ions or the oxidation state of stannous and pertechnetate ions. (author)

  9. Immune Cells in Blood Recognize Tumors

    Science.gov (United States)

    NCI scientists have developed a novel strategy for identifying immune cells circulating in the blood that recognize specific proteins on tumor cells, a finding they believe may have potential implications for immune-based therapies.

  10. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... MD. Bone marrow transplantation (BMT) and peripheral blood stem cell transplantation (PBSCT) are most commonly used in the treatment of cancers like leukemia and lymphoma to restore stem cells ...

  11. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation (BMT) and peripheral blood stem cell transplantation (PBSCT) are most commonly used in the treatment of cancers like leukemia and lymphoma to restore stem cells ...

  12. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... cell donation experience at the National Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation ( ... About Cord Blood Banking - Duration: 49:19. Children's Health 25,312 views 49:19 23. Stem Cells - ...

  13. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... Duration: 3:35. hemaquebec1998 667 views 3:35 Bone Marrow/Stem Cell ... Jeff, peripheral blood stem cell (PBSC) donor, explains the donation process - Duration: 3:28. Be The Match 22,203 ...

  14. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... cell donation experience at the National Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation ( ... About Cord Blood Banking - Duration: 49:19. Children's Health 26,239 views 49:19 23. Stem Cells - ...

  15. Deep Learning in Label-free Cell Classification

    OpenAIRE

    Claire Lifan Chen; Ata Mahjoubfar; Li-Chia Tai; Ian K. Blaby; Allen Huang; Kayvan Reza Niazi; Bahram Jalali

    2016-01-01

    Label-free cell analysis is essential to personalized genomics, cancer diagnostics, and drug development as it avoids adverse effects of staining reagents on cellular viability and cell signaling. However, currently available label-free cell assays mostly rely only on a single feature and lack sufficient differentiation. Also, the sample size analyzed by these assays is limited due to their low throughput. Here, we integrate feature extraction and deep learning with high-throughput quantitati...

  16. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation (BMT) and peripheral blood stem cell transplantation (PBSCT) ... Medicine Clinics 225,676 views 6:18 Alicia's bone marrow donation - Duration: 8:33. ... Peripheral Blood Stem Cell Transplant - Duration: 15:50. Dartmouth-Hitchcock 2,764 views ...

  17. Cadmium uptake by rat red blood cells

    International Nuclear Information System (INIS)

    Rat red blood cells were employed to study the uptake of cadmium (109Cd). Suspensions of red blood cells were exposed to Cd concentrations (both bound and free) observed following in vivo Cd administration. Cd uptake was biphasic with an initial rapid phase (0C was one-fourth of that at 370C. The metabolic inhibitors: sodium fluoride (1mM), potassium cyanide (1mM) and carbonyl cyanide-m-chlorophenyl hydrazone (2μM) and the Na+-K+-ATPase inhibitor, ouabain (1mM) did not reduce Cd (50μM) uptake into red blood cells. This suggests that the uptake of Cd into red blood cells was not an active process. Incubation of Cd (10μM) with an equimolar concentration of Zn did not alter uptake of Cd into red blood cells, but at 5 and 10 times higher concentrations of Zn, Cd uptake was enhanced 5-fold. Mercury at one-tenth and equimolar concentrations of Cd increased Cd uptake by red blood cells 2-fold. N-Ethylmaleimide (0.5-5mM), which irreversibly inactivates membrane sulfhydryl groups, decreased Cd uptake. The data indicate that Cd uptake into rat red blood cells occurs by passive transport and that alterations of sulfhydryls of red blood cell membrane may modulate the process. (author)

  18. 77 FR 6463 - Revisions to Labeling Requirements for Blood and Blood Components, Including Source Plasma...

    Science.gov (United States)

    2012-02-08

    ... 20993-0002, (301) 796-9148. SUPPLEMENTARY INFORMATION: In the FR Doc. 2011-33554, appearing on page 7 in the Federal Register of Tuesday, January 3, 2012 (77 FR 7), the following correction is made: 1. On... Requirements for Blood and Blood Components, Including Source Plasma; Correction AGENCY: Food and...

  19. The transport pathway of labelled Mycena osmundicola and assimilated labelled materials in the embryonic cells of Gastrodia elata

    International Nuclear Information System (INIS)

    Mycena osmundicola Lange. was labelled by 3H-glucose and the seeds of Gastrodia elata B1. were sown on the saprophytic leaves of labelled M. osmundicola. By means of autoradiography, it is found that the labelled M. osmundicola infected embryonic cells of G. elata only through the suspensor cells. In the embryonic cells of G. elata the assimilated labelled materials entered into the cells by the wall of cell. After the protocorm formed, the assimilated labelled materials were transferred by the vascular tissue of G. elata

  20. Immunospecific red cell binding of iodine 125-labeled immunoglobulin G erythrocyte autoantibodies

    International Nuclear Information System (INIS)

    The primary interaction of autoantibodies with red cells has been studied by using labeled autoantibodies. Immunoglobulin G red cell autoantibodies obtained from IgG antiglobulin-positive normal blood donors were labeled with radioactive iodine and compared with alloanti-D with respect to their properties and binding behavior. Iodine 125-labeled IgG autoantibody migrated as a single homogeneous peak with the same relative mobility as human IgG on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric focusing pattern of labeled autoantibodies varied from donor to donor but was similar to that of alloanti-D, consisting of multiple IgG populations with isoelectric points in the neutral to alkaline range. 125I-autoantibody bound to all human red cells of common Rh phenotypes. Evidence for immunospecific antibody binding of the labeled autoantibody was based on variation in equilibrium binding to nonhuman and human red cells of common and rare phenotypes, enhanced binding after red cell protease modification, antiglobulin reactivity of cell-bound IgG comparable to that of cell-bound anti-D, and saturation binding in autoantibody excess. Scatchard analysis of two 125I-autoantibody preparations yielded site numbers of 41,500 and 53,300 with equilibrium constants of 3.7 and 2.1 X 10(8) L X mol-1. Dog, rabbit, rhesus monkey, and baboon red cells were antigen(s) negative by quantitative adsorption studies adsorbing less than 3% of the labeled autoantibody. Reduced ability of rare human D--red blood cells to adsorb the autoantibody and identification of donor autoantibodies that bind to Rh null red blood cells indicated that eluates contained multiple antibody populations of complex specificities in contrast to anti-D, which consists of a monospecific antibody population. Another difference is that less than 70% of the autoantibody IgG was adsorbed by maximum binding red blood cells as compared with greater than 85% for alloanti-D

  1. Immunospecific red cell binding of iodine /sup 125/-labeled immunoglobulin G erythrocyte autoantibodies

    Energy Technology Data Exchange (ETDEWEB)

    Masouredis, S.P.; Branks, M.J.; Garratty, G.; Victoria, E.J.

    1987-09-01

    The primary interaction of autoantibodies with red cells has been studied by using labeled autoantibodies. Immunoglobulin G red cell autoantibodies obtained from IgG antiglobulin-positive normal blood donors were labeled with radioactive iodine and compared with alloanti-D with respect to their properties and binding behavior. Iodine /sup 125/-labeled IgG autoantibody migrated as a single homogeneous peak with the same relative mobility as human IgG on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric focusing pattern of labeled autoantibodies varied from donor to donor but was similar to that of alloanti-D, consisting of multiple IgG populations with isoelectric points in the neutral to alkaline range. /sup 125/I-autoantibody bound to all human red cells of common Rh phenotypes. Evidence for immunospecific antibody binding of the labeled autoantibody was based on variation in equilibrium binding to nonhuman and human red cells of common and rare phenotypes, enhanced binding after red cell protease modification, antiglobulin reactivity of cell-bound IgG comparable to that of cell-bound anti-D, and saturation binding in autoantibody excess. Scatchard analysis of two /sup 125/I-autoantibody preparations yielded site numbers of 41,500 and 53,300 with equilibrium constants of 3.7 and 2.1 X 10(8) L X mol-1. Dog, rabbit, rhesus monkey, and baboon red cells were antigen(s) negative by quantitative adsorption studies adsorbing less than 3% of the labeled autoantibody. Reduced ability of rare human D--red blood cells to adsorb the autoantibody and identification of donor autoantibodies that bind to Rh null red blood cells indicated that eluates contained multiple antibody populations of complex specificities in contrast to anti-D, which consists of a monospecific antibody population. Another difference is that less than 70% of the autoantibody IgG was adsorbed by maximum binding red blood cells as compared with greater than 85% for alloanti-D.

  2. Accelerated stem cell labeling with ferucarbotran and protamine

    OpenAIRE

    Golovko, Daniel M.; Henning, Tobias; Bauer, Jan S.; Settles, Marcus; Frenzel, Thomas; Mayerhofer, Artur; Rummeny, Ernst J.; Daldrup-Link, Heike E.

    2009-01-01

    Objective To develop and characterize a clinically applicable, fast and efficient method for stem cell labeling with ferucarbotran and protamine for depiction with clinical MRI. Methods The hydrodynamic diameter, zeta potential and relaxivities of ferucarbotran and varying concentrations of protamine were measured. Once the optimized ratio was found, human mesenchymal stem cells (MSCs) were labeled at varying incubation times (1–24 h). Viability was assessed via Trypan blue exclusion testing....

  3. Radiolabeled red blood cells: status, problems, and prospects

    International Nuclear Information System (INIS)

    Radionuclidic labels for red cells can be divided into two main categories - cohort or pulse labels, and random labels. The random labels are incorporated into circulating cells of all ages and the labeling process is usually carried out in vitro. The red cell labels in predominant use involve random labeling and employ technetium-99m, chromium-51, indium-111, and gallium-68, roughly in that order. The extent of usefulness depends on the properties of the label such as the half-life, decay mode, and in-vivo stability, etc. Labeled cells can be used for red cell survival measurements when the half-life of the radionuclide is sufficiently long. The major portion of this article deals with random labels

  4. Radiolabeled red blood cells: status, problems, and prospects

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, S.C.

    1983-01-01

    Radionuclidic labels for red cells can be divided into two main categories - cohort or pulse labels, and random labels. The random labels are incorporated into circulating cells of all ages and the labeling process is usually carried out in vitro. The red cell labels in predominant use involve random labeling and employ technetium-99m, chromium-51, indium-111, and gallium-68, roughly in that order. The extent of usefulness depends on the properties of the label such as the half-life, decay mode, and in-vivo stability, etc. Labeled cells can be used for red cell survival measurements when the half-life of the radionuclide is sufficiently long. The major portion of this article deals with random labels.

  5. Instant magnetic labeling of tumor cells by ultrasound in vitro

    International Nuclear Information System (INIS)

    Magnetic labeling of living cells creates opportunities for numerous biomedical applications. Here we describe an instantly cell magnetic labeling method based on ultrasound. We present a detailed study on the ultrasound performance of a simple and efficient labeling protocol for H-22 cells in vitro. High frequency focus ultrasound was investigated as an alternative method to achieve instant cell labeling with the magnetic particles without the need for adjunct agents or initiating cell cultures. Mean diameter of 168 nm dextran-T40 coated superparamagnetic iron oxide (SPIO) nanoparticles were prepared by means of classical coprecipitation in solution in our laboratory. H-22 tumor cells suspended in phosphate-buffered saline (PBS, pH=7.2) were exposed to ultrasound at 1.37 MHz for up to 120 s in the presence of SPIOs. The cellular uptake of iron oxide nanoparticles was detected by prussion blue staining. The viability of cells was determined by a trypan blue exclusion test. At 2 W power and 60 s ultrasound exposure in presence of 410 μg/ml SPIOs, H-22 cell labeling efficiency reached 69.4±6.3% and the labeled cells exhibited an iron content of 10.38±2.43 pg per cell. Furthermore, 95.2±3.2% cells remained viable. The results indicated that the ultrasound protocol could be potentially applied to label cells with large-sized magnetic particles. We also calculated the shear stress at the 2 W power and 1.37 MHz used in experiments. The results showed that the shear stress threshold for ultrasonically induced H-22 cell reparable sonoporation was 697 Pa. These findings provide a quantitative guidance in designing ultrasound protocols for cell labeling. - Highlights: → High frequency focus ultrasound can be used as a safe method for instant magnetic labeling of cells. → 8-16 times increased efficiency can be gained by ultrasound versus that by transfection agents. → Calculation of shear stress around cells provide a quantitative design for ultrasound protocols.

  6. Instant magnetic labeling of tumor cells by ultrasound in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Mo Runyang, E-mail: mmrryycn@snnu.edu.cn [Shaanxi Key Laboratory of Ultrasonic, College of Physics and Information Technology, Shaanxi Normal University, No. 199 of South Changan Road, Xi' an 710062 (China); Yang Jian, E-mail: cjr.yangjian@vip.163.com [Department of Diagnostic Radiology, The First Hospital of Medical School, Xi' an Jiaotong University, No. 277 of West Yanta Road, Shannxi Province, Xi' an 710062 (China); Wu, Ed X. [Laboratory of Biomedical Imaging and Signal Processing, University of Hong Kong, Pokfulam, Hong Kong (Hong Kong); Lin Shuyu [Shaanxi Key Laboratory of Ultrasonic, College of Physics and Information Technology, Shaanxi Normal University, No. 199 of South Changan Road, Xi' an 710062 (China)

    2011-09-15

    Magnetic labeling of living cells creates opportunities for numerous biomedical applications. Here we describe an instantly cell magnetic labeling method based on ultrasound. We present a detailed study on the ultrasound performance of a simple and efficient labeling protocol for H-22 cells in vitro. High frequency focus ultrasound was investigated as an alternative method to achieve instant cell labeling with the magnetic particles without the need for adjunct agents or initiating cell cultures. Mean diameter of 168 nm dextran-T40 coated superparamagnetic iron oxide (SPIO) nanoparticles were prepared by means of classical coprecipitation in solution in our laboratory. H-22 tumor cells suspended in phosphate-buffered saline (PBS, pH=7.2) were exposed to ultrasound at 1.37 MHz for up to 120 s in the presence of SPIOs. The cellular uptake of iron oxide nanoparticles was detected by prussion blue staining. The viability of cells was determined by a trypan blue exclusion test. At 2 W power and 60 s ultrasound exposure in presence of 410 {mu}g/ml SPIOs, H-22 cell labeling efficiency reached 69.4{+-}6.3% and the labeled cells exhibited an iron content of 10.38{+-}2.43 pg per cell. Furthermore, 95.2{+-}3.2% cells remained viable. The results indicated that the ultrasound protocol could be potentially applied to label cells with large-sized magnetic particles. We also calculated the shear stress at the 2 W power and 1.37 MHz used in experiments. The results showed that the shear stress threshold for ultrasonically induced H-22 cell reparable sonoporation was 697 Pa. These findings provide a quantitative guidance in designing ultrasound protocols for cell labeling. - Highlights: > High frequency focus ultrasound can be used as a safe method for instant magnetic labeling of cells. > 8-16 times increased efficiency can be gained by ultrasound versus that by transfection agents. > Calculation of shear stress around cells provide a quantitative design for ultrasound protocols.

  7. Measurement of cell mediated cytotoxicity by post-labeling surviving target cells

    International Nuclear Information System (INIS)

    The 51Cr release assay (CRA) is the commonly accepted technique for measurement of cell mediated cytotoxicity. This assay shows some disadvantages when mononucleated cells of human peripheral blood (MNC) are used as effector and target cells. The uptake of 51Cr by PHA stimulated lymphocytes is low compared to the spontaneous release. In an attempt to develop a cytotoxicity assay suitable for human lymphocytes we used 14C-TdR to label target cells surviving after contact with effector cells. Cytotoxic lymphocytes were generated by incubation of MNC with irradiated allogeneic MNC for 6 days. On day 6 the effector cells are irradiated and cocultured with PHA stimulated target cells. Twenty-four hours later 14C-TdR is added. After an additional 24 h the cultures are harvested and 14C-TdR taken up by target cells is measured. It is shown that the effector cells are still cytotoxic after irradiation. These cells do not take up 14C-TdR. Cell-free supernatants do not influence the uptake of 14C-TdR by target cells. The results obtained with this assay correlate very well those obtained by the CRA, if the spontaneous release does not exceed 30%. (author)

  8. Labeling of lectin receptors during the cell cycle.

    Science.gov (United States)

    Garrido, J

    1976-12-01

    Labeling of lectin receptors during the cell cycle. (Localizabión de receptores para lectinas durante el ciclo celular). Arch. Biol. Med. Exper. 10: 100-104, 1976. The topographic distribution of specific cell surface receptors for concanavalin A and wheat germ agglutinin was studied by ultrastructural labeling in the course of the cell cycle. C12TSV5 cells were synchronized by double thymidine block or mechanical selection (shakeoff). They were labeled by means of lectin-peroxidase techniques while in G1 S, G2 and M phases of the cycle. The results obtained were similar for both lectins employed. Interphase cells (G1 S, G2) present a stlihtly discontinous labeling pattern that is similar to the one observed on unsynchronized cells of the same line. Cells in mitosis, on the contrary, present a highly discontinous distribution of reaction product. This pattern disappears after the cells enters G1 and is not present on mitotic cells fixed in aldehyde prior to labeling. PMID:1030938

  9. 77 FR 7 - Revisions to Labeling Requirements for Blood and Blood Components, Including Source Plasma

    Science.gov (United States)

    2012-01-03

    ... Federal Register of July 30, 2003 (68 FR 44678), FDA published a proposed rule that proposed revisions to... International Society of Blood Transfusion Code (ISBT) 128 In the Federal Register of August 30, 1985 (50 FR... Register of November 27, 1998 (63 FR 65600), we announced the availability of the draft standard...

  10. IBCIS:Intelligent blood cell identification system

    Institute of Scientific and Technical Information of China (English)

    Adnan Khashman

    2008-01-01

    The analysis of blood cells in microscope images can provide useful information concerning the health of patients.There are three major blood cell types,namely,erythrocytes (red),leukocytes (white),and platelets.Manual classification is time consuming and susceptible to error due to the different morphological features of the cells.This paper presents an intelligent system that simulates a human visual inspection and classification of the three blood cell types.The proposed system comprises two phases:The image preprocessing phase where blood cell features are extracted via global pattern averaging,and the neural network arbitration phase where training is the first and then classification is carried out.Experimental results suggest that the proposed method performs well in identifying blood cell types regardless of their irregular shapes,sizes and orientation,thus providing a fast,simple and efficient rotational and scale invariant blood cell identification system which can be used in automating laboratory reporting.

  11. Blood-Forming Stem Cell Transplants

    Science.gov (United States)

    ... Health Professionals Questions to Ask about Your Treatment Research Blood-Forming Stem Cell Transplants On This Page What are bone marrow ... are evaluating BMT and PBSCT in clinical trials (research studies) for the treatment ... are the donor’s stem cells matched to the patient’s stem cells in allogeneic ...

  12. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... cell donation experience at the National Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation ( ... About Cord Blood Banking - Duration: 49:19. Children's Health 25,665 views 49:19 Susan Solomon: The ...

  13. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... cell donation experience at the National Institutes of Health Clinical Center in Bethesda, MD. Bone marrow transplantation ( ... About Cord Blood Banking - Duration: 49:19. Children's Health 25,496 views 49:19 Susan Solomon: The ...

  14. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... blood stem cell (PBSC) donor, explains the donation process - Duration: 3:28. Be The Match 22,464 views 3:28 Pain Control: Support for People with Cancer - Duration: 11:58. ...

  15. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... Queue __count__/__total__ Find out why Close Becoming a Blood Stem Cell Donor NCIcancertopics Subscribe Subscribed Unsubscribe ... later? Sign in to add this video to a playlist. Sign in Share More Report Need to ...

  16. Polyelectrolyte coating of ferumoxytol nanoparticles for labeling of dendritic cells

    Science.gov (United States)

    Celikkin, Nehar; Jakubcová, Lucie; Zenke, Martin; Hoss, Mareike; Wong, John Erik; Hieronymus, Thomas

    2015-04-01

    Engineered magnetic nanoparticles (MNPs) are emerging to be used as cell tracers, drug delivery vehicles, and contrast agents for magnetic resonance imaging (MRI) for enhanced theragnostic applications in biomedicine. In vitro labeling of target cell populations with MNPs and their implantation into animal models and patients shows promising outcomes in monitoring successful cell engraftment, differentiation and migration by using MRI. Dendritic cells (DCs) are professional antigen-presenting cells that initiate adaptive immune responses. Thus, DCs have been the focus of cellular immunotherapy and are increasingly applied in clinical trials. Here, we addressed the coating of different polyelectrolytes (PE) around ferumoxytol particles using the layer-by-layer technique. The impact of PE-coated ferumoxytol particles for labeling of DCs and Flt3+ DC progenitors was then investigated. The results from our studies revealed that PE-coated ferumoxytol particles can be readily employed for labeling of DC and DC progenitors and thus are potentially suitable as contrast agents for MRI tracking.

  17. Polyelectrolyte coating of ferumoxytol nanoparticles for labeling of dendritic cells

    International Nuclear Information System (INIS)

    Engineered magnetic nanoparticles (MNPs) are emerging to be used as cell tracers, drug delivery vehicles, and contrast agents for magnetic resonance imaging (MRI) for enhanced theragnostic applications in biomedicine. In vitro labeling of target cell populations with MNPs and their implantation into animal models and patients shows promising outcomes in monitoring successful cell engraftment, differentiation and migration by using MRI. Dendritic cells (DCs) are professional antigen-presenting cells that initiate adaptive immune responses. Thus, DCs have been the focus of cellular immunotherapy and are increasingly applied in clinical trials. Here, we addressed the coating of different polyelectrolytes (PE) around ferumoxytol particles using the layer-by-layer technique. The impact of PE-coated ferumoxytol particles for labeling of DCs and Flt3+ DC progenitors was then investigated. The results from our studies revealed that PE-coated ferumoxytol particles can be readily employed for labeling of DC and DC progenitors and thus are potentially suitable as contrast agents for MRI tracking

  18. 99mTc-red blood cells SPECT and planar scintigraphy in the diagnosis of hepatic hemangiomas.

    Science.gov (United States)

    Artiko, M V; Sobić-Saranović, P D; Perisić-Savić, S M; Stojković, V M; Radoman, B I; Knezević, S J; Petrović, S N; Obradović, B V; Milović, V

    2008-01-01

    The aim of the study is the assessment of the value of SPECT (single photon emission computerized tomography) using 99mTc-labeled red blood cells in the detection of liver hemangioma, in comparison to planar imaging. With planar red blood cell scintigraphy, sensitivity of the method was 76%, specificity 98%, positive predictive value 98% and negative predictive value 79%. With SPECT, sensitivity of the method was 95%, specificity 98%, positive predictive value 98% and negative predictive value 94%. The smallest lesion detected by planar red blood cell scintigraphy was 1.2 cm, and with SPECT red blood cell scintigraphy 0.8 cm. The use of 99mTc-labeled red blood cells SPECT improved the sensitivity much more in smaller lesions (0.8 to 2 cm), than in bigger ones (2-5 cm). SPECT with radiolabeled red blood cells significantlyy improves the results of scintigraphic findings, especially in the small lesions. PMID:19245136

  19. Adeno associated viral-mediated intraosseous labeling of bone marrow derived cells for CNS tracking.

    Science.gov (United States)

    Selenica, Maj-Linda B; Reid, Patrick; Pena, Gabriela; Alvarez, Jennifer; Hunt, Jerry B; Nash, Kevin R; Morgan, Dave; Gordon, Marcia N; Lee, Daniel C

    2016-05-01

    Inflammation, including microglial activation in the CNS, is an important hallmark in many neurodegenerative diseases. Microglial stimuli not only impact the brain microenvironment by production and release of cytokines and chemokines, but also influence the activity of bone marrow derived cells and blood born macrophage populations. In many diseases including brain disorders and spinal cord injury, researchers have tried to harbor the neuroprotective and repair properties of these subpopulations. Hematopoietic bone marrow derived cells (BMDCs) are of great interest, especially during gene therapy because certain hematopoietic cell subpopulations traffic to the sites of injury and inflammation. The aim of this study was to develop a method of labeling endogenous bone marrow derived cells through intraosseous impregnation of recombinant adeno-associated virus (rAAV) or lentivirus. We utilized rAAV serotype 9 (rAAV-9) or lentivirus for gene delivery of green florescence protein (GFP) to the mouse bone marrow cells. Flow cytometry showed that both viruses were able to efficiently transduce mouse bone marrow cells in vivo. However, the rAAV9-GFP viral construct transduced BMDCs more efficiently than the lentivirus (11.2% vs. 6.8%), as indicated by cellular GFP expression. We also demonstrate that GFP labeled cells correspond to bone marrow cells of myeloid origin using CD11b as a marker. Additionally, we characterized the ability of bone marrow derived, GFP labeled cells to extravasate into the brain parenchyma upon acute and subchronic neuroinflammatory stimuli in the mouse CNS. Viral mediated over expression of chemokine (C-C motif) ligand 2 (CCL2) or intracranial injection of lipopolysaccharide (LPS) recruited GFP labeled BMDCs from the periphery into the brain parenchyma compared to vehicle treated mice. Altogether our findings demonstrate a useful method of labeling endogenous BMDCs via viral transduction and the ability to track subpopulations throughout the body

  20. Method for evaluating the potential of 14C labeled plant polyphenols to cross the blood-brain barrier using accelerator mass spectrometry

    International Nuclear Information System (INIS)

    Bioactive compounds in botanicals may be beneficial in preventing age-related neurodegenerative diseases, but for many compounds conventional methods may be inadequate to detect if these compounds cross the blood-brain barrier or to track the pharmacokinetics in the brain. By combining a number of unique technologies it has been possible to utilize the power of AMS to study the pharmacokinetics of bioactive compounds in the brain at very low concentrations. 14C labeled compounds can be biosynthesized by plant cell suspension cultures co-incubated with radioisotopically-labeled sucrose and isolated and separated into a series of bioactive fractions. To study the pharmacokinetics and tissue distribution of 14C labeled plant polyphenols, rats were implanted with jugular catheters, subcutaneous ultrafiltration probes and brain microdialysis probes. Labeled fractions were dosed orally. Interstitial fluid (ISF) and brain microdialysate samples were taken in tandem with blood samples. It was often possible to determine 14C in blood and ISF with a β-counter. However, brain microdialysate samples 14C levels on the order of 107 atoms/sample required AMS technology. The Brain MicrodialysateAUC/SerumAUC ranged from .021- to .029, with the higher values for the glycoside fractions. By using AMS in combination with traditional methods, it is possible to study uptake by blood, distribution to ISF and determine the amount of a dose which can reach the brain and follow the pharmacokinetics in the brain.

  1. Method for evaluating the potential of {sup 14}C labeled plant polyphenols to cross the blood-brain barrier using accelerator mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Janle, Elsa M., E-mail: janle@purdue.ed [Purdue University, Department of Foods and Nutrition, 700 West State Street, West Lafayette, IN 47907-0259 (United States); Lila, Mary Ann [University of Illinois, Department of Natural Resources and Environmental Sciences Urbana IL (United States); Grannan, Michael; Wood, Lauren; Higgins, Aine [Purdue University, Department of Foods and Nutrition, 700 West State Street, West Lafayette, IN 47907-0259 (United States); Yousef, Gad G.; Rogers, Randy B. [University of Illinois, Department of Natural Resources and Environmental Sciences Urbana IL (United States); Kim, Helen [University of Alabama at Birmingham, Department of Pharmacology, Birmingham AB (United States); Jackson, George S. [Purdue University, Department of Physics, West Lafayette, IN (United States); Weaver, Connie M. [Purdue University, Department of Foods and Nutrition, 700 West State Street, West Lafayette, IN 47907-0259 (United States)

    2010-04-15

    Bioactive compounds in botanicals may be beneficial in preventing age-related neurodegenerative diseases, but for many compounds conventional methods may be inadequate to detect if these compounds cross the blood-brain barrier or to track the pharmacokinetics in the brain. By combining a number of unique technologies it has been possible to utilize the power of AMS to study the pharmacokinetics of bioactive compounds in the brain at very low concentrations. {sup 14}C labeled compounds can be biosynthesized by plant cell suspension cultures co-incubated with radioisotopically-labeled sucrose and isolated and separated into a series of bioactive fractions. To study the pharmacokinetics and tissue distribution of {sup 14}C labeled plant polyphenols, rats were implanted with jugular catheters, subcutaneous ultrafiltration probes and brain microdialysis probes. Labeled fractions were dosed orally. Interstitial fluid (ISF) and brain microdialysate samples were taken in tandem with blood samples. It was often possible to determine {sup 14}C in blood and ISF with a beta-counter. However, brain microdialysate samples {sup 14}C levels on the order of 10{sup 7} atoms/sample required AMS technology. The Brain Microdialysate{sub AUC}/Serum{sub AUC} ranged from .021- to .029, with the higher values for the glycoside fractions. By using AMS in combination with traditional methods, it is possible to study uptake by blood, distribution to ISF and determine the amount of a dose which can reach the brain and follow the pharmacokinetics in the brain.

  2. 21 CFR 864.9245 - Automated blood cell separator.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated blood cell separator. 864.9245 Section... Blood and Blood Products § 864.9245 Automated blood cell separator. (a) Identification. An automated blood cell separator is a device that uses a centrifugal or filtration separation principle...

  3. Single-cell measurement of red blood cell oxygen affinity

    CERN Document Server

    Caprio, Di; Higgins, John M; Schonbrun, Ethan

    2015-01-01

    Oxygen is transported throughout the body by hemoglobin in red blood cells. While the oxygen affinity of blood is well understood and is routinely assessed in patients by pulse oximetry, variability at the single-cell level has not been previously measured. In contrast, single-cell measurements of red blood cell volume and hemoglobin concentration are taken millions of times per day by clinical hematology analyzers and are important factors in determining the health of the hematologic system. To better understand the variability and determinants of oxygen affinity on a cellular level, we have developed a system that quantifies the oxygen saturation, cell volume and hemoglobin concentration for individual red blood cells in high-throughput. We find that the variability in single-cell saturation peaks at an oxygen partial pressure of 2.5%, which corresponds to the maximum slope of the oxygen-hemoglobin dissociation curve. In addition, single-cell oxygen affinity is positively correlated with hemoglobin concentr...

  4. Blood Tfh Cells Come with Colors

    Science.gov (United States)

    Schmitt, Nathalie; Ueno, Hideki

    2014-01-01

    Blood CXCR5+ CD4+ T cells share phenotypic and functional similarities with T follicular helper cells. Studies by He et al. (2013) and Locci et al. (2013) in this issue of Immunity provide insight into their ontogeny and functionally distinct subsets. PMID:24138878

  5. Blood cell morphology : controversies and alternatives

    NARCIS (Netherlands)

    Meer, Wim van der

    2006-01-01

    In this thesis we describe controversial morphologic features in both microscopic and automated differentiation of blood cells. In addition, we have investigated alternative methods to overcome these shortcomings. Furthermore we describe the variance of microscopic counting of band cells and variant

  6. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... 074 views 8:21 Bone Marrow/Stem Cell Transplant - Duration: 7:24. tannermom80 99,818 views 7: ... 253 views 6:18 Peripheral Blood Stem Cell Transplant - Duration: 15:50. Dartmouth-Hitchcock 2,689 views ...

  7. Haemopoietic progenitor cells in human peripheral blood

    International Nuclear Information System (INIS)

    The purpose of the investigation reported is to purify haemopoietic progenitor cells from human peripheral blood using density gradient centrifugation in order to isolate a progenitor cell fraction without immunocompetent cells. The purification technique of peripheral blood flow colony forming unit culture (CFU-c) by means of density gradient centrifugation and a combined depletion of various rosettes is described. The results of several 'in vitro' characteristics of purified CFU-c suspensions and of the plasma clot diffusion chamber culture technique are presented. Irradiation studies revealed that for both human bone marrow and peripheral blood the CFU-c were less radioresistant than clusters. Elimination of monocytes (and granulocytes) from the test suspensions induced an alteration in radiosensitivity pararmeters. The results obtained with the different techniques are described by analysing peripheral progenitor cell activity in myeloproliferative disorders. (Auth.)

  8. Deterministic Aperiodic Sickle Cell Blood Flows

    Science.gov (United States)

    Atsaves, Louis; Harris, Wesley

    2013-11-01

    In this paper sickle cell blood flow in the capillaries is modeled as a hydrodynamical system. The hydrodynamical system consists of the axisymmetric unsteady, incompressible Navier-Stokes equations and a set of constitutive equations for oxygen transport. Blood cell deformation is not considered in this paper. The hydrodynamical system is reduced to a system of non-linear partial differential equations that are then transformed into a system of three autonomous non-linear ordinary differential equations and a set of algebraic equations. We examine the hydrodynamical system to discern stable/unstable, periodic/nonperiodic, reversible/irreversible properties of the system. The properties of the solutions are driven in large part by the coefficients of the governing system of equations. These coefficients depend on the physiological properties of the sickle cell blood. The chaotic nature of the onset of crisis in sickle cell patients is identified. Research Assistant.

  9. Accelerated stem cell labeling with ferucarbotran and protamine

    Energy Technology Data Exchange (ETDEWEB)

    Golovko, Daniel M.; Henning, Tobias; Bauer, Jan S. [Department of Radiology and Biomedical Imaging, University of California San Francisco, San Francisco, CA (United States); Settles, Marcus; Rummeny, Ernst J. [Technical University Munich, Department of Radiology, Munich (Germany); Frenzel, Thomas [Bayer Schering Pharma AG, Berlin (Germany); Mayerhofer, Artur [Ludwig-Maximilians-Universitaet, Institute of Cell Biology, Munich (Germany); Daldrup-Link, Heike E. [Department of Radiology and Biomedical Imaging, University of California San Francisco, San Francisco, CA (United States); UCSF Medical Center, Contrast Agent Research Group, Department of Radiology, San Francisco, CA (United States)

    2010-03-15

    To develop and characterize a clinically applicable, fast and efficient method for stem cell labeling with ferucarbotran and protamine for depiction with clinical MRI. The hydrodynamic diameter, zeta potential and relaxivities of ferucarbotran and varying concentrations of protamine were measured. Once the optimized ratio was found, human mesenchymal stem cells (MSCs) were labeled at varying incubation times (1-24 h). Viability was assessed via Trypan blue exclusion testing. 150,000 labeled cells in Ficoll solution were imaged with T1-, T2- and T2*-weighted sequences at 3 T, and relaxation rates were calculated. Varying the concentrations of protamine allows for easy modification of the physicochemical properties. Simple incubation with ferucarbotran alone resulted in efficient labeling after 24 h of incubation while assisted labeling with protamine resulted in similar results after only 1 h. Cell viability remained unaffected. R2 and R2* relaxation rates were drastically increased. Electron microscopy confirmed intracellular iron oxide uptake in lysosomes. Relaxation times correlated with results from ICP-AES. Our results show internalization of ferucarbotran can be accelerated in MSCs with protamine, an approved heparin antagonist and potentially clinically applicable uptake-enhancing agent. (orig.)

  10. Accelerated stem cell labeling with ferucarbotran and protamine

    International Nuclear Information System (INIS)

    To develop and characterize a clinically applicable, fast and efficient method for stem cell labeling with ferucarbotran and protamine for depiction with clinical MRI. The hydrodynamic diameter, zeta potential and relaxivities of ferucarbotran and varying concentrations of protamine were measured. Once the optimized ratio was found, human mesenchymal stem cells (MSCs) were labeled at varying incubation times (1-24 h). Viability was assessed via Trypan blue exclusion testing. 150,000 labeled cells in Ficoll solution were imaged with T1-, T2- and T2*-weighted sequences at 3 T, and relaxation rates were calculated. Varying the concentrations of protamine allows for easy modification of the physicochemical properties. Simple incubation with ferucarbotran alone resulted in efficient labeling after 24 h of incubation while assisted labeling with protamine resulted in similar results after only 1 h. Cell viability remained unaffected. R2 and R2* relaxation rates were drastically increased. Electron microscopy confirmed intracellular iron oxide uptake in lysosomes. Relaxation times correlated with results from ICP-AES. Our results show internalization of ferucarbotran can be accelerated in MSCs with protamine, an approved heparin antagonist and potentially clinically applicable uptake-enhancing agent. (orig.)

  11. Interfacial Polymerization for Colorimetric Labeling of Protein Expression in Cells

    Science.gov (United States)

    Lilly, Jacob L.; Sheldon, Phillip R.; Hoversten, Liv J.; Romero, Gabriela; Balasubramaniam, Vivek; Berron, Brad J.

    2014-01-01

    Determining the location of rare proteins in cells typically requires the use of on-sample amplification. Antibody based recognition and enzymatic amplification is used to produce large amounts of visible label at the site of protein expression, but these techniques suffer from the presence of nonspecific reactivity in the biological sample and from poor spatial control over the label. Polymerization based amplification is a recently developed alternative means of creating an on-sample amplification for fluorescence applications, while not suffering from endogenous labels or loss of signal localization. This manuscript builds upon polymerization based amplification by developing a stable, archivable, and colorimetric mode of amplification termed Polymer Dye Labeling. The basic concept involves an interfacial polymer grown at the site of protein expression and subsequent staining of this polymer with an appropriate dye. The dyes Evans Blue and eosin were initially investigated for colorimetric response in a microarray setting, where both specifically stained polymer films on glass. The process was translated to the staining of protein expression in human dermal fibroblast cells, and Polymer Dye Labeling was specific to regions consistent with desired protein expression. The labeling is stable for over 200 days in ambient conditions and is also compatible with modern mounting medium. PMID:25536421

  12. Performance Evaluation and Labeling Comprehension of a New Blood Glucose Monitoring System with Integrated Information Management

    Science.gov (United States)

    List, Susan M; Starks, Nykole; Baum, John; Greene, Carmine; Pardo, Scott; Parkes, Joan L; Schachner, Holly C; Cuddihy, Robert

    2011-01-01

    Background This study evaluated performance and product labeling of CONTOUR® USB, a new blood glucose monitoring system (BGMS) with integrated diabetes management software and a universal serial bus (USB) port, in the hands of untrained lay users and health care professionals (HCPs). Method Subjects and HCPs tested subject's finger stick capillary blood in parallel using CONTOUR USB meters; deep finger stick blood was tested on a Yellow Springs Instruments (YSI) glucose analyzer for reference. Duplicate results by both subjects and HCPs were obtained to assess system precision. System accuracy was assessed according to International Organization for Standardization (ISO) 15197:2003 guidelines [within ±15 mg/dl of mean YSI results (samples system features and ease-of-use were evaluated by subject questionnaires. Results All subjects who completed the study (N = 74) successfully performed blood glucose measurements, connected the meter to a laptop computer, and used key features of the system. The system was accurate; 98.6% (146/148) of subject results and 96.6% (143/148) of HCP results exceeded ISO 15197:2003 criteria. All subject and HCP results were clinically accurate (97.3%; zone A) or associated with benign errors (2.7%; zone B). The majority of subjects rated features of the BGMS as “very good” or “excellent.” Conclusions CONTOUR USB exceeded ISO 15197:2003 system performance criteria in the hands of untrained lay users. Subjects understood the product labeling, found the system easy to use, and successfully performed blood glucose testing. PMID:22027308

  13. Tomographic sensing and localization of fluorescently labeled circulating cells in mice in vivo

    Science.gov (United States)

    Zettergren, Eric; Swamy, Tushar; Runnels, Judith; Lin, Charles P.; Niedre, Mark

    2012-07-01

    Sensing and enumeration of specific types of circulating cells in small animals is an important problem in many areas of biomedical research. Microscopy-based fluorescence in vivo flow cytometry methods have been developed previously, but these are typically limited to sampling of very small blood volumes, so that very rare circulating cells may escape detection. Recently, we described the development of a ‘diffuse fluorescence flow cytometer’ (DFFC) that allows sampling of much larger blood vessels and therefore circulating blood volumes in the hindlimb, forelimb or tail of a mouse. In this work, we extend this concept by developing and validating a method to tomographically localize circulating fluorescently labeled cells in the cross section of a tissue simulating optical flow phantom and mouse limb. This was achieved using two modulated light sources and an array of six fiber-coupled detectors that allowed rapid, high-sensitivity acquisition of full tomographic data sets at 10 Hz. These were reconstructed into two-dimensional cross-sectional images using Monte Carlo models of light propagation and the randomized algebraic reconstruction technique. We were able to obtain continuous images of moving cells in the sample cross section with 0.5 mm accuracy or better. We first demonstrated this concept in limb-mimicking optical flow photons with up to four flow channels, and then in the tails of mice with fluorescently labeled multiple myeloma cells. This approach increases the overall diagnostic utility of our DFFC instrument.

  14. Tomographic sensing and localization of fluorescently labeled circulating cells in mice in vivo

    International Nuclear Information System (INIS)

    Sensing and enumeration of specific types of circulating cells in small animals is an important problem in many areas of biomedical research. Microscopy-based fluorescence in vivo flow cytometry methods have been developed previously, but these are typically limited to sampling of very small blood volumes, so that very rare circulating cells may escape detection. Recently, we described the development of a ‘diffuse fluorescence flow cytometer’ (DFFC) that allows sampling of much larger blood vessels and therefore circulating blood volumes in the hindlimb, forelimb or tail of a mouse. In this work, we extend this concept by developing and validating a method to tomographically localize circulating fluorescently labeled cells in the cross section of a tissue simulating optical flow phantom and mouse limb. This was achieved using two modulated light sources and an array of six fiber-coupled detectors that allowed rapid, high-sensitivity acquisition of full tomographic data sets at 10 Hz. These were reconstructed into two-dimensional cross-sectional images using Monte Carlo models of light propagation and the randomized algebraic reconstruction technique. We were able to obtain continuous images of moving cells in the sample cross section with 0.5 mm accuracy or better. We first demonstrated this concept in limb-mimicking optical flow photons with up to four flow channels, and then in the tails of mice with fluorescently labeled multiple myeloma cells. This approach increases the overall diagnostic utility of our DFFC instrument. (paper)

  15. Studies on sequestration of neuraminidase-treated red blood cells

    International Nuclear Information System (INIS)

    The effects of reduction in the surface charge of red blood cells (RBCs) on regional blood flow and RBC distribution were studied in rats anesthetized with pentobarbital sodium. RBCs were treated with neuraminidase to reduce their electrophoretic mobility by 56%. Normal and neuraminidase-treated RBCs labeled with 51Cr or 111In were injected into a femoral vein while an equal volume of blood was simultaneously withdrawn from a femoral artery. More than 70% of the neuraminidase-treated RBCs injected disappeared from the circulating blood in 30 min compared with less than 2% of normal RBCs. The relative distributions of neuraminidase-treated RBCs to normal RBCs, as determined from radioactivity counting, were significantly greater than 1 in the spleen (5.65 +/- 0.97, mean +/- SD), the liver (2.84 +/- 0.21), the lung (1.48 +/- 0.31), and the kidney (1.49 +/- 0.27), indicating a preferential trapping of neuraminidase-treated RBCs in these regions. This ratio was approximately 1 in all other organs. Regional blood flows in tissues were determined with 15-micron microspheres in the control period and after the infusion of neuraminidase-treated RBCs (experimental). Experimental-to-control blood flow ratios were 0.40 +/- 0.05 in the spleen, 0.66 +/- 0.06 in the liver, 0.78 +/- 0.03 in the lung, and 0.78 +/- 0.09 in the kidneys; this ratio was approximately 1 in all other organs. An experimental-to-control blood flow ratio less than 1 indicates a reduction in blood flow; this occurred in the same organs as those with trapping of neuraminidase-treated RBCs

  16. Quantum dots for labeling adipose tissue-derived stem cells.

    Science.gov (United States)

    Yukawa, Hiroshi; Mizufune, Shogo; Mamori, Chiharu; Kagami, Yukimasa; Oishi, Koichi; Kaji, Noritada; Okamoto, Yukihiro; Takeshi, Manabu; Noguchi, Hirofumi; Baba, Yoshinobu; Hamaguchi, Michinari; Hamajima, Nobuyuki; Hayashi, Shuji

    2009-01-01

    Adipose tissue-derived stem cells (ASCs) have a self-renewing ability and can be induced to differentiate into various types of mesenchymal tissue. Because of their potential for clinical application, it has become desirable to label the cells for tracing transplanted cells and for in vivo imaging. Quantum dots (QDs) are novel inorganic probes that consist of CdSe/ZnS-core/shell semiconductor nanocrystals and have recently been explored as fluorescent probes for stem cell labeling. In this study, negatively charged QDs655 were applied for ASCs labeling, with the cationic liposome, Lipofectamine. The cytotoxicity of QDs655-Lipofectamine was assessed for ASCs. Although some cytotoxicity was observed in ASCs transfected with more than 2.0 nM of QDs655, none was observed with less than 0.8 nM. To evaluate the time dependency, the fluorescent intensity with QDs655 was observed until 24 h after transfection. The fluorescent intensity gradually increased until 2 h at the concentrations of 0.2 and 0.4 nM, while the intensity increased until 4 h at 0.8 nM. The ASCs were differentiated into both adipogenic and osteogenic cells with red fluorescence after transfection with QDs655, thus suggesting that the cells retain their potential for differentiation even after transfected with QDs655. These data suggest that QDs could be utilized for the labeling of ASCs. PMID:19775521

  17. Effects of broccoli extract on biodistribution and labeling blood components with 99mTc-GH

    International Nuclear Information System (INIS)

    Purpose: people consume vegetables without the knowledge of the side effects of the biological and chemical contents and interactions between radiopharmaceuticals and herbal extract. To this end, current study is focused on the effects of broccoli extract on biodistribution of radiolabeled glucoheptonate (99mTc-GH) and radiolabeling of blood components. Methods: GH was labeled with 99mTc. Quality control studies were done utilizing TLC method. Biodistribution studies were performed on male rats which were treated via gavage with either broccoli extract or SF as control group for 15 days. Blood samples were withdrawn from rats' heart. Radiolabeling of blood constituents performed incubating with GH, SnCl2 and 99m Tc. Results: radiochemical yield of 99mTc-GH is 98.46±1.48 % (n=8). Biodistribution studies have shown that according to the control, the treated group with broccoli has approximately 10 times less uptake in kidney. The percentage of the radioactivity ratios of the blood components is found to be same in both groups. Conclusions: although there is no considerable effect on the radiolabeling of blood components, there is an outstanding change on the biodistribution studies especially on kidneys. The knowledge of this change on kidney uptake may contribute to reduce the risk of misdiagnosis and/or repetition of the examinations in Nuclear Medicine. (author)

  18. Automated red blood cell analysis compared with routine red blood cell morphology by smear review

    OpenAIRE

    Dr.Poonam Radadiya; Dr.Nandita Mehta; Dr.Hansa Goswami; Dr.R.N.Gonsai

    2015-01-01

    The RBC histogram is an integral part of automated haematology analysis and is now routinely available on all automated cell counters. This histogram and other associated complete blood count (CBC) parameters have been found abnormal in various haematological conditions and may provide major clues in the diagnosis and management of significant red cell disorders. Performing manual blood smears is important to ensure the quality of blood count results an...

  19. Fluorophore-conjugated iron oxide nanoparticle labeling and analysis of engrafting human hematopoietic stem cells

    DEFF Research Database (Denmark)

    Maxwell, Dustin J; Bonde, Jesper; Hess, David A;

    2008-01-01

    The use of nanometer-sized iron oxide particles combined with molecular imaging techniques enables dynamic studies of homing and trafficking of human hematopoietic stem cells (HSC). Identifying clinically applicable strategies for loading nanoparticles into primitive HSC requires strictly defined...... culture conditions to maintain viability without inducing terminal differentiation. In the current study, fluorescent molecules were covalently linked to dextran-coated iron oxide nanoparticles (Feridex) to characterize human HSC labeling to monitor the engraftment process. Conjugating fluorophores to the...... vivo. Transplantation of purified primary human cord blood lineage-depleted and CD34(+) cells into immunodeficient mice allowed detection of labeled human HSC in the recipient bones. Flow cytometry was used to precisely quantitate the cell populations that had sequestered the nanoparticles and to...

  20. Label Free Detection of CD4+ and CD8+ T Cells Using the Optofluidic Ring Resonator

    Directory of Open Access Journals (Sweden)

    John T. Gohring

    2010-06-01

    Full Text Available We have demonstrated label free detection of CD4+ and CD8+ T-Lymphocyte whole cells and CD4+ T-Lymphocyte cell lysis using the optofluidic ring resonator (OFRR sensor. The OFRR sensing platform incorporates microfluidics and photonics in a setup that utilizes small sample volume and achieves a fast detection time. In this work, white blood cells were isolated from healthy blood and the concentrations were adjusted to match T-Lymphocyte levels of individuals infected with HIV. Detection was accomplished by immobilizing CD4 and CD8 antibodies on the inner surface of the OFRR. Sensing results show excellent detection of CD4+ and CD8+ T-Lymphocyte cells at medically significant concentrations with a detection time of approximately 30 minutes. This work will lead to a rapid and low-cost sensing device that can provide a CD4 and CD8 count as a measure of HIV progression.

  1. Kinetics of cell labeling and thymidine replacement after continuous infusion of halogenated pyrimidines in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez, R.; Ritter, M.A.; Fowler, J.F.; Kinsella, T.J. (Univ. of Wisconsin Medical School, Madison, WI (United States))

    1994-04-30

    The authors present experiments on an in vivo human tumor xenograft continuously exposed to a fixed serum concentration of halogenated pyrimidines so as to study the kinetics of cell labeling and thymidine replacement. Human colon tumor (HCT-116) cells were injected subcutaneously into nude mice. After 10 days, most animals (>90%) developed measurable tumor nodules with a volume doubling time of 5 [+-] 1 days. Once the tumors reached a cross-sectional area of 0.25-0.30 cm[sup 2], miniosmotic pumps were implanted to deliver a dose of 100 mg/kg/day of IdUrd (iododeoxyuridine) by continuous infusion. After an IdUrd exposure time of 1-7 days, blood and tumor tissue were collected. The steady state serum IdUrd concentration was 0.95 [+-] 0.1 [mu]M, which is a clinically relevant concentration for a prolonged continuous intravenous infusion. The tumor cell potential doubling time (T[sub pot]) was 25. The percent IdUrd thymidine replacement and the fraction of cells labeled followed exponential saturation kinetics with a halflife of 33 and 27 h, respectively. After 5 days of exposure, the thymidine replacement in tumor cells was 2.0% and the fraction of tumor cells labeled was 94%. Immunohistochemical staining of IdUrd labeled tumor tissues showed an exposure-dependent gradient of cellular labeling that was initially highest in regions close to blood vessels. After 4 days of exposure at 100 mg/kg/day, there was an increase in the fraction of cells in G[sub 0] + G[sub 1] and a decrease in the S phase population, suggesting a block between G[sub 1] and S phase. They conclude that the in vivo kinetics of IdUrd thymidine replacement and fraction of cells labeled after continuous exposure followed exponential saturation kinetics with a halflife of approximately the potential doubling time of the tumor cell population. Some form of prolonged, or briefly interrupted, continuous infusion should be considered for clinical administration. 48 refs., 5 figs.

  2. Whole Blood Cell Staining Device

    Science.gov (United States)

    Sams, Clarence F.; Clift, Vaughan L.; McDonald, Kelly E.

    2000-01-01

    An apparatus and method for staining particular cell markers is disclosed. The apparatus includes a flexible tube that is reversibly pinched into compartments with one or more clamps. Each compartment of the tube contains a separate reagent and is in selective fluid communication with adjoining compartments.

  3. The preparation of 11C-labelled fluoromethane for the study of regional cerebral blood flow using positron emission tomography

    International Nuclear Information System (INIS)

    Fluoromethane, previously labelled with 18F and used as a tracer in the measurement of regional cerebral blood flow, was 11C-labelled by the reaction of 11C-methyl iodide with tetraethylammonium fluoride. Sufficient quantities of radiotracer were prepared with a minimum amount of handling from 15 min target irradiations in the 14N(p, α)11C reaction. Total synthesis time was 25 min from end-of-bombardment, allowing serial blood flow measurements 30 min apart. The use of 11C-fluoromethane as a cerebral blood flow tracer in positron emission tomography is discussed. (orig.)

  4. Label-free haemogram using wavelength modulated Raman spectroscopy for identifying immune-cell subset

    Science.gov (United States)

    Ashok, Praveen C.; Praveen, Bavishna B.; Campbell, Elaine C.; Dholakia, Kishan; Powis, Simon J.

    2014-03-01

    Leucocytes in the blood of mammals form a powerful protective system against a wide range of dangerous pathogens. There are several types of immune cells that has specific role in the whole immune system. The number and type of immune cells alter in the disease state and identifying the type of immune cell provides information about a person's state of health. There are several immune cell subsets that are essentially morphologically identical and require external labeling to enable discrimination. Here we demonstrate the feasibility of using Wavelength Modulated Raman Spectroscopy (WMRS) with suitable machine learning algorithms as a label-free method to distinguish between different closely lying immune cell subset. Principal Component Analysis (PCA) was performed on WMRS data from single cells, obtained using confocal Raman microscopy for feature reduction, followed by Support Vector Machine (SVM) for binary discrimination of various cell subset, which yielded an accuracy >85%. The method was successful in discriminating between untouched and unfixed purified populations of CD4+CD3+ and CD8+CD3+ T lymphocyte subsets, and CD56+CD3- natural killer cells with a high degree of specificity. It was also proved sensitive enough to identify unique Raman signatures that allow clear discrimination between dendritic cell subsets, comprising CD303+CD45+ plasmacytoid and CD1c+CD141+ myeloid dendritic cells. The results of this study clearly show that WMRS is highly sensitive and can distinguish between cell types that are morphologically identical.

  5. Cell differentiation mediated by co-culture of human umbilical cord blood stem cells with murine hepatic cells.

    Science.gov (United States)

    Stecklum, Maria; Wulf-Goldenberg, Annika; Purfürst, Bettina; Siegert, Antje; Keil, Marlen; Eckert, Klaus; Fichtner, Iduna

    2015-02-01

    In the present study, purified human cord blood stem cells were co-cultivated with murine hepatic alpha mouse liver 12 (AML12) cells to compare the effect on endodermal stem cell differentiation by either direct cell-cell interaction or by soluble factors in conditioned hepatic cell medium. With that approach, we want to mimic in vitro the situation of preclinical transplantation experiments using human cells in mice. Cord blood stem cells, cultivated with hepatic conditioned medium, showed a low endodermal differentiation but an increased connexin 32 (Cx32) and Cx43, and cytokeratin 8 (CK8) and CK19 expression was monitored by reverse transcription polymerase chain reaction (RT-PCR). Microarray profiling indicated that in cultivated cord blood cells, 604 genes were upregulated 2-fold, with the highest expression for epithelial CK19 and epithelial cadherin (E-cadherin). On ultrastructural level, there were no major changes in the cellular morphology, except a higher presence of phago(ly)some-like structures observed. Direct co-culture of AML12 cells with cord blood cells led to less incisive differentiation with increased sex-determining region Y-box 17 (SOX17), Cx32 and Cx43, as well as epithelial CK8 and CK19 expressions. On ultrastructural level, tight cell contacts along the plasma membranes were revealed. FACS analysis in co-cultivated cells quantified dye exchange on low level, as also proved by time relapse video-imaging of labelled cells. Modulators of gap junction formation influenced dye transfer between the co-cultured cells, whereby retinoic acid increased and 3-heptanol reduced the dye transfer. The study indicated that the cell-co-cultured model of human umbilical cord blood cells and murine AML12 cells may be a suitable approach to study some aspects of endodermal/hepatic cell differentiation induction. PMID:25270685

  6. A label-free and portable graphene FET aptasensor for children blood lead detection

    Science.gov (United States)

    Wang, Chenyu; Cui, Xinyi; Li, Ying; Li, Hongbo; Huang, Lei; Bi, Jun; Luo, Jun; Ma, Lena Q.; Zhou, Wei; Cao, Yi; Wang, Baigeng; Miao, Feng

    2016-02-01

    Lead is a cumulative toxicant, which can induce severe health issues, especially in children’s case due to their immature nervous system. While realizing large-scale monitoring of children blood lead remains challenging by utilizing traditional methods, it is highly desirable to search for alternative techniques or novel sensing materials. Here we report a label-free and portable aptasensor based on graphene field effect transistor (FET) for effective children blood lead detection. With standard solutions of different Pb2+ concentrations, we obtained a dose-response curve and a detection limitation below 37.5 ng/L, which is three orders lower than the safe blood lead level (100 μg/L). The devices also showed excellent selectivity over other metal cations such as, Na+, K+, Mg2+, and Ca2+, suggesting the capability of working in a complex sample matrix. We further successfully demonstrated the detection of Pb2+ ions in real blood samples from children by using our aptasensors, and explored their potential applications for quantification. Our results underscore such graphene FET aptasensors for future applications on fast detection of heavy metal ions for health monitoring and disease diagnostics.

  7. A label-free and portable graphene FET aptasensor for children blood lead detection

    Science.gov (United States)

    Wang, Chenyu; Cui, Xinyi; Li, Ying; Li, Hongbo; Huang, Lei; Bi, Jun; Luo, Jun; Ma, Lena Q.; Zhou, Wei; Cao, Yi; Wang, Baigeng; Miao, Feng

    2016-01-01

    Lead is a cumulative toxicant, which can induce severe health issues, especially in children’s case due to their immature nervous system. While realizing large-scale monitoring of children blood lead remains challenging by utilizing traditional methods, it is highly desirable to search for alternative techniques or novel sensing materials. Here we report a label-free and portable aptasensor based on graphene field effect transistor (FET) for effective children blood lead detection. With standard solutions of different Pb2+ concentrations, we obtained a dose-response curve and a detection limitation below 37.5 ng/L, which is three orders lower than the safe blood lead level (100 μg/L). The devices also showed excellent selectivity over other metal cations such as, Na+, K+, Mg2+, and Ca2+, suggesting the capability of working in a complex sample matrix. We further successfully demonstrated the detection of Pb2+ ions in real blood samples from children by using our aptasensors, and explored their potential applications for quantification. Our results underscore such graphene FET aptasensors for future applications on fast detection of heavy metal ions for health monitoring and disease diagnostics. PMID:26906251

  8. Detection of pulmonary hemorrhage with technetium-labeled red cells

    Energy Technology Data Exchange (ETDEWEB)

    Winzelberg, G.G.; Laman, D.; Sachs, M.; Miller, W.H.

    1981-10-01

    Noninvasive techniques to aid in the diagnosis of massive pulmonary hemoptysis would be helpful in guiding more-invasive procedures such as bronchial artery angiography, which carries a risk of transverse myelitis. A patient was studied with technetium-labeled red cells and successfully detected a site of intermittent hemorrhage from the lung.

  9. Detection of pulmonary hemorrhage with technetium-labeled red cells

    International Nuclear Information System (INIS)

    Noninvasive techniques to aid in the diagnosis of massive pulmonary hemoptysis would be helpful in guiding more-invasive procedures such as bronchial artery angiography, which carries a risk of transverse myelitis. A patient was studied with technetium-labeled red cells and successfully detected a site of intermittent hemorrhage from the lung

  10. Tracing the distribution of labelled aflatoxin and ochratoxin in blood and some organs of white pekin ducklings

    International Nuclear Information System (INIS)

    Labelling of some mycotoxins (ochratoxin and aflatoxin) with radioactive iodine has been performed. The factors affecting the labelling yield such as reaction time, concentration of oxidizing agent (N-bromosuccinimide), pH and concentration of substrate were studied. Separation and purification of the labelled product using ITLC and gel chromatography on sephadex G-25 column have been done. The purified labelled products were orally administrated to white Pekin ducklings to study the bio-distribution in blood and some organs. The present results cleared that high concentrations of ochratoxin and aflatoxin were found in intestinal contents. The labelled ochratoxin reached high concentration in the kidney whereas the labelled aflatoxin reached high concentration in the liver

  11. 21 CFR 660.30 - Reagent Red Blood Cells.

    Science.gov (United States)

    2010-04-01

    ... identify human blood-group antibodies. (b) Source. Reagent Red Blood Cells shall be prepared from human... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Reagent Red Blood Cells. 660.30 Section 660.30...) BIOLOGICS ADDITIONAL STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells §...

  12. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... Duration: 3:35. hemaquebec1998 667 views 3:35 Bone Marrow/Stem Cell Transplant - Duration: 7:24. tannermom80 99,818 views 7:24 Peripheral Blood Stem Cell Transplant - Duration: 15:50. Dartmouth-Hitchcock 2,689 views 15:50 ... Working... Sign in to add this to Watch Later Add to Loading playlists...

  13. Red blood cell transfusion in septic shock

    DEFF Research Database (Denmark)

    Rosland, Ragnhild G; Hagen, Marte U; Haase, Nicolai;

    2014-01-01

    BACKGROUND: Treating anaemia with red blood cell (RBC) transfusion is frequent, but controversial, in patients with septic shock. Therefore we assessed characteristics and outcome associated with RBC transfusion in this group of high risk patients. METHODS: We did a prospective cohort study at 7...

  14. Colour measurement and white blood cell recognition

    CERN Document Server

    Gelsema, E S

    1972-01-01

    As a part of a collaboration with NEMCH aimed at the automation of the differential white blood cell count, studies have been made of the different possibilities for using colour to help in the recognition process. Results are presented comparing data obtained with a microspectrophotometer and with a simulated three-colour scanner.

  15. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... be donors at http://www.marrow.org . Category Science & Technology License Standard YouTube License Show more Show ... Monks 3,700 views 4:41 Stem Cell Basics - How Blood is Made. - Duration: 10:58. Vernon ...

  16. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... blood stem cell (PBSC) donor, explains the donation process - Duration: 3:28. Be The Match 23,393 ... Copyright Creators Advertise Developers +YouTube Terms Privacy Policy & Safety Send feedback Try something new! Loading... Working... Sign ...

  17. GPI-anchored influenza hemagglutinin induces hemifusion to both red blood cell and planar bilayer membranes

    OpenAIRE

    1995-01-01

    Under fusogenic conditions, fluorescent dye redistributed from the outer monolayer leaflet of red blood cells (RBCs) to cells expressing glycophosphatidylinositol-anchored influenza virus hemagglutinin (GPI- HA) without transfer of aqueous dye. This suggests that hemifusion, but not full fusion, occurred (Kemble, G. W., T. Danieli, and J. M. White. 1994. Cell. 76:383-391). We extended the evidence for hemifusion by labeling the inner monolayer leaflets of RBCs with FM4-64 and observing that t...

  18. Sorting white blood cells in microfabricated arrays

    Science.gov (United States)

    Castelino, Judith Andrea Rose

    Fractionating white cells in microfabricated arrays presents the potential for detecting cells with abnormal adhesive or deformation properties. A possible application is separating nucleated fetal red blood cells from maternal blood. Since fetal cells are nucleated, it is possible to extract genetic information about the fetus from them. Separating fetal cells from maternal blood would provide a low cost noninvasive prenatal diagnosis for genetic defects, which is not currently available. We present results showing that fetal cells penetrate further into our microfabricated arrays than adult cells, and that it is possible to enrich the fetal cell fraction using the arrays. We discuss modifications to the array which would result in further enrichment. Fetal cells are less adhesive and more deformable than adult white cells. To determine which properties limit penetration, we compared the penetration of granulocytes and lymphocytes in arrays with different etch depths, constriction size, constriction frequency, and with different amounts of metabolic activity. The penetration of lymphocytes and granulocytes into constrained and unconstrained arrays differed qualitatively. In constrained arrays, the cells were activated by repeated shearing, and the number of cells stuck as a function of distance fell superexponentially. In unconstrained arrays the number of cells stuck fell slower than an exponential. We attribute this result to different subpopulations of cells with different sticking parameters. We determined that penetration in unconstrained arrays was limited by metabolic processes, and that when metabolic activity was reduced penetration was limited by deformability. Fetal cells also contain a different form of hemoglobin with a higher oxygen affinity than adult hemoglobin. Deoxygenated cells are paramagnetic and are attracted to high magnetic field gradients. We describe a device which can separate cells using 10 μm magnetic wires to deflect the paramagnetic

  19. Labelling of live cells using fluorescent aptamers: binding reversal with DNA nucleases

    OpenAIRE

    Terazono Hideyuki; Anzai Yu; Soloviev Mikhail; Yasuda Kenji

    2010-01-01

    Abstract A reversible cell labelling method has been developed for non-destructive and non-invasive cell labelling and purification. Our method uses high affinity single strand DNA (ssDNA) aptamers against surface exposed target molecules on cells. The aptamers are subsequently removed from the cell surface using DNase nuclease treatment. We exemplified our method by labelling human acute lymphoblastic leukemia cells with Qdot-ssDNA aptamers, and restoring them to the label-free condition by ...

  20. Investigations into agents for improving cell labeling with positron- and gamma-emitting radionuclides

    International Nuclear Information System (INIS)

    It was possible to label leukocytes with Co-oxine, but a large proportion of the radioactivity was eluted from the cells upon washing. Ruthenium oxine labeled platelets efficiently in plasma while negligible proportion of radioactivity was eluted from the cells. Three factors influence the labeling efficiency of the cells: duration of the incubation periods; cell concentration; and ACD concentration

  1. A Chemical Probe that Labels Human Pluripotent Stem Cells

    Directory of Open Access Journals (Sweden)

    Nao Hirata

    2014-03-01

    Full Text Available A small-molecule fluorescent probe specific for human pluripotent stem cells would serve as a useful tool for basic cell biology research and stem cell therapy. Screening of fluorescent chemical libraries with human induced pluripotent stem cells (iPSCs and subsequent evaluation of hit molecules identified a fluorescent compound (Kyoto probe 1 [KP-1] that selectively labels human pluripotent stem cells. Our analyses indicated that the selectivity results primarily from a distinct expression pattern of ABC transporters in human pluripotent stem cells and from the transporter selectivity of KP-1. Expression of ABCB1 (MDR1 and ABCG2 (BCRP, both of which cause the efflux of KP-1, is repressed in human pluripotent stem cells. Although KP-1, like other pluripotent markers, is not absolutely specific for pluripotent stem cells, the identified chemical probe may be used in conjunction with other reagents.

  2. Polyelectrolyte coating of ferumoxytol nanoparticles for labeling of dendritic cells

    Energy Technology Data Exchange (ETDEWEB)

    Celikkin, Nehar; Jakubcová, Lucie; Zenke, Martin [Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074 Aachen (Germany); Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Pauwelsstrasse 20, 52074 Aachen (Germany); Hoss, Mareike [Institute of Pathology, Electron Microscopy Facility, RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074 Aachen (Germany); Wong, John Erik, E-mail: John.Wong@avt.rwth-aachen.de [Chemical Process Engineering, RWTH Aachen University, Turmstrasse 46, 52056 Aachen (Germany); DWI – Leibniz Institute for Interactive Materials Research, Forckenbeckstrasse 50, Aachen (Germany); Hieronymus, Thomas, E-mail: thomas.hieronymus@rwth-aachen.de [Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University Hospital, Pauwelsstrasse 30, 52074 Aachen (Germany); Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, Pauwelsstrasse 20, 52074 Aachen (Germany)

    2015-04-15

    Engineered magnetic nanoparticles (MNPs) are emerging to be used as cell tracers, drug delivery vehicles, and contrast agents for magnetic resonance imaging (MRI) for enhanced theragnostic applications in biomedicine. In vitro labeling of target cell populations with MNPs and their implantation into animal models and patients shows promising outcomes in monitoring successful cell engraftment, differentiation and migration by using MRI. Dendritic cells (DCs) are professional antigen-presenting cells that initiate adaptive immune responses. Thus, DCs have been the focus of cellular immunotherapy and are increasingly applied in clinical trials. Here, we addressed the coating of different polyelectrolytes (PE) around ferumoxytol particles using the layer-by-layer technique. The impact of PE-coated ferumoxytol particles for labeling of DCs and Flt3{sup +} DC progenitors was then investigated. The results from our studies revealed that PE-coated ferumoxytol particles can be readily employed for labeling of DC and DC progenitors and thus are potentially suitable as contrast agents for MRI tracking.

  3. Evaluation of copper-labeled Cu(II) bis(thiosemicarbazone) complexes as blood flow tracers for positron emission tomography

    Energy Technology Data Exchange (ETDEWEB)

    Bott, A.J.

    1990-01-01

    Positron emmision tomography (PET) is an imaging technique not widely available for clinical diagnosis due to the costs of producing positron-emitting radiolabels. The development of radiotracers labeled with generator-produced positron emitters would facilitate the use of PET by eliminating the need for an in-house cyclotron. Copper-62 is a generator-produced positron emitter potentially useful for labeling PET radiopharmaceuticals. Copper-62 labeled Cu(II) pyruvaldehyde bis(N[sup 4]-methylthiosemicarbazone), Cu(PTSM), is a proposed PET perfusion tracer of the brain, heart, and kidneys. After IV injection in animals, copper-labeled Cu(PTSM) affords high initial uptake followed by prolonged retention of radiocopper in these organs. This retention is thought to be a result of reductive decomposition of the copper-labeled copper(II) complex by intracellular glutathione, GSH. To validate copper-62 labeled Cu(PTSM) as a myocardial and renal perfusion tracer, the regional radiocopper level afforded by intravenous copper-67 labeled Cu(PTSM) was compared to the absolute blood flow measured with labeled microspheres in normal dog kidneys and surgically infarcted myocardia. In the heart and randomly sectioned kidneys, an excellent correlation resulted. In kidneys dissected to separate anatomical regions, radiocopper levels increased montonically with increasing blood flows for individual dogs, but a linear correlation was observed when data from 12 animals was combined. Should the distribution of Cu(PTSM) vary with naturally occuring GSH fluctuations, the clinical utility of this radiotracer would be limited. Therefore, to validate further copper-62 labeled Cu(PTSM), the biodistribution of copper-67 labeled (PTSM) was determined in GSH-depleted rats. Relatively large GSH reductions in the experimental animals caused only slight changes in the distribution of Cu(PTSM). Another 23 copper-67 labeled compounds were tested.

  4. Evaluation of copper-labeled Cu(II) bis(thiosemicarbazone) complexes as blood flow tracers for positron emission tomography

    International Nuclear Information System (INIS)

    Positron emmision tomography (PET) is an imaging technique not widely available for clinical diagnosis due to the costs of producing positron-emitting radiolabels. The development of radiotracers labeled with generator-produced positron emitters would facilitate the use of PET by eliminating the need for an in-house cyclotron. Copper-62 is a generator-produced positron emitter potentially useful for labeling PET radiopharmaceuticals. Copper-62 labeled Cu(II) pyruvaldehyde bis(N4-methylthiosemicarbazone), Cu(PTSM), is a proposed PET perfusion tracer of the brain, heart, and kidneys. After IV injection in animals, copper-labeled Cu(PTSM) affords high initial uptake followed by prolonged retention of radiocopper in these organs. This retention is thought to be a result of reductive decomposition of the copper-labeled copper(II) complex by intracellular glutathione, GSH. To validate copper-62 labeled Cu(PTSM) as a myocardial and renal perfusion tracer, the regional radiocopper level afforded by intravenous copper-67 labeled Cu(PTSM) was compared to the absolute blood flow measured with labeled microspheres in normal dog kidneys and surgically infarcted myocardia. In the heart and randomly sectioned kidneys, an excellent correlation resulted. In kidneys dissected to separate anatomical regions, radiocopper levels increased montonically with increasing blood flows for individual dogs, but a linear correlation was observed when data from 12 animals was combined. Should the distribution of Cu(PTSM) vary with naturally occuring GSH fluctuations, the clinical utility of this radiotracer would be limited. Therefore, to validate further copper-62 labeled Cu(PTSM), the biodistribution of copper-67 labeled (PTSM) was determined in GSH-depleted rats. Relatively large GSH reductions in the experimental animals caused only slight changes in the distribution of Cu(PTSM). Another 23 copper-67 labeled compounds were tested

  5. Kinetics of indium-111-labeled leukemic cells in patients with acute non-lymphocytic leukemia

    International Nuclear Information System (INIS)

    The kinetics of autologous leukemic cells labeled with In-111 oxine were studied in 5 patients with acute myeloblastic leukemia (AML) and one patient with acute premyelocytic leukemia (APL), and kinetics of OKM1 monoclonal antibody-treated leukemic cells were studied in one patient with acute monoblastic leukemia (AMoL). Recoveries of 33.7 +- 23.3%(range, 22.0 to 48.1%) were achieved at 10min after injection of In-111 oxine labeled leukemic cells in AML and APL patients. However, in a patient with AMoL recovery of 12.3% was only achieved at 10min after injection of OKM1-treated leukemic cells. Clearance of the activity from blood was rapid up to one in all patients. The clearance curve of the activity in 5 AML patients showed a hump or a plateau from one to 5hr after injection of labeled leukemic cells. In APL patient and AMoL patient, however, this hump or plateau was not noted. In AML and APL patients the activity over the spleen was higher than that of over the liver at from 30min to 3hr after and showed a plateau or gradual rising thereafter. In a patient with AMoL, the hepatic activity was higher than the splenic activity at 30min after, but thereafter the latter became higher than the former. Liver activity curves showed transient fall at 3hr after and then gradual uprising in all patients. In a patient with APL, high activity was noted over the kidneys. This rose to a maximum after 3hr and then decreased rapidly. Since In-111 oxine stays firmly attached to the cells in spite of the possibility of radiation damage in a long-term survey, it seems an ideal label for studying leukemic cell kinetics

  6. Red blood cell replacement, or nanobiotherapeutics with enhanced red blood cell functions?

    Science.gov (United States)

    Chang, Thomas Ming Swi

    2015-06-01

    Why is this important? Under normal circumstances, donor blood is the best replacement for blood. However, there are exceptions: During natural epidemics (e.g., HIV, Ebola, etc.) or man-made epidemics (terrorism, war, etc.), there is a risk of donor blood being contaminated, and donors being disqualified because they have contracted disease. Unlike red blood cells (RBCs), blood substitutes can be sterilized to remove infective agents. Heart attack and stroke are usually caused by obstruction of arterial blood vessels. Unlike RBCs, which are particulate, blood substitutes are in the form of a solution that can perfuse through obstructed vessels with greater ease to reach the heart and brain, as has been demonstrated in animal studies. Severe blood loss from injuries sustained during accidents, disasters, or war may require urgent blood transfusion that cannot wait for transportation to the hospital for blood group testing. Unlike RBCs, blood substitutes do not have specific blood groups, and can be administered on the spot. RBCs have to be stored under refrigeration for up to 42 days, and are thus difficult to transport and store in times of disaster and at the battlefront. Blood substitutes can be stored at room temperature for more than 1 year, compared to the RBC shelf life of 1 day, at room temperature. In cases of very severe hemorrhagic shock, there is usually a safety window of 60 min for blood replacement, beyond which there could be problems related to irreversible shock. Animal studies show that a particular type of blood substitute, with enhanced RBC enzymes, may be able to prolong the duration of the safety window. PMID:26096663

  7. Optimization of an Enrichment process for Circulating tumor cells from the blood of Head and Neck Cancer patients through depletion of normal cells

    OpenAIRE

    Yang, Liying; Lang, James C.; Balasubramanian, Priya; Jatana, Kris R.; Schuller, David; Agrawal, Amit; Zborowski, Maciej; Chalmers, Jeffrey J.

    2009-01-01

    The optimization of a purely negative depletion, enrichment process for circulating tumor cells, CTC's, in the peripheral blood of Head and Neck cancer patients is presented. The enrichment process uses a red cell lysis step followed by immunomagnetic labeling, and subsequent depletion, of CD45 positive cells. A number of relevant variables are quantified, or attempted to be quantified, which control the performance of the enrichment process. Six different immunomagnetic labeling combinations...

  8. Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy

    Science.gov (United States)

    Kemper, Björn; Bauwens, Andreas; Vollmer, Angelika; Ketelhut, Steffi; Langehanenberg, Patrik; Müthing, Johannes; Karch, Helge; von Bally, Gert

    2010-05-01

    Digital holographic microscopy (DHM) enables quantitative multifocus phase contrast imaging for nondestructive technical inspection and live cell analysis. Time-lapse investigations on human brain microvascular endothelial cells demonstrate the use of DHM for label-free dynamic quantitative monitoring of cell division of mother cells into daughter cells. Cytokinetic DHM analysis provides future applications in toxicology and cancer research.

  9. Effects of Passiflora edulis flavicarpa on the radiolabeling of blood constituents, morphology of red blood cells and on the biodistribution of sodium pertechnetate in rats

    International Nuclear Information System (INIS)

    The aim of this study was to evaluate possible effects of Passiflora edulis flavicarpa (P. flavicarpa) extract on the labeling of blood constituents with 99mTc, on the morphology of red blood cells, and on the biodistribution of sodium pertechnetate (sodium 99mTc). Male Wistar rats were treated with either P. flavicarpa extract or 0.9% NaCl. After that, radiolabeling of blood constituents, morphological analysis of red blood cells and biodistribution of sodium 99mTc was evaluated. Radiolabeling of blood constituents and shape of red blood cells were not modified, but a significant (p99mTc was observed after treatment with P. flavicarpa extract. Although our results were obtained with animals, they could contribute to reduce the risk of misdiagnosis and/or repetition of the examinations in nuclear medicine

  10. Development of radiochemical method of analysis of binding of tritium labeled drotaverine hydrochloride with human blood serum albumin

    International Nuclear Information System (INIS)

    Full text: The albumin, being a basic functional linkage of numerous endogenous and exogenous substances is the most important protein of blood plasma. At the diseases connected to liver disfunction, collected in blood metabolite reduce connecting ability of albumino. The aim of the present research was a development of radiochemical method of determination of ability of albumin to bind the tritium labeled preparation drotaverine hydrochloride (no - spa). We had developed a micromethod of definition of connecting ability of albumin, allowing to analyse 20 mkl of blood serum. The method consists in incubation of tritium labeled drotaverine hydrochloride with blood serum in vitro, the following fractionation of serum proteins by gel - filtration on a microcolumn with Sephadex G-25, and direct measurement of the radioactivity connected to fraction of proteins of blood serum. The method has been tested on a series of blood serum of control group of healthy people and on a series of blood serum of patients with hepatitis B. We received quantitative characteristics of binding of drotaverine hydrochloride with albumin of patients with hepatitis B. It was preliminary established that binding ability of serum albumin of children with various forms of acute virus hepatitis tends to decrease in comparison with group of the control. Advantage of the developed radiochemical method is high precision and the high sensitivity of detection of infringement of binding ability of albumin. Application of tritium labeled drotaverine hydrochloride allows to measure directly levels of binding of a preparation with albumin

  11. The effect of an extract from Ganoderma lucidum (reishi) on the labeling of blood constituents with technetium-99m and on the survival of Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Agostinho, Raquel Terra [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil); Santos Filho, Sebastiao David; Fonseca, Adenilson de Souza da; Bernardo-Filho, Mario [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Inst. de Biologia Roberto Alcantara Gomes. Dept. de Biofisica e Biometria; Missailidis, Sotiris [The Open University, Milton Keynes (United Kingdom). Dept. of Chemistry and Analytical Sciences

    2008-12-15

    This study evaluated effects of an aqueous extract of Ganoderma lucidum (reishi) on the labeling of blood constituents with technetium-99m ({sup 99m}Tc) and on the survival of cultures of Escherichia coli treated with stannous chloride. Blood samples from Wistar rats were treated with reishi extract, radiolabeling procedure was performed, plasma (P), blood cells (BC) and insoluble (IF) and soluble (SF) fractions of P and BC were separated. The radioactivity was counted for the determination of the percentages of radioactivity (%ATI). Cultures of Escherichia coli AB1157 were treated with stannous chloride in the presence and absence of reishi extract. Blood samples and bacterial cultures treated with NaCl 0.9% were used as controls. Data indicated that reishi extract altered significantly (p<0.05) the %ATI of P, BC, IF-P, SF-P, IF-BC and SF-BC, as well as increased the survival of bacterial cultures treated with stannous chloride. Our results suggest that reishi extract could present a redox/chelating action, altering the labeling of blood constituents with {sup 99}mTc and protecting bacterial cultures against oxidative damage induced by stannous chloride. (author)

  12. Blood cells and endothelial barrier function.

    Science.gov (United States)

    Rodrigues, Stephen F; Granger, D Neil

    2015-01-01

    The barrier properties of endothelial cells are critical for the maintenance of water and protein balance between the intravascular and extravascular compartments. An impairment of endothelial barrier function has been implicated in the genesis and/or progression of a variety of pathological conditions, including pulmonary edema, ischemic stroke, neurodegenerative disorders, angioedema, sepsis and cancer. The altered barrier function in these conditions is often linked to the release of soluble mediators from resident cells (e.g., mast cells, macrophages) and/or recruited blood cells. The interaction of the mediators with receptors expressed on the surface of endothelial cells diminishes barrier function either by altering the expression of adhesive proteins in the inter-endothelial junctions, by altering the organization of the cytoskeleton, or both. Reactive oxygen species (ROS), proteolytic enzymes (e.g., matrix metalloproteinase, elastase), oncostatin M, and VEGF are part of a long list of mediators that have been implicated in endothelial barrier failure. In this review, we address the role of blood borne cells, including, neutrophils, lymphocytes, monocytes, and platelets, in the regulation of endothelial barrier function in health and disease. Attention is also devoted to new targets for therapeutic intervention in disease states with morbidity and mortality related to endothelial barrier dysfunction. PMID:25838983

  13. Responder individuality in red blood cell alloimmunization.

    Science.gov (United States)

    Körmöczi, Günther F; Mayr, Wolfgang R

    2014-11-01

    Many different factors influence the propensity of transfusion recipients and pregnant women to form red blood cell alloantibodies (RBCA). RBCA may cause hemolytic transfusion reactions, hemolytic disease of the fetus and newborn and may be a complication in transplantation medicine. Antigenic differences between responder and foreign erythrocytes may lead to such an immune answer, in part with suspected specific HLA class II associations. Biochemical and conformational characteristics of red blood cell (RBC) antigens, their dose (number of transfusions and pregnancies, absolute number of antigens per RBC) and the mode of exposure impact on RBCA rates. In addition, individual circumstances determine the risk to form RBCA. Responder individuality in terms of age, sex, severity of underlying disease, disease- or therapy-induced immunosuppression and inflammation are discussed with respect to influencing RBC alloimmunization. For particular high-risk patients, extended phenotype matching of transfusion and recipient efficiently decreases RBCA induction and associated clinical risks. PMID:25670932

  14. Assessment of the effect of Bacopa monnieri (L. Wettst. extract on the labeling of blood elements with technetium-99m and on the morphology of red blood cells Avaliação do efeito do extrato de Bacopa monnieri (L. Wettst. na marcação de elementos sanguíneos com tecnécio-99m e na morfologia de células vermelhas do sangue

    Directory of Open Access Journals (Sweden)

    Kakali De

    2009-09-01

    Full Text Available Bacopa monnieri (L. Wettst. (BM, a traditional Ayurvedic medicine, used for centuries as a memory enhancing, anti-inflammatory, antipyretic, sedative and antiepileptic agent. BM extract have been extensively investigated by several authors for their neuropharmacological effects. In nuclear medicine, red blood cells (RBC labeled with technetium-99m (99mTc have several clinical applications. However, data have demonstrated that synthetic or natural drugs could modify the labeling of RBC with 99mTc. As Bacopa monnieri is extensively used in medicine, we evaluated its influence on the labeling of RBC and plasma proteins using technetium-99m (99mTc. This labeling procedure depends on a reducing agent and usually stannous chloride is used. Blood was incubated with BM extracts. Stannous chloride solution and 99mTc were added. Blood was centrifuged and plasma (P and blood cells (BC were isolated. Samples of P or BC were also precipitated, centrifuged and insoluble fraction (IF and soluble fraction (SF were separated. The percentage of radioactivity (%ATI in BC, IF-BC and IF-P were calculated. The %ATI significantly decreased on BC from 95.53±0.45 to 35.41±0.44, on IF-P from 80.20±1.16 to 7.40±0.69 and on IF-BC from 73.31±1.76 to 21.26±1.40. The morphology study of RBC revealed important morphological alterations due to treatment with BM extracts. We suggest that the BM extract effect could be explained by an inhibition of the stannous and pertechnetate ions or oxidation of the stannous ion or by damages induced in the plasma membrane.Bacopa monnieri (L. Wettst. (BM, uma planta tradicional da medicina ayurvédica, é usada por séculos para problemas de memória, antiinflamatória, antitérmica, sedativa e como agente anti-epiléptico. O extrato BM têm sido extensivamente investigada por diversos autores por seus efeitos neurofarmacológicas. Na medicina nuclear, os glóbulos vermelhos (RBC marcados com tecnécio-99m (99mTc tem várias aplica

  15. Adherence of radiopharmaceuticals and labeled cells to intravenous tubing

    International Nuclear Information System (INIS)

    A survey of 67 nuclear medicine departments revealed no agreement on which radiolabeled agents could be injected through intravenous lines (IVs) and which required direct venipuncture. Labeled cells and several common radiopharmaceuticals were tested for adherence to intravenous tubing. Residual activity remaining in the tubing after an adequate flush was less than 1% of the injected dose in each case. Administration of radiolabeled agents through existing IVs is an acceptable alternative to direct venipuncture in many cases

  16. Ultra-fast stem cell labelling using cationised magnetoferritin

    Science.gov (United States)

    Correia Carreira, S.; Armstrong, J. P. K.; Seddon, A. M.; Perriman, A. W.; Hartley-Davies, R.; Schwarzacher, W.

    2016-03-01

    Magnetic cell labelling with superparamagnetic iron oxide nanoparticles (SPIONs) facilitates many important biotechnological applications, such as cell imaging and remote manipulation. However, to achieve adequate cellular loading of SPIONs, long incubation times (24 hours and more) or laborious surface functionalisation are often employed, which can adversely affect cell function. Here, we demonstrate that chemical cationisation of magnetoferritin produces a highly membrane-active nanoparticle that can magnetise human mesenchymal stem cells (hMSCs) using incubation times as short as one minute. Magnetisation persisted for several weeks in culture and provided significant T2* contrast enhancement during magnetic resonance imaging. Exposure to cationised magnetoferritin did not adversely affect the membrane integrity, proliferation and multi-lineage differentiation capacity of hMSCs, which provides the first detailed evidence for the biocompatibility of magnetoferritin. The combination of synthetic ease and flexibility, the rapidity of labelling and absence of cytotoxicity make this novel nanoparticle system an easily accessible and versatile platform for a range of cell-based therapies in regenerative medicine.Magnetic cell labelling with superparamagnetic iron oxide nanoparticles (SPIONs) facilitates many important biotechnological applications, such as cell imaging and remote manipulation. However, to achieve adequate cellular loading of SPIONs, long incubation times (24 hours and more) or laborious surface functionalisation are often employed, which can adversely affect cell function. Here, we demonstrate that chemical cationisation of magnetoferritin produces a highly membrane-active nanoparticle that can magnetise human mesenchymal stem cells (hMSCs) using incubation times as short as one minute. Magnetisation persisted for several weeks in culture and provided significant T2* contrast enhancement during magnetic resonance imaging. Exposure to cationised

  17. Dopamine metabolism in red blood cells in schizophrena

    International Nuclear Information System (INIS)

    A method was developed for the separation by thin-layer chromatography of 14C-labelled 3-methoxy, 4-hydroxyphenethylamine, 3-hydroxy, 4-methoxyphenethylamine and 3,4-dimethoxyphenethylamine (DMPEA) after incubation of dopamine with catechol-O-methyltransferese (COMT) in lysates of human red blood cells (RBC). 14C-methyl-S-adenosyl-methionine was used as the methyl donor. Total COMT activity with noradrenaline or dopamine as substrates, respectively, and the pattern of 14C-methylated metabolites of dopamine were measured in RBC of 47 schizophrenic patients and in 34 control subjects. There were no differences between patients and controls. DMPEA was not formed by RBC in schizophrenic patients (or in controls), a finding which argues against the ''pnk spot''/DMPEA hypothesis of schizophrenia. The methods used seem suitable for studies of other human disorders where COMT might be involved. (author)

  18. Erythropoietin reduces storage lesions and decreases apoptosis indices in blood bank red blood cells

    OpenAIRE

    Oscar Andrés Penuela; Fernando Palomino; Lina Andrea Gómez

    2015-01-01

    ABSTRACT Background: Recent evidence shows a selective destruction of the youngest circulating red blood cells (neocytolysis) trigged by a drop in erythropoietin levels. Objective: The aim of this study was to evaluate the effect of recombinant human erythropoietin beta on the red blood cell storage lesion and apoptosis indices under blood bank conditions. Methods: Each one of ten red blood cell units preserved in additive solution 5 was divided in two volumes of 100 mL and assigned to one...

  19. Preparation and evaluation of [201Tl](III)-DTPA complex for cell labeling

    International Nuclear Information System (INIS)

    Due to interesting physical properties and wide availability of 201Tl as a SPECT radionuclide, the incorporation of this nuclide into DTPA for cell labeling was targeted. Thallium-201 (T1/2 = 3.04 d) in Tl+ form was converted to Tl3+ cation in the presence of O3/6M HCl and di-isopropyl ether, controlled by RTLC/gel electrophoresis methods. The final evaporated activity reacted with cDTPA in normal saline to yield [201Tl](III)DTPA at room temperature after 0.5 hour, followed by solid phase extraction purification using C18 Sep-Pak column (radiochemical yield >95%). Radiochemical purity of more than 99% was obtained using RTLC with specific activity of about 260 GBq/mmol. The stability of the tracer was checked in the final product in the presence of human serum at 37 deg C up to 3 days. The partition coefficient was also measured. The labeled compound was used in red blood cell (RBC) labeling. The cell uptake ratio was determined at 4, 25 and 37 deg C up to 3 hours. (author)

  20. Label-free electronic detection of target cells

    Science.gov (United States)

    Esfandyarpour, Rahim; Javanmard, Mehdi; Harris, James; Davis, Ronald W.

    2014-03-01

    In this manuscript we describe an electronic label-free method for detection of target cells, which has potential applications ranging from pathogen detection for food safety all the way to detection of circulating tumor cells for cancer diagnosis. The nanoelectronic platform consists of a stack of electrodes separated by a 30nm thick insulating layer. Cells binding to the tip of the sensor result in a decrease in the impedance at the sensing tip due to an increase in the fringing capacitance between the electrodes. As a proof of concept we demonstrate the ability to detect Saccharomyces Cerevisae cells with high specificity using a sensor functionalized with Concanavalin A. Ultimately we envision using this sensor in conjunction with a technology for pre-concentration of target cells to develop a fully integrated micro total analysis system.

  1. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice

    CERN Document Server

    Kim, Kyoohyun; Park, Inwon; Kim, Pilhan; Park, YongKeun

    2016-01-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mouse were also investigated.

  2. 21 CFR 864.7100 - Red blood cell enzyme assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Red blood cell enzyme assay. 864.7100 Section 864... enzyme assay. (a) Identification. Red blood cell enzyme assay is a device used to measure the activity in... kinase or 2,3-diphosphoglycerate. A red blood cell enzyme assay is used to determine the enzyme...

  3. Synthesis of fluorine-18 radio-labeled serum albumins for PET blood pool imaging

    International Nuclear Information System (INIS)

    We sought to develop a practical, reproducible and clinically translatable method of radiolabeling serum albumins with fluorine-18 for use as a PET blood pool imaging agent in animals and man. Fluorine-18 radiolabeled fluoronicotinic acid-2,3,5,6-tetrafluorophenyl ester, [18F]F-Py-TFP was prepared first by the reaction of its quaternary ammonium triflate precursor with [18F]tetrabutylammonium fluoride ([18F]TBAF) according to a previously published method for peptides, with minor modifications. The incubation of [18F]F-Py-TFP with rat serum albumin (RSA) in phosphate buffer (pH 9) for 15 min at 37–40 °C produced fluorine-18-radiolabeled RSA and the product was purified using a mini-PD MiniTrap G-25 column. The overall radiochemical yield of the reaction was 18–35% (n = 30, uncorrected) in a 90-min synthesis. This procedure, repeated with human serum albumin (HSA), yielded similar results. Fluorine-18-radiolabeled RSA demonstrated prolonged blood retention (biological half-life of 4.8 hours) in healthy awake rats. The distribution of major organ radioactivity remained relatively unchanged during the 4 hour observation periods either by direct tissue counting or by dynamic PET whole-body imaging except for a gradual accumulation of labeled metabolic products in the bladder. This manual method for synthesizing radiolabeled serum albumins uses fluorine-18, a widely available PET radionuclide, and natural protein available in both pure and recombinant forms which could be scaled up for widespread clinical applications. These preclinical biodistribution and PET imaging results indicate that [18F]RSA is an effective blood pool imaging agent in rats and might, as [18F]HSA, prove similarly useful as a clinical imaging agent

  4. Influence of biflorin on the labelling of red blood cells, plasma protein, cell protein, and lymphocytes with technetium-99m: in vitro study Influência da biflorina na marcação do tecnécio-99m em células vermelhas do sangue, proteínas do plasma, proteínas celulares e em linfócitos: estudos in vitro

    Directory of Open Access Journals (Sweden)

    Thiago M. Aquino

    2007-06-01

    Full Text Available In this paper we report the results of an in vitro study involving the influence of biflorin (an o-quinone isolated from Capraria biflora L. that has potent antimicrobial activity on the Tc-99m labeling of red blood cells, plasma protein, cells protein, and lymphocytes. Blood was withdrawn from Wistar rats and incubated with various concentrations of biflorin, and solutions of stannous chloride and Tc-99m were added. Plasma (P and red blood cells (RBC were isolated, precipitated, and centrifuged, and soluble (SF and insoluble (IF fractions were isolated. The results show that the highest concentration (100% of biflorin is able to reduce the uptake of Tc-99m (%ATI on RBC and the fixation on IF-P. To study the influence of biflorin on 99mTc lymphocyte labeling, human blood was submitted to a technique with Ficoll-Hypac and centrifuged, and white cells were isolated. Lymphocytes (2.5 mL; 1.0 x 10(6 cells/mL were obtained and a 0.2 mL solution was incubated with biflorin (0.1 mL. Solutions of stannous chloride and 99mTc were added. Lymphocytes were separated and the %ATI bound in these cells was evaluated. A reduction in %ATI (from 97.85 ± 0.99 to 88.86 ± 5 was observed for RBC and for IF-P (73.24 ± 5.51 to 20.72 ± 6.95. In this case the results showed no decrease in %ATI for the lymphocytes with biflorin.Neste artigo relatam-se os resultados de um estudo in vitro envolvendo a influência da biflorina (uma o-quinona isolada de Capraria biflora L. que possui uma potente atividade antimicrobiana na marcação do Tc-99m em células vermelhas do sangue, proteínas do plasma, proteínas celulares e em linfócitos. O sangue foi coletado de ratos Wistar e incubado com várias concentrações de biflorina, e soluções de cloreto estanoso (SnCl2 adicionando-se Tc-99m. O plasma (P e as células vermelhas do sangue (CVS foram isolados, precipitados e centrifugados, isolando-se as frações solúveis (FS e insolúveis (FI. A maior concentração de

  5. The aging of the red blood cell. A multifactor process.

    Science.gov (United States)

    Danon, D; Marikovsky, Y

    1988-01-01

    Red blood cell (rbc) senescence is associated with loss of surface sialic acid, which is the principal carrier of surface negative charge and determines the electrokinetic behavior of old rbcs. Loss of sialic acid in an old rbc is demonstrated in its decreased electric mobility and lower negative charge density, determined topographically with cationic particle labeling. Surface sialic acid determines also the mutual attraction--repulsion forces, as demonstrated in enhanced aggluinability with cationic molecules, lectins, and blood group antibodies. Loss of sialic acid accompanies ATP-depletion in vitro; thus, a T-antigen site is unmasked. Macrophages have specific receptors to the site as to newly exposed galactose and N-acetyl galactosamine sugars. Furthermore, the involvement of complement molecules in the recognition of old RBCs by macrophages has been shown. This is possibly due to loss of sialic acid or at least a regrouping--relocation of surface anionic sites due to cell shape changes from discocytes to crenated forms, which accompany both in vivo and in vitro rbc aging. In turn, shape changes are apparently controlled by the cytoskeletal network underlying the rbc membrane, which undergoes structural alteration with physiologic aging in changing the dimensions of oligomeric spectrin and the thickness of the spectrin-actin cytoskeletal assembly. PMID:3052636

  6. Automated red blood cell analysis compared with routine red blood cell morphology by smear review

    Directory of Open Access Journals (Sweden)

    Dr.Poonam Radadiya

    2015-01-01

    Full Text Available The RBC histogram is an integral part of automated haematology analysis and is now routinely available on all automated cell counters. This histogram and other associated complete blood count (CBC parameters have been found abnormal in various haematological conditions and may provide major clues in the diagnosis and management of significant red cell disorders. Performing manual blood smears is important to ensure the quality of blood count results and to make presumptive diagnosis. In this article we have taken 100 samples for comparative study between RBC histograms obtained by automated haematology analyzer with peripheral blood smear. This article discusses some morphological features of dimorphism and the ensuing characteristic changes in their RBC histograms.

  7. Mechanosensing Dynamics of Red blood Cells

    Science.gov (United States)

    Wan, Jiandi

    2015-11-01

    Mechanical stress-induced deformation of human red blood cells (RBCs) plays important physiopathological roles in oxygen delivery, blood rheology, transfusion, and malaria. Recent studies demonstrate that, in response to mechanical deformation, RBCs release adenosine-5'-triphosphate (ATP), suggesting the existence of mechanotransductive pathways in RBCs. Most importantly, the released ATP from RBCs regulates vascular tone and impaired release of ATP from RBCs has been linked to diseases such as type II diabetes and cystic fibrosis. To date, however, the mechanisms of mechanotransductive release of ATP from RBCs remain unclear. Given that RBCs experience shear stresses continuously during the circulation cycle and the released ATP plays a central role in vascular physiopathology, understanding the mechanotransductive release of ATP from RBCs will provide not only fundamental insights to the role of RBCs in vascular homeostasis but also novel therapeutic strategies for red cell dysfunction and vascular disease. This talk describes the main research in my group on integrating microfluidic-based approaches to study the mechanosensing dynamics of RBCs. Specifically, I will introduce a micro?uidic approach that can probe the dynamics of shear-induced ATP release from RBCs with millisecond resolution and provide quantitative understandings of the mechanosensitive ATP release processes in RBCs. Furthermore, I will also describe our recent findings about the roles of the Piezo1 channel, a newly discovered mechanosensitive cation channel in the mechanotransductive ATP release in RBCs. Last, possible functions of RBCs in the regulation of cerebral blood flow will be discussed.

  8. Red blood cell in simple shear flow

    Science.gov (United States)

    Chien, Wei; Hew, Yayu; Chen, Yeng-Long

    2013-03-01

    The dynamics of red blood cells (RBC) in blood flow is critical for oxygen transport, and it also influences inflammation (white blood cells), thrombosis (platelets), and circulatory tumor migration. The physical properties of a RBC can be captured by modeling RBC as lipid membrane linked to a cytoskeletal spectrin network that encapsulates cytoplasm rich in hemoglobin, with bi-concave equilibrium shape. Depending on the shear force, RBC elasticity, membrane viscosity, and cytoplasm viscosity, RBC can undergo tumbling, tank-treading, or oscillatory motion. We investigate the dynamic state diagram of RBC in shear and pressure-driven flow using a combined immersed boundary-lattice Boltzmann method with a multi-scale RBC model that accurately captures the experimentally established RBC force-deformation relation. It is found that the tumbling (TU) to tank-treading (TT) transition occurs as shear rate increases for cytoplasm/outer fluid viscosity ratio smaller than 0.67. The TU frequency is found to be half of the TT frequency, in agreement with experiment observations. Larger viscosity ratios lead to the disappearance of stable TT phase and unstable complex dynamics, including the oscillation of the symmetry axis of the bi-concave shape perpendicular to the flow direction. The dependence on RBC bending rigidity, shear modulus, the order of membrane spectrin network and fluid field in the unstable region will also be discussed.

  9. The tumour distribution of bromodeoxyuridine labelled S-phase cells is found to be strongly dose dependant

    International Nuclear Information System (INIS)

    Bromodeoxyurdine (BrdU) is used extensively to measure the fraction of S-phase cells in tumours. Unlike endogenous markers of proliferation, such as PCNA and Ki-67, BrdU is exogenously administered and reaches the tumour via vasculature where it must then distribute throughout the tissue in order to label S-phase cells. Interest in using BrdU labelling of histological sections to evaluate the distribution of the effect of different treatment modalities on tumours led us to study the ability of BrdU to distribute within tissue. The study used SiHa (human cervix squamous cell carcinoma) xenografts, a tumour that exhibits cords of cells extending up to 150 μm away from blood vessels. A quantitative microscopy-based technique was employed to determine the distribution of S-phase labelled cells relative to the vasculature over a dose range of 25-2000 mg/kg BrdU. Detection of BrdU incorporation in DNA was carried out immunohistochemically and vasculature was identified using perfusion of carbocyanine, a fluorescent perivascular stain. Analysis of BrdU labelling distribution in the tissue found that a dose of 1000 mg/kg was required to label cells furthest from vasculature. Dosing at lower levels resulted in only the cells close to blood vessels being labelled. This result is surprising since 100 mg/kg BrdU is commonly used in flow cytometry studies. Results were compared with penetration seen in vitro using multilayered cell culture, a three-dimensional tissue culture model of solid tumours. Using multilayered cell culture, an exposure of 100 μM BrdU for 1 hour was required for labelling of S-phase cells 150 μm into the tissue, while cells adjacent to the edge of the tissue could be adequately labelled with just 5 μM BrdU for 1 hour. The AUC for a 100 mg/kg BrdU dose in mice was found to be ∼30 μM x h

  10. The white blood cell scan in orthopedics

    Energy Technology Data Exchange (ETDEWEB)

    Propst-Proctor, S.L.; Dillingham, M.F.; McDougall, I.R.; Goodwin, D.

    1982-08-01

    A new nuclear scanning technique was found more specific for bone, joint, and soft tissue infections than any previously described scanning technique. The leukocyte scan, whereby a patient's own cells are labeled with a radioactive tagging agent (/sup 111/In oxine), can distinguish an active infectious process from other pain-inducing conditions. Ninety-seven /sup 111/In labeled autologous leukocyte scans were performed in 88 patients. The findings in 17 of 40 patients scanned for possible acute osteomyelitis, six of nine for suspected septic arthritis, and six for possible soft tissue infections, were positive. Subsequent clinical courses verified the infectious nature of these processes in all patients. Patients who had chronic osteomyelitis (14), bony metastases (four patients), heterotopic ossification (three), and degenerative arthritis (two) demonstrated negative findings. Of the seven patients scanned for acute long-bone fractures, one demonstrated positive findings. Nine scans demonstrated positive findings without determined causes. The leukocyte scan is a useful addition to the diagnostic tools of the orthopedic surgeon.

  11. The white blood cell scan in orthopedics

    International Nuclear Information System (INIS)

    A new nuclear scanning technique was found more specific for bone, joint, and soft tissue infections than any previously described scanning technique. The leukocyte scan, whereby a patient's own cells are labeled with a radioactive tagging agent (111In oxine), can distinguish an active infectious process from other pain-inducing conditions. Ninety-seven 111In labeled autologous leukocyte scans were performed in 88 patients. The findings in 17 of 40 patients scanned for possible acute osteomyelitis, six of nine for suspected septic arthritis, and six for possible soft tissue infections, were positive. Subsequent clinical courses verified the infectious nature of these processes in all patients. Patients who had chronic osteomyelitis (14), bony metastases (four patients), heterotopic ossification (three), and degenerative arthritis (two) demonstrated negative findings. Of the seven patients scanned for acute long-bone fractures, one demonstrated positive findings. Nine scans demonstrated positive findings without determined causes. The leukocyte scan is a useful addition to the diagnostic tools of the orthopedic surgeon

  12. Optical analysis of red blood cell suspension

    Science.gov (United States)

    Szołna, Alicja A.; Grzegorzewski, Bronisław

    2008-12-01

    The optical properties of suspensions of red blood cells (RBCs) were studied. Fresh human venues blood was obtained from adult healthy donors. RBCs were suspended in isotonic salt solution, and in autologous plasma. Suspensions with haematocrit 0.25 - 3% were investigated. Novel technique was proposed to determine the scattering coefficient μs for the suspensions. The intensity of He-Ne laser light transmitted through a wedge-shape container filled with a suspension was recorded. To find the dependence of the intensity on the thickness of the sample the container was moved horizontally. The dependence of μs on the haematocrit was determined for RBCs suspended in the isotonic salt solution. RBCs suspended in plasma tend to form rouleaux. For the RBCs suspended in plasma, the scattering coefficient as a function of time was obtained. It is shown that this technique can be useful in the study of rouleaux formation.

  13. 111In oxine labeled red cells for detection of simulated lower gastrointestinal bleeding in an animal model

    International Nuclear Information System (INIS)

    111In oxine in vitro labeled red cells were evaluated in rabbits for the ability to detect gastrointestinal (Gl) bleeding. A mean labeling efficiency of 81% (+- 15.5%) was achieved. Biodistribution and translocation data demonstrated 81% of the activity within the blood pool at four hours after intravenous injection, falling to 29% by 72 hours. Peak urine excretion occurred after 60 to 150 minutes. Normal Gl excretion was less than 1% over 72 hours. Simulated lower Gl bleeding was imaged at 4, 12, and 72 hours, and amounts as small as 2 ml (1% blood volume) were seen. In rabbits the total body dose of injected 111In is 0.15 mGy/MBq (0.56 rad/mCi), and the critical organ is the spleen, which received 0.49 mGy/MBq (1.82 rad/mCi). 111In oxine labeled red cells provide a sustained blood pool label wthout significant accumulation in the Gl tract, and may have a potential use in the detection of intermittent Gl bleeding in humans

  14. Uncaria tomentosa extract: evaluation of effects on the in vitro and in vivo labeling of blood constituents with technetium-99m

    Energy Technology Data Exchange (ETDEWEB)

    Moreno, Silvana Ramos Farias; Olej, Beni; Arnobio, Adriano; Caldas, Luiz Querino de Araujo [Universidade Federal Fluminense (UFF), Niteroi, RJ (Brazil)]. E-mail: srfmoreno@hotmail.com; Carvalho, Jorge Jose de; Nascimento, Ana Lucia [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Dept. de Histologia e Embriologia; Rocha, Emely Kazan [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Dept. de Biologia Celular e Genetica; Bernardo-Filho, Mario [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Dept. de Biofisica e Biometria; Honeycut, Hayden [University of North Carolina, Chapel Hill, NC (United States). School of Pharmacy

    2008-12-15

    The influence (in vivo and in vitro) of an Uncaria tomentosa extract (Cats claw) on the labeling of red blood cells (RBCs) and plasma and cellular proteins with technetium-99m (Tc-99m) was evaluated. For the in vivo treatment, animals were treated with Cats claw. For the in vitro treatment, heparinized blood was incubated with Cats claw before the addition of stannous chloride (SnCl{sub 2}) and Tc-99m. Samples of plasma (P) and RBCs were separated and also precipitated with trichloroacetic acid. The soluble and insoluble fractions of P and RBCs were isolated. The analysis of the results of the in vivo study, indicates that there is no significant alteration on the uptake of Tc-99m by the blood constituents, but it significantly decrease (p<0.05) the labeling of blood constituents by in vitro methods. These effects could be due to chelation of stannous and /or pertechnetate ions and blockage of the Tc-99m bindings sites. (author)

  15. Immunophenotyping of hematopoietic progenitor cells: Comparison between cord blood and adult mobilized blood grafts

    OpenAIRE

    2011-01-01

    AIM: To study the immunophenotype of hematopoietic progenitor cells from cord blood (CB) grafts (n = 39) in comparison with adult apheresis grafts (AG, n = 229) and pre-apheresis peripheral blood (PAPB) samples (n = 908) using flow cytometry analysis.

  16. Arterial Blood, Rather Than Venous Blood, is a Better Source for Circulating Melanoma Cells

    Directory of Open Access Journals (Sweden)

    Mizue Terai

    2015-11-01

    Interpretation: Our data indicate that arterial blood specimens might be a better source of circulating uveal melanoma cells. Although less conveniently processed, perhaps arterial blood should be evaluated as sample source for measurement of CTCs.

  17. In-Depth Profiling of the Peripheral Blood Mononuclear Cells Proteome for Clinical Blood Proteomics

    OpenAIRE

    Saša Končarević; Christopher Lößner; Karsten Kuhn; Thorsten Prinz; Ian Pike; Hans-Dieter Zucht

    2014-01-01

    Peripheral blood mononuclear cells (PBMCs) are an easy accessible cellular part of the blood organ and, along with platelets, represent the only site of active gene expression in blood. These cells undergo immunophenotypic changes in various diseases and represent a peripheral source of monitoring gene expression and posttranslational modifications relevant to many diseases. Little is known about the source of many blood proteins and we hypothesise that release from PBMCs through active and p...

  18. Magnetophoretic separation of blood cells at the microscale

    CERN Document Server

    Furlani, E P

    2006-01-01

    We present a method and model for the direct and continuous separation of red and white blood cells in plasma. The method is implemented at the microscale using a microfluidic system that consists of an array of integrated soft-magnetic elements embedded beneath a microfluidic channel. The microsystem is passive, and is activated via application of a bias field that magnetizes the elements. Once magnetized, the elements produce a nonuniform magnetic field distribution in the microchannel, which gives rise to a force on blood cells as they pass through the microsystem. In whole blood, white blood cells behave as diamagnetic microparticles while red blood cells exhibit diamagnetic or paramagnetic behavior depending on the oxygenation of their hemoglobin. We develop a mathematical model for predicting the motion of blood cells in the microsystem that takes into account the dominant magnetic, fluidic and buoyant forces on the cells. We use the model to study red/white blood cell transport, and our analysis indica...

  19. Nanoparticle-labeled stem cells: a novel therapeutic vehicle

    Directory of Open Access Journals (Sweden)

    Abir O El-Sadik

    2010-03-01

    Full Text Available Abir O El-Sadik1, Afaf El-Ansary2, Sherif M Sabry31Stem Cell Unit, Anatomy Department, College of Medicine, Health Science Colleges; 2Biochemistry Department, Science College, King Saud University; 3Anatomy Department, Faculty of Medicine, Cairo University, Cairo, EgyptAbstract: Nanotechnology has been described as a general purpose technology. It has already generated a range of inventions and innovations. Development of nanotechnology will provide clinical medicine with a range of new diagnostic and therapeutic opportunities such as medical imaging, medical diagnosis, drug delivery, and cancer detection and management. Nanoparticles such as manganese, polystyrene, silica, titanium oxide, gold, silver, carbon, quantum dots, and iron oxide have received enormous attention in the creation of new types of analytical tools for biotechnology and life sciences. Labeling of stem cells with nanoparticles overcame the problems in homing and fixing stem cells to their desired site and guiding extension of stem cells to specific directions. Although the biologic effects of some nanoparticles have already been assessed, information on toxicity and possible mechanisms of various particle types remains inadequate. The aim of this review is to give an overview of the mechanisms of internalization and distribution of nanoparticles inside stem cells, as well as the influence of different types of nanoparticles on stem cell viability, proliferation, differentiation, and cytotoxicity, and to assess the role of nanoparticles in tracking the fate of stem cells used in tissue regeneration.Keywords: nanoparticles, stem cells, uptake, differentiation, cytotoxicity, tracking

  20. Isolation of mesenchymal stem cells from equine umbilical cord blood

    DEFF Research Database (Denmark)

    Koch, Thomas Gadegaard; Heerkens, Tammy; Thomsen, Preben Dybdahl;

    2007-01-01

    Background: There are no published studies on stem cells from equine cord blood although commercial storage of equine cord blood for future autologous stem cell transplantations is available. Mesenchymal stem cells (MSC) have been isolated from fresh umbilical cord blood of humans collected non......-invasively at the time of birth and from sheep cord blood collected invasively by a surgical intrauterine approach. Mesenchymal stem cells isolation percentage from frozen-thawed human cord blood is low and the future isolation percentage of MSCs from cryopreserved equine cord blood is therefore expectedly low....... The hypothesis of this study was that equine MSCs could be isolated from fresh whole equine cord blood. Results: Cord blood was collected from 7 foals immediately after foaling. The mononuclear cell fraction was isolated by Ficoll density centrifugation and cultured in a DMEM low glucose based media...

  1. Protection of plasmid DNA by a Ginkgo biloba extract from the effects of stannous chloride and the action on the labeling of blood elements with technetium-99m

    Directory of Open Access Journals (Sweden)

    Moreno S.R.F.

    2004-01-01

    Full Text Available Ginkgo biloba extract (EGb is a phytotherapeutic agent used for the treatment of ischemic and neurological disorders. Because the action of this important extract is not fully known, assays using different biological systems need to be performed. Red blood cells (RBC are labeled with technetium-99m (Tc-99m and used in nuclear medicine. The labeling depends on a reducing agent, usually stannous chloride (SnCl2. We assessed the effect of different concentrations of EGb on the labeling of blood constituents with Tc-99m, as sodium pertechnetate (3.7 MBq, and on the mobility of a plasmid DNA treated with SnCl2 (1.2 µg/ml at room temperature. Blood was incubated with EGb before the addition of SnCl2 and Tc-99m. Plasma (P and RBC were separated and precipitated with trichloroacetic acid, and soluble (SF-P and SF-RBC and insoluble (IF-P and IF-RBC fractions were isolated. The plasmid was incubated with Egb, SnCl2 or EGb plus SnCl2 and agarose gel electrophoresis was performed. The gel was stained with ethidium bromide and the DNA bands were visualized by fluorescence in an ultraviolet transilluminator system. EGb decreased the labeling of RBC, IF-P and IF-RBC. The supercoiled form of the plasmid was modified by treatment with SnCl2 and protected by 40 mg/ml EGb. The effect of EGb on the tested systems may be due to its chelating action with the stannous ions and/or pertechnetate or to the capability to generate reactive oxygen species that could oxidize the stannous ion.

  2. Dielectric Constant of Suspensions of Blood Cells

    Science.gov (United States)

    Mendelson, Kenneth; Ackmann, James

    1996-03-01

    Measurements of the complex dielectric constant of suspensions of blood cells have recently been reported by Ackmann, et al.(J. J. Ackmann, et al., Ann. Biomed. Eng. 24), 58 (1996). At frequencies below 100 kHz, the real part of the dielectric constant (ɛ') goes through a maximum at a blood cell volume fraction of about 70%. Effective medium approximations do not agree well with this behavior. As a more realistic model, we are studying the grain consolidation model of Roberts and Schwartz(J. N. Roberts and L. M. Schwartz, Phys. Rev. B 31), 5990 (1985). We have used a finite element method to calculate the dielectric constant of this model for a cubic array of spheres. The simulations agree remarkably well with experiment. They suggest, however, that ɛ' may be showing oscillations rather than a simple maximum. Comparison of the simulated and experimental points suggests that this is not an artifact of the periodic array used in the model. Furthermore the simulations indicate that the maximum (or oscillations) disappears at low conductivities of the suspending fluid.

  3. Creation of Primary Cell Lines from Lineage-Labeled Mouse Models of Cancer

    Science.gov (United States)

    Rhim, Andrew D.

    2015-01-01

    Frequently, it is necessary to isolate pure populations of cancer cells for downstream assays, such as transcriptional analysis, signaling studies, and the creation of noncontaminated primary cell lines. Genetic lineage labeling with fluorescent reporter alleles allows for the identification of epithelial-derived cells within tumors. This protocol describes a method to isolate lineage-labeled pancreatic epithelial cells for ex vivo analysis, but it can be adapted for any type of lineage-labeled tumor. PMID:25934932

  4. Creation of Primary Cell Lines from Lineage-Labeled Mouse Models of Cancer

    OpenAIRE

    Rhim, Andrew D.

    2015-01-01

    Frequently, it is necessary to isolate pure populations of cancer cells for downstream assays, such as transcriptional analysis, signaling studies, and the creation of noncontaminated primary cell lines. Genetic lineage labeling with fluorescent reporter alleles allows for the identification of epithelial-derived cells within tumors. This protocol describes a method to isolate lineage-labeled pancreatic epithelial cells for ex vivo analysis, but it can be adapted for any type of lineage-label...

  5. Alterations in cell surface area and deformability of individual human red blood cells in stored blood

    CERN Document Server

    Park, HyunJoo; Lee, SangYun; Kim, Kyoohyun; Sohn, Yong-Hak; Jang, Seongsoo; Park, YongKeun

    2015-01-01

    The functionality and viability of stored human red blood cells (RBCs) is an important clinical issue in transfusion. To systematically investigate changes in stored whole blood, the hematological properties of individual RBCs were quantified in blood samples stored for various periods with and without a preservation solution called CPDA-1. With 3-D quantitative phase imaging techniques, the optical measurements of the 3-D refractive index (RI) distributions and membrane fluctuations were done at the individual cell level. From the optical measurements, the morphological (volume, surface area and sphericity), biochemical (hemoglobin content and concentration), and mechanical parameters (dynamic membrane fluctuation) were simultaneously quantified to investigate the functionalities and their progressive alterations in stored RBCs. Our results show that the stored RBCs without CPDA-1 had a dramatic morphological transformation from discocytes to spherocytes within 2 weeks which was accompanied with significant ...

  6. Label-free detection of circulating melanoma cells by in vivo photoacoustic flow cytometry

    Science.gov (United States)

    Wang, Xiaoling; Yang, Ping; Liu, Rongrong; Niu, Zhenyu; Suo, Yuanzhen; He, Hao; Gao, Wenyuan; Tang, Shuo; Wei, Xunbin

    2016-03-01

    Melanoma is a malignant tumor of melanocytes. Melanoma cells have high light absorption due to melanin highly contained in melanoma cells. This property is employed for the detection of circulating melanoma cell by in vivo photoacoustic flow cytometry (PAFC), which is based on photoacoustic effect. Compared to in vivo flow cytometry based on fluorescence, PAFC can employ high melanin content of melanoma cells as endogenous biomarkers to detect circulating melanoma cells in vivo. We have developed in vitro experiments to prove the ability of PAFC system of detecting photoacoustic signals from melanoma cells. For in vivo experiments, we have constructed a model of melanoma tumor bearing mice by inoculating highly metastatic murine melanoma cancer cells, B16F10 with subcutaneous injection. PA signals are detected in the blood vessels of mouse ears in vivo. The raw signal detected from target cells often contains some noise caused by electronic devices, such as background noise and thermal noise. We choose the Wavelet denoising method to effectively distinguish the target signal from background noise. Processing in time domain and frequency domain would be combined to analyze the signal after denoising. This algorithm contains time domain filter and frequency transformation. The frequency spectrum image of the signal contains distinctive features that can be used to analyze the property of target cells or particles. The processing methods have a great potential for analyzing signals accurately and rapidly. By counting circulating melanoma cells termly, we obtain the number variation of circulating melanoma cells as melanoma metastasized. Those results show that PAFC is a noninvasive and label-free method to detect melanoma metastases in blood or lymph circulation.

  7. HaloTag protein-mediated specific labeling of living cells with quantum dots

    International Nuclear Information System (INIS)

    Quantum dots emerge as an attractive alternative to small molecule fluorophores as fluorescent tags for in vivo cell labeling and imaging. This communication presents a method for specific labeling of live cells using quantum dots. The labeling is mediated by HaloTag protein expressed at the cell surface which forms a stable covalent adduct with its ligand (HaloTag ligand). The labeling can be performed in one single step with quantum dot conjugates that are functionalized with HaloTag ligand, or in two steps with biotinylated HaloTag ligand first and followed by streptavidin coated quantum dots. Live cell fluorescence imaging indicates that the labeling is specific and takes place at the cell surface. This HaloTag protein-mediated cell labeling method should facilitate the application of quantum dots for live cell imaging

  8. Aptamer-mediated indirect quantum dot labeling and fluorescent imaging of target proteins in living cells

    International Nuclear Information System (INIS)

    Protein labeling for dynamic living cell imaging plays a significant role in basic biological research, as well as in clinical diagnostics and therapeutics. We have developed a novel strategy in which the dynamic visualization of proteins within living cells is achieved by using aptamers as mediators for indirect protein labeling of quantum dots (QDs). With this strategy, the target protein angiogenin was successfully labeled with fluorescent QDs in a minor intactness model, which was mediated by the aptamer AL6-B. Subsequent living cell imaging analyses indicated that the QDs nanoprobes were selectively bound to human umbilical vein endothelial cells, gradually internalized into the cytoplasm, and mostly localized in the lysosome organelle, indicating that the labeled protein retained high activity. Compared with traditional direct protein labeling methods, the proposed aptamer-mediated strategy is simple, inexpensive, and provides a highly selective, stable, and intact labeling platform that has shown great promise for future biomedical labeling and intracellular protein dynamic analyses. (paper)

  9. Multiplexed labeling system for high-throughput cell sorting.

    Science.gov (United States)

    Shin, Seung Won; Park, Kyung Soo; Song, In Hyun; Shin, Woo Jung; Kim, Byung Woo; Kim, Dong-Ik; Um, Soong Ho

    2016-09-01

    Flow cytometry and fluorescence activated cell sorting techniques were designed to realize configurable classification and separation of target cells. A number of cell phenotypes with different functionalities have recently been revealed. Before simultaneous selective capture of cells, it is desirable to label different samples with the corresponding dyes in a multiplexing manner to allow for a single analysis. However, few methods to obtain multiple fluorescent colors for various cell types have been developed. Even when restricted laser sources are employed, a small number of color codes can be expressed simultaneously. In this study, we demonstrate the ability to manifest DNA nanostructure-based multifluorescent colors formed by a complex of dyes. Highly precise self-assembly of fluorescent dye-conjugated oligonucleotides gives anisotropic DNA nanostructures, Y- and tree-shaped DNA (Y-DNA and T-DNA, respectively), which may be used as platforms for fluorescent codes. As a proof of concept, we have demonstrated seven different fluorescent codes with only two different fluorescent dyes using T-DNA. This method provides maximum efficiency for current flow cytometry. We are confident that this system will provide highly efficient multiplexed fluorescent detection for bioanalysis compared with one-to-one fluorescent correspondence for specific marker detection. PMID:27181032

  10. The effect of varying type and volume of sedimenting agents on leukocyte harvesting and labelling in sickle cell patients

    International Nuclear Information System (INIS)

    Leukocyte labelling in patients with sickle cell anaemia has been reported as difficult if not impossible due to the slow erythrocyte sedimentation rate (ESR) in these patients. This study investigated standard sedimentation methods in patients with sickle cell disease (n=16) and compared the results obtained with those following changes in the amount and type of sedimenting agent used. Labelling with either 111In-oxine or 99Tcm-exametazime was attempted in only five patients. Replacement of the commonly used 6% Hetastarch (Hespan) with Dextran or Haemaccel did not improve leukocyte harvesting, even when the proportions used of these agents were increased. In most cases where standard procedures for leukocyte collection did not lead to harvesting of viable samples, it was possible to collect reasonably pure samples by increasing the proportion of Hespan used. It is possible to obtain adequate leukocyte labelling in the majority of sickle cell patients using a minor modification of standard techniques. In this group of patients a ratio of 8 ml of Hespan to 16 ml of blood should be used for cell separation. If this fails then donor cells, anti-granulocyte antibody labelling or HIG should be considered. (author)

  11. Red Blood Cells Estimation Using Hough Transform Technique

    Directory of Open Access Journals (Sweden)

    Nasrul Humaimi Mahmood

    2012-05-01

    Full Text Available The number of red blood cells contributes more to clinical diagnosis with respect to blood diseases. Theaim of this research is to produce a computer vision system that can detect and estimate the number of redblood cells in the blood sample image. Morphological is a very powerful tool in image processing, and it isbeen used to segment and extract the red blood cells from the background and other cells. The algorithmused features such as shape of red blood cells for counting process, and Hough transform is introduced inthis process. The result presented here is based on images with normal blood cells. The tested data consistsof 10 samples and produced the accurate estimation rate closest to 96% from manual counting.

  12. Improving Cerebral Blood Flow Quantification for Arterial Spin Labeled Perfusion MRI by Removing Residual Motion Artifacts and Global Signal Fluctuations

    OpenAIRE

    Wang, Ze

    2012-01-01

    Denoising is critical to improving the quality and stability of cerebral blood flow (CBF) quantification in arterial spin labeled (ASL) perfusion MRI due to the intrinsic low signal-to-noise-ratio (SNR) of ASL data. Previous studies have been focused on reducing the spatial or temporal noise using standard filtering techniques, and less attention has been paid to two global nuisance effects, the residual motion artifacts and the global signal fluctuations. Since both nuisances affect the whol...

  13. Efficient in vitro labeling rabbit neural stem cell with paramagnetic Gd-DTPA and fluorescent substance

    International Nuclear Information System (INIS)

    Objectives: The aim of this study is to label rabbit neural stem cells (NSCs) by using standard contrast agents (Gd-DTPA) in combination with PKH26 and in vitro track them with MR imaging. Materials and methods: NSCs from prenatal brains of rabbits were cultured and propagated. Intracellular uptake of Gd-DTPA was achieved by using a non-liposomal lipid transfection reagent (Effectene) as the transfection agent. After labeling with Gd-DTPA, cells were incubated with cellular membrane fluorescent dye PKH26. The labeling effectiveness and the longevity of Gd-DTPA maintenance were measured on a 1.5 T MR scanner. The influence of labeling on the cellular biological behaviors was assessed by cellular viability, proliferation and differentiation assessment. Results: The labeling efficiency of Gd-DTPA was up to 90%. The signal intensity on T1-weighted imaging and T1 values of labeled cells were significantly higher than those of unlabeled cells (P 3. Cellular uptake of Gd-DTPA was maintained until 15 days after initially labeling. There was no significant difference in the cellular viability and proliferation between the labeled and unlabeled NSCs (P > 0.05). Normal glial and neuronal differentiation remained in labeled NSCs like unlabeled NSCs. Conclusion: Highly efficient labeling NSCs with Gd-DTPA could be achieved by using Effectene. This method of labeling NSCs allows for tracking cells with MR imaging, and without alterations of cellular biological behaviors.

  14. Superparamagnetic iron oxide nanoparticles for direct labeling of stem cells and in vivo MRI tracking.

    Science.gov (United States)

    Kim, Saejeong J; Lewis, Bobbi; Steiner, Mark-Steven; Bissa, Ursula V; Dose, Christian; Frank, Joseph A

    2016-01-01

    To develop effective stem cell therapies, it is important to track therapeutic cells non-invasively and monitor homing to areas of pathology. The purpose of this study was to design and evaluate the labeling efficiency of commercially available dextran-coated superparamagnetic iron oxide nanoparticles, FeraTrack Direct (FTD), in various stem and immune cells; assess the cytotoxicity and tolerability of the FTD in stem cells; and monitor stem cell homing using FTD-labeled bone-marrow-derived mesenchymal stromal cells (BMSCs) and neural stem cells (NSCs) in a tumor model by in vivo MRI. BMSCs, NSCs, hematopoietic stem cells (HSCs), T-lymphocytes, and monocytes were labeled effectively with FTD without the need for transfection agents, and Prussian blue (PB) staining and transmission electron microscopy (TEM) confirmed intracellular uptake of the agent. The viability, proliferation, and functionality of the labeled cells were minimally or not affected after labeling. When 10(6) FTD-labeled BMSCs or NSCs were injected into C6 glioma bearing nude mice, the cells homing to the tumors were detected as hypointense regions within the tumor using 3 T clinical MRI up to 10 days post injection. Histological analysis confirmed the homing of injected cells to the tumor by the presence of PB positive cells that are not macrophages. Labeling of stem cells or immune cells with FTD was non-toxic, and should facilitate the translation of this agent to clinical trials for evaluation of trafficking of cells by MRI. PMID:26234504

  15. IgG red blood cell autoantibodies in autoimmune hemolytic anemia bind to epitopes on red blood cell membrane band 3 glycoprotein

    International Nuclear Information System (INIS)

    Red blood cell (RBC) autoantibodies from patients with IgG warm-type autoimmune hemolytic anemia were labeled with iodine 125 and their RBC binding behavior characterized. Epitope-bearing RBC membrane polypeptides were identified after autoantibody immunoprecipitation of labeled membranes and immunoblotting. Immunoaffinity isolation of labeled membrane proteins with 12 different IgG hemolytic autoantibodies with protein A-agarose revealed a major polypeptide at Mr 95 to 110 kd, which coelectrophoresed on sodium dodecylsulfate-polyacrylamide gel electrophoresis with a membrane component isolated with sheep IgG anti-band 3. Immunoprecipitation studies with chymotrypsinized RBCs resulted in the recovery of two labeled membrane polypeptides with molecular weights characteristically resulting from the chymotryptic fragmentation of band 3. Immunoblotting with sheep IgG anti-band 3 of the immunoprecipitated polypeptides confirmed that hemolytic autoantibody binding led to recovery of band 3 or its fragments. Two 125I-labeled IgG hemolytic autoantibodies showed binding behavior consistent with epitope localization on band 3. The labeled RBC autoantibodies bound immunospecifically to all types of human RBC tested, including those of rare Rh type (Rh-null, D--) at a site density of approximately 10(6) per RBC. The 125I-IgG in two labeled autoantibodies was 84% and 92% adsorbable by human and higher nonhuman primate RBCs. Antigen-negative animal RBC bound less than 10%, consistent with immunospecific RBC binding. IgG-1 was the major subclass in five autoantibodies tested; one of six fixed complement; and autoantibody IgG appeared polyclonal by isoelectric focusing. We conclude that IgG eluted from RBCs of patients with autoimmune hemolytic anemia consists predominantly of a single totally RBC-adsorbable antibody population that binds to antigenic determinants on band 3

  16. Efficient nano iron particle-labeling and noninvasive MR imaging of mouse bone marrow-derived endothelial progenitor cells

    Directory of Open Access Journals (Sweden)

    Zhen-Yu Jia

    2011-03-01

    efficiently labeled with SPIO and imaged with 7.0-T MRI. They may thus be traced by MRI following transplantation for blood vessel disorders and cancer treatment.Keywords: endothelial progenitor cells, cell labeling, Resovist, magnetic resonance imaging

  17. Phenotype and Functions of Memory Tfh cells in Human Blood

    Science.gov (United States)

    Schmitt, Nathalie; Bentebibel, Salah-Eddine; Ueno, Hideki

    2014-01-01

    Our understanding of the origin and functions of human blood CXCR5+ CD4+ T cells found in human blood has changed dramatically in the past years. These cells are currently considered to represent a circulating memory compartment of T follicular helper (Tfh)-lineage cells. Recent studies have shown that blood memory Tfh cells are composed of phenotypically and functionally distinct subsets. Here we review the current understanding of human blood memory Tfh cells and the subsets within this compartment. We present a strategy to define these subsets based on cell surface profiles. Finally, we discuss how increased understanding of the biology of blood memory Tfh cells may contribute insight into the pathogenesis of autoimmune diseases and the mode of action of vaccines. PMID:24998903

  18. CdSe/ZnS Quantum Dots-Labeled Mesenchymal Stem Cells for Targeted Fluorescence Imaging of Pancreas Tissues and Therapy of Type 1 Diabetic Rats

    Science.gov (United States)

    Liu, Haoqi; Tang, Wei; Li, Chao; Lv, Pinlei; Wang, Zheng; Liu, Yanlei; Zhang, Cunlei; Bao, Yi; Chen, Haiyan; Meng, Xiangying; Song, Yan; Xia, Xiaoling; Pan, Fei; Cui, Daxiang; Shi, Yongquan

    2015-06-01

    Mesenchymal stem cells (MSCs) have been used for therapy of type 1 diabetes mellitus. However, the in vivo distribution and therapeutic effects of transplanted MSCs are not clarified well. Herein, we reported that CdSe/ZnS quantum dots-labeled MSCs were prepared for targeted fluorescence imaging and therapy of pancreas tissues in rat models with type 1 diabetes. CdSe/ZnS quantum dots were synthesized, their biocompatibility was evaluated, and then, the appropriate concentration of quantum dots was selected to label MSCs. CdSe/ZnS quantum dots-labeled MSCs were injected into mouse models with type 1 diabetes via tail vessel and then were observed by using the Bruker In-Vivo F PRO system, and the blood glucose levels were monitored for 8 weeks. Results showed that prepared CdSe/ZnS quantum dots owned good biocompatibility. Significant differences existed in distribution of quantum dots-labeled MSCs between normal control rats and diabetic rats ( p monitored for 8 weeks after quantum dots-labeled MSC injection. Statistical differences existed between the blood glucose levels of the diabetic rat control group and MSC-injected diabetic rat group ( p < 0.01), and the MSC-injected diabetic rat group displayed lower blood glucose levels. In conclusion, CdSe/ZnS-labeled MSCs can target in vivo pancreas tissues in diabetic rats, and significantly reduce the blood glucose levels in diabetic rats, and own potential application in therapy of diabetic patients in the near future.

  19. Cost effectiveness of cord blood versus bone marrow and peripheral blood stem cells

    Directory of Open Access Journals (Sweden)

    Thomas Bart

    2010-10-01

    Full Text Available Thomas BartSwiss Blood Stem Cells, Bern, SwitzerlandAbstract: Umbilical cord blood (CB has become, since its first successful use more than two decades ago, an increasingly important source of blood stem cells. In this light, an overview of current usage of CB in the field of unrelated hematopoietic blood stem cell transplantation (HSCT is given. The three main sources of hematopoietic stem cells: bone marrow (BM, peripheral blood stem cells (PBSC, and cord blood (CB are compared as regards their current quantitative usage in HSCT. A cost analysis of the named three hematopoietic blood stem cell (HSC sources, taking into account various factors, is undertaken. The health economical comparison shows significant differences between CB on the one side, and BM and PBSC on the other. The consequences for the public health side and propositions for a possible health care policy, especially regarding future resource allocation towards the different choices for HSCT products, are discussed. An outlook on the possible future usage of BM, PBSC, and CB and its implications on health systems, donor registries, and CB banks is given.Keywords: health economy, cord blood, hematopoietic stem cell transplantation

  20. Leucocyte filtration of salvaged blood during cardiac surgery : effect on red blood cell function in concentrated blood compared with diluted blood

    NARCIS (Netherlands)

    Gu, Y. John; de Vries, Adrianus J.; Hagenaars, J. Ans M.; van Oeveren, Willem

    2009-01-01

    Objective: Leucocyte filtration of salvaged blood has been suggested to prevent patients from receiving activated leucocytes during autotransfusion in cardiac surgery. This study examines whether leucocyte filtration of salvaged blood affects the red blood cell (RBC) function and whether there is a

  1. Retrograde Labeling of Adult Rat Retinal Ganglion Cells with the Flurogold

    Institute of Scientific and Technical Information of China (English)

    WeiHuang; YannianHui; 等

    2002-01-01

    Purpose:To study the densities and distribution of retinal ganglion cells(RGC) in adult rat retinae with flurogold(FG) labeling retogradely.Methods:FG was injected to the superior colliculid(SC) and dorsal lateral geniculate nuclei(dLGN) in adult rats and the retinae were examined by fluorescence microscopy at various periods of time.Results:FG-labelled RGC were observed in the retina as early as 3 days after application of FG.The labeled cells gradually increased in density,reached 95% of the maximal number on days 7 and the maximal nuber on days 30.The density of labeled cells was higher in the posterior pole than in the peripheral area.The fluorescence intensity in labeled cells maintained up to 60 days.Conclusion:The FG retrograde labeling method is reliable and effective for quantity of RGC.Eye Science 2000;46:29-33.

  2. Bystander apoptosis in human cells mediated by irradiated blood plasma

    Energy Technology Data Exchange (ETDEWEB)

    Vinnikov, Volodymyr, E-mail: vlad.vinnikov@mail.ru [Grigoriev Institute for Medical Radiology of the National Academy of Medical Science of Ukraine (Ukraine); Lloyd, David; Finnon, Paul [Centre for Radiation, Chemical and Environmental Hazards of the Health Protection Agency of the United Kingdom (United Kingdom)

    2012-03-01

    Following exposure to high doses of ionizing radiation, due to an accident or during radiotherapy, bystander signalling poses a potential hazard to unirradiated cells and tissues. This process can be mediated by factors circulating in blood plasma. Thus, we assessed the ability of plasma taken from in vitro irradiated human blood to produce a direct cytotoxic effect, by inducing apoptosis in primary human peripheral blood mononuclear cells (PBM), which mainly comprised G{sub 0}-stage lymphocytes. Plasma was collected from healthy donors' blood irradiated in vitro to 0-40 Gy acute {gamma}-rays. Reporter PBM were separated from unirradiated blood with Histopaque and held in medium with the test plasma for 24 h at 37 Degree-Sign C. Additionally, plasma from in vitro irradiated and unirradiated blood was tested against PBM collected from blood given 4 Gy. Apoptosis in reporter PBM was measured by the Annexin V test using flow cytometry. Plasma collected from unirradiated and irradiated blood did not produce any apoptotic response above the control level in unirradiated reporter PBM. Surprisingly, plasma from irradiated blood caused a dose-dependent reduction of apoptosis in irradiated reporter PBM. The yields of radiation-induced cell death in irradiated reporter PBM (after subtracting the respective values in unirradiated reporter PBM) were 22.2 {+-} 1.8% in plasma-free cultures, 21.6 {+-} 1.1% in cultures treated with plasma from unirradiated blood, 20.2 {+-} 1.4% in cultures with plasma from blood given 2-4 Gy and 16.7 {+-} 3.2% in cultures with plasma from blood given 6-10 Gy. These results suggested that irradiated blood plasma did not cause a radiation-induced bystander cell-killing effect. Instead, a reduction of apoptosis in irradiated reporter cells cultured with irradiated blood plasma has implications concerning oncogenic risk from mutated cells surviving after high dose in vivo irradiation (e.g. radiotherapy) and requires further study.

  3. A photonic crystal hydrogel suspension array for the capture of blood cells from whole blood

    Science.gov (United States)

    Zhang, Bin; Cai, Yunlang; Shang, Luoran; Wang, Huan; Cheng, Yao; Rong, Fei; Gu, Zhongze; Zhao, Yuanjin

    2016-02-01

    Diagnosing hematological disorders based on the separation and detection of cells in the patient's blood is a significant challenge. We have developed a novel barcode particle-based suspension array that can simultaneously capture and detect multiple types of blood cells. The barcode particles are polyacrylamide (PAAm) hydrogel inverse opal microcarriers with characteristic reflection peak codes that remain stable during cell capture on their surfaces. The hydrophilic PAAm hydrogel scaffolds of the barcode particles can entrap various plasma proteins to capture different cells in the blood, with little damage to captured cells.Diagnosing hematological disorders based on the separation and detection of cells in the patient's blood is a significant challenge. We have developed a novel barcode particle-based suspension array that can simultaneously capture and detect multiple types of blood cells. The barcode particles are polyacrylamide (PAAm) hydrogel inverse opal microcarriers with characteristic reflection peak codes that remain stable during cell capture on their surfaces. The hydrophilic PAAm hydrogel scaffolds of the barcode particles can entrap various plasma proteins to capture different cells in the blood, with little damage to captured cells. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr06368j

  4. Stem Cell Transplant (Peripheral Blood, Bone Marrow, and Cord Blood Transplants)

    Science.gov (United States)

    ... are studied in cloning and other types of research. These stem cells are blood-forming stem cells. Stem cells mostly ... Preventing and managing GVHD are major priorities for research. Chronic ... 90 to 600 days after the stem cell transplant. A rash on the palms of the ...

  5. Critical study of the in vitro labelling of circulating blood platelets

    International Nuclear Information System (INIS)

    At present it is possible by chromium-51 labelling in vitro to measure the platelet lifetime in man under normal conditions and in most thrombopenic states. Unfortunately this technique cannot be used for autologous platelet labelling in subjects possessing less than 80.000 platelets per mm3, while isologous labelling cannot distinguish between corpuscular and extracorpuscular anomalies. Moreover in most cases the aspect of the platelet survival curve obtained with 51Cr labelling provides no decisive argument as to the role of senescence and that of 'consumption' in platelet destruction. It seems that the use of indium-111 for the study of platelet survival parameters can supply certain positive elements: - kinetic study of autologous platelets in very low content thrombopenias since the number of platelets put to incubate seems to have little effect on the labelling yield. - Simultaneous isologue-autologue study by 51Cr - 111In double labelling to distinguish between corpuscular and extracorpuscular mechanisms. - Possible static and/or dynamic scintigraphic visualization of thromboses and/or atheromatous lesions with indium-113m - labelled platelets in the hope of obtaining some additional information on platelet 'consumption', at least in pathological cases

  6. Detection of the cancer marker CD146 expression in melanoma cells with semiconductor quantum dot label.

    Science.gov (United States)

    Zheng, Hong; Chen, Guangchun; DeLouise, Lisa A; Lou, Ziyang

    2010-08-01

    The use of highly specific and highly sensitive quantum dots immunofluorescent label is a promising approach for biomedical imaging in cancer cells. Human melanoma cell adhesion molecule CD146, overexpressed on the surface of melanoma cells, is an important target for melanoma diagnostics. We synthesized PEG-COOH capped highly fluorescent CdSe/ZnS QDs and conjugated them with streptavidin to prepare QD-SA label. Then, we used QD-SA to link with biotinylated goat anti-mouse IgG and mouse anti-human CD146 to label CD146 overexpressed on live and fixed cells by FACS and Confocal microscopy. Labeling of target cells was shown to have high brightness, photostability, and specificity. Advantages of QD conjugates over FITC conjugates are discussed. The results indicate that construction based on QD-SA label, biotinylated IgG and CD146 antibody can be successfully used for detection of melanoma cells for biomedical applications. PMID:21323102

  7. Hormones that Stimulate the Growth of Blood Cells.

    Science.gov (United States)

    Golde, David W.; Gasson, Judith C.

    1988-01-01

    Describes the nature and action of hematopoietic proteins which regulate the production of specific sets of blood cells. Discusses the production of these hematopoietins by recombinant-DNA methods in an effort to enable physicians to treat patients by eliciting production of specific types of blood cells. (CW)

  8. Multifactorial aspects of antibody-mediated blood cell destruction

    NARCIS (Netherlands)

    R. Kapur

    2014-01-01

    The research described in this thesis focuses on diseases of antibody-mediated blood cell destruction via FcγRs on phagocytes, in particular regarding platelets in fetal or neonatal alloimmune thrombocytopenia (FNAIT) and red blood cells (RBC) in hemolytic disease of the fetus and newborn (HDFN). Di

  9. Light scattering by aggregated red blood cells

    Science.gov (United States)

    Tsinopoulos, Stephanos V.; Sellountos, Euripides J.; Polyzos, Demosthenes

    2002-03-01

    In low flow rates, red blood cells (RBCs) fasten together along their axis of symmetry and form a so-called rouleaux. The scattering of He-Ne laser light by a rouleau consisting of n (2 less-than-or-equal n less-than-or-equal 8) average-sized RBCs is investigated. The interaction problem is treated numerically by means of an advanced axisymmetric boundary element--fast Fourier transform methodology. The scattering problem of one RBC was solved first, and the results showed that the influence of the RBC's membrane on the scattering patterns is negligible. Thus the rouleau is modeled as an axisymmetric, homogeneous, low-contrast dielectric cylinder, on the surface of which appears, owing to aggregated RBCs, a periodic roughness along the direction of symmetry. The direction of the incident laser light is considered to be perpendicular to the scatterer's axis of symmetry. The differential scattering cross sections in both perpendicular and parallel scattering planes and for all the scattering angles are calculated and presented in detail.

  10. Novel Cell Preservation Technique to Extend Bovine In Vitro White Blood Cell Viability

    OpenAIRE

    Laurin, Emilie L.; McKenna, Shawn L. B.; Sanchez, Javier; Bach, Horacio; Rodriguez-Lecompte, Juan Carlos; Chaffer, Marcelo; Keefe, Greg P

    2015-01-01

    Although cell-mediated immunity based diagnostics can be integral assays for early detection of various diseases of dairy cows, processing of blood samples for these tests is time-sensitive, often within 24 hours of collection, to maintain white blood cell viability. Therefore, to improve utility and practicality of such assays, the objective of this study was to assess the use of a novel white blood cell preservation technology in whole bovine blood. Blood samples from ten healthy cows were ...

  11. Isolation of mesenchymal stem cells from equine umbilical cord blood

    Directory of Open Access Journals (Sweden)

    Thomsen Preben D

    2007-05-01

    Full Text Available Abstract Background There are no published studies on stem cells from equine cord blood although commercial storage of equine cord blood for future autologous stem cell transplantations is available. Mesenchymal stem cells (MSC have been isolated from fresh umbilical cord blood of humans collected non-invasively at the time of birth and from sheep cord blood collected invasively by a surgical intrauterine approach. Mesenchymal stem cells isolation percentage from frozen-thawed human cord blood is low and the future isolation percentage of MSCs from cryopreserved equine cord blood is therefore expectedly low. The hypothesis of this study was that equine MSCs could be isolated from fresh whole equine cord blood. Results Cord blood was collected from 7 foals immediately after foaling. The mononuclear cell fraction was isolated by Ficoll density centrifugation and cultured in a DMEM low glucose based media at 38.5°C in humidified atmosphere containing 5% CO2. In 4 out of 7 samples colonies with MSC morphology were observed. Cellular morphology varied between monolayers of elongated spindle-shaped cells to layered cell clusters of cuboidal cells with shorter cytoplasmic extensions. Positive Alizarin Red and von Kossa staining as well as significant calcium deposition and alkaline phosphatase activity confirmed osteogenesis. Histology and positive Safranin O staining of matrix glycosaminoglycans illustrated chondrogenesis. Oil Red O staining of lipid droplets confirmed adipogenesis. Conclusion We here report, for the first time, the isolation of mesenchymal-like stem cells from fresh equine cord blood and their differentiation into osteocytes, chondrocytes and adipocytes. This novel isolation of equine cord blood MSCs and their preliminary in vitro differentiation positions the horse as the ideal pre-clinical animal model for proof-of-principle studies of cord blood derived MSCs.

  12. Preparation of labelled lipids by the use of plant cell cultures

    International Nuclear Information System (INIS)

    The preparation of some radioacitvely labelled lipids by the use of plant cell cultures is discussed and further applications of the new method are suggested. Cell suspension cultures of rape (Brassica napus) and soya (Glycine max) have been used for the preparation of lipids labelled with radioisotopes. Radioactive acetic acid as well as various long-chain fatty acids are readily incorporated into the neutral and ionic lipids of plant cell cultures. In addition, 14C-labelled glycerol, ethanolamine and choline are well utilized by the cells. Randomly labelled lipids have been obtained by incubating cell suspension cultures of rape and soya with [1-14C] acetic acid, and uniformly labelled lipids have been isolated from cultures that had been incubated with a mixture of [1-14C] acetic acid plus [2-14C] acetic acid. The use of techniques of plant cell cultures for the preparation of lipds labelled with stable or radioactive isotopesappears particularly rewarding because the uptake of precursors by the cells and their incorporation into various lipid compounds proceeds rapidly and often quanitatively.This new approach should be useful also for the biosynthesis of lipids whose acyl moieties contain a spn radical, a fluorescent group, or a light-sensitive label. Thus, plant cell cultures constitute valuable new tools for the biosynthetic preparation of a great variety of labelled lipids. (A.G.)

  13. Blood Thixotropy in Patients with Sickle Cell Anaemia: Role of Haematocrit and Red Blood Cell Rheological Properties

    OpenAIRE

    Vent-Schmidt, Jens; Waltz, Xavier; Romana, Marc; Hardy-Dessources, Marie-Dominique; Lemonne, Nathalie; Billaud, Marie; Etienne-Julan, Maryse; Connes, Philippe

    2014-01-01

    We compared the blood thixotropic/shear-thinning properties and the red blood cells’ (RBC) rheological properties between a group of patients with sickle cell anaemia (SS) and healthy individuals (AA). Blood thixotropy was determined by measuring blood viscosity with a capillary viscometer using a “loop” protocol: the shear rate started at 1 s−1 and increased progressively to 922 s−1 and then re-decreased to the initial shear rate. Measurements were performed at native haematocrit for the two...

  14. A Simulation of Blood Cells in Branching Capillaries

    CERN Document Server

    Isfahani, Amir H G; Freund, Jonathan B

    2008-01-01

    The multi-cellular hydrodynamic interactions play a critical role in the phenomenology of blood flow in the microcirculation. A fast algorithm has been developed to simulate large numbers of cells modeled as elastic thin membranes. For red blood cells, which are the dominant component in blood, the membrane has strong resistance to surface dilatation but is flexible in bending. Our numerical method solves the boundary integral equations built upon Green's functions for Stokes flow in periodic domains. This fluid dynamics video is an example of the capabilities of this model in handling complex geometries with a multitude of different cells. The capillary branch geometries have been modeled based upon observed capillary networks. The diameter of the branches varies between 10-20 mum. A constant mean pressure gradient drives the flow. For the purpose of this fluid dynamics video, the red blood cells are initiated as biconcave discs and white blood cells and platelets are initiated as spheres and ellipsoids resp...

  15. Labeling of human mesenchymal stem cell: Comparison between paramagnetic and superparamagnetic agents

    Science.gov (United States)

    Yang, Chung-Yi; Tai, Ming-Fong; Chen, Shin-Tai; Wang, Yi-Ting; Chen, Ya-Fang; Hsiao, Jong-Kai; Wang, Jaw-Lin; Liu, Hon-Man

    2009-04-01

    Paramagnetic and superparamagnetic substances are used to trace stem cell in living organisms under magnetic resonance imaging (MRI). We compared paramagnetic and superparamagnetic substance for their labeling efficiency by using clinically widely used gadolinium chelates and iron oxide nanoparticles. Without the aid of transfection agent, human mesenchymal stem cells were labeled with each agent separately in different concentration and the optimized concentration was determined by maintaining same cell viability as unlabeled cells. Iron oxide nanoparticle labeling has a detecting threshold of 12 500 cells in vitro, while gadolinium chelates labeling could be detected for at least 50 000 cells. In life animal study, we found there is an eightfold sensitivity in cells labeled with iron oxide superparamagnetic nanoparticles; however, the magnetic susceptibility artifact would obscure the detail of adjacent anatomical structures. We conclude that labeling stem cells with superparamagnetic substance is more efficacious. However, the cells labeled by superparamagnetic nanoparticles might interfere with the interpretation of anatomical structure. These findings would be beneficial to applications of magnetic substances toward stem cell biology and tissue engineering.

  16. Interpretation of automated blood cell counts

    OpenAIRE

    Zühre Kaya

    2013-01-01

    Complete blood count (CBC) tests are rapid, inexpensiveand universally available, and often aid primary clinicianswith decision making about patients with severaldisorders. Thus the rapid availability of the results of CBCcould provide considerable advantage for both patientsand clinicians. Furthermore, physicians can also avoidunnecessary peripheral blood smear examination usingCBC parameters. Many hematology analyzers, which enabledus simultaneously, measure several different CBCparameters,...

  17. Analysis of blood clearance and labeled metabolites for the estrogen receptor tracer [F-18]-16α-fluorestradiol (FES)

    International Nuclear Information System (INIS)

    [F-18] 16α-Fluoroestradiol (FES) has been shown to be a tracer of estrogen receptor content in breast tumors; however, quantitative analysis of FES images is complicated by the rapid metabolism of the tracer in vivo. To optimize FES PET imaging studies and to provide an input function for the quantitative analysis of the tracer FES uptake in breast tumors, we studied the clearance and metabolism of FES in 15 breast cancer patients. FES clearance, protein binding, and metabolite production and limited assays to determine the identity of labeled metabolites were performed. These studies show that FES was rapidly cleared from the blood and metabolized; at 20 min only 20% of the circulating radioactivity was unmetabolized FES, and much of this was protein bound. The detectable metabolites in either blood or urine are conjugation products, largely the glucuronide and the sulfate of FES, and these are excreted through the kidneys at a rate comparable to their introduction into the circulation. After 20 min postinjection the blood levels of radioactivity remain fairly constant. Our results, the first report on human metabolites, are in close agreement with previous animal studies of FES metabolism. These studies show that because FES clearance is rapid and metabolite background is nearly constant, imaging starting at 20 to 30 min after injection may provide good visualization of estrogen-containing tissues. Labeled metabolites need to be accounted for in quantifying FES uptake

  18. Uptake of Retrograde Tracers by Intact Optic Nerve Axons: A New Way to Label Retinal Ganglion Cells

    OpenAIRE

    Liang, Yu-Xiang; Yang, Jian; Yuan, Ti-Fei; So, Kwok-Fai

    2015-01-01

    Retrograde labelling of retinal ganglion cells with optic nerve transection often leads to degeneration of ganglion cells in prolonged experiments. Here we report that an intact optic nerve could uptake retrograde tracers applied onto the surface of the nerve, leading to high efficiency labelling of ganglion cells in the retina with long-term survival of cells. This method labelled a similar number of ganglion cells (2289±174 at 2 days) as the retrograde labeling technique from the superior c...

  19. Effects of broccoli extract on biodistribution and labeling blood components with {sup 99m}Tc-GH

    Energy Technology Data Exchange (ETDEWEB)

    Cekic, Betul; Muftuler, Fazilet Zumrut Biber; Kilcar, Ayfer Yurt; Ichedef, Cigdem; Unak, Perihan [Ege University, Izmir (Turkey). Inst. of Nuclear Sciences. Dept. of Nuclear Applications

    2011-09-15

    Purpose: people consume vegetables without the knowledge of the side effects of the biological and chemical contents and interactions between radiopharmaceuticals and herbal extract. To this end, current study is focused on the effects of broccoli extract on biodistribution of radiolabeled glucoheptonate ({sup 99m}Tc-GH) and radiolabeling of blood components. Methods: GH was labeled with {sup 99m}Tc. Quality control studies were done utilizing TLC method. Biodistribution studies were performed on male rats which were treated via gavage with either broccoli extract or SF as control group for 15 days. Blood samples were withdrawn from rats' heart. Radiolabeling of blood constituents performed incubating with GH, SnCl{sub 2} and {sup 99m} Tc. Results: radiochemical yield of {sup 99m}Tc-GH is 98.46{+-}1.48 % (n=8). Biodistribution studies have shown that according to the control, the treated group with broccoli has approximately 10 times less uptake in kidney. The percentage of the radioactivity ratios of the blood components is found to be same in both groups. Conclusions: although there is no considerable effect on the radiolabeling of blood components, there is an outstanding change on the biodistribution studies especially on kidneys. The knowledge of this change on kidney uptake may contribute to reduce the risk of misdiagnosis and/or repetition of the examinations in Nuclear Medicine. (author)

  20. NMR water-proton spin-lattice relaxation time of human red blood cells and red blood cell suspensions

    International Nuclear Information System (INIS)

    NMR water-proton spin-lattice relaxation times were studied as probes of water structure in human red blood cells and red blood cell suspensions. Normal saline had a relaxation time of about 3000 ms while packed red blood cells had a relaxation time of about 500 ms. The relaxation time of a red blood cell suspension at 50% hematocrit was about 750 ms showing that surface charges and polar groups of the red cell membrane effectively structure extracellular water. Incubation of red cells in hypotonic saline increases relaxation time whereas hypertonic saline decreases relaxation time. Relaxation times varied independently of mean corpuscular volume and mean corpuscular hemoglobin concentration in a sample population. Studies with lysates and resealed membrane ghosts show that hemoglobin is very effective in lowering water-proton relaxation time whereas resealed membrane ghosts in the absence of hemoglobin are less effective than intact red cells. 9 refs.; 3 figs.; 1 table

  1. Deep coverage mouse red blood cell proteome: a first comparison with the human red blood cell

    DEFF Research Database (Denmark)

    Pasini, Erica M; Kirkegaard, Morten; Salerno, Doris;

    2008-01-01

    Mice have close genetic/physiological relationships to humans, breed rapidly, and can be genetically modified, making them the most used mammal in biomedical research. Because the red blood cell (RBC) is the sole gas transporter in vertebrates, diseases of the RBC are frequently severe; much...... identified, and polypeptides migrating with anomalous apparent molecular weights were grouped into putatively ubiquitinated or partially degraded complexes. Overall there was close concordance between mouse and human proteomes, confirming the unexpected RBC complexity. Several novel findings in the human...

  2. Electrostatically Stabilized Magnetic Nanoparticles - An Optimized Protocol to Label Murine T Cells for in vivo MRI.

    Science.gov (United States)

    Wuerfel, Eva; Smyth, Maureen; Millward, Jason M; Schellenberger, Eyk; Glumm, Jana; Prozorovski, Timour; Aktas, Orhan; Schulze-Topphoff, Ulf; Schnorr, Jörg; Wagner, Susanne; Taupitz, Matthias; Infante-Duarte, Carmen; Wuerfel, Jens

    2011-01-01

    We present a novel highly efficient protocol to magnetically label T cells applying electrostatically stabilized very small superparamagnetic iron oxide particles (VSOP). Our long-term aim is to use magnetic resonance imaging (MRI) to investigate T cell dynamics in vivo during the course of neuroinflammatory disorders such as experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. Encephalitogenic T cells were co-incubated with VSOP, or with protamine-complexed VSOP (VProt), respectively, at different conditions, optimizing concentrations and incubation times. Labeling efficacy was determined by atomic absorption spectrometry as well as histologically, and evaluated on a 7 T MR system. Furthermore, we investigated possible alterations of T cell physiology caused by the labeling procedure. T cell co-incubation with VSOP resulted in an efficient cellular iron uptake. T2 times of labeled cells dropped significantly, resulting in prominent hypointensity on T2*-weighted scans. Optimal labeling efficacy was achieved by VProt (1 mM Fe/ml, 8 h incubation; T2 time shortening of ∼80% compared to untreated cells). Although VSOP promoted T cell proliferation and altered the ratio of T cell subpopulations toward a CD4(+) phenotype, no effects on CD4 T cell proliferation or phenotypic stability were observed by labeling in vitro differentiated Th17 cells with VProt. Yet, high concentrations of intracellular iron oxide might induce alterations in T cell function, which should be considered in cell tagging studies. Moreover, we demonstrated that labeling of encephalitogenic T cells did not affect pathogenicity; labeled T cells were still capable of inducing EAE in susceptible recipient mice. PMID:22203815

  3. Therapeutic Potential of Umbilical Cord Blood Stem Cells on Brain Damage of a Model of Stroke

    Directory of Open Access Journals (Sweden)

    Mohammad Reza Nikravesh

    2011-11-01

    Full Text Available Introduction: Human cord blood-derived stem cells are a rich source of stem cells as well as precursors. With regard to the researchers have focused on the therapeutic potential of stem cell in the neurological disease such as stroke, the aim of this study was the investiga-tion of the therapeutic effects of human cord blood-derived stem cells in cerebral ischemia on rat. Methods: This study was carried out on young rats. Firstly, to create a laboratory model of ischemic stroke, carotid artery of animals was occluded for 30 minutes. Then, umbilical cord blood cells were isolated and labeled using bromodeoxyuridine and 2×105 cells were injected into the experimental group via the tail vein. Rats with hypoxic condi-tions were used as a sham group. A group of animals did not receive any injection or sur-geries were used as a control. Results: Obtained results were evaluated based on behavior-al responses and immunohistochemistry, with emphasis on areas of putamen and caudate nucleus in the control, sham and experimental groups. Our results indicated that behavioral recovery was observed in the experimental group compared to the either the sham or the control group. However, histological studies demonstrated a low percent of tissue injury in the experimental group in comparison with the sham group. Conclusion: Stem cell trans-plantation is beneficial for the brain tissue reparation after hypoxic ischemic cell death.

  4. CdSe/ZnS Quantum Dots-Labeled Mesenchymal Stem Cells for Targeted Fluorescence Imaging of Pancreas Tissues and Therapy of Type 1 Diabetic Rats

    Science.gov (United States)

    Liu, Haoqi; Tang, Wei; Li, Chao; Lv, Pinlei; Wang, Zheng; Liu, Yanlei; Zhang, Cunlei; Bao, Yi; Chen, Haiyan; Meng, Xiangying; Song, Yan; Xia, Xiaoling; Pan, Fei; Cui, Daxiang; Shi, Yongquan

    2015-06-01

    Mesenchymal stem cells (MSCs) have been used for therapy of type 1 diabetes mellitus. However, the in vivo distribution and therapeutic effects of transplanted MSCs are not clarified well. Herein, we reported that CdSe/ZnS quantum dots-labeled MSCs were prepared for targeted fluorescence imaging and therapy of pancreas tissues in rat models with type 1 diabetes. CdSe/ZnS quantum dots were synthesized, their biocompatibility was evaluated, and then, the appropriate concentration of quantum dots was selected to label MSCs. CdSe/ZnS quantum dots-labeled MSCs were injected into mouse models with type 1 diabetes via tail vessel and then were observed by using the Bruker In-Vivo F PRO system, and the blood glucose levels were monitored for 8 weeks. Results showed that prepared CdSe/ZnS quantum dots owned good biocompatibility. Significant differences existed in distribution of quantum dots-labeled MSCs between normal control rats and diabetic rats ( p pancreas of rats in the diabetes group, and was about 32 %, while that in the normal control group rats was about 18 %. The blood glucose levels were also monitored for 8 weeks after quantum dots-labeled MSC injection. Statistical differences existed between the blood glucose levels of the diabetic rat control group and MSC-injected diabetic rat group ( p pancreas tissues in diabetic rats, and significantly reduce the blood glucose levels in diabetic rats, and own potential application in therapy of diabetic patients in the near future.

  5. Quantification of depletion-induced adhesion of Red Blood Cells

    OpenAIRE

    Steffen, Patrick; Verdier, Claude; Wagner, Christian

    2013-01-01

    Red blood cells (RBC) are known to form aggregates in the forms of rouleaux due to the presence of plasma proteins under physiological conditions. Rouleaux formation can be also induced in vitro by the addition of macromolecules to the RBC solution. Current data on the adhesion strength between red blood cells in their natural discocyte shapes mostly rely on indirect measurements like flow chamber experiments, but on the single cell level data is lacking. Here we present measurements on the d...

  6. Interpretation of automated blood cell counts

    Directory of Open Access Journals (Sweden)

    Zühre Kaya

    2013-09-01

    Full Text Available Complete blood count (CBC tests are rapid, inexpensiveand universally available, and often aid primary clinicianswith decision making about patients with severaldisorders. Thus the rapid availability of the results of CBCcould provide considerable advantage for both patientsand clinicians. Furthermore, physicians can also avoidunnecessary peripheral blood smear examination usingCBC parameters. Many hematology analyzers, which enabledus simultaneously, measure several different CBCparameters, are available for early diagnosis. Herein theimpact of both pre and post analytic variations on the interpretationof the CBC results with case reports are reviewedin the light of the latest literature.Key words: Complete blood count, interpretation

  7. An enzyme-linked immunoabsorbent assay for estimating red cell survival of transfused red cells-validation using CR-51 labeling

    International Nuclear Information System (INIS)

    The survival time of transfused red cells antigenically distinct from the recipient's red cells was determined using an indirect enzyme linked antiglobulin test. These results were then compared to those determined by Cr-51 labeling. Three patients with hypoproliferative anemias and one patient (2 studies) with traumatic hemolytic anemia caused by a prosthetic heart valve were studied. Survival times were performed by transfusing a 5cc aliquot of Cr-51 labeled cells along with the remaining unit. One hour post transfusion, a blood sample was drawn and used as the 100% value. Subsequent samples drawn over a 2-3 week period were then compared to the initial sample to determine percent survival for both methods. The ELISA method for measuring red cell survival in antigenically distinct cells is in close agreement with the Cr-51 method. Although CR-51 labeling is the accepted method for red cell survival determination the ELISA method can be used when radioisotopes are unavailable or contraindicated or when the decision to estimate red cell survival is made after transfusion

  8. Labeling of mesenchymal stem cells for MRI with single-cell sensitivity.

    Science.gov (United States)

    Ariza de Schellenberger, Angela; Kratz, Harald; Farr, Tracy D; Löwa, Norbert; Hauptmann, Ralf; Wagner, Susanne; Taupitz, Matthias; Schnorr, Jörg; Schellenberger, Eyk A

    2016-01-01

    Sensitive cell detection by magnetic resonance imaging (MRI) is an important tool for the development of cell therapies. However, clinically approved contrast agents that allow single-cell detection are currently not available. Therefore, we compared very small iron oxide nanoparticles (VSOP) and new multicore carboxymethyl dextran-coated iron oxide nanoparticles (multicore particles, MCP) designed by our department for magnetic particle imaging (MPI) with discontinued Resovist(®) regarding their suitability for detection of single mesenchymal stem cells (MSC) by MRI. We achieved an average intracellular nanoparticle (NP) load of >10 pg Fe per cell without the use of transfection agents. NP loading did not lead to significantly different results in proliferation, colony formation, and multilineage in vitro differentiation assays in comparison to controls. MRI allowed single-cell detection using VSOP, MCP, and Resovist(®) in conjunction with high-resolution T2*-weighted imaging at 7 T with postprocessing of phase images in agarose cell phantoms and in vivo after delivery of 2,000 NP-labeled MSC into mouse brains via the left carotid artery. With optimized labeling conditions, a detection rate of ~45% was achieved; however, the experiments were limited by nonhomogeneous NP loading of the MSC population. Attempts should be made to achieve better cell separation for homogeneous NP loading and to thus improve NP-uptake-dependent biocompatibility studies and cell detection by MRI and future MPI. Additionally, using a 7 T MR imager equipped with a cryocoil resulted in approximately two times higher detection. In conclusion, we established labeling conditions for new high-relaxivity MCP, VSOP, and Resovist(®) for improved MRI of MSC with single-cell sensitivity. PMID:27110112

  9. Efficient in vitro labeling rabbit neural stem cell with paramagnetic Gd-DTPA and fluorescent substance

    Energy Technology Data Exchange (ETDEWEB)

    Shen Jun, E-mail: junshenjun@hotmail.co [Department of Radiology, Second Affiliated Hospital, Sun Yat-sen University, 107 Yanjiang Road West, Guangzhou 510120, Guangdong (China); Cheng Lina; Zhong Xiaomei; Duan Xiaohui; Guo Ruomi; Hong Guobing [Department of Radiology, Second Affiliated Hospital, Sun Yat-sen University, 107 Yanjiang Road West, Guangzhou 510120, Guangdong (China)

    2010-09-15

    Objectives: The aim of this study is to label rabbit neural stem cells (NSCs) by using standard contrast agents (Gd-DTPA) in combination with PKH26 and in vitro track them with MR imaging. Materials and methods: NSCs from prenatal brains of rabbits were cultured and propagated. Intracellular uptake of Gd-DTPA was achieved by using a non-liposomal lipid transfection reagent (Effectene) as the transfection agent. After labeling with Gd-DTPA, cells were incubated with cellular membrane fluorescent dye PKH26. The labeling effectiveness and the longevity of Gd-DTPA maintenance were measured on a 1.5 T MR scanner. The influence of labeling on the cellular biological behaviors was assessed by cellular viability, proliferation and differentiation assessment. Results: The labeling efficiency of Gd-DTPA was up to 90%. The signal intensity on T1-weighted imaging and T1 values of labeled cells were significantly higher than those of unlabeled cells (P < 0.05). The minimal number of detectable cells for T1-weighted imaging was 5 x 10{sup 3}. Cellular uptake of Gd-DTPA was maintained until 15 days after initially labeling. There was no significant difference in the cellular viability and proliferation between the labeled and unlabeled NSCs (P > 0.05). Normal glial and neuronal differentiation remained in labeled NSCs like unlabeled NSCs. Conclusion: Highly efficient labeling NSCs with Gd-DTPA could be achieved by using Effectene. This method of labeling NSCs allows for tracking cells with MR imaging, and without alterations of cellular biological behaviors.

  10. MR imaging features of gadofluorine-labeled matrix-associated stem cell implants in cartilage defects.

    Directory of Open Access Journals (Sweden)

    Hossein Nejadnik

    Full Text Available OBJECTIVES: The purpose of our study was to assess the chondrogenic potential and the MR signal effects of GadofluorineM-Cy labeled matrix associated stem cell implants (MASI in pig knee specimen. MATERIALS AND METHODS: Human mesenchymal stem cells (hMSCs were labeled with the micelle-based contrast agent GadofluorineM-Cy. Ferucarbotran-labeled hMSCs, non-labeled hMSCs and scaffold only served as controls. Chondrogenic differentiation was induced and gene expression and histologic evaluation were performed. The proportions of spindle-shaped vs. round cells of chondrogenic pellets were compared between experimental groups using the Fisher's exact test. Labeled and unlabeled hMSCs and chondrocytes in scaffolds were implanted into cartilage defects of porcine femoral condyles and underwent MR imaging with T1- and T2-weighted SE and GE sequences. Contrast-to-noise ratios (CNR between implants and adjacent cartilage were determined and analyzed for significant differences between different experimental groups using the Kruskal-Wallis test. Significance was assigned for p0.017. However, hMSC differentiation into chondrocytes was superior for unlabeled and GadofluorineM-Cy-labeled cells compared with Ferucarbotran-labeled cells, as evidenced by a significantly higher proportion of spindle cells in chondrogenic pellets (p<0.05. GadofluorineM-Cy-labeled hMSCs and chondrocytes showed a positive signal effect on T1-weighted images and a negative signal effect on T2-weighted images while Ferucarbotran-labeled cells provided a negative signal effect on all sequences. CNR data for both GadofluorineM-Cy-labeled and Ferucarbotran-labeled hMSCs were significantly different compared to unlabeled control cells on T1-weighted SE and T2*-weighted MR images (p<0.017. CONCLUSION: hMSCs can be labeled by simple incubation with GadofluorineM-Cy. The labeled cells provide significant MR signal effects and less impaired chondrogenesis compared to Ferucarbotran-labeled h

  11. Modified procedure for labelling target cells in a europium release assay of natural killer cell activity.

    Science.gov (United States)

    Pacifici, R; Di Carlo, S; Bacosi, A; Altieri, I; Pichini, S; Zuccaro, P

    1993-05-01

    Lanthanide europium chelated to diethylenetriaminopentaacetate (EuDTPA) can be used to label target cells such as tumor cells and lymphocytes (Blomberg et al., 1986a,b; Granberg et al., 1988). This procedure has permitted the development of new non-radioactive methods for the detection of target cell cytolysis by natural killer (NK) cells (Blomberg et al., 1986a,b), cytotoxic T lymphocytes (CTL) (Granberg et al., 1988) or complement-mediated cytolysis (Cui et al., 1992). However, we had no success with this method because of a lack of comparability between human NK cell activity simultaneously measured by a classical 51Cr release assay (Seaman et al., 1981) and EuDTPA release assay (Blomberg et al., 1986a). Furthermore, cell division and cell viability were significantly impaired by the suggested concentrations of EuCl3. In this paper, we present a modified non-cytotoxic method for target cell labelling with EuDTPA while cells are growing in culture medium. PMID:8486925

  12. Net haemoglobin increase from reinfusion of refrigerated vs. frozen red blood cells after autologous blood transfusions

    DEFF Research Database (Denmark)

    Ashenden, M; Mørkeberg, Jakob Sehested

    2011-01-01

    freezing. Nevertheless, frozen storage allowed haemoglobin to fully recover before reinfusion, while the haemoglobin was 10% lower in the refrigerated group compared with baseline. After reinfusion, the haemoglobin levels were 11·5% higher than the baseline values in the group reinfused with frozen blood......BACKGROUND AND OBJECTIVES  Two main blood storage procedures can be used for storing red blood cells: refrigeration and freezing. Nevertheless, the efficiency of these procedures measured as the increase in haemoglobin after reinfusion compared with baseline has never been examined. The main...... objective was to examine which storage procedure yielded the largest increase in circulating haemoglobin after reinfusion compared to baseline. MATERIALS AND METHODS  Equal volumes of blood from 15 men were withdrawn and stored either frozen or refrigerated as packed red blood cells. Serial measures...

  13. Variability of the thymidine labeling index in squamous cell carcinoma of the head and neck

    International Nuclear Information System (INIS)

    Tritiated thymidine (3HTdR) labeling is the standard technique for determining the kinetic activity of tumors. This method has been used to label multiple sections of tumor specimens obtained from seven patients with advanced squamous cell carcinoma of the head and neck. Considerable variability was observed in the labeling index in different sites from the same specimen. To reduce the large sampling error due to heterogeneity, we recommend that an average value be determined from multiple sections when employing this technique

  14. Quantification of Depletion-Induced Adhesion of Red Blood Cells

    Science.gov (United States)

    Steffen, P.; Verdier, C.; Wagner, C.

    2013-01-01

    Red blood cells (RBCs) are known to form aggregates in the form of rouleaux due to the presence of plasma proteins under physiological conditions. The formation of rouleaux can also be induced in vitro by the addition of macromolecules to the RBC suspension. Current data on the adhesion strength between red blood cells in their natural discocyte shapes mostly originate from indirect measurements such as flow chamber experiments, but data is lacking at the single cell level. Here, we present measurements on the dextran-induced aggregation of red blood cells using atomic force microscopy-based single cell force spectroscopy. The effects of dextran concentration and molecular weight on the interaction energy of adhering RBCs were determined. The results on adhesion energy are in excellent agreement with a model based on the depletion effect and previous experimental studies. Furthermore, our method allowed to determine the adhesion force, a quantity that is needed in theoretical investigations on blood flow.

  15. Vesicle-associated microRNAs are released from blood cells on incubation of blood samples.

    Science.gov (United States)

    Köberle, Verena; Kakoschky, Bianca; Ibrahim, Ahmed Atef; Schmithals, Christian; Peveling-Oberhag, Jan; Zeuzem, Stefan; Kronenberger, Bernd; Waidmann, Oliver; Pleli, Thomas; Piiper, Albrecht

    2016-03-01

    MicroRNAs (miRNAs) circulating extracellularly in the blood are currently intensively studied as novel disease markers. However, the preanalytical factors influencing the levels of the extracellular miRNAs are still incompletely explored. In particular, it is unknown, whether the incubation of blood samples as occurring in clinical routine can lead to a release of miRNAs from blood cells and thus alter the extracellular miRNA levels before the preparation of serum or plasma from the blood cells. Using a set of marker miRNAs and quantitative RT-PCR, we found that the levels of extracellular miRNA-1, miRNA-16, and miRNA-21 were increased in EDTA and serum collection tubes incubated for 1-3 hours at room temperature and declined thereafter; the levels of the liver-specific miRNA-122 declined monophasically. These events occurred in the absence of significant hemolysis. When the blood was supplemented with Ribonuclease A inhibitor, the levels of miRNA-1, miRNA-16, and miRNA-21 increased substantially during the initial 3 hours of incubation and those of miRNA-122 remained unchanged, indicating that the release of blood cell-derived miRNAs occurred during the initial 3 hours of incubation of the blood tubes, but not at later time points. Separation of 5-hour preincubated blood into vesicle and nonvesicle fractions revealed a selective increase in the portion of vesicle-associated miRNAs. Together, these data indicate that the release of vesicle-associated miRNAs from blood cells can occur in blood samples within the time elapsing in normal clinical practice until their processing without significant hemolysis. This becomes particularly visible on the inhibition of miRNA degradation by Ribonuclease A inhibitor. PMID:26608461

  16. Simultaneous measurement of NK cell cytotoxicity against two target cell lines labelled with fluorescent lanthanide chelates.

    Science.gov (United States)

    Lövgren, J; Blomberg, K

    1994-07-12

    We describe a cytotoxicity assay which permits the simultaneous measurement of natural killer cell activity against two different cell lines. The target cell lines are labelled either with a fluorescent europium chelate or with a fluorescent terbium chelate and cell death is quantified by measuring the chelate release. K-562, Molt4 and Daudi cell lines have been used as targets. The release of the two chelates from the target cells can be detected with the help of time resolved fluorometry. As the measurements are made after background fluorescence has decayed no additional steps are needed to correct for the background from the medium. The assay procedure used for measurement of cytotoxicity against two target cell lines is very similar to the widely used 51Cr release assay. PMID:8034979

  17. WHITE BLOOD CELLS IN POLISH ATHLETES OF VARIOUS SPORTS DISCIPLINES

    OpenAIRE

    Joanna Orysiak; Konrad Witek; Piotr Zmijewski; Jan Gajewski

    2012-01-01

    The purpose of this study was to examine the diversity of white blood cell (WBC) counts and their subsets (neutrophils, lymphocytes and monocytes) among competitive athletes of different sports disciplines. The blood samples were collected from 608 healthy, medically examined athletes (181 females and 427 males) aged 20.1 ± 5.1 years, who represented five sport disciplines: canoeing, judo, rowing, swimming and volleyball. All blood samples were taken from the antecubital vein in the morning, ...

  18. Transfusion management of patients with red blood cell antibodies

    OpenAIRE

    Bujandrić Nevenka B.; Grujić Jasmina N.; Krga-Milanović Mirjana M.

    2013-01-01

    Introduction. Red blood cell antibodies may cause a positive result of pre-transfusion blood compatibility testing (crossmatch test). It can be a problem to provide suitable blood units for patients with clinically significant antibodies to high-frequency antigens as well as for those with multiple alloantibody specificities. This study was aimed at identifying transfused patients in the population of South-Backa who had developed clinically significant red...

  19. The effect of 51Cr-labelling on cell morphology, in vitro, when evaluating the cytotoxicity of endodontic filling materials

    International Nuclear Information System (INIS)

    Human periodontal ligament fibroblasts and L 929 cell line labelled with chromium 51 were examined byelectron microscope to evaluate the effect of the chromium labeling on the cell ultrastructure. The cells were labeled with chromium 12-20 hours before the start of the experiment. After two and four hours of incubation at 37 degree C and 100% humidity, the cells were examined by scanning and transmission electron microscopy. The result showed that the chromium labeling did not cause any morphological changes. (author)

  20. Monoclonal antibodies and coupling reagents to cell membrane proteins for leukocyte labeling

    International Nuclear Information System (INIS)

    Current gamma-emitting agents for tagging leukocytes, In-111 oxine or tropolone, label all cell types indiscriminantly, and nuclear localization in lymphocytes results in radiation damage. Coupling reagents and murine monoclonal antibodies (Mab) specific for cell surface antigens of human leukocytes were tried as cell labeling agents to avoid nuclear localization. 10/sup 8/ mixed human leukocytes in Hepes buffer were added to tubes coated with 5 mg of dry cyclic dianhydride of DTPA for 15 minutes at room temperature. After washing, 0.1 ml of In-111 Cl in ACD (pH 6.8) was added. After 30 minutes, a cell labeling yield of 23% was obtained. Washing the cells in an elutriation centrifuge showed that this label was irreversible. Mab for cell surface antigens of human granulocytes were labeled with 300 μCi of I-125 using the Iodobead technic and unbound activity was removed by gel column chromatography. 1-10 μg were added to 10/sup 8/ mixed leukocytes in 0.5 ml plasma or saline for 1 hr. With Mab anti-leu M4 (clone G7 E11), an IgM, the cell labeling yield was 21%, irreversible, and specific for granulocytes. With anti-human leukocyte Mab NEI-042 (clone 9.4), and IgG2a, and anti-granulocyte Mab MAS-065 (clone FMCl1) an IgG1, the cell labeling was relatively unstable. Labeling of leukocyte subpopulations with Mab is feasible, and the binding of multivalent IgM is stronger than that of other immunoglobulins. DTPA cyclic anhydride is firmly bound to cell membranes, but the labeling is non-specific

  1. Flow cytometric measurement of RNA synthesis based on bromouridine labelling and combined with measurement of DNA content or cell surface antigen

    DEFF Research Database (Denmark)

    Jensen, P O; Larsen, J; Larsen, J K

    1993-01-01

    RNA synthesis can be analysed in nuclei or cells labelled with 5-bromouridine (BrUrd) and stained using cross-reacting anti-bromodeoxyuridine (BrdUrd) antibody. Flow cytometric dual parameter analysis of BrUrd incorporation and DNA content in nuclear suspensions of human blood lymphocytes showed ...... synthesis in HL-60 and K-562 cells was measured simultaneous with CD13 expression....

  2. Differentiation of cytotoxicity using target cells labelled with europium and samarium by electroporation.

    Science.gov (United States)

    Bohlen, H; Manzke, O; Engert, A; Hertel, M; Hippler-Altenburg, R; Diehl, V; Tesch, H

    1994-07-12

    We report the simultaneous use of europium-DTPA (Eu-DTPA) and samarium-DTPA (Sm-DTPA) in cytotoxicity experiments to analyze simultaneously LAK and NK cell lysis and to differentiate between specific target lysis and bystander killing. The target cells were either labelled with Eu-DTPA or Sm-DTPA chelates by electroporation, which permits the use of target cell lines or primary leukemic B cells (B-CLL) that cannot be labelled by the conventional dextran-sulphate method. The release of europium and samarium reaches a maximum at comparable time intervals (2-3 h). Due to the shorter counting interval within the samarium window the labelling efficiency is about ten times less efficient compared to europium. Using europium as label for the LAK target Daudi and samarium as label for the NK sensitive cell line K562 the differentiation of LAK versus NK activity can be performed in a single culture assay. Also, the killing of B cells and bystander cells by cytotoxic T cells was analyzed in a system where T cells were redirected to B cells through CD3 x CD19 bispecific antibodies. In fact, no bystander killing was noted when bispecific antibodies were used to bridge cytotoxic T cells to the B cells. This approach provides a simple non-radioactive method for evaluating cytotoxicity against two different cells in a single culture well. PMID:8034986

  3. Impaired myocardial blood flow reserve in subjects with metabolic syndrome analyzed using positron emission tomography and N-13 labeled ammonia

    International Nuclear Information System (INIS)

    Coronary vasomotor response might be impaired in metabolic syndrome (MS); however, the precise abnormality has not been elucidated. The aim of this study was to assess coronary-vasomotor response in MS subjects using N-13 labeled ammonia and positron emission tomography. Myocardial blood flow (MBF) was measured at rest and during adenosine infusion in MS subjects (n = 13, MS group) with no definite evidence of heart disease and in subjects without MS (n = 14, non-MS group). Coronary vascular resistance (CVR) was calculated by dividing the mean aortic blood pressure by MBF. Myocardial blood flow reserve (MFR) was calculated as the ratio of the MBF during adenosine infusion to that during rest. Blood chemical parameters were measured to evaluate their relationship with MFR. During adenosine infusion, MBF was lower (p = 0.0085) and CVR higher (p = 0.0128) in the MS group than in the non-MS group and MFR was significantly lower in the MS group than in the non-MS group (2.13 ± 0.99 vs. 3.38 ± 0.95, p = 0.0027). Multivariate analysis demonstrated that the homeostasis model assessment-insulin resistance (p < 0.05) and the presence of hypertension (p < 0.05) were independent determinants of MFR. The results indicate that MFR was impaired in MS subjects, suggesting that an abnormal coronary microvascular response occurred in these subjects. This abnormality may have been partially due to insulin resistance and hypertension. (orig.)

  4. Impaired myocardial blood flow reserve in subjects with metabolic syndrome analyzed using positron emission tomography and N-13 labeled ammonia

    Energy Technology Data Exchange (ETDEWEB)

    Teragawa, Hiroki; Kihara, Yasuki [Hiroshima University Graduate School of Biomedical Sciences, Department of Cardiovascular Medicine, Hiroshima (Japan); Morita, Koichi; Tamaki, Nagara [Hokkaido University Graduate School of Medicine, Department of Nuclear Medicine, Sapporo (Japan); Shishido, Hiroki; Otsuka, Nobuaki; Hirokawa, Yutaka [Hiroshima Heiwa Clinic, Hiroshima (Japan); Chayama, Kazuaki [Hiroshima University Graduate School of Biomedical Sciences, Department of Molecular Science and Medicine, Hiroshima (Japan)

    2010-02-15

    Coronary vasomotor response might be impaired in metabolic syndrome (MS); however, the precise abnormality has not been elucidated. The aim of this study was to assess coronary-vasomotor response in MS subjects using N-13 labeled ammonia and positron emission tomography. Myocardial blood flow (MBF) was measured at rest and during adenosine infusion in MS subjects (n = 13, MS group) with no definite evidence of heart disease and in subjects without MS (n = 14, non-MS group). Coronary vascular resistance (CVR) was calculated by dividing the mean aortic blood pressure by MBF. Myocardial blood flow reserve (MFR) was calculated as the ratio of the MBF during adenosine infusion to that during rest. Blood chemical parameters were measured to evaluate their relationship with MFR. During adenosine infusion, MBF was lower (p = 0.0085) and CVR higher (p = 0.0128) in the MS group than in the non-MS group and MFR was significantly lower in the MS group than in the non-MS group (2.13 {+-} 0.99 vs. 3.38 {+-} 0.95, p = 0.0027). Multivariate analysis demonstrated that the homeostasis model assessment-insulin resistance (p < 0.05) and the presence of hypertension (p < 0.05) were independent determinants of MFR. The results indicate that MFR was impaired in MS subjects, suggesting that an abnormal coronary microvascular response occurred in these subjects. This abnormality may have been partially due to insulin resistance and hypertension. (orig.)

  5. 111Indium labeling of hepatocytes for analysis of short-term biodistribution of transplanted cells.

    Science.gov (United States)

    Gupta, S; Lee, C D; Vemuru, R P; Bhargava, K K

    1994-03-01

    Hepatocyte transplantation is useful for ex vivo gene therapy and liver repopulation. Methods for hepatic reconstitution have recently been developed but optimization of hepatocyte transplantation systems is necessary. To develop systems for noninvasive assessment of the biodistribution of transplanted cells, we labeled hepatocytes with 111indium-oxine. Our initial studies showed that hepatocytes incorporated 111indium-oxine with an efficiency of approximately 20%. After labeling, cell viability was unchanged and 111indium was present in hepatocytes after overnight culture, as well as after intrasplenic transplantation. Transplanted cells were successfully localized by means of scintigraphic imaging. The scintigraphic patterns of cell distribution were different when hepatocytes were transplanted by means of either spleen or internal jugular vein, which deposit cells into separate vascular beds. Quantitative analysis of the biodistribution of 111indium-labeled hepatocytes indicated that within 2 hr of intrasplenic transplantation, cells were predominantly localized in liver and spleen, and occasionally in lungs. To determine whether the rate of intrasplenic cell injection influenced translocation of hepatocytes, we transplanted cells in normal rats. Despite intrasplenic cell injection at a variety of rates, organ-specific distribution of 111indium-labeled hepatocytes remained unchanged. Labeling with 111indium did not affect long-term survival of transplanted hepatocytes. These results indicate that 111indium-labeling of hepatocytes should greatly assist noninvasive analysis in the short-term of the biodistribution of transplanted hepatocytes. PMID:8119703

  6. GABAergic and glycinergic pathways to goldfish retinal ganglion cells: an ultrastructural double label study

    International Nuclear Information System (INIS)

    An ultrastructural double label has been employed to compare GABAergic and glycinergic systems in the inner plexiform layer (IPL) of the goldfish retina. Electron microscope autoradiography of 3H-GABA and 3H-glycine uptake was combined with retrograde HRP-labeling of ganglion cells. When surveyed for distribution, GABAergic and glycinergic synapses were found onto labeled ganglion cells throughout the IPL. This reinforces previous physiological work that described GABAergic and glycinergic influences on a variety of ganglion cells in goldfish and carp; These physiological effects often reflect direct inputs

  7. Nanodiamonds with silicon vacancy defects for non-toxic photostable fluorescent labeling of neural precursor cells

    CERN Document Server

    Merson, Tobias D; Aharonovich, Igor; Turbic, Alisa; Kilpatrick, Trevor J; Turnley, Ann M

    2013-01-01

    Nanodiamonds (NDs) containing silicon vacancy (SiV) defects were evaluated as a potential biomarker for the labeling and fluorescent imaging of neural precursor cells (NPCs). SiV-containing NDs were synthesized using chemical vapor deposition and silicon ion implantation. Spectrally, SiV-containing NDs exhibited extremely stable fluorescence and narrow bandwidth emission with an excellent signal to noise ratio exceeding that of NDs containing nitrogen-vacancy (NV) centers. NPCs labeled with NDs exhibited normal cell viability and proliferative properties consistent with biocompatibility. We conclude that SiVcontaining NDs are a promising biomedical research tool for cellular labeling and optical imaging in stem cell research.

  8. Indium-111 oxine labelling affects the cellular integrity of haematopoietic progenitor cells

    Energy Technology Data Exchange (ETDEWEB)

    Nowak, Bernd; Reinartz, Patrick; Schaefer, Wolfgang M.; Buell, Ulrich [University Hospital, RWTH Aachen University, Department of Nuclear Medicine, Aachen (Germany); Weber, Christian; Schober, Andreas; Zeiffer, Ute; Liehn, Elisa A.; Hundelshausen, Philipp von [University Hospital, RWTH Aachen University, Department of Molecular Cardiovascular Research, Aachen (Germany)

    2007-05-15

    Cell-based therapy by transplantation of progenitor cells has emerged as a promising development for organ repair, but non-invasive imaging approaches are required to monitor the fate of transplanted cells. Radioactive labelling with {sup 111}In-oxine has been used in preclinical trials. This study aimed to validate {sup 111}In-oxine labelling and subsequent in vivo and ex vivo detection of haematopoietic progenitor cells. Murine haematopoietic progenitor cells (10{sup 6}, FDCPmix) were labelled with 0.1 MBq (low dose) or 1.0 MBq (high dose) {sup 111}In-oxine and compared with unlabelled controls. Cellular retention of {sup 111}In, viability and proliferation were determined up to 48 h after labelling. Labelled cells were injected into the cavity of the left or right cardiac ventricle in mice. Scintigraphic images were acquired 24 h later. Organ samples were harvested to determine the tissue-specific activity. Labelling efficiency was 75 {+-} 14%. Cellular retention of incorporated {sup 111}In after 48 h was 18 {+-} 4%. Percentage viability after 48 h was 90 {+-} 1% (control), 58 {+-} 7% (low dose) and 48 {+-} 8% (high dose) (p<0.0001). Numbers of viable cells after 48 h (normalised to 0 h) were 249 {+-} 51% (control), 42 {+-} 8% (low dose) and 32 {+-} 5% (high dose) (p<0.0001). Cells accumulated in the spleen (86.6 {+-} 27.0% ID/g), bone marrow (59.1 {+-} 16.1% ID/g) and liver (30.3 {+-} 9.5% ID/g) after left ventricular injection, whereas most of the cells were detected in the lungs (42.4 {+-} 21.8% ID/g) after right ventricular injection. Radiolabelling of haematopoietic progenitor cells with {sup 111}In-oxine is feasible, with high labelling efficiency but restricted stability. The integrity of labelled cells is significantly affected, with substantially reduced viability and proliferation and limited migration after systemic transfusion. (orig.)

  9. Cell Surface Proteome of Dental Pulp Stem Cells Identified by Label-Free Mass Spectrometry

    Science.gov (United States)

    Niehage, Christian; Karbanová, Jana; Steenblock, Charlotte

    2016-01-01

    Multipotent mesenchymal stromal cells (MSCs) are promising tools for regenerative medicine. They can be isolated from different sources based on their plastic-adherence property. The identification of reliable cell surface markers thus becomes the Holy Grail for their prospective isolation. Here, we determine the cell surface proteomes of human dental pulp-derived MSCs isolated from single donors after culture expansion in low (2%) or high (10%) serum-containing media. Cell surface proteins were tagged on intact cells using cell impermeable, cleavable sulfo-NHS-SS-biotin, which allows their enrichment by streptavidin pull-down. For the proteomic analyses, we first compared label-free methods to analyze cell surface proteomes i.e. composition, enrichment and proteomic differences, and we developed a new mathematical model to determine cell surface protein enrichment using a combinatorial gene ontology query. Using this workflow, we identified 101 cluster of differentiation (CD) markers and 286 non-CD cell surface proteins. Based on this proteome profiling, we identified 14 cell surface proteins, which varied consistently in abundance when cells were cultured under low or high serum conditions. Collectively, our analytical methods provide a basis for identifying the cell surface proteome of dental pulp stem cells isolated from single donors and its evolution during culture or differentiation. Our data provide a comprehensive cell surface proteome for the precise identification of dental pulp-derived MSC populations and their isolation for potential therapeutic intervention. PMID:27490675

  10. Cell Surface Proteome of Dental Pulp Stem Cells Identified by Label-Free Mass Spectrometry.

    Science.gov (United States)

    Niehage, Christian; Karbanová, Jana; Steenblock, Charlotte; Corbeil, Denis; Hoflack, Bernard

    2016-01-01

    Multipotent mesenchymal stromal cells (MSCs) are promising tools for regenerative medicine. They can be isolated from different sources based on their plastic-adherence property. The identification of reliable cell surface markers thus becomes the Holy Grail for their prospective isolation. Here, we determine the cell surface proteomes of human dental pulp-derived MSCs isolated from single donors after culture expansion in low (2%) or high (10%) serum-containing media. Cell surface proteins were tagged on intact cells using cell impermeable, cleavable sulfo-NHS-SS-biotin, which allows their enrichment by streptavidin pull-down. For the proteomic analyses, we first compared label-free methods to analyze cell surface proteomes i.e. composition, enrichment and proteomic differences, and we developed a new mathematical model to determine cell surface protein enrichment using a combinatorial gene ontology query. Using this workflow, we identified 101 cluster of differentiation (CD) markers and 286 non-CD cell surface proteins. Based on this proteome profiling, we identified 14 cell surface proteins, which varied consistently in abundance when cells were cultured under low or high serum conditions. Collectively, our analytical methods provide a basis for identifying the cell surface proteome of dental pulp stem cells isolated from single donors and its evolution during culture or differentiation. Our data provide a comprehensive cell surface proteome for the precise identification of dental pulp-derived MSC populations and their isolation for potential therapeutic intervention. PMID:27490675

  11. flowCL: ontology-based cell population labelling in flow cytometry

    Science.gov (United States)

    Courtot, Mélanie; Meskas, Justin; Diehl, Alexander D.; Droumeva, Radina; Gottardo, Raphael; Jalali, Adrin; Taghiyar, Mohammad Jafar; Maecker, Holden T.; McCoy, J. Philip; Ruttenberg, Alan; Scheuermann, Richard H.; Brinkman, Ryan R.

    2015-01-01

    Motivation: Finding one or more cell populations of interest, such as those correlating to a specific disease, is critical when analysing flow cytometry data. However, labelling of cell populations is not well defined, making it difficult to integrate the output of algorithms to external knowledge sources. Results: We developed flowCL, a software package that performs semantic labelling of cell populations based on their surface markers and applied it to labelling of the Federation of Clinical Immunology Societies Human Immunology Project Consortium lyoplate populations as a use case. Conclusion: By providing automated labelling of cell populations based on their immunophenotype, flowCL allows for unambiguous and reproducible identification of standardized cell types. Availability and implementation: Code, R script and documentation are available under the Artistic 2.0 license through Bioconductor (http://www.bioconductor.org/packages/devel/bioc/html/flowCL.html). Contact: rbrinkman@bccrc.ca Supplementary information: Supplementary data are available at Bioinformatics online. PMID:25481008

  12. Tumor-initiating label-retaining cancer cells in human gastrointestinal cancers undergo asymmetric cell division.

    Science.gov (United States)

    Xin, Hong-Wu; Hari, Danielle M; Mullinax, John E; Ambe, Chenwi M; Koizumi, Tomotake; Ray, Satyajit; Anderson, Andrew J; Wiegand, Gordon W; Garfield, Susan H; Thorgeirsson, Snorri S; Avital, Itzhak

    2012-04-01

    Label-retaining cells (LRCs) have been proposed to represent adult tissue stem cells. LRCs are hypothesized to result from either slow cycling or asymmetric cell division (ACD). However, the stem cell nature and whether LRC undergo ACD remain controversial. Here, we demonstrate label-retaining cancer cells (LRCCs) in several gastrointestinal (GI) cancers including fresh surgical specimens. Using a novel method for isolation of live LRCC, we demonstrate that a subpopulation of LRCC is actively dividing and exhibits stem cells and pluripotency gene expression profiles. Using real-time confocal microscopic cinematography, we show live LRCC undergoing asymmetric nonrandom chromosomal cosegregation LRC division. Importantly, LRCCs have greater tumor-initiating capacity than non-LRCCs. Based on our data and that cancers develop in tissues that harbor normal-LRC, we propose that LRCC might represent a novel population of GI stem-like cancer cells. LRCC may provide novel mechanistic insights into the biology of cancer and regenerative medicine and present novel targets for cancer treatment. PMID:22331764

  13. Deep diving in the blood stem cell-ome

    OpenAIRE

    Kalaitzidis, Demetrios; Scadden, David T.

    2014-01-01

    Defining the functional distinctions between cells comprising the bone marrow has yielded fundamental insights into lineage ordering and drivers of blood cell production. A novel, highly granular and multi-dimensional molecular characterization of functional subsets of hematopoietic stem- and progenitor cells recently published in Cell Stem Cell (Cabezas-Wallscheid et al, 2014) will serve as a landmark and treasure trove for unanticipated insights into basic biology and the development of fut...

  14. Mechanisms Linking Red Blood Cell Disorders and Cardiovascular Diseases

    Directory of Open Access Journals (Sweden)

    Ioana Mozos

    2015-01-01

    Full Text Available The present paper aims to review the main pathophysiological links between red blood cell disorders and cardiovascular diseases, provides a brief description of the latest studies in this area, and considers implications for clinical practice and therapy. Anemia is associated with a special risk in proatherosclerotic conditions and heart disease and became a new therapeutic target. Guidelines must be updated for the management of patients with red blood cell disorders and cardiovascular diseases, and targets for hemoglobin level should be established. Risk scores in several cardiovascular diseases should include red blood cell count and RDW. Complete blood count and hemorheological parameters represent useful, inexpensive, widely available tools for the management and prognosis of patients with coronary heart disease, heart failure, hypertension, arrhythmias, and stroke. Hypoxia and iron accumulation cause the most important cardiovascular effects of sickle cell disease and thalassemia. Patients with congenital chronic hemolytic anemia undergoing splenectomy should be monitored, considering thromboembolic and cardiovascular risk.

  15. Scattering pulse of label free fine structure cells to determine the size scale of scattering structures

    Science.gov (United States)

    Zhang, Lu; Chen, Xingyu; Zhang, Zhenxi; Chen, Wei; Zhao, Hong; Zhao, Xin; Li, Kaixing; Yuan, Li

    2016-04-01

    Scattering pulse is sensitive to the morphology and components of each single label-free cell. The most direct detection result, label free cell's scattering pulse is studied in this paper as a novel trait to recognize large malignant cells from small normal cells. A set of intrinsic scattering pulse calculation method is figured out, which combines both hydraulic focusing theory and small particle's scattering principle. Based on the scattering detection angle ranges of widely used flow cytometry, the scattering pulses formed by cell scattering energy in forward scattering angle 2°-5° and side scattering angle 80°-110° are discussed. Combining the analysis of cell's illuminating light energy, the peak, area, and full width at half maximum (FWHM) of label free cells' scattering pulses for fine structure cells with diameter 1-20 μm are studied to extract the interrelations of scattering pulse's features and cell's morphology. The theoretical and experimental results show that cell's diameter and FWHM of its scattering pulse agree with approximate linear distribution; the peak and area of scattering pulse do not always increase with cell's diameter becoming larger, but when cell's diameter is less than about 16 μm the monotone increasing relation of scattering pulse peak or area with cell's diameter can be obtained. This relationship between the features of scattering pulse and cell's size is potentially a useful but very simple criterion to distinguishing malignant and normal cells by their sizes and morphologies in label free cells clinical examinations.

  16. Functional endothelial cells derived from embryonic stem cells labeled with HIV transactivator peptide-conjugated superparamagnetic nanoparticles

    Institute of Scientific and Technical Information of China (English)

    GAO Bin; FU Wei-guo; DONG Zhi-hui; FANG Zheng-dong; LIU Zhen-jie; SI Yi; ZHANG Xiang-man; WANG Yu-qi

    2011-01-01

    Background The development of regenerative therapies using derivatives of embryonic stem (ES) cells would be facilitated by a non-invasive method to monitor transplanted cells in vivo,for example,magnetic resonance imaging of cells labeled with superparamagnetic iron oxide (SPIO) nanoparticles.Although ES cells have been labeled with SPIO particles,the potential adverse effects of the label have not been fully examined.The objective of this study was to determine whether SPIO labeling affects murine ES cell viability,proliferation,or ability to differentiate into functional endothelial cells (ECs).Methods Cross-linked iron oxide (CLIO,an SPIO) was conjugated with human immunodeficiency virus transactivator of transcription (HIV-Tat) peptides,and murine ES cells were labeled with either CLiO-Tat,CLIO,or HIV-Tat.After labeling,ES cells were cultured for 4 days and FIk-1+ ES cells identified and sorted by immunocytochemistry and fluorescence activated cell sorting (FACS).FIk-1+ cells were raplated on fibronectin-coated dishes,and ECs were obtained by culturing these for 4 weeks in endothelial cell growth medium supplemented with vascular endothelial growth factor (VEGF).ES cell viability was determined using trypan blue exclusion,and the proportion of SPIO+ cells was evaluated using Prussian blue staining and transmission electron microscopy.After differentiation,the behavior and phenotype of ECs were analyzed by reverse transcription-polymerase chain reaction,flow cytometry,immunocytochemistry,Dil-labeled acetylated low-density lipoprotein (AcLDL) uptake,and Matrigel tube formation assay.Results CLIO-Tat was a highly effective label for ES cells,with >96% of cells incorporating the particles,and it did not alter the viability of the labeled cells.ECs derived from CLIO-Tat+ ES cells were very similar to murine aortic ECs in their morphology,expression of endothelial cell markers,ability to form vascular-like channels,and scavenging of AcLDL from the culture medium

  17. Multifactorial aspects of antibody-mediated blood cell destruction

    OpenAIRE

    Schoot, van der, B.H.; Vidarsson, G.; Kapur, R.

    2014-01-01

    The research described in this thesis focuses on diseases of antibody-mediated blood cell destruction via FcγRs on phagocytes, in particular regarding platelets in fetal or neonatal alloimmune thrombocytopenia (FNAIT) and red blood cells (RBC) in hemolytic disease of the fetus and newborn (HDFN). Diagnostically, for HDFN laboratory tests are in place in order to predict risk for severe fetal RBC destruction and thereby initiate appropriate treatments. This test is sensitive, but has relativel...

  18. Raman spectroscopy of stored red blood cells: evaluating clinically-relevant biochemical markers in donated blood

    Science.gov (United States)

    Atkins, Chad G.; Buckley, Kevin; Chen, Deborah; Schulze, H. G.; Devine, Dana V.; Blades, Michael W.; Turner, Robin F. B.

    2015-07-01

    Modern transfusion medicine relies on the safe, secure, and cost-effective delivery of donated red blood cells (RBCs). Once isolated, RBCs are suspended in a defined additive solution and stored in plastic blood bags in which, over time, they undergo chemical, physiological, and morphological changes that may have a deleterious impact on some patients. Regulations limit the storage period to 42 days and the cells do not routinely undergo analytical testing before use. In this study, we use Raman spectroscopy to interrogate stored RBCs and we identify metabolic and cell-breakdown products, such as haemoglobin and membrane fragments, that build-up in the blood bags as the cells age. Our work points the way to the development of an instrument which could quickly and easily assess the biochemical nature of stored RBC units before they are transfused.

  19. A Discrete-Element Approach for Blood Cell Adhesion

    Science.gov (United States)

    Chesnutt, Jennifer; Marshall, Jeffrey

    2006-11-01

    An efficient computational model for simulation of the individual dynamics of adhering blood cells is discussed. Each cell is represented as a discrete particle so that the model can extend existing discrete-element approaches for dense particulate fluid flows to account for receptor-ligand binding of particles, elliptical particle shape, and deformation of the particles due to shear forces. Capabilities of the method in simulating large numbers of particles are illustrated through simulations of the formation of red blood cell rouleaux in shear flow. The effects of several factors, such as aspect ratio of the elliptical particle, shear rate, strength of the cell adhesion force, and hematocrit are investigated. Comparison of the discrete-element results with results of a level-set approach which computes the entire flow field about a small number of cells is used to develop an improved model of the effect of nearby red blood cells on the cell drag force expression. The method is also being applied to examine the influence of red blood cells on other components of the blood, such as platelet dispersion and activation in high shear regions.

  20. Erythropoietin reduces storage lesions and decreases apoptosis indices in blood bank red blood cells

    Directory of Open Access Journals (Sweden)

    Oscar Andrés Penuela

    2016-02-01

    Full Text Available ABSTRACT Background: Recent evidence shows a selective destruction of the youngest circulating red blood cells (neocytolysis trigged by a drop in erythropoietin levels. Objective: The aim of this study was to evaluate the effect of recombinant human erythropoietin beta on the red blood cell storage lesion and apoptosis indices under blood bank conditions. Methods: Each one of ten red blood cell units preserved in additive solution 5 was divided in two volumes of 100 mL and assigned to one of two groups: erythropoietin (addition of 665 IU of recombinant human erythropoietin and control (isotonic buffer solution was added. The pharmacokinetic parameters of erythropoietin were estimated and the following parameters were measured weekly, for six weeks: Immunoreactive erythropoietin, hemolysis, percentage of non-discocytes, adenosine triphosphate, glucose, lactate, lactate dehydrogenase, and annexin-V/esterase activity. The t-test or Wilcoxon's test was used for statistical analysis with significance being set for a p-value 6 weeks under blood bank conditions, with persistent supernatant concentrations of erythropoietin during the entire storage period. Adenosine triphosphate was higher in the Erythropoietin Group in Week 6 (4.19 ± 0.05 µmol/L vs. 3.53 ± 0.02 µmol/L; p-value = 0.009. The number of viable cells in the Erythropoietin Group was higher than in the Control Group (77% ± 3.8% vs. 71% ± 2.3%; p-value <0.05, while the number of apoptotic cells was lower (9.4% ± 0.3% vs. 22% ± 0.8%; p-value <0.05. Conclusions: Under standard blood bank conditions, an important proportion of red blood cells satisfy the criteria of apoptosis. Recombinant human erythropoietin beta seems to improve storage lesion parameters and mitigate apoptosis.

  1. Supernatant of Bone Marrow Mesenchymal Stromal Cells Induces Peripheral Blood Mononuclear Cells Possessing Mesenchymal Features

    OpenAIRE

    Hu, Gang; Xu, Jun-jun; Deng, Zhi-Hong; Feng, Jie; Jin, Yan

    2011-01-01

    Increasing evidence shows that some cells from peripheral blood fibroblast-like mononuclear cells have the capacity to differentiate into mesenchymal lineages. However, the insufficiency of these cells in the circulation challenges the cell isolation and subsequently limits the clinical application of these cells. In the present study, the peripheral blood mononuclear cells (pbMNCs) were isolated from wound animals and treated with the supernatant of bone marrow mesenchymal stromal cells (bmM...

  2. Explicit kinetic heterogeneity: mathematical models for interpretation of deuterium labeling of heterogeneous cell populations.

    Directory of Open Access Journals (Sweden)

    Vitaly V Ganusov

    2010-02-01

    Full Text Available Estimation of division and death rates of lymphocytes in different conditions is vital for quantitative understanding of the immune system. Deuterium, in the form of deuterated glucose or heavy water, can be used to measure rates of proliferation and death of lymphocytes in vivo. Inferring these rates from labeling and delabeling curves has been subject to considerable debate with different groups suggesting different mathematical models for that purpose. We show that the three most common models, which are based on quite different biological assumptions, actually predict mathematically identical labeling curves with one parameter for the exponential up and down slope, and one parameter defining the maximum labeling level. By extending these previous models, we here propose a novel approach for the analysis of data from deuterium labeling experiments. We construct a model of "kinetic heterogeneity" in which the total cell population consists of many sub-populations with different rates of cell turnover. In this model, for a given distribution of the rates of turnover, the predicted fraction of labeled DNA accumulated and lost can be calculated. Our model reproduces several previously made experimental observations, such as a negative correlation between the length of the labeling period and the rate at which labeled DNA is lost after label cessation. We demonstrate the reliability of the new explicit kinetic heterogeneity model by applying it to artificially generated datasets, and illustrate its usefulness by fitting experimental data. In contrast to previous models, the explicit kinetic heterogeneity model 1 provides a novel way of interpreting labeling data; 2 allows for a non-exponential loss of labeled cells during delabeling, and 3 can be used to describe data with variable labeling length.

  3. Lentivirus-mediated bifunctional cell labeling for in vivo melanoma study.

    Science.gov (United States)

    Day, Chi-Ping; Carter, John; Bonomi, Carrie; Esposito, Dominic; Crise, Bruce; Ortiz-Conde, Betty; Hollingshead, Melinda; Merlino, Glenn

    2009-06-01

    Lentiviral vectors (LVs) are capable of labeling a broad spectrum of cell types, achieving stable expression of transgenes. However, for in vivo studies, the duration of marker gene expression has been highly variable. We have developed a series of LVs harboring different promoters for expressing reporter gene in mouse cells. Long-term culture and colony formation of several LV-labeled mouse melanoma cells showed that promoters derived from mammalian house-keeping genes, especially those encoding RNA polymerase II (Pol2) and ferritin (FerH), provided the highest consistency for reporter expression. For in vivo studies, primary B16BL6 mouse melanoma were infected with LVs whose luciferase-green fluorescence protein fusion gene (Luc/GFP) was driven by either Pol2 or FerH promoters. When transplanted into syngeneic C57BL/6 mice, Luc/GFP-labeled B16BL6 mouse melanoma cells can be monitored by bioluminescence imaging in vivo, and GFP-positive cells can be isolated from the tumors by fluorescence-activated cell sorter. Pol2-Luc/GFP labeling, while lower in activity, was more sustainable than FerH-Luc/GFP labeling in B16BL6 over consecutive passages into mice. We conclude that Pol-2-Luc/GFP labeling allows long-term in vivo monitoring and tumor cell isolation in immunocompetent mouse melanoma models. PMID:19175523

  4. In Vivo Imaging and Tracking of Technetium-99m Labeled Bone Marrow Mesenchymal Stem Cells in Equine Tendinopathy.

    Science.gov (United States)

    Dudhia, Jayesh; Becerra, Patricia; Valdés, Miguel A; Neves, Francisco; Hartman, Neil G; Smith, Roger K W

    2015-01-01

    Recent advances in the application of bone marrow mesenchymal stem cells (BMMSC) for the treatment of tendon and ligament injuries in the horse suggest improved outcome measures in both experimental and clinical studies. Although the BMMSC are implanted into the tendon lesion in large numbers (usually 10 - 20 million cells), only a relatively small number survive (horses. Tc-99m is a short-lived (t1/2 of 6.01 hr) isotope that emits gamma rays and can be internalized by cells in the presence of the lipophilic compound hexamethylpropyleneamine oxime (HMPAO). These properties make it ideal for use in nuclear medicine clinics for the diagnosis of many different diseases. The fate of the labeled cells can be followed in the short term (up to 36 hr) by gamma scintigraphy to quantify both the number of cells retained in the lesion and distribution of the cells into lungs, thyroid and other organs. This technique is adapted from the labeling of blood leukocytes and could be utilized to image implanted BMMSC in other organs. PMID:26709915

  5. Dynamic quantitative microscopy and nanoscopy of red blood cells in sickle cell disease

    Science.gov (United States)

    Shaked, Natan T.; Satterwhite, Lisa L.; Telen, Marilyn J.; Truskey, George A.; Wax, Adam

    2012-03-01

    We have applied wide-field digital interferometric techniques to quantitatively image sickle red blood cells (RBCs) [1] in a noncontact label-free manner, and measure the nanometer-scale fluctuations in their thickness as an indication of their stiffness. The technique can simultaneously measure the fluctuations for multiple spatial points on the RBC and thus yields a map describing the stiffness of each RBC in the field of view. Using this map, the local rigidity regions of the RBC are evaluated quantitatively. Since wide-field digital interferometry is a quantitative holographic imaging technique rather than one-point measurement, it can be used to simultaneously evaluate cell transverse morphology plus thickness in addition to its stiffness profile. Using this technique, we examine the morphology and dynamics of RBCs from individuals who suffer from sickle cell disease, and find that the sickle RBCs are significantly stiffer than healthy RBCs. Furthermore, we show that the technique is sensitive enough to distinguish various classes of sickle RBCs, including sickle RBCs with visibly-normal morphology, compared to the stiffer crescent-shaped sickle RBCs.

  6. A Framework for White Blood Cell Segmentation in Microscopic Blood Images Using Digital Image Processing

    OpenAIRE

    Seman Zainina; Abdul Kahar Badrul; Sadeghian Farnoosh; Ramli Abdul; Saripan M-Iqbal

    2009-01-01

    Abstract Evaluation of blood smear is a commonly clinical test these days. Most of the time, the hematologists are interested on white blood cells (WBCs) only. Digital image processing techniques can help them in their analysis and diagnosis. For example, disease like acute leukemia is detected based on the amount and condition of the WBC. The main objective of this paper is to segment the WBC to its two dominant elements: nucleus and cytoplasm. The segmentation is conducted using a proposed ...

  7. Magnetic targeting of iron-oxide-labeled fluorescent hepatoma cells to the liver

    International Nuclear Information System (INIS)

    The purpose of this study was to determine whether an external magnet field can induce preferential trafficking of magnetically labeled Huh7 hepatoma cells to the liver following liver cell transplantation. Huh7 hepatoma cells were labeled with anionic magnetic nanoparticles (AMNP) and tagged with a fluorescent membrane marker (PKH67). Iron-uptake was measured by magnetophoresis. Twenty C57Bl6 mice received an intrasplenic injection of 2 x 106 labeled cells. An external magnet (0.29 T; 25 T/m) was placed over the liver of 13 randomly selected animals (magnet group), while the remaining 7 animals served as controls. MRI (1.5 T) and confocal fluorescence microscopy (CFM) were performed 10 days post-transplantation. The presence and location of labeled cells within the livers were compared in the magnet group and controls, and confronted with histological analysis representing the standard of reference. Mean iron content per cell was 6 pg. Based on histology, labeled cells were more frequently present within recipient livers in the magnet group (p < 0.01) where their distribution was preferentially peri-vascular (p<0.05). MRI and CFM gave similar results for the overall detection of transplanted cells (kappa=0.828) and for the identification of peri-vascular cells (kappa=0.78). Application of an external magnet can modify the trafficking of transplanted cells, especially by promoting the formation of perivascular aggregates. (orig.)

  8. Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis

    DEFF Research Database (Denmark)

    Harkness, Linda; Prokhorova, Tatyana A; Kassem, Moustapha;

    2012-01-01

    The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass...... spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we...

  9. Immunoglobulin and enzyme-conjugated dextran polymers enhance u-PAR staining intensity of carcinoma cells in peripheral blood smears

    DEFF Research Database (Denmark)

    Werther, K; Normark, M; Hansen, B F;

    1999-01-01

    shown between the expression of tumor cell proteases and tumor invasion. Therefore, phenotypic characterization of disseminated carcinoma cells for expression of protease activators might define the invasive potential of the cells. We present an immunocytochemically enhanced staining method that allows......The presence of disseminated carcinoma cells in bone marrow and peripheral blood has prognostic importance in patients with carcinomas. Much evidence indicates that dissemination of tumor cells may depend on activation of a variety of degradative enzymes. A strong positive correlation has been...... phenotyping of disseminated carcinoma cells in bone marrow and peripheral blood smears. In the first step, the cells were incubated with antibodies against urokinase plasminogen activator receptor (u-PAR) and subsequently with secondary antibodies conjugated to peroxidase-labeled dextran polymers. A brown...

  10. Labeling of mesenchymal stem cells for MRI with single-cell sensitivity

    Directory of Open Access Journals (Sweden)

    Ariza de Schellenberger A

    2016-04-01

    Full Text Available Angela Ariza de Schellenberger,1 Harald Kratz,1 Tracy D Farr,2,3 Norbert Löwa,4 Ralf Hauptmann,1 Susanne Wagner,1 Matthias Taupitz,1 Jörg Schnorr,1 Eyk A Schellenberger1 1Department of Radiology, 2Department of Experimental Neurology, Center for Stroke Research Berlin, Charité – Universitätsmedizin Berlin, Berlin, Germany; 3School of Life Sciences, University of Nottingham, Medical School, Nottingham, UK; 4Department of Biomagnetic Signals, Physikalisch-Technische Bundesanstalt Berlin, Berlin, Germany Abstract: Sensitive cell detection by magnetic resonance imaging (MRI is an important tool for the development of cell therapies. However, clinically approved contrast agents that allow single-cell detection are currently not available. Therefore, we compared very small iron oxide nanoparticles (VSOP and new multicore carboxymethyl dextran-coated iron oxide nanoparticles (multicore particles, MCP designed by our department for magnetic particle imaging (MPI with discontinued Resovist® regarding their suitability for detection of single mesenchymal stem cells (MSC by MRI. We achieved an average intracellular nanoparticle (NP load of >10 pg Fe per cell without the use of transfection agents. NP loading did not lead to significantly different results in proliferation, colony formation, and multilineage in vitro differentiation assays in comparison to controls. MRI allowed single-cell detection using VSOP, MCP, and Resovist® in conjunction with high-resolution T2*-weighted imaging at 7 T with postprocessing of phase images in agarose cell phantoms and in vivo after delivery of 2,000 NP-labeled MSC into mouse brains via the left carotid artery. With optimized labeling conditions, a detection rate of ~45% was achieved; however, the experiments were limited by nonhomogeneous NP loading of the MSC population. Attempts should be made to achieve better cell separation for homogeneous NP loading and to thus improve NP

  11. [Introduction and prospect of peripheral blood stem cell transplantation].

    Science.gov (United States)

    Nakanishi, Y

    1995-12-01

    The number of hematopoietic stem cells circulating in peripheral blood increases remarkably during the recovery of marrow function after myelosuppressive chemotherapy. In peripheral blood stem cell transplantation, these stem cells are collected and cryopreserved, and then used to restore marrow function after myelodisruptive (high-dose) anticancer therapy, Marrow recovery is faster with this procedure than with autologous bone marrow transplantation. Recently, this procedure has been used after high-dose chemotherapy for chemosensitive solid tumors such as breast cancer. We used high-dose chemotherapy with etoposide and carboplatin, followed by peripheral blood stem cell transplantation, to treat 5 patients with intrathoracic malignant tumors, including small cell lung cancer Neutrophils recovered (> 500 microliters) with 9 to 11 days and platelets recovered (> 5,000 microliters) within 8 to 13 days after the transplantation. No other serious complication was seen. Current topics regarding this procedure, problems to be solved, and prospects for further development are discussed. PMID:8752478

  12. Quantification of depletion-induced adhesion of Red Blood Cells

    CERN Document Server

    Steffen, Patrick; Wagner, Christian

    2012-01-01

    Red blood cells (RBC) are known to form aggregates in the forms of rouleaux due to the presence of plasma proteins under physiological conditions. Rouleaux formation can be also induced in vitro by the addition of macromolecules to the RBC solution. Current data on the adhesion strength between red blood cells in their natural discocyte shapes mostly rely on indirect measurements like flow chamber experiments, but on the single cell level data is lacking. Here we present measurements on the dextran induced aggregation of red blood cells by use of atomic force microscopy based single cell force spectroscopy (SCFS). The effects of dextran concentration and molecular weight on the interaction energy of adhering RBCs was determined. The results are in good agreement with a model based on the depletion effect and former experimental studies.

  13. Effect of red blood cell aggregation and sedimentation on optical coherence tomography signals from blood samples

    International Nuclear Information System (INIS)

    In this work, Monte Carlo simulation is used to obtain model optical coherence tomography (OCT) signals from a horizontally orientated blood layer at different stages of red blood cell (RBC) aggregation and sedimentation processes. The parameters for aggregating and sedimenting blood cells were chosen based on the data available from the literature and our earlier experimental studies. We consider two different cases: a suspension of washed RBCs in physiological solution (where aggregation does not take place) and RBCs in blood plasma (which provides necessary conditions for aggregation). Good agreement of the simulation results with the available experimental data shows that the chosen optical parameters are reasonable. The dependence of the numbers of photons contributing to the OCT signal on the number of experienced scattering events was analysed for each simulated signal. It was shown that the maxima of these dependences correspond to the peaks in the OCT signals related to the interfaces between the layers of blood plasma and blood cells. Their positions can be calculated from the optical thicknesses of the layers, and the absorption and scattering coefficients of the media

  14. Following red blood cells in a pulmonary capillary

    CERN Document Server

    Mauroy, Benjamin

    2007-01-01

    The red blood cells or erythrocytes are biconcave shaped cells and consist mostly in a membrane delimiting a cytosol with a high concentration in hemoglobin. This membrane is highly deformable and allows the cells to go through narrow passages like the capillaries which diameters can be much smaller than red blood cells one. They carry oxygen thanks to hemoglobin, a complex molecule that have very high affinity for oxygen. The capacity of erythrocytes to load and unload oxygen is thus a determinant factor in their efficacy. In this paper, we will focus on the pulmonary capillary where red blood cells capture oxygen. We propose a camera method in order to numerically study the behavior of the red blood cell along a whole capillary. Our goal is to understand how erythrocytes geometrical changes along the capillary can affect its capacity to capture oxygen. The first part of this document presents the model chosen for the red blood cells along with the numerical method used to determine and follow their shapes a...

  15. Multicolor protein labeling in living cells using mutant β-lactamase-tag technology.

    Science.gov (United States)

    Watanabe, Shuji; Mizukami, Shin; Hori, Yuichiro; Kikuchi, Kazuya

    2010-12-15

    Protein labeling techniques using small molecule probes have become important as practical alternatives to the use of fluorescent proteins (FPs) in live cell imaging. These labeling techniques can be applied to more sophisticated fluorescence imaging studies such as pulse-chase imaging. Previously, we reported a novel protein labeling system based on the combination of a mutant β-lactamase (BL-tag) with coumarin-derivatized probes and its application to specific protein labeling on cell membranes. In this paper, we demonstrated the broad applicability of our BL-tag technology to live cell imaging by the development of a series of fluorescence labeling probes for this technology, and the examination of the functions of target proteins. These new probes have a fluorescein or rhodamine chromophore, each of which provides enhanced photophysical properties relative to coumarins for the purpose of cellular imaging. These probes were used to specifically label the BL-tag protein and could be used with other small molecule fluorescent probes. Simultaneous labeling using our new probes with another protein labeling technology was found to be effective. In addition, it was also confirmed that this technology has a low interference with respect to the functions of target proteins in comparison to GFP. Highly specific and fast covalent labeling properties of this labeling technology is expected to provide robust tools for investigating protein functions in living cells, and future applications can be improved by combining the BL-tag technology with conventional imaging techniques. The combination of probe synthesis and molecular biology techniques provides the advantages of both techniques and can enable the design of experiments that cannot currently be performed using existing tools. PMID:20961132

  16. Blood thixotropy in patients with sickle cell anaemia: role of haematocrit and red blood cell rheological properties.

    Directory of Open Access Journals (Sweden)

    Jens Vent-Schmidt

    Full Text Available We compared the blood thixotropic/shear-thinning properties and the red blood cells' (RBC rheological properties between a group of patients with sickle cell anaemia (SS and healthy individuals (AA. Blood thixotropy was determined by measuring blood viscosity with a capillary viscometer using a "loop" protocol: the shear rate started at 1 s-1 and increased progressively to 922 s-1 and then re-decreased to the initial shear rate. Measurements were performed at native haematocrit for the two groups and at 25% and 40% haematocrit for the AA and SS individuals, respectively. RBC deformability was determined by ektacytometry and RBC aggregation properties by laser backscatter versus time. AA at native haematocrit had higher blood thixotropic index than SS at native haematocrit and AA at 25% haematocrit. At 40% haematocrit, SS had higher blood thixotropic index than AA. While RBC deformability and aggregation were lower in SS than in AA, the strength of RBC aggregates was higher in the former population. Our results showed that 1 anaemia is the main modulator of blood thixtropy and 2 the low RBC deformability and high RBC aggregates strength cause higher blood thixotropy in SS patients than in AA individuals at 40% haematocrit, which could impact blood flow in certain vascular compartments.

  17. IMAGING RED BLOOD CELL DYNAMICS BY QUANTITATIVE PHASE MICROSCOPY

    OpenAIRE

    Popescu, Gabriel; Park, YoungKeun; Choi, Wonshik; Dasari, Ramachandra R.; Michael S. Feld; Badizadegan, Kamran

    2008-01-01

    Red blood cells (RBCs) play a crucial role in health and disease, and structural and mechanical abnormalities of these cells have been associated with important disorders such as Sickle cell disease and hereditary cytoskeletal abnormalities. Although several experimental methods exist for analysis of RBC mechanical properties, optical methods stand out as they enable collecting mechanical and dynamic data from live cells without physical contact and without the need for exogenous contrast age...

  18. Subcutaneous adipose tissue blood flow in the forefoot during 24 hours. Labeling pattern and reproducibility

    DEFF Research Database (Denmark)

    Jelnes, Rolf; Bülow, J; Tønnesen, K H

    1987-01-01

    (range: 3-90 days). The patients were studied under two different conditions. Firstly, during the day in the erect position, awake (sitting, standing and quiet walking) and secondly, during night hours in the supine position, asleep. The coefficient of variation of nocturnal adipose tissue blood flow was...

  19. Edwardsiella tarda Endocarditis Confirmed by Indium-111 White Blood Cell Scan: An Unusual Pathogen and Diagnostic Modality.

    Science.gov (United States)

    Litton, Kayleigh M; Rogers, Bret A

    2016-01-01

    Edwardsiella tarda is a freshwater marine member of the family Enterobacteriaceae which often colonizes fish, lizards, snakes, and turtles but is an infrequent human pathogen. Indium-111- ((111)In-) labeled white blood cell (WBC) scintigraphy is an imaging modality which has a wide range of reported sensitivity and specificity (from 60 to 100% and from 68 to 92%, resp.) for diagnosing acute and chronic infection. We describe a case of suspected E. tarda prosthetic aortic valve and mitral valve endocarditis with probable vegetations and new mitral regurgitation on transthoracic and transesophageal echocardiograms which was supported with the use of (111)In-labeled WBC scintigraphy. PMID:26885418

  20. In vivo ultrasound and photoacoustic monitoring of mesenchymal stem cells labeled with gold nanotracers.

    Directory of Open Access Journals (Sweden)

    Seung Yun Nam

    Full Text Available Longitudinal monitoring of cells is required in order to understand the role of delivered stem cells in therapeutic neovascularization. However, there is not an imaging technique that is capable of quantitative, longitudinal assessment of stem cell behaviors with high spatial resolution and sufficient penetration depth. In this study, in vivo and in vitro experiments were performed to demonstrate the efficacy of ultrasound-guided photoacoustic (US/PA imaging to monitor mesenchymal stem cells (MSCs labeled with gold nanotracers (Au NTs. The Au NT labeled MSCs, injected intramuscularly in the lower limb of the Lewis rat, were detected and spatially resolved. Furthermore, our quantitative in vitro cell studies indicate that US/PA imaging is capable of high detection sensitivity (1×10⁴ cells/mL of the Au NT labeled MSCs. Finally, Au NT labeled MSCs captured in the PEGylated fibrin gel system were imaged in vivo, as well as in vitro, over a one week time period, suggesting that longitudinal cell tracking using US/PA imaging is possible. Overall, Au NT labeling of MSCs and US/PA imaging can be an alternative approach in stem cell imaging capable of noninvasive, sensitive, quantitative, longitudinal assessment of stem cell behaviors with high spatial and temporal resolutions at sufficient depths.

  1. The homeostasis of Plasmodium falciparum-infected red blood cells.

    Directory of Open Access Journals (Sweden)

    Jakob M A Mauritz

    2009-04-01

    Full Text Available The asexual reproduction cycle of Plasmodium falciparum, the parasite responsible for severe malaria, occurs within red blood cells. A merozoite invades a red cell in the circulation, develops and multiplies, and after about 48 hours ruptures the host cell, releasing 15-32 merozoites ready to invade new red blood cells. During this cycle, the parasite increases the host cell permeability so much that when similar permeabilization was simulated on uninfected red cells, lysis occurred before approximately 48 h. So how could infected cells, with a growing parasite inside, prevent lysis before the parasite has completed its developmental cycle? A mathematical model of the homeostasis of infected red cells suggested that it is the wasteful consumption of host cell hemoglobin that prevents early lysis by the progressive reduction in the colloid-osmotic pressure within the host (the colloid-osmotic hypothesis. However, two critical model predictions, that infected cells would swell to near prelytic sphericity and that the hemoglobin concentration would become progressively reduced, remained controversial. In this paper, we are able for the first time to correlate model predictions with recent experimental data in the literature and explore the fine details of the homeostasis of infected red blood cells during five model-defined periods of parasite development. The conclusions suggest that infected red cells do reach proximity to lytic rupture regardless of their actual volume, thus requiring a progressive reduction in their hemoglobin concentration to prevent premature lysis.

  2. A microfluidics-based technique for automated and rapid labeling of cells for flow cytometry

    International Nuclear Information System (INIS)

    Flow cytometry is a powerful technique capable of simultaneous multi-parametric analysis of heterogeneous cell populations for research and clinical applications. In recent years, the flow cytometer has been miniaturized and made portable for application in clinical- and resource-limited settings. The sample preparation procedure, i.e. labeling of cells with antibodies conjugated to fluorescent labels, is a time consuming (∼45 min) and labor-intensive procedure. Microfluidics provides enabling technologies to accomplish rapid and automated sample preparation. Using an integrated microfluidic device consisting of a labeling and washing module, we demonstrate a new protocol that can eliminate sample handling and accomplish sample and reagent metering, high-efficiency mixing, labeling and washing in rapid automated fashion. The labeling module consists of a long microfluidic channel with an integrated chaotic mixer. Samples and reagents are precisely metered into this device to accomplish rapid and high-efficiency mixing. The mixed sample and reagents are collected in a holding syringe and held for up to 8 min following which the mixture is introduced into an inertial washing module to obtain ‘analysis-ready’ samples. The washing module consists of a high aspect ratio channel capable of focusing cells to equilibrium positions close to the channel walls. By introducing the cells and labeling reagents in a narrow stream at the center of the channel flanked on both sides by a wash buffer, the elution of cells into the wash buffer away from the free unbound antibodies is accomplished. After initial calibration experiments to determine appropriate ‘holding time’ to allow antibody binding, both modules were used in conjunction to label MOLT-3 cells (T lymphoblast cell line) with three different antibodies simultaneously. Results confirm no significant difference in mean fluorescence intensity values for all three antibodies labels (p < 0.01) between the

  3. Development of a Microfluidic-Based Optical Sensing Device for Label-Free Detection of Circulating Tumor Cells (CTCs Through Their Lactic Acid Metabolism

    Directory of Open Access Journals (Sweden)

    Tzu-Keng Chiu

    2015-03-01

    Full Text Available This study reports a microfluidic-based optical sensing device for label-free detection of circulating tumor cells (CTCs, a rare cell species in blood circulation. Based on the metabolic features of cancer cells, live CTCs can be quantified indirectly through their lactic acid production. Compared with the conventional schemes for CTC detection, this label-free approach could prevent the biological bias due to the heterogeneity of the surface antigens on cancer cells. In this study, a microfluidic device was proposed to generate uniform water-in-oil cell-encapsulating micro-droplets, followed by the fluorescence-based optical detection of lactic acid produced within the micro-droplets. To test its feasibility to quantify cancer cells, experiments were carried out. Results showed that the detection signals were proportional to the number of cancer cells within the micro-droplets, whereas such signals were insensitive to the existence and number of leukocytes within. To further demonstrate its feasibility for cancer cell detection, the cancer cells with known cell number in a cell suspension was detected based on the method. Results revealed that there was no significant difference between the detected number and the real number of cancer cells. As a whole, the proposed method opens up a new route to detect live CTCs in a label-free manner.

  4. Controllable labelling of stem cells with a novel superparamagnetic iron oxide-loaded cationic nanovesicle for MR imaging

    International Nuclear Information System (INIS)

    To investigate the feasibility of highly efficient and controllable stem cell labelling for cellular MRI. A new class of cationic, superparamagnetic iron oxide nanoparticle (SPION)-loaded nanovesicles was synthesised to label rat bone marrow mesenchymal stem cells without secondary transfection agents. The optimal labelling conditions and controllability were assessed, and the effect of labelling on cell viability, proliferation activity and multilineage differentiation was determined. In 18 rats, focal ischaemic cerebral injury was induced and the rats randomly injected with 1 x 106 cells labelled with 0-, 8- or 20-mV nanovesicles (n = 6 each). In vivo MRI was performed to follow grafted cells in contralateral striata, and results were correlated with histology. Optimal cell labelling conditions involved a concentration of 3.15 μg Fe/mL nanovesicles with 20-mV positive charge and 1-h incubation time. Labelling efficiency showed linear change with an increase in the electric potentials of nanovesicles. Labelling did not affect cell viability, proliferation activity or multilineage differentiation capacity. The distribution and migration of labelled cells could be detected by MRI. Histology confirmed that grafted cells retained the label and remained viable. Stem cells can be effectively and safely labelled with cationic, SPION-loaded nanovesicles in a controllable way for cellular MRI. (orig.)

  5. A smart core-sheath nanofiber that captures and releases red blood cells from the blood

    Science.gov (United States)

    Shi, Q.; Hou, J.; Zhao, C.; Xin, Z.; Jin, J.; Li, C.; Wong, S.-C.; Yin, J.

    2016-01-01

    A smart core-sheath nanofiber for non-adherent cell capture and release is demonstrated. The nanofibers are fabricated by single-spinneret electrospinning of poly(N-isopropylacrylamide) (PNIPAAm), polycaprolactone (PCL) and nattokinase (NK) solution blends. The self-assembly of PNIPAAm and PCL blends during the electrospinning generates the core-sheath PCL/PNIPAAm nanofibers with PNIPAAm as the sheath. The PNIPAAm-based core-sheath nanofibers are switchable between hydrophobicity and hydrophilicity with temperature change and enhance stability in the blood. When the nanofibers come in contact with blood, the NK is released from the nanofibers to resist platelet adhesion on the nanofiber surface, facilitating the direct capture and isolation of red blood cells (RBCs) from the blood above phase-transition temperature of PNIPAAm. Meanwhile, the captured RBCs are readily released from the nanofibers with temperature stimuli in an undamaged manner. The release efficiency of up to 100% is obtained while maintaining cellular integrity and function. This work presents promising nanofibers to effectively capture non-adherent cells and release for subsequent molecular analysis and diagnosis of single cells.A smart core-sheath nanofiber for non-adherent cell capture and release is demonstrated. The nanofibers are fabricated by single-spinneret electrospinning of poly(N-isopropylacrylamide) (PNIPAAm), polycaprolactone (PCL) and nattokinase (NK) solution blends. The self-assembly of PNIPAAm and PCL blends during the electrospinning generates the core-sheath PCL/PNIPAAm nanofibers with PNIPAAm as the sheath. The PNIPAAm-based core-sheath nanofibers are switchable between hydrophobicity and hydrophilicity with temperature change and enhance stability in the blood. When the nanofibers come in contact with blood, the NK is released from the nanofibers to resist platelet adhesion on the nanofiber surface, facilitating the direct capture and isolation of red blood cells (RBCs) from

  6. Sex hormone drives blood stem cell reproduction

    OpenAIRE

    Calvanese, Vincenzo; Lee, Lydia K.; Mikkola, Hanna K. A.

    2014-01-01

    Stem cells ensure the maintenance of tissue homeostasis throughout life by tightly regulating their self-renewal and differentiation. In a recent study published in Nature, Nakada et al, 2014 unveil an unexpected endocrine mechanism that regulates hematopoietic stem cell (HSC) self-renewal.

  7. Development of human connective tissue mast cells from purified blood monocytes.

    Science.gov (United States)

    Czarnetzki, B M; Figdor, C G; Kolde, G; Vroom, T; Aalberse, R; de Vries, J E

    1984-01-01

    Highly purified subfractions of human peripheral blood monocytes, when cultured in the presence of 30% L cell supernatant and 30% horse serum, assumed all the characteristics that define human connective tissue mast cells. After three weeks of culture, 75% of the cells developed metachromasia and granular chloroacetate esterase staining, and their intracellular histamine levels increased from 0.0 to 50.5 ng/10(6) cells. On electron microscopy, the cells developed intracytoplasmic granules with all the features typical for mature and immature mast cells. Cultured cells bound 55 pg 125I-IgE/10(6) cells, while labelling was negligible with cells prior to culture and with heat-denatured 125I-IgE. Fluorescent staining with anti-IgE increased slightly as well, while staining with monoclonal anti-monocyte and anti-HLA-Dr markers decreased. Purified lymphocytes did not assume mast cell characteristics, and lymphokines did not induce or enhance in vitro mast cell development or IgE binding. The data therefore further support the concept that connective tissue mast cells arise from the monocytoid lineage. Images Figure 1 PMID:6698581

  8. Spatial distributions of red blood cells significantly alter local haemodynamics.

    Directory of Open Access Journals (Sweden)

    Joseph M Sherwood

    Full Text Available Although bulk changes in red blood cell concentration between vessels have been well characterised, local distributions are generally overlooked. Red blood cells aggregate, deform and migrate within vessels, forming heterogeneous distributions which have considerable effect on local haemodynamics. The present study reports data on the local distribution of human red blood cells in a sequentially bifurcating microchannel, representing the branching geometry of the microvasculature. Imaging methodologies with simple extrapolations are used to infer three dimensional, time-averaged velocity and haematocrit distributions under a range of flow conditions. Strong correlation between the bluntness of the velocity and haematocrit profiles in the parent branch of the geometry is observed and red blood cell aggregation has a notable effect on the observed trends. The two branches of the first bifurcation show similar characteristics in terms of the shapes of the profiles and the extent of plasma skimming, despite the difference in geometric configuration. In the second bifurcation, considerable asymmetry between the branches in the plasma skimming relationship is observed, and elucidated by considering individual haematocrit profiles. The results of the study highlight the importance of considering local haematocrit distributions in the analysis of blood flow and could lead to more accurate computational models of blood flow in microvascular networks. The experimental approaches developed in this work provide a foundation for further examining the characteristics of microhaemodynamics.

  9. Labeling and Imaging Mesenchymal Stem Cells with Quantum Dots

    Science.gov (United States)

    Mesenchymal stem cells (MSCs) are multipotent cells with the potential to differentiate into bone, cartilage, adipose and muscle cells. Adult derived MSCs are being actively investigated because of their potential to be utilized for therapeutic cell-based transplantation. Methods...

  10. Hyaluronic Acid-Human Blood Hydrogels for Stem Cell Transplantation

    OpenAIRE

    Connie Y. Chang; Chan, Angel; Armstrong, Patrick; Luo, Hong-Chang; Higuchi, Takahiro; Strehin, Iossif; Vakrou, Styliani; Lin, Xiaoping; Brown, Sophia; O’Rourke, Brian; Abraham, Theodore P.; Wahl, Richard; Steenbergen, Charles; ELISSEEFF, JENNIFER; Abraham, M. Roselle

    2012-01-01

    Tissue engineering-based approaches have the potential to improve stem cell engraftment by increasing cell delivery to the myocardium. Our objective was to develop and characterize a naturally-derived, autologous, biodegradable hydrogel in order to improve acute stem cell retention in the myocardium. HA-blood hydrogels(HA-Bl) were synthesized by mixing in a 1:1(v/v) ratio, lysed whole blood and hyaluronic acid(HA), whose carboxyl groups were functionalized with N-hydroxysuccinimide(NHS) to yi...

  11. 111In)oxine labelling of polymorphonuclear leucocytes: doubts concerning elution and effects on cell behaviour

    Energy Technology Data Exchange (ETDEWEB)

    Sheehan, N.J.; Brown, K.A.; Camacho, A.; Dumonde, D.C.

    1985-01-01

    Polymorphonuclear leucocytes (PMN) from normal human subjects were labelled with (111In)oxine (20 muCi 10(8) cells). In the presence of 20% autologous serum (AS), dissociation of 111In from the cells resulted in mean losses of radioactivity of 13% at 3 h and 30% at 24 h. Adherence of 111In-labelled PMN to cultured porcine endothelial monolayers was increased by 40.7 +/- 31.6% after 60 min incubation in 20% AS at 37 degrees C when compared with unlabelled cells. Phagocytosis and intracellular killing of Candida albicans were unaltered by labelling. Elution of 111In from labelled PMN together with enhanced adhesiveness may have important implications for the study of PMN kinetics and the investigation of inflammatory disease.

  12. Determination of adipose tissue blood flow with local 133Xe clearance. Evaluation of a new labelling technique

    DEFF Research Database (Denmark)

    Simonsen, Lene; Enevoldsen, Lotte Hahn; Bülow, Jens

    2003-01-01

    Adipose tissue blood flow was measured in six healthy, non-obese subjects with the xenon wash-out technique after labelling of the tissue by either injection of 133Xe dissolved in isotonic sodium chloride (water depot) or injection of 133Xe in gas form (gas depot). The wash-out rates were...... registered from four depots simultaneously. Two depots were placed above the umbilicus, and two depots were placed below the umbilicus in the abdominal, subcutaneous adipose tissue. A water depot and a gas depot were placed in the two positions, respectively. It was not possible to demonstrate any difference...... between the wash-out rates registered from the two depot types, and it was also not possible to demonstrate any difference between the changes in wash-out rates induced by an oral glucose load. Similarly, the tissue distribution of the water and the gas depots appeared to be similar as registered by a...

  13. White blood cells of peripheral blood with ConA-positive glycotopes in patients with chronic leukemia

    Directory of Open Access Journals (Sweden)

    G. S. Maslak

    2015-09-01

     Tumor growth progression of blood cells occurs due to changes in their genetic apparatus, which affects not only the cells morphological characteristics, but also their functional activity which to a greater extent depends on the membrane surface structures, a significant part of which is of glycoprotein nature. Complex type N-glycans are components of surface glycoproteins in the most of leukocytes. Thus, the study of changes in carbohydrate determinants of glycoproteins on the surface of leucocytes in tumorigenesis can help to reveal the mechanisms of this process. The aim of our study was to investigate the monocytes and granulocytes cytoplasmic membrane N-glycosylation in patients with chronic leukemia. The object of the study were blood cells of patients with chronic lymphocytic leukemia (n = 12 and polycythemia vera (n = 15 aged 58–66 years. Healthy hematologic volunteers (n = 15 aged 55 to 65 years were in the control group. N-glycan exposure on monocytes and granulocytes was investigated by flow cytometer Beckman Сoulter EPICS with Canavalia ensiformis lectin – Con A conjugated with fluorescent labels. The number of dead cells was monitored by means of binding them with propidium iodide. The result has been analyzed with FC Express. According to our data, levels of ConA-positive monocytes and granulocytes were 9,9 ± 1,0% and 32,7 ± 3,2%, respectively, in peripheral blood of healthy persons. The level of ConA-positive monocytes decreased to 31,0 ± 2,3% and the number of ConA-binding granulocytes increased to 66,7 ± 3,8% in patients with chronic lymphocytic leukemia compared with the norm. The number of ConA-positive monocytes decreased 3.3 times, and the level of granulocytes interacting with Canavalia ensiformis lectin slightly increased relative to control in polycythemia vera patients. There is significant increase in Con A-positive epitopes on granulocytes in patients with chronic lymphocytic leukemia and polycythemia vera compared with the

  14. On-chip Extraction of Intracellular Molecules in White Blood Cells from Whole Blood

    Science.gov (United States)

    Choi, Jongchan; Hyun, Ji-Chul; Yang, Sung

    2015-10-01

    The extraction of virological markers in white blood cells (WBCs) from whole blood—without reagents, electricity, or instruments—is the most important first step for diagnostic testing of infectious diseases in resource-limited settings. Here we develop an integrated microfluidic chip that continuously separates WBCs from whole blood and mechanically ruptures them to extract intracellular proteins and nucleic acids for diagnostic purposes. The integrated chip is assembled with a device that separates WBCs by using differences in blood cell size and a mechanical cell lysis chip with ultra-sharp nanoblade arrays. We demonstrate the performance of the integrated device by quantitatively analyzing the levels of extracted intracellular proteins and genomic DNAs. Our results show that compared with a conventional method, the device yields 120% higher level of total protein amount and similar levels of gDNA (90.3%). To demonstrate its clinical application to human immunodeficiency virus (HIV) diagnostics, the developed chip was used to process blood samples containing HIV-infected cells. Based on PCR results, we demonstrate that the chip can extract HIV proviral DNAs from infected cells with a population as low as 102/μl. These findings suggest that the developed device has potential application in point-of-care testing for infectious diseases in developing countries.

  15. Bos taurus papillomavirus activity in peripheral blood mononuclear cells: demonstrating a productive infection.

    Science.gov (United States)

    Melo, T C; Araldi, R P; Pessoa, N S D; de-Sá-Júnior, P L; Carvalho, R F; Beçak, W; Stocco, R C

    2015-01-01

    Bovine papillomavirus (BPV) is an oncogenic virus with mucous and epithelial tropism. Possible productive virus infection in other tissues, such as blood, has been hypothesized. In order to investigate this possibility, three samples of skin papillomas and blood were collected from bovines with BPV infection and five samples of peripheral blood and one sample of normal tissue were collected from a calf without BPV infection. Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood and examined by reverse transcription-polymerase chain reaction, immunofluorescence, in situ hybridization, and electron microscopy. The tissue samples were examined for histopathological and immunohistochemical features. The skin papillomas showed the presence of DNA sequences of BPV-2, BPV-11, and a putative virus type. The blood samples showed DNA sequences of BPV-1, 2, and 4 simultaneously. Immunohistochemistry showed BPV L1 protein in both epithelium and stroma and BPV E2 protein in koilocytes. In situ hybridization confirmed the presence of BPV DNA in PBMCs and immunofluorescence showed nuclear labeling of E2 and L1 BPV proteins in PBMCs. The transcription analysis revealed transcripts of BPV-1 L1, BPV-2 L2, and BPV-4 E7 in blood and papilloma samples of BPV-infected cattle. The comet assay revealed high levels of host cell DNA damage upon BPV infection. Electron microscopy analysis of PBMCs identified the presence of particles in the cytoplasm that are consistent with papillomavirus in size and shape. The productive infection of PBMCs with BPV has been previously discussed and this study provides evidence indicating that PBMCs are a target of BPV. PMID:26681018

  16. Ibogaine labeling with 99mTc-tricarbonyl: synthesis and transport at the mouse blood-brain barrier.

    Science.gov (United States)

    Tournier, Nicolas; André, Pascal; Blondeel, Sandy; Rizzo-Padoin, Nathalie; du Moulinet d'Hardemarre, Amaury; Declèves, Xavier; Scherrmann, Jean-Michel; Cisternino, Salvatore

    2009-12-01

    The (99m)Tc-tricarbonyl core may be used as an ideal tool for gamma-labeling ligands in noninvasive SPECT imaging. However, most (99m)Tc-tricarbonyl-labeled agents have difficulty crossing the blood-brain barrier (BBB). We radiolabeled the neuroactive indole ibogaine with (99m)Tc-tricarbonyl and measured its transport into the mouse brain by in situ brain perfusion. We measured the interactions of [(99m)Tc(CO)(3)-ibogaine](+) and (99m)Tc-tricarbonyl with the main BBB efflux transporters P-gp and BCRP in vitro and in vivo. Ibogaine was radiolabeled (yield: over 95%). [(99m)Tc(CO)(3)-ibogaine](+) entered the brain (K(in)) poorly (0.18 microL/g/s), at about the same rate as (99m)Tc-tricarbonyl (0.16 microL/g/s) and [(99m)Tc-sestamibi](+) (0.10 microL/g/s). The CNS tracer [(99m)Tc-HMPAO](0) entered the brain approximately 70-times higher than [(99m)Tc(CO)(3)-ibogaine](+). In vitro studies revealed that neither [(99m)Tc(CO)(3)-ibogaine](+) nor (99m)Tc-tricarbonyl ion were substrates for P-gp or BCRP. But lowering the membrane dipole potential barrier with phloretin enhanced the brain transport of [(99m)Tc(OH(2))(3)(CO)(3)](+) approximately 3-fold. Thus, ibogaine directly labeled with (99m)Tc-tricarbonyl is not suitable for CNS imaging because of its poor uptake. Brain transport is not restricted by efflux transporters but is reduced by its lipophilicity and interaction with the membrane-positive dipole potential. PMID:19492342

  17. A role for activated endothelial cells in red blood cell clearance: implications for vasopathology

    DEFF Research Database (Denmark)

    Fens, Marcel H A M; van Wijk, Richard; Andringa, Grietje;

    2012-01-01

    Background Phosphatidylserine exposure by red blood cells is acknowledged as a signal that initiates phagocytic removal of the cells from the circulation. Several disorders and conditions are known to induce phosphatidylserine exposure. Removal of phosphatidylserine-exposing red blood cells gener...

  18. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... are most commonly used in the treatment of cancers like leukemia and lymphoma to restore stem cells ... use of BMT and PBSCT, see http://www.cancer.gov/cancertopics/fa... If you are interested in ...

  19. Transfusion management of patients with red blood cell antibodies

    Directory of Open Access Journals (Sweden)

    Bujandrić Nevenka B.

    2013-01-01

    Full Text Available Introduction. Red blood cell antibodies may cause a positive result of pre-transfusion blood compatibility testing (crossmatch test. It can be a problem to provide suitable blood units for patients with clinically significant antibodies to high-frequency antigens as well as for those with multiple alloantibody specificities. This study was aimed at identifying transfused patients in the population of South-Backa who had developed clinically significant red blood cell alloantibodies. Material and methods. We analyzed the records of crossmatch results and antibody screening performed at the Blood Transfusion Institute of Vojvodina during 2012. Results. Antibodies were found in 103 patients: A 63 patients with single antibodies: 1 16 with antibodies of unknown specificity (3 autoantibodies, 13 alloantibodies; 2 39 with clinically significant antibodies (23 from Rh system (2 anti-C, 2 anti-D, 12 anti-E, 7 anti-c, 4 anti-K, 3 anti-Fya, 7 anti-Jka, 2 anti-S; 3 8 with usually not significant antibodies (6 anti-M, 1 anti-A1, 1 anti- Cw; B 40 patients developed multiple antibodies: 1 all patients had at least one clinically significant antibody from various blood group system (44 Rh, 13 Kell, 7 Kidd, 7 MNSs (S, s; 2 3 patients had usually not significant antibodies (1 Lewis, 2 Lutheran; 3 3 patients occasionally had clinically significant antibody (3 anti- Yta; 4 3 patients had antibodies of unknown specificity (2 autoantibodies, 1alloantibody. Antibodies detected in the majority of patients (65-63.1% had a specificity of Rh and/or the Kell system. Conclusions. The main goal of pre-transfusion blood compatibility testing is to detect clinically significant antibodies. The provision of antigen negative blood units for those patients is a special challenge for blood establishments. Database with a sufficient number of typed blood donors can help to resolve this problem.

  20. Filtration parameters influencing circulating tumor cell enrichment from whole blood.

    Directory of Open Access Journals (Sweden)

    Frank A W Coumans

    Full Text Available Filtration can achieve circulating tumor cell (CTC enrichment from blood. Key parameters such as flow-rate, applied pressure, and fixation, vary largely between assays and their influence is not well understood. Here, we used a filtration system, to monitor these parameters and determine their relationships. Whole blood, or its components, with and without spiked tumor cells were filtered through track-etched filters. We characterize cells passing through filter pores by their apparent viscosity; the viscosity of a fluid that would pass with the same flow. We measured a ratio of 5·10(4∶10(2∶1 for the apparent viscosities of 15 µm diameter MDA-231 cells, 10 µm white cells and 90 fl red cells passing through a 5 µm pore. Fixation increases the pressure needed to pass cells through 8 µm pores 25-fold and halves the recovery of spiked tumor cells. Filtration should be performed on unfixed samples at a pressure of ∼10 mbar for a 1 cm(2 track-etched filter with 5 µm pores. At this pressure MDA-231 cells move through the filter in 1 hour. If fixation is needed for sample preservation, a gentle fixative should be selected. The difference in apparent viscosity between CTC and blood cells is key in optimizing recovery of CTC.

  1. In vivo MRI discrimination between live and lysed iron-labelled cells using balanced steady state free precession

    International Nuclear Information System (INIS)

    The goal of this study was to evaluate the ability of balanced steady state free precession (b-SSFP) magnetic resonance imaging sequence to distinguish between live and lysed iron-labelled cells. Human breast cancer cells were labelled with iron oxide nanoparticles. Cells were lysed using sonication. Imaging was performed at 3 T. The timing parameters for b-SSFP and the number of iron-labelled cells in samples were varied to optimise the b-SSFP signal difference between live and lysed iron-labelled cell samples. For in vivo experiments, cells were mixed with Matrigel and implanted into nude mice. Three mice implanted with live labelled cancer cells were irradiated to validate this method. Lysed iron-labelled cells have a significantly higher signal compared with live, intact iron-labelled cells in bSSFP images. The contrast between live and dead cells can be maximised by careful optimisation of timing parameters. A change in the b-SSFP signal was measured 6 days after irradiation, reflecting cell death in vivo. Histology confirmed the presence of dead cells in the implant. Our results show that the b-SSFP sequence can be optimised to allow for the discrimination of live iron-labelled cells and lysed iron-labelled cells in vitro and in vivo. (orig.)

  2. CD34+ stem cells from umbilical cord blood

    Directory of Open Access Journals (Sweden)

    Carlo Pafumi

    2011-10-01

    Full Text Available We describe the relation between umbilical cord clamping time and two different enrichment system of CD34+ stem cells from umbilical cord blood with the proliferative ability and bone marrow reconstitution of the stem cells obtained. After an obstetrician performed the cord blood collection, the purification of stem cells was performed either with a combination of monoclonal antibodies (negative selections using the Stem Sep method, or with a positive cells selection based on their surface CD34 antigens using the Mini Macs system. An excellent recovery of haematopoietic progenitors [Burst Forming Unit Erythroids (BFUE; Colony Forming Unit Granulocytes and Macrophages (CFU-GM; and Colony Forming Unit Granulocytes, Erythroids, Monocytes and Macrophages (CFU-GME], inversely related to the increase in clamping time, was performed with the Mini Macs system (54% of colonies, with 90% purity. With Stem Sep method, haematopoietic progenitor’s recovery was 35% (with 80% purity. By applying early clamping of umbilical cord blood we obtained a greater number of CD34+ cells and their clonogenic activity was increased with enrichment. This is a useful technique considering that the number of CD34+ stem cells usually contained from a unit of placental blood is enough for the transplant to a child, but not for an adult. Thus, using these methods, we can get a larger number of CD34+ stem cells which reduces the risk of Graft versus Host Disease also in adult patients, producing survival rates similar to those obtained with transplantation of bone marrow from unrelated donors.

  3. CD34+ stem cells from umbilical cord blood

    Directory of Open Access Journals (Sweden)

    Alfio D’Agati

    2011-09-01

    Full Text Available We describe the relation between umbilical cord clamping time and two different enrichment system of CD34+ stem cells from umbilical cord blood with the proliferative ability and bone marrow reconstitution of the stem cells obtained. After an obstetrician performed the cord blood collection, the purification of stem cells was performed either with a combination of monoclonal antibodies (negative selections using the Stem Sep method, or with a positive cells selection based on their surface CD34 antigens using the Mini Macs system. An excellent recovery of haematopoietic progenitors [Burst Forming Unit Erythroids (BFUE; Colony Forming Unit Granulocytes and Macrophages (CFU-GM; and Colony Forming Unit Granulocytes, Erythroids, Monocytes and Macrophages (CFU-GME], inversely related to the increase in clamping time, was performed with the Mini Macs system (54% of colonies, with 90% purity. With Stem Sep method, haematopoietic progenitor’s recovery was 35% (with 80% purity. By applying early clamping of umbilical cord blood we obtained a greater number of CD34+ cells and their clonogenic activity was increased with enrichment. This is a useful technique considering that the number of CD34+ stem cells usually contained from a unit of placental blood is enough for the transplant to a child, but not for an adult. Thus, using these methods, we can get a larger number of CD34+ stem cells which reduces the risk of Graft versus Host Disease also in adult patients, producing survival rates similar to those obtained with transplantation of bone marrow from unrelated donors.

  4. Peripheral blood derived cells and angiogenesis in cardiovascular disease

    OpenAIRE

    Post, S

    2009-01-01

    Patients suffering from myocardial infarction (MI), atherosclerosis and Hereditary Hemorrhagic Telangiectasia type 1 (HHT-1) all have diseased and dysfunctional blood vessels. Cardiovascular repair in these diseases occurs not only locally, but also peripheral blood (progenitor) cells and cytokines/growth factors positively contribute to repair of malfunctioning tissue. In this thesis several aspects of cardiovascular repair have been explored. First, we show that in MI patients relatively la...

  5. In-vitro red blood cell partitioning of doxycycline

    OpenAIRE

    Deshmukh, P.V.; Badgujar, P.C.; Gatne, M. M.

    2009-01-01

    Objective: In-vitro red blood cell (RBC) partitioning of doxycycline was studied to determine whether doxycycline penetrates RBC and its concentration was assayed keeping in view its high lipophilicity. Materials and Methods: Standardization of doxycycline was performed in whole blood and plasma of cattle by microbiological assay using Bacillus subtillis ATCC 6633 as indicator organizm. Actual concentration of the drug was obtained by comparing zone inhibition with standard graph and the exte...

  6. Bacterial glycosidases for the production of universal red blood cells

    DEFF Research Database (Denmark)

    Liu, Qiyong P; Sulzenbacher, Gerlind; Yuan, Huaiping;

    2007-01-01

    Enzymatic removal of blood group ABO antigens to develop universal red blood cells (RBCs) was a pioneering vision originally proposed more than 25 years ago. Although the feasibility of this approach was demonstrated in clinical trials for group B RBCs, a major obstacle in translating this...... alpha-N-acetylgalactosaminidase family reveals an unusual catalytic mechanism involving NAD+. The enzymatic conversion processes we describe hold promise for achieving the goal of producing universal RBCs, which would improve the blood supply while enhancing the safety of clinical transfusions....

  7. The migration of synthetic magnetic nanoparticle labeled dendritic cells into lymph nodes with optical imaging

    Directory of Open Access Journals (Sweden)

    Su H

    2013-10-01

    Full Text Available Hang Su,1,* Yongbin Mou,1,* Yanli An,2 Wei Han,1 Xiaofeng Huang,1 Guohua Xia,3 Yanhong Ni,1 Yu Zhang,4 Jianmin Ma,1 Qingang Hu1,5 1Center Laboratory of Stomatology, Stomatological Hospital Affiliated Medical School, Nanjing University, Nanjing, People's Republic of China; 2Jiangsu Key Lab of Molecular and Function Imaging, Department of Radiology; 3Department of Hematology, Zhongda Hospital, Medical School, 4State Key Laboratory of Molecule and Bimolecular Electronics, Jiangsu Provincial Laboratory for Biomaterials and Devices; Southeast University, Nanjing, People's Republic of China; 5Leeds Dental Institute, Faculty of Medicine and health, University of Leeds, Leeds, United Kingdom*These authors contributed equally to this workBackground: The successful biotherapy of carcinoma with dendritic cell (DC vaccines pivotally relies on DCs’ migratory capability into lymph tissues and activation of T cells. Accurate imaging and evaluation of DC migration in vivo have great significance during antitumor treatment with DC vaccine. We herein examined the behavior of DCs influenced by synthetic superparamagnetic iron oxide (SPIO nanoparticle labeling.Methods: γ-Fe2O3 nanoparticles were prepared and DCs, which were induced from bone marrow monocytes of enhanced green fluorescent protein (EGFP transgenic mice, were labeled. The endocytosis of the SPIO, surface molecules, cell apoptosis and fluorescence intensity of EGFP-DCs were displayed by Prussian blue staining and flow cytometry (FCM, respectively. After EGFP-DCs, labeled with SPIO, were injected into footpads (n = 5 for 24 hours, the mice were examined in vivo by optical imaging (OPI. Meanwhile, confocal imaging and FCM were applied, respectively, to detect the migration of labeled DCs into draining lymph nodes.Results: Nearly 100% of cells were labeled by the SPIO, in which the intracellular blue color gradually deepened and the iron contents rose with the increase of labeling iron concentrations

  8. Self-Assembled Superparamagnetic Iron Oxide Nanoclusters for Universal Cell Labeling and MRI

    Science.gov (United States)

    Chen, Shuzhen; Zhang, Jun; Jiang, Shengwei; Lin, Gan; Luo, Bing; Yao, Huan; Lin, Yuchun; He, Chengyong; Liu, Gang; Lin, Zhongning

    2016-05-01

    Superparamagnetic iron oxide (SPIO) nanoparticles have been widely used in a variety of biomedical applications, especially as contrast agents for magnetic resonance imaging (MRI) and cell labeling. In this study, SPIO nanoparticles were stabilized with amphiphilic low molecular weight polyethylenimine (PEI) in an aqueous phase to form monodispersed nanocomposites with a controlled clustering structure. The iron-based nanoclusters with a size of 115.3 ± 40.23 nm showed excellent performance on cellular uptake and cell labeling in different types of cells, moreover, which could be tracked by MRI with high sensitivity. The SPIO nanoclusters presented negligible cytotoxicity in various types of cells as detected using MTS, LDH, and flow cytometry assays. Significantly, we found that ferritin protein played an essential role in protecting stress from SPIO nanoclusters. Taken together, the self-assembly of SPIO nanoclusters with good magnetic properties provides a safe and efficient method for universal cell labeling with noninvasive MRI monitoring capability.

  9. SMIM1 underlies the Vel blood group and influences red blood cell traits

    DEFF Research Database (Denmark)

    Cvejic, Ana; Haer-Wigman, Lonneke; Stephens, Jonathan C;

    2013-01-01

    The blood group Vel was discovered 60 years ago, but the underlying gene is unknown. Individuals negative for the Vel antigen are rare and are required for the safe transfusion of patients with antibodies to Vel. To identify the responsible gene, we sequenced the exomes of five individuals negative...... and expression of the Vel antigen on SMIM1-transfected cells confirm SMIM1 as the gene underlying the Vel blood group. An expression quantitative trait locus (eQTL), the common SNP rs1175550 contributes to variable expression of the Vel antigen (P = 0.003) and influences the mean hemoglobin concentration of red...... blood cells (RBCs; P = 8.6 × 10(-15)). In vivo, zebrafish with smim1 knockdown showed a mild reduction in the number of RBCs, identifying SMIM1 as a new regulator of RBC formation. Our findings are of immediate relevance, as the homozygous presence of the deletion allows the unequivocal identification...

  10. Separation of cancer cells from white blood cells by pinched flow fractionation

    DEFF Research Database (Denmark)

    Jensen, Marie Pødenphant; Ashley, Neil; Koprowska, Kamila; Mir, Kalim U.; Zalkovskij, Maksim; Bilenberg, Brian; Bodmer, Walter; Kristensen, Anders; Marie, Rodolphe

    2015-01-01

    In this paper, the microfluidic size-separation technique pinched flow fractionation (PFF) is used to separate cancer cells from white blood cells (WBCs). The cells are separated at efficiencies above 90% for both cell types. Circulating tumor cells (CTCs) are found in the blood of cancer patients...... challenged by the size overlap between cancer cells and the 106 times more abundant WBCs. The size overlap prevents high efficiency separation, however we demonstrate that cell deformability can be exploited in PFF devices to gain higher efficiencies than expected from the size distribution of the cells....... and can form new tumors. CTCs are rare cells in blood, but they are important for the understanding of metastasis. There is therefore a high interest in developing a method for the enrichment of CTCs from blood samples, which also enables further analysis of the separated cells. The separation is...

  11. Acute hydrodynamic damage induced by SPLITT fractionation and centrifugation in red blood cells.

    Science.gov (United States)

    Urbina, Adriana; Godoy-Silva, Ruben; Hoyos, Mauricio; Camacho, Marcela

    2016-05-01

    Though blood bank processing traditionally employs centrifugation, new separation techniques may be appealing for large scale processes. Split-flow fractionation (SPLITT) is a family of techniques that separates in absence of labelling and uses very low flow rates and force fields, and is therefore expected to minimize cell damage. However, the hydrodynamic stress and possible consequent damaging effects of SPLITT fractionation have not been yet examined. The aim of this study was to investigate the hydrodynamic damage of SPLITT fractionation to human red blood cells, and to compare these effects with those induced by centrifugation. Peripheral whole blood samples were collected from healthy volunteers. Samples were diluted in a buffered saline solution, and were exposed to SPLITT fractionation (flow rates 1-10ml/min) or centrifugation (100-1500g) for 10min. Cell viability, shape, diameter, mean corpuscular hemoglobin, and membrane potential were measured. Under the operating conditions employed, both SPLITT and centrifugation maintained cell viability above 98%, but resulted in significant sublethal damage, including echinocyte formation, decreased cell diameter, decreased mean corpuscular hemoglobin, and membrane hyperpolarization which was inhibited by EGTA. Wall shear stress and maximum energy dissipation rate showed significant correlation with lethal and sublethal damage. Our data do not support the assumption that SPLITT fractionation induces very low shear stress and is innocuous to cell function. Some changes in SPLITT channel design are suggested to minimize cell damage. Measurement of membrane potential and cell diameter could provide a new, reliable and convenient basis for evaluation of hydrodynamic effects on different cell models, allowing identification of optimal operating conditions on different scales. PMID:27023157

  12. Aggregation of Red Blood Cells: From Rouleaux to Clot Formation

    OpenAIRE

    Wagner, C.; Steffen, P.; Svetina, S

    2013-01-01

    Red blood cells are known to form aggregates in the form of rouleaux. This aggregation process is believed to be reversible, but there is still no full understanding on the binding mechanism. There are at least two competing models, based either on bridging or on depletion. We review recent experimental results on the single cell level and theoretical analyses of the depletion model and of the influence of the cell shape on the binding strength. Another important aggregation mechanism is caus...

  13. Labeling human embryonic stem-cell-derived cardiomyocytes for tracking with MR imaging

    International Nuclear Information System (INIS)

    Human embryonic stem cells (hESC) can generate cardiomyocytes (CM), which offer promising treatments for cardiomyopathies in children. However, challenges for clinical translation result from loss of transplanted cell from target sites and high cell death. An imaging technique that noninvasively and repetitively monitors transplanted hESC-CM could guide improvements in transplantation techniques and advance therapies. To develop a clinically applicable labeling technique for hESC-CM with FDA-approved superparamagnetic iron oxide nanoparticles (SPIO) by examining labeling before and after CM differentiation. Triplicates of hESC were labeled by simple incubation with 50 μg/ml of ferumoxides before or after differentiation into CM, then imaged on a 7T MR scanner using a T2-weighted multi-echo spin-echo sequence. Viability, iron uptake and T2-relaxation times were compared between groups using t-tests. hESC-CM labeled before differentiation demonstrated significant MR effects, iron uptake and preserved function. hESC-CM labeled after differentiation showed no significant iron uptake or change in MR signal (P < 0.05). Morphology, differentiation and viability were consistent between experimental groups. hESC-CM should be labeled prior to CM differentiation to achieve a significant MR signal. This technique permits monitoring delivery and engraftment of hESC-CM for potential advancements of stem cell-based therapies in the reconstitution of damaged myocardium. (orig.)

  14. Labeling human embryonic stem-cell-derived cardiomyocytes for tracking with MR imaging

    Energy Technology Data Exchange (ETDEWEB)

    Castaneda, Rosalinda T.; Daldrup-Link, Heike [Lucile Packard Children' s Hospital, Stanford School of Medicine, Pediatric Radiology, Stanford, CA (United States); Boddington, Sophie; Wendland, Mike; Mandrussow, Lydia [University of California, Department of Radiology and Biomedical Imaging, UCSF Medical Center, San Francisco, CA (United States); Henning, Tobias D. [University Hospital of Cologne, Department of Radiology and Neuroradiology, Cologne (Germany); Liu, Siyuan [National Institutes of Health, Language Section, Voice, Speech and Language Branch, National Institute on Deafness and Other Communication Disorders, Bethesda, MD (United States)

    2011-11-15

    Human embryonic stem cells (hESC) can generate cardiomyocytes (CM), which offer promising treatments for cardiomyopathies in children. However, challenges for clinical translation result from loss of transplanted cell from target sites and high cell death. An imaging technique that noninvasively and repetitively monitors transplanted hESC-CM could guide improvements in transplantation techniques and advance therapies. To develop a clinically applicable labeling technique for hESC-CM with FDA-approved superparamagnetic iron oxide nanoparticles (SPIO) by examining labeling before and after CM differentiation. Triplicates of hESC were labeled by simple incubation with 50 {mu}g/ml of ferumoxides before or after differentiation into CM, then imaged on a 7T MR scanner using a T2-weighted multi-echo spin-echo sequence. Viability, iron uptake and T2-relaxation times were compared between groups using t-tests. hESC-CM labeled before differentiation demonstrated significant MR effects, iron uptake and preserved function. hESC-CM labeled after differentiation showed no significant iron uptake or change in MR signal (P < 0.05). Morphology, differentiation and viability were consistent between experimental groups. hESC-CM should be labeled prior to CM differentiation to achieve a significant MR signal. This technique permits monitoring delivery and engraftment of hESC-CM for potential advancements of stem cell-based therapies in the reconstitution of damaged myocardium. (orig.)

  15. Counting White Blood Cells from a Blood Smear Using Fourier Ptychographic Microscopy

    OpenAIRE

    Chung, Jaebum; Ou, Xiaoze; Kulkarni, Rajan P.; Yang, Changhuei

    2015-01-01

    White blood cell (WBC) count is a valuable metric for assisting with diagnosis or prognosis of various diseases such as coronary heart disease, type 2 diabetes, or infection. Counting WBCs can be done either manually or automatically. Automatic methods are capable of counting a large number of cells to give a statistically more accurate reading of the WBC count of a sample, but the specialized equipment tends to be expensive. Manual methods are inexpensive since they only involve a convention...

  16. Magnetic labeling and in vitro MR imaging of rat bone marrow mesenchymal stem cells

    International Nuclear Information System (INIS)

    Objective: To label rat bone marrow mesenchymal stem cells with feridex combined with poly-l-lysine (PLL), and to determine the feasibility of detection of magnetically labeled stem cells with MR imaging. Methods: Feridex were incubated with PLL for 1 hour to obtain a complex of feridex-PLL. Mesenchymal stem cells isolated from the bone marrows of Wistar rats were cultured and expanded. By the 4th passage, cells were co-incubated overnight with the feridex-PLL complex. Prussian blue staining for demonstrating intracytoplastic nanoparticles and trypan-blue exclusion test for cell viability were performed respectively at 24 h, 1 w, 2 w, 3 w after labeling. MR imaging of cell suspensions was performed by using T1WI, T2WI and T2* WI sequences at a clinical 1.5 T MR system. Results: Numerous intracytoplastic iron particles were stained with Prussian blue. With division of stern cells, the stained particles were seen decreased gradually. Trypan blue exclusion test at 24 h, 1 w, 2 w and 3 w showed that the viability of the labeled cells was 91.00%, 93.00%, 91.75%, and 92.50%, not significantly different with that of nonlabeled cells (P>0.05). For 103, 104 and l05 cells, T2 signal intensity decreased by 63.75%, 82.31% and 91.92% respectively, T2* signal intensity decreased by 68.24%, 83.01%, and 93.94% respectively. For 105 labeled cells, T2* signal intensity decreased by 93.75%, 75.92%, 41.75% and 8.83 % respectively at 24 h, 1 w, 2 w and 3 w after labeling. Conclusion: Magnetic labeling of rat bone marrow stem cells with feridex-PLL complex is feasible, efficient and safe. T2* WI is the most sensitive sequence to detect the labeled cells. The degree of T2 signal decreasing may be related to the cell count and division phase. (authors)

  17. Depletion induced clustering of red blood cells in microchannels

    Science.gov (United States)

    Wagner, Christian; Brust, Mathias; Podgorski, Thomas; Coupier, Gwennou

    2012-11-01

    The flow properties of blood are determined by the physical properties of its main constituents, the red blood cells (RBC's). At low shear rates RBC's form aggregates, so called rouleaux. Higher shear rates can break them up and the viscosity of blood shows a shear thinning behavior. The physical origin of the rouleaux formation is not yet fully resolved and there are two competing models available. One predicts that the adhesion is induced by bridging of the plasma (macromolecular) proteins in-between two RBC's. The other is based on the depletion effect and thus predicts the absence of macromolecules in-between the cells of a rouleaux. Recent single cell force measurements by use of an AFM support strongly the depletion model. By varying the concentration of Dextran at different molecular weights we can control the adhesions strength. Measurements at low hematocrit in a microfluidic channel show that the number of size of clusters is determined by the depletion induced adhesion strength.

  18. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... a Patient-Centered Approach - Duration: 4:12. NCIcancertopics 3,087 views 4:12 The Truth About Cord ... 19 Stem cell donation: Step by step - Duration: 3:35. hemaquebec1998 1,127 views 3:35 Two ...

  19. Becoming a Blood Stem Cell Donor

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    Full Text Available ... 13:41. BOOKparty! 1,367 views 13:41 Bone marrow transplantation, donation procedure (HD, ENG subtitles) - Duration: 8:21. Marcin Ostajewski 155,257 views 8:21 Pain Control: Support for People with Cancer - Duration: 11:58. ... Bone Marrow/Stem Cell Transplant - Duration: 7:24. tannermom80 99,818 views 7: ...

  20. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... be donors at http://www.marrow.org . Category Science & Technology License Standard YouTube License Show more Show ... 41. Annabelle Monks 3,487 views 4:41 Science Friction: Stem Cell Research - Duration: 54:44. Irishstemcell ...

  1. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... be donors at http://www.marrow.org . Category Science & Technology License Standard YouTube License Show more Show ... views 4:25 Susan Solomon: The promise of research with stem cells - Duration: 14:59. TED 55, ...

  2. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... playlist. Sign in Share More Report Need to report the video? Sign in to report inappropriate content. Sign in Transcript 6,983 views ... Stem Cell Therapy Injections - Duration: 6:18. Caring Medical Regenerative Medicine Clinics 234,106 views 6:18 ...

  3. Becoming a Blood Stem Cell Donor

    Medline Plus

    Full Text Available ... views 4:25 Susan Solomon: The promise of research with stem cells - Duration: 14:59. TED 54, ... 1:04 Pain Control: Support for People with Cancer - Duration: 11:58. NCIcancertopics 1,987 views 11: ...

  4. Red blood cell-derived microparticles: An overview.

    Science.gov (United States)

    Westerman, Maxwell; Porter, John B

    2016-07-01

    The red blood cell (RBC) is historically the original parent cell of microparticles (MPs). In this overview, we describe the discovery and the early history of red cell-derived microparticles (RMPs) and present an overview of the evolution of RMP. We report the formation, characteristics, effects of RMP and factors which may affect RMP evaluation. The review examines RMP derived from both normal and pathologic RBC. The pathologic RBC studies include sickle cell anemia (SCA), sickle cell trait (STr), thalassemia intermedia (TI), hereditary spherocytosis (HS), hereditary elliptocytosis (HE), hereditary stomatocytosis (HSt) and glucose-6-phosphate dehydrogenase deficiency (G6PD). PMID:27282583

  5. Variability of the thymidine labeling index in squamous cell carcinoma of the head and neck

    Energy Technology Data Exchange (ETDEWEB)

    Greenberg, B.; Woo, L.; Blatchford, S.; Aguirre, M.; Garewal, H.

    1988-06-01

    Tritiated thymidine (/sup 3/HTdR) labeling is the standard technique for determining the kinetic activity of tumors. This method has been used to label multiple sections of tumor specimens obtained from seven patients with advanced squamous cell carcinoma of the head and neck. Considerable variability was observed in the labeling index in different sites from the same specimen. To reduce the large sampling error due to heterogeneity, we recommend that an average value be determined from multiple sections when employing this technique.

  6. Magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes

    Science.gov (United States)

    Busato, Alice; Bonafede, Roberta; Bontempi, Pietro; Scambi, Ilaria; Schiaffino, Lorenzo; Benati, Donatella; Malatesta, Manuela; Sbarbati, Andrea; Marzola, Pasquina; Mariotti, Raffaella

    2016-01-01

    Purpose Recent findings indicate that the beneficial effects of adipose stem cells (ASCs), reported in several neurodegenerative experimental models, could be due to their paracrine activity mediated by the release of exosomes. The aim of this study was the development and validation of an innovative exosome-labeling protocol that allows to visualize them with magnetic resonance imaging (MRI). Materials and methods At first, ASCs were labeled using ultrasmall superparamagnetic iron oxide nanoparticles (USPIO, 4–6 nm), and optimal parameters to label ASCs in terms of cell viability, labeling efficiency, iron content, and magnetic resonance (MR) image contrast were investigated. Exosomes were then isolated from labeled ASCs using a standard isolation protocol. The efficiency of exosome labeling was assessed by acquiring MR images in vitro and in vivo as well as by determining their iron content. Transmission electron microscopy images and histological analysis were performed to validate the results obtained. Results By using optimized experimental parameters for ASC labeling (200 µg Fe/mL of USPIO and 72 hours of incubation), it was possible to label 100% of the cells, while their viability remained comparable to unlabeled cells; the detection limit of MR images was of 102 and 2.5×103 ASCs in vitro and in vivo, respectively. Exosomes isolated from previously labeled ASCs retain nanoparticles, as demonstrated by transmission electron microscopy images. The detection limit by MRI was 3 µg and 5 µg of exosomes in vitro and in vivo, respectively. Conclusion We report a new approach for labeling of exosomes by USPIO that allows detection by MRI while preserving their morphology and physiological characteristics. PMID:27330291

  7. Nanoparticle encapsulation in red blood cells enables blood-pool magnetic particle imaging hours after injection

    International Nuclear Information System (INIS)

    Magnetic particle imaging (MPI) is a new medical imaging approach that is based on the nonlinear magnetization response of super-paramagnetic iron oxide nanoparticles (SPIOs) injected into the blood stream. To date, real-time MPI of the bolus passage of an approved MRI SPIO contrast agent injected into the tail vein of living mice has been demonstrated. However, nanoparticles are rapidly removed from the blood stream by the mononuclear phagocyte system. Therefore, imaging applications for long-term monitoring require the repeated administration of bolus injections, which complicates quantitative comparisons due to the temporal variations in concentration. Encapsulation of SPIOs into red blood cells (RBCs) has been suggested to increase the blood circulation time of nanoparticles. This work presents first evidence that SPIO-loaded RBCs can be imaged in the blood pool of mice several hours after injection using MPI. This finding is supported by magnetic particle spectroscopy performed to quantify the iron concentration in blood samples extracted from the mice 3 and 24 h after injection of SPIO-loaded RBCs. Based on these results, new MPI applications can be envisioned, such as permanent 3D real-time visualization of the vessel tree during interventional procedures, bleeding monitoring after stroke, or long-term monitoring and treatment control of cardiovascular diseases. (paper)

  8. CD163 positive subsets of blood dendritic cells

    DEFF Research Database (Denmark)

    Maniecki, Maciej Bogdan; Møller, Holger Jon; Moestrup, Søren Kragh; Møller, Bjarne Kuno

    CD163 and CD91 are scavenging receptors with highly increased expression during the differentiation of monocytes into the anti-inflammatory macrophage phenotype. In addition, CD91 is expressed in monocyte-derived dendritic cells (MoDCs), where the receptor is suggested to be important for...... internalization of CD91-targeted antigens to be presented on the dendritic cell surface for T-cell stimulation. Despite their overlap in functionality, the expression of CD91 and CD163 has never been compared and the expression of CD163 in the monocyte-dendritic cell lineage is not yet characterized. CD163...... expression in dendritic cells (DCs) was investigated using multicolor flow cytometry in peripheral blood from 31 healthy donors and 15 HIV-1 patients in addition to umbilical cord blood from 5 newborn infants. Total RNA was isolated from MACS purified DCs and CD163 mRNA was determined with real-time reverse...

  9. The DNA methylome of human peripheral blood mononuclear cells

    DEFF Research Database (Denmark)

    Li, Yingrui; Zhu, Jingde; Tian, Geng;

    2010-01-01

    DNA methylation plays an important role in biological processes in human health and disease. Recent technological advances allow unbiased whole-genome DNA methylation (methylome) analysis to be carried out on human cells. Using whole-genome bisulfite sequencing at 24.7-fold coverage (12.3-fold per...... strand), we report a comprehensive (92.62%) methylome and analysis of the unique sequences in human peripheral blood mononuclear cells (PBMC) from the same Asian individual whose genome was deciphered in the YH project. PBMC constitute an important source for clinical blood tests world-wide. We found...

  10. Detection of hepatitis B virus DNA in mononuclear blood cells.

    OpenAIRE

    Pontisso, P; Poon, M C; Tiollais, P.; Brechot, C

    1984-01-01

    The Southern transfer hybridisation technique was used to test mononuclear blood cells for hepatitis B virus DNA. Viral DNA sequences were detected in mononuclear cells of 10 out of 16 patients with hepatitis B virus infection and in none of 21 normal controls. Blood contamination was excluded by the absence of hepatitis B virus DNA in the corresponding serum samples in all cases. Free monomeric hepatitis B virus DNA was found in three patients positive for hepatitis Be antigen (HBeAg) and on...

  11. Procedure for the fluorographic determination of radioactivity of labelled macromolecules and cells

    International Nuclear Information System (INIS)

    The invention aims at a procedure for the quantitative determination of radioactivity of labelled macromolecules, viruses and cells in water suspensions by means of fluorographic methods. It is applicable to routine diagnosis in medicine and veterinary medicine as well as to studies in molecular biology and biochemistry. The procedure is characterized by fixing the labelled macromolecules as circular spots on the filter as evidenced by film blackening

  12. Mesenchymal stem cell in vitro labeling by hybrid fluorescent magnetic polymeric particles for application in cell tracking.

    Science.gov (United States)

    Supokawej, Aungkura; Nimsanor, Natakarn; Sanvoranart, Tanwarat; Kaewsaneha, Chariya; Hongeng, Suradej; Tangboriboonrat, Pramuan; Jangpatarapongsa, Kulachart

    2015-12-01

    Mesenchymal stem cells (MSCs) are a type of adult stem cell that contains multi-differentiation and proliferative properties and that shows high treatment implications for many clinical problems. The outcome of stem cell transplantation is still limited due to many factors, especially their survival and their interaction with the microenvironment after transplantation. Molecular imaging is a challenging technique that has been used to overcome this limitation and is based on the concept of labeling cells with tractable, visible, and non-toxic materials to track the cells after transplantation. In this study, magnetic polymeric nanoparticles (MPNPs) were used to directly label Wharton's jelly-derived MSCs (WJ-MSCs). After labeling, the growth rate and the viability of the MSCs as well as the time of exposure were determined. The 3D images of WJ-MSCs labeled with MPNPs for 24 h were created using confocal microscopy. The results showed that, after incubation with fluorescent MPNPs for over 8 h, the growth rate and cell viability of the WJ-MSCs was similar to those of the control. Three-dimensional imaging revealed that the fluorescent MPNPs could infiltrate into the cells and spread into the cytoplasm, which suggests that the synthesized fluorescent MPNPs could possibly label MSCs for cell tracking study and be further developed for in vivo applications. PMID:25893425

  13. Electron microscope analysis of young and old red blood cells stained with colloidal iron for surface charge evaluation.

    Science.gov (United States)

    Marikovsky, Y; Danon, D

    1969-10-01

    Human and rabbit red blood cells, separated into "young" and "old" age groups by differential flotation on phthalate esters, were fixed with glutaraldehyde and labeled with colloidal ferric oxide. Electron micrographs of thin sections of young cells showed a uniform and dense depostion of positive iron particles. Old cells showed particles deposited irregularly, leaving unlabeled gaps on the membrane surface. Red cells incubated with 10 units/ml receptor-destroying enzyme (RDE) demonstrate a reduced labeling, similar to that of old cells. After neuraminic acid had been removed from red cells by 20 units/ml RDE, no iron particles were found on membrane surfaces. The different labeling of young, old, and RDE-treated human and rabbit red cells was correlated with their electric mobility and agglutinability by poly-L-lysine. The contradiction between the apparent similarity in charge density of human and rabbit red cells as estimated by density of iron particles and the markedly lower electric mobility of rabbit red cells is discussed. PMID:4186411

  14. Blood cells and endothelial barrier function

    OpenAIRE

    Rodrigues, Stephen F.; Granger, D Neil

    2015-01-01

    The barrier properties of endothelial cells are critical for the maintenance of water and protein balance between the intravascular and extravascular compartments. An impairment of endothelial barrier function has been implicated in the genesis and/or progression of a variety of pathological conditions, including pulmonary edema, ischemic stroke, neurodegenerative disorders, angioedema, sepsis and cancer. The altered barrier function in these conditions is often linked to the release of solub...

  15. Nanostructured Substrates for Capturing Circulating Tumor Cells in Whole Blood

    Science.gov (United States)

    Tseng, Hsian-Rong

    2009-03-01

    Over the past decade, circulating tumor cells (CTCs) has become an emerging ``biomarker'' for detecting early-stage cancer metastasis, predicting patient prognosis, as well as monitoring disease progression and therapeutic outcomes. However, isolation of CTCs has been technically challenging due to the extremely low abundance (a few to hundreds per ml) of CTCs among a high number of hematologic cells (109 per mL) in the blood. Our joint research team at UCLA has developed a new cell capture technology for quantification of CTCs in whole blood samples. Similar to most of the existing approaches, epithelial cell adhesion molecule antibody (anti-EpCAM) was grafted onto the surfaces to distinguish CTCs from the surrounding hematologic cells. The uniqueness of our technology is the use of nanostructured surfaces, which facilitates local topographical interactions between CTCs and substrates at the very first cell/substrate contacting time point. We demonstrated the ability of these nanostructured substrates to capture CTCs in whole blood samples with significantly improved efficiency and selectivity. The successful demonstration of this cell capture technology using brain, breast and prostate cancer cell lines encouraged us to test this approach in clinical setting. We have been able to bond our first validation study with a commercialized technology based on the use of immunomagnetic nanoparticles. A group of clinically well-characterized prostate cancer patients at UCLA hospital have been recruited and tested in parallel by these two technologies.

  16. Consequences of the magnetic field, sonic and radiofrequency waves and intense pulsed light on the labeling of blood constituents with technetium-99m

    International Nuclear Information System (INIS)

    Sources of magnetic field, radiofrequency and audible sonic waves and pulsed light have been used in physiotherapy to treat different disorders. In nuclear medicine, blood constituents(Bl-Co) are labeled with technetium-99m (99mTc) are used. This study evaluated the consequences of magnetic field, radiofrequency and audible sonic waves and intense pulsed light sources on the labeling of Bl-Co with 99mTc. Blood from Wistar rats was exposed to the cited sources. The labeling of Bl-Co with 99mTc was performed. Blood not exposed to the physical agents was used(controls). Data showed that the exposure to the different studied sources did not alter significantly (p>0.05) the labeling of Bl-Co. Although the results were obtained with animals, the data suggest that no alteration on examinations performed with Bl-Co labeled with 99mTc after exposition to the cited agents. The biological consequences associated with these agents would be not capable to interfere with some properties of the Bl-Co. (author)

  17. Consequences of the magnetic field, sonic and radiofrequency waves and intense pulsed light on the labeling of blood constituents with technetium-99m

    Energy Technology Data Exchange (ETDEWEB)

    Meyer, Patricia Froes; Costa, Iris do Ceu Clara; Brandao-Neto, Jose; Medeiros, Aldo da Cunha [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil). Programa de Pos-graduacao em Ciencias da Saude; Santos-Filho, Sebastiao David; Adenilson de Souza da Fonseca; Bernardo-Filho, Mario [Universidade do Estado do Rio de Janeiro (UERJ), RJ (Brazil). Inst. de Biologia Roberto Alcantara Gomes. Lab. de Radiofarmacia Experimental; Ariel Ronzio, Oscar [Universidad de Buenos Aires (Argentina); Bonelli, Ludmila [Universidade Salgado de Oliveira, Belo Horizonte, MG (Brazil)

    2007-09-15

    Sources of magnetic field, radiofrequency and audible sonic waves and pulsed light have been used in physiotherapy to treat different disorders. In nuclear medicine, blood constituents(Bl-Co) are labeled with technetium-99m ({sup 99m}Tc) are used. This study evaluated the consequences of magnetic field, radiofrequency and audible sonic waves and intense pulsed light sources on the labeling of Bl-Co with {sup 99m}Tc. Blood from Wistar rats was exposed to the cited sources. The labeling of Bl-Co with {sup 99m}Tc was performed. Blood not exposed to the physical agents was used(controls). Data showed that the exposure to the different studied sources did not alter significantly (p>0.05) the labeling of Bl-Co. Although the results were obtained with animals, the data suggest that no alteration on examinations performed with Bl-Co labeled with {sup 99m}Tc after exposition to the cited agents. The biological consequences associated with these agents would be not capable to interfere with some properties of the Bl-Co. (author)

  18. Labeling cells in microtiter plates for determination of [3H]thymidine uptake.

    Science.gov (United States)

    Shevach, E M

    2001-05-01

    A number of protocols in Current Protocols in Immunology use as their end-point the determination of cell proliferation by determining the incorporation of [(3)H]thymidine into cellular DNA. This appendix presents a protocol in which the radioactive label is added during the last 4 to 24 hr of the culture. A semiautomated cell harvesting apparatus is then used to lyse the cells with water and precipitate the labeled DNA on glass fiber filters. The filter pads are then dried and counted by standard liquid scintillation counting techniques in a scintillation counter. PMID:18432656

  19. Monitoring Protein Synthesis in Living Cells with Fluorescent Labeled tRNA FRET Pairs

    OpenAIRE

    Barhoom, Sima; Farrel, Ian; Dahary, Dvir; Ehrlich, Marcelo; Cooperman, Barry S.; Elroy-Stein, Orna; Smilansky, Zeev

    2013-01-01

    We introduce Protein Synthesis Monitoring (PSM) – a technique to monitor protein synthesis in live cells. In PSM, we transfect cells with tRNA labeled as FRET donors and acceptors. A FRET signal is generated only when a donor- and an acceptor-labeled tRNA come in close contact (< 7nM), as they do on the ribosome during elongation. The intensity of the FRET signal correlates with the number of ribosomes engaged in protein synthesis, providing a real-time, live-cell assay for measuring rates of...

  20. Near-infrared imaging of adoptive immune cell therapy in breast cancer model using cell membrane labeling.

    Directory of Open Access Journals (Sweden)

    Fatma M Youniss

    Full Text Available The overall objective of this study is to non-invasively image and assess tumor targeting and retention of directly labeled T-lymphocytes following their adoptive transfer in mice. T-lymphocytes obtained from draining lymph nodes of 4T1 (murine breast cancer cell sensitized BALB/C mice were activated in-vitro with Bryostatin/Ionomycin for 18 hours, and were grown in the presence of Interleukin-2 for 6 days. T-lymphocytes were then directly labeled with 1,1-dioctadecyltetramethyl indotricarbocyanine Iodide (DiR, a lipophilic near infrared fluorescent dye that labels the cell membrane. Assays for viability, proliferation, and function of labeled T-lymphocytes showed that they were unaffected by DiR labeling. The DiR labeled cells were injected via tail vein in mice bearing 4T1 tumors in the flank. In some cases labeled 4T1 specific T-lymphocytes were injected a week before 4T1 tumor cell implantation. Multi-spectral in vivo fluorescence imaging was done to subtract the autofluorescence and isolate the near infrared signal carried by the T-lymphocytes. In recipient mice with established 4T1 tumors, labeled 4T1 specific T-lymphocytes showed marked tumor retention, which peaked 6 days post infusion and persisted at the tumor site for up to 3 weeks. When 4T1 tumor cells were implanted 1-week post-infusion of labeled T-lymphocytes, T-lymphocytes responded to the immunologic challenge and accumulated at the site of 4T1 cell implantation within two hours and the signal persisted for 2 more weeks. Tumor accumulation of labeled 4T1 specific T-lymphocytes was absent in mice bearing Meth A sarcoma tumors. When lysate of 4T1 specific labeled T-lymphocytes was injected into 4T1 tumor bearing mice the near infrared signal was not detected at the tumor site. In conclusion, our validated results confirm that the near infrared signal detected at the tumor site represents the DiR labeled 4T1 specific viable T-lymphocytes and their response to immunologic challenge

  1. Label-Free Quantitative Proteomics and N-terminal Analysis of Human Metastatic Lung Cancer Cells

    OpenAIRE

    Min, Hophil; Han, Dohyun; Kim, Yikwon; Cho, Jee Yeon; Jin, Jonghwa; Kim, Youngsoo

    2014-01-01

    Proteomic analysis is helpful in identifying cancer-associated proteins that are differentially expressed and fragmented that can be annotated as dysregulated networks and pathways during metastasis. To examine meta-static process in lung cancer, we performed a proteomics study by label-free quantitative analysis and N-terminal analysis in 2 human non-small-cell lung cancer cell lines with disparate metastatic potentials—NCI-H1703 (primary cell, stage I) and NCI-H1755 (metastatic cell, stage ...

  2. Cytochemical characteristics of blood cells from Brazilian tortoises (Testudines: Testudinidae).

    Science.gov (United States)

    Martins, G S; Alevi, K C C; Azeredo-Oliveira, M T V; Bonini-Domingos, C R

    2016-01-01

    The hematology of wild and captive animals is essential for obtaining details about species and represents a simple method of diagnosing disease and determining prognosis. Few studies have described the morphology of chelonian blood cells, which are more common in sea and freshwater turtle species. Thus, in order to further our understanding and recognition of different chelonian cells types, the present study aimed to describe blood cells from the two species of Brazilian tortoises, Chelonoidis carbonarius and C. denticulatus. Cytochemical analysis of tortoise blood tissue with Panótico®, made it possible to describe all the of the chelonian cell types (with the exception of thrombocytes): erythrocytes, agranular leukocytes (monocytes and lymphocytes), and granular leukocytes (eosinophils, heterophils, basophils, and azurophils). These data are of high importance for establishing hematological profiles of Brazilian tortoises and reptiles. Therefore, based on our results and on comparative analyses with data from the literature for other reptile species, we can conclude that the blood cells described for Brazilian tortoises are found in all species of reptiles that have been analyzed thus far, and may be characterized and used as a comparative parameter between different groups to evaluate the health status of these animals. PMID:27050968

  3. Thrombin regulates the function of human blood dendritic cells

    International Nuclear Information System (INIS)

    Thrombin is the key enzyme in the coagulation cascade and activates endothelial cells, neutrophils and monocytes via protease-activated receptors (PARs). At the inflammatory site, immune cells have an opportunity to encounter thrombin. However little is known about the effect of thrombin for dendritic cells (DC), which are efficient antigen-presenting cells and play important roles in initiating and regulating immune responses. The present study revealed that thrombin has the ability to stimulate blood DC. Plasmacytoid DC (PDC) and myeloid DC (MDC) isolated from PBMC expressed PAR-1 and released MCP-1, IL-10, and IL-12 after thrombin stimulation. Unlike blood DC, monocyte-derived DC (MoDC), differentiated in vitro did not express PAR-1 and were unresponsive to thrombin. Effects of thrombin on blood DC were significantly diminished by the addition of anti-PAR-1 Ab or hirudin, serine protease inhibitor. Moreover, thrombin induced HLA-DR and CD86 expression on DC and the thrombin-treated DC induced allogenic T cell proliferation. These findings indicate that thrombin plays a role in the regulation of blood DC functions

  4. Prevention of diabetic microangiopathy by prophylactic transplant of mobilized peripheral blood mononuclear cells

    Institute of Scientific and Technical Information of China (English)

    Bin ZHOU; Xiao-cang CAO; Zhi-hong FANG; Cui-lin ZHENG; Zhi-bo HAN; He REN; Man-chiu POON; Zhong-chao HAN

    2007-01-01

    Aim: To investigate whether the prophylactic local delivery of mobilized periph-eral blood mononuclear cells (M-PBMNC) could prevent peripheral microangio-pathy in diabetic nude mice. Methods: Diabetic nude mice were induced with intraperitoneal injections of streptozotocin. With the time course of diabetes, we detected the capillary and arteriole density of mice adductor muscles by immuno-histopathy. In situ apoptosis was detected by using TdT-mediated dUTP nick end labeling (TUNEL) methods. M-PBMNC were labeled and locally delivered to the adductor muscles. Mononuclear cells were also isolated and cultured in vitro for the detection and counting of endothelial progenitor cells(EPC). Results: Rarefication of capillaries and arterioles, enhanced apoptosis in adductor muscles,and reduced circulating EPC in diabetic nude mice. Prophylactic local delivery of M-PBMNC halted the progression of microvascular rarefaction in hind-limb skel-etal muscles by inhibiting apoptosis. We detected the survival, migration and incorporation of transplanted M-PBMNC into the murine vasculature in vivo. In addition, more EPC were available from M-PBMNC than non-mobilized cells.Conclusion: These results suggested that the prophylactic local delivery of M-PBMNC may represent a novel approach for the treatment of microvascular complications in diabetics.

  5. Selective amine labeling of cell surface proteins guided by coiled-coil assembly.

    Science.gov (United States)

    Yano, Yoshiaki; Furukawa, Nami; Ono, Satoshi; Takeda, Yuki; Matsuzaki, Katsumi

    2016-11-01

    Covalent labeling of target proteins in living cells is useful for both fluorescence live-cell imaging and the subsequent biochemical analyses of the proteins. Here, we report an efficient method for the amine labeling of membrane proteins on the cell surface, guided by a noncovalent coiled-coil interaction. A carboxyl sulfosuccinimidyl ester introduced at the C-terminus of the coiled-coil probe reacted with target proteins under mild labeling conditions ([probe] = 150 nM, pH 7.4, 25°C) for 20 min. Various fluorescent moieties with different hydrophobicities are available for covalent labeling with high signal/background labeling ratios. Using this method, oligomeric states of glycophorin A (GpA) were compared in mammalian CHO-K1 cells and sodium dodecyl sulfate (SDS) micelles. In the cell membranes, no significant self-association of GpA was detected, whereas SDS-PAGE suggested partial dimerization of the proteins. Membrane cholesterol was found to be an important factor that suppressed the dimerization of GpA. Thus, the covalent functionality enables direct comparison of the oligomeric state of membrane proteins under various conditions. © 2015 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106: 484-490, 2016. PMID:26285787

  6. Macrophage phagocytosis alters the MRI signal of ferumoxytol-labeled mesenchymal stromal cells in cartilage defects

    Science.gov (United States)

    Nejadnik, Hossein; Lenkov, Olga; Gassert, Florian; Fretwell, Deborah; Lam, Isaac; Daldrup-Link, Heike E.

    2016-05-01

    Human mesenchymal stem cells (hMSCs) are a promising tool for cartilage regeneration in arthritic joints. hMSC labeling with iron oxide nanoparticles enables non-invasive in vivo monitoring of transplanted cells in cartilage defects with MR imaging. Since graft failure leads to macrophage phagocytosis of apoptotic cells, we evaluated in vitro and in vivo whether nanoparticle-labeled hMSCs show distinct MR signal characteristics before and after phagocytosis by macrophages. We found that apoptotic nanoparticle-labeled hMSCs were phagocytosed by macrophages while viable nanoparticle-labeled hMSCs were not. Serial MRI scans of hMSC transplants in arthritic joints of recipient rats showed that the iron signal of apoptotic, nanoparticle-labeled hMSCs engulfed by macrophages disappeared faster compared to viable hMSCs. This corresponded to poor cartilage repair outcomes of the apoptotic hMSC transplants. Therefore, rapid decline of iron MRI signal at the transplant site can indicate cell death and predict incomplete defect repair weeks later. Currently, hMSC graft failure can be only diagnosed by lack of cartilage defect repair several months after cell transplantation. The described imaging signs can diagnose hMSC transplant failure more readily, which could enable timely re-interventions and avoid unnecessary follow up studies of lost transplants.

  7. flowCL: ontology-based cell population labelling in flow cytometry

    OpenAIRE

    Courtot, Mélanie; Meskas, Justin; Diehl, Alexander D.; Droumeva, Radina; Gottardo, Raphael; Jalali, Adrin; Taghiyar, Mohammad Jafar; Maecker, Holden T; McCoy, J. Philip; Ruttenberg, Alan; Scheuermann, Richard H.; Brinkman, Ryan R

    2014-01-01

    Motivation: Finding one or more cell populations of interest, such as those correlating to a specific disease, is critical when analysing flow cytometry data. However, labelling of cell populations is not well defined, making it difficult to integrate the output of algorithms to external knowledge sources.

  8. Discrimination of bromodeoxyuridine labelled and unlabelled mitotic cells in flow cytometric bromodeoxyuridine/DNA analysis

    DEFF Research Database (Denmark)

    Jensen, P O; Larsen, J K; Christensen, I J;

    1994-01-01

    Bromodeoxyuridine (BrdUrd) labelled and unlabelled mitotic cells, respectively, can be discriminated from interphase cells using a new method, based on immunocytochemical staining of BrdUrd and flow cytometric four-parameter analysis of DNA content, BrdUrd incorporation, and forward and orthogona...

  9. Quantum dot labeling using positive charged peptides in human hematopoetic and mesenchymal stem cells.

    Science.gov (United States)

    Ranjbarvaziri, Sarah; Kiani, Sahar; Akhlaghi, Aliasghar; Vosough, Ahmad; Baharvand, Hossein; Aghdami, Nasser

    2011-08-01

    Quantum dots (QDs), as new and promising fluorescent probes, hold great potential in long term non-invasive bio-imaging, however there are many uncovered issues regarding their competency. In the present study, different QDs (525, 585 and 800 nm) were used to label CD133, CD34, CD14 and mesenchymal stem cells (MSCs) using positively charged peptides. Results demonstrated highly efficient internalization with the possible involvement of macropinocytosis. As indicated by LDH release and the TUNEL assay, no measurable effects on cell viability were detected at a concentration of 10 nM. QDs did not have any deleterious effects on normal cell functionality where both labeled CD133(+) cells and MSCs remarkably differentiated along multiple lineages with the use of the colony forming assay and adipo/osteo induction, respectively. Our results regarding QD maintenance revealed that these nano-particles are not properly stable and various excretion times have been observed depending on particle size and cell type. In vitro co-culture system and transplantation of labeled cells to an animal model showed that QDs leaked out from labeled cells and the released nano-particles were able to re-enter adjacent cells over time. These data suggest that before any utilization of QDs in bio-imaging and related applications, an efficient intra-cellular delivery technique should be considered to preserve QDs for a prolonged time as well as eliminating their leakage. PMID:21549422

  10. Identification of miRNA targets with stable isotope labeling by amino acids in cell culture

    DEFF Research Database (Denmark)

    Vinther, Jeppe; Hedegaard, Mads Marquardt; Gardner, Paul Phillip;

    2006-01-01

    miRNAs are small noncoding RNAs that regulate gene expression. We have used stable isotope labeling by amino acids in cell culture (SILAC) to investigate the effect of miRNA-1 on the HeLa cell proteome. Expression of 12 out of 504 investigated proteins was repressed by miRNA-1 transfection. This ...

  11. Human umbilical cord blood-derived mesenchymal stem cells promote regeneration of crush-injured rat sciatic nerves

    Institute of Scientific and Technical Information of China (English)

    Mi-Ae Sung; Jong-Ho Lee; Hun Jong Jung; Jung-Woo Lee; Jin-Yong Lee; Kang-Mi Pang; Sang Bae Yoo; Mohammad S. Alrashdan; Soung-Min Kim; Jeong Won Jahng

    2012-01-01

    Several studies have demonstrated that human umbilical cord blood-derived mesenchymal stem cells can promote neural regeneration following brain injury. However, the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells in guiding peripheral nerve regeneration remain poorly understood. This study was designed to investigate the effects of human umbilical cord blood-derived mesenchymal stem cells on neural regeneration using a rat sciatic nerve crush injury model. Human umbilical cord blood-derived mesenchymal stem cells (1 × 106) or a PBS control were injected into the crush-injured segment of the sciatic nerve. Four weeks after cell injection, brain-derived neurotrophic factor and tyrosine kinase receptor B mRNA expression at the lesion site was increased in comparison to control. Furthermore, sciatic function index, Fluoro Gold-labeled neuron counts and axon density were also significantly increased when compared with control. Our results indicate that human umbilical cord blood-derived mesenchymal stem cells promote the functional recovery of crush-injured sciatic nerves.

  12. Liver Label Retaining Cancer Cells Are Relatively Resistant to the Reported Anti-Cancer Stem Cell Drug Metformin

    OpenAIRE

    Xin, Hong-Wu; Ambe, Chenwi M.; Miller, Tyler C.; Chen, Jin-Qiu; Wiegand, Gordon W.; Anderson, Andrew J.; Ray, Satyajit; Mullinax, John E.; Hari, Danielle M; Koizumi, Tomotake; Godbout, Jessica D.; Goldsmith, Paul K.; Stojadinovic, Alexander; Rudloff, Udo; Thorgeirsson, Snorri S.

    2016-01-01

    Background & Aims: Recently, we reported that liver Label Retaining Cancer Cells (LRCC) can initiate tumors with only 10 cells and are relatively resistant to the targeted drug Sorafenib, a standard of practice in advanced hepatocellular carcinoma (HCC). LRCC are the only cancer stem cells (CSC) isolated alive according to a stem cell fundamental function, asymmetric cell division. Metformin has been reported to preferentially target many other types of CSC of different organs, including live...

  13. Microfluidic structures for flow cytometric analysis of hydrodynamically focussed blood cells fabricated by ultraprecision micromachining.

    Science.gov (United States)

    Kummrow, A; Theisen, J; Frankowski, M; Tuchscheerer, A; Yildirim, H; Brattke, K; Schmidt, M; Neukammer, J

    2009-04-01

    We present three-dimensional microfluidic structures with integrated optical fibers, mirrors and electrodes for flow cytometric analysis of blood cells. Ultraprecision milling technique was used to fabricate different flow cells featuring single-stage and two-stage cascaded hydrodynamic focusing of particles by a sheath flow. Two dimensional focussing of the sample fluid was proven by fluorescence imaging in horizontal and vertical directions and found to agree satisfactorily with finite element calculations. Focussing of the sample stream down to 5 microm at a particle velocity of 3 m s(-1) is accessible while maintaining stable operation for sample flow rates of up to 20 microL min(-1). In addition to fluorescence imaging, the micro-flow cells were characterised by measurements of pulse shapes and pulse height distributions of monodisperse microspheres. We demonstrated practical use of the microstructures for cell differentiation employing light scatter to distinguish platelets and red blood cells. Furthermore, T-helper lymphocytes labelled by monoclonal antibodies were identified by measuring side scatter and fluorescence. PMID:19294310

  14. Related Hematopoietic Stem Cell Transplantation (HSCT) for Genetic Diseases of Blood Cells

    Science.gov (United States)

    2016-05-11

    Stem Cell Transplantation; Bone Marrow Transplantation; Peripheral Blood Stem Cell Transplantation; Allogeneic Transplantation,; Genetic Diseases; Thalassemia; Pediatrics; Diamond-Blackfan Anemia; Combined Immune Deficiency; Wiskott-Aldrich Syndrome; Chronic Granulomatous Disease; X-linked Lymphoproliferative Disease; Metabolic Diseases

  15. Laser capture microdissection and genetic analysis of carbon-labeled Kupffer cells

    Institute of Scientific and Technical Information of China (English)

    Stephan Gehring; Edmond Sabo; Maryann E San Martin; Elizabeth M Dickson; Chao-Wen Cheng; Stephen H Gregory

    2009-01-01

    AIM: To develop a method of labeling and microdissecting mouse Kupffer cells within an extraordinarily short period of time using laser capture microdissection (LCM). METHODS: Tissues are complex structures comprised of a heterogeneous population of interconnected cells. LCM offers a method of isolating a single cell type from specific regions of a tissue section. LCM is an essential approach used in conjunction with molecular analysis to study the functional interaction of cells in their native tissue environment. The process of labeling and acquiring cells by LCM prior to mRNA isolation can be elaborate, thereby subjecting the RNA to considerable degradation. Kupffer cell labeling is achieved by injecting India ink intravenously, thus circumventing the need for in vitro staining. The significance of this novel approach was validated using a cholestatic liver injury model. RESULTS: mRNA extracted from the microdissected cell population displayed marked increases in colonystimulating factor-1 receptor and Kupffer cell receptor message expression, which demonstrated Kupffer cell enrichment. Gene expression by Kupffer cells derived from bile-duct-ligated, versus sham-operated, mice was compared. Microarray analysis revealed a significant (2.5-fold, q value < 10) change in 493 genes. Based on this fold-change and a standardized PubMed search, 10 genes were identified that were relevant to the ability of Kupffer cells to suppress liver injury.CONCLUSION: The methodology outlined herein provides an approach to isolating high quality RNA from Kupffer cells, without altering the tissue integrity.

  16. Membranotropic photobiomodulation on red blood cell deformability

    Science.gov (United States)

    Luo, Gang-Yue; Zhao, Yan-Ping; Liu, Timon C.; Liu, Song-Hao

    2007-05-01

    To assess modulation of laser on erythrocyte permeability and deformability via cell morphology changes, healthy human echinocytes with shrinking size and high plasmic viscosity due to cellular dehydration were treated with 1 mW, 2 mW, 3 mW, and 5 mW laser power exposure respectively. Image analyzing system on single intact erythrocyte was applied for measuring comprehensive cell morphological parameters (surface area, external membrane perimeter, circle index and elongation index) that were determined by the modulation of erythrocyte water permeability and deformability to detect relationship between erythrocyte water permeability alteration and deformability. Our preliminary experiment showed that exposure under light dose of 5 mW for 5 min could induce more active erythrocyte swelling and deformation. water channel aquaporin-1(AQP-1) was inhibited by the incubation of HgCl II in the presence and absence of 5 mW laser irradiation. The result suggested that osmotic water permeability is a primary factor in the procedure of erythrocyte deformability. In addition, no modulation of laser(5mW) on erythrocyte deformability had been found when the echinocytes were cultured with GDP-β-S (G protein inhibitor).

  17. In vivo phenotypic characterisation of nucleoside label-retaining cells in mouse periosteum

    Directory of Open Access Journals (Sweden)

    HM Cherry

    2014-03-01

    Full Text Available Periosteum is known to contain cells that, after isolation and culture-expansion, display properties of mesenchymal stromal/stem cells (MSCs. However, the equivalent cells have not been identified in situ mainly due to the lack of specific markers. Postnatally, stem cells are slow-cycling, long-term nucleoside-label-retaining cells. This study aimed to identify and characterise label-retaining cells in mouse periosteum in vivo. Mice received iodo-deoxy-uridine (IdU via the drinking water for 30 days, followed by a 40-day washout period. IdU+ cells were identified by immunostaining in conjunction with MSC and lineage markers. IdU-labelled cells were detected throughout the periosteum with no apparent focal concentration, and were negative for the endothelial marker von Willebrand factor and the pan-haematopoietic marker CD45. Subsets of IdU+ cells were positive for the mesenchymal/stromal markers vimentin and cadherin-11. IdU+ cells expressed stem cell antigen-1, CD44, CD73, CD105, platelet-derived growth factor receptor-α and p75, thereby displaying an MSC-like phonotype. Co-localisation was not detectable between IdU and the pericyte markers CD146, alpha smooth muscle actin or NG2, nor did IdU co-localise with β-galactosidase in a transgenic mouse expressing this reporter gene in pericytes and smooth muscle cells. Subsets of IdU+ cells expressed the osteoblast-lineage markers Runx2 and osteocalcin. The IdU+ cells expressing osteocalcin were lining the bone and were negative for the MSC marker p75. In conclusion, mouse periosteum contains nucleoside-label-retaining cells with a phenotype compatible with MSCs that are distinct from pericytes and osteoblasts. Future studies characterising the MSC niche in vivo could reveal novel therapeutic targets for promoting bone regeneration/repair.

  18. Macromolecular Dynamics in Red Blood Cells Investigated Using Neutron Spectroscopy

    CERN Document Server

    Stadler, Andreas Maximilian; Demmel, Franz; Artmann, Gerhard; 10.1098/rsif.2010.0306

    2011-01-01

    We present neutron scattering measurements on the dynamics of hemoglobin (Hb) in human red blood cells in vivo. Global and internal Hb dynamics were measured in the ps to ns time- and {\\AA} length-scale using quasielastic neutron backscattering spectroscopy. We observed the cross-over from global Hb short-time to long-time self-diffusion. Both short- and long-time diffusion coefficients agree quantitatively with predicted values from hydrodynamic theory of non-charged hard-sphere suspensions when a bound water fraction of around 0.23g H2O/ g Hb is taken into account. The higher amount of water in the cells facilitates internal protein fluctuations in the ps time-scale when compared to fully hydrated Hb powder. Slower internal dynamics of Hb in red blood cells in the ns time-range were found to be rather similar to results obtained with fully hydrated protein powders, solutions and E. coli cells.

  19. Aggregation of Red Blood Cells: From Rouleaux to Clot Formation

    CERN Document Server

    Wagner, C; Svetina, S

    2013-01-01

    Red blood cells are known to form aggregates in the form of rouleaux. This aggregation process is believed to be reversible, but there is still no full understanding on the binding mechanism. There are at least two competing models, based either on bridging or on depletion. We review recent experimental results on the single cell level and theoretical analyses of the depletion model and of the influence of the cell shape on the binding strength. Another important aggregation mechanism is caused by activation of platelets. This leads to clot formation which is life saving in the case of wound healing but also a major cause of death in the case of a thrombus induced stroke. We review historical and recent results on the participation of red blood cells in clot formation.

  20. Aggregation of red blood cells: From rouleaux to clot formation

    Science.gov (United States)

    Wagner, Christian; Steffen, Patrick; Svetina, Saša

    2013-06-01

    Red blood cells are known to form aggregates in the form of rouleaux. This aggregation process is believed to be reversible, but there is still no full understanding on the adhesion mechanism. There are at least two competing models, based either on bridging or on depletion. We review recent experimental results on the single cell level and theoretical analyses of the depletion model and of the influence of the cell shape on the adhesion strength. Another important aggregation mechanism is caused by activation of platelets. This leads to clot formation which is life-saving in the case of wound healing, but also a major cause of death in the case of a thrombus induced stroke. We review historical and recent results on the participation of red blood cells in clot formation.

  1. RBCs and Parasites Segmentation from Thin Smear Blood Cell Images

    Directory of Open Access Journals (Sweden)

    Vishal V. Panchbhai

    2012-09-01

    Full Text Available Manually examine the blood smear for the detection of malaria parasite consumes lot of time for trend pathologists. As the computational power increases, the role of automatic visual inspection becomes more important. An automated system is therefore needed to complete as much work as possible for the identification of malaria parasites. The given scheme based on used of RGB color space, G layer processing, and segmentation of Red Blood Cells (RBC as well as cell parasites by auto-thresholding with offset value and use of morphological processing. The work compare with the manual results obtained from the pathology lab, based on total RBC count and cells parasite count. The designed system successfully detects malaria parasites and RBC cells in thin smear image.

  2. Concise review: programming human pluripotent stem cells into blood.

    Science.gov (United States)

    Easterbrook, Jennifer; Fidanza, Antonella; Forrester, Lesley M

    2016-06-01

    Blood disorders are treated with cell therapies including haematopoietic stem cell (HSC) transplantation as well as platelet and red blood cell transfusions. However the source of cells is entirely dependent on donors, procedures are susceptible to transfusion-transmitted infections and serious complications can arise in recipients due to immunological incompatibility. These problems could be alleviated if it was possible to produce haematopoietic cells in vitro from an autologous and renewable cell source. The production of haematopoietic cells in the laboratory from human induced pluripotent stem cells (iPSCs) may provide a route to realize this goal but it has proven challenging to generate long-term reconstituting HSCs. To date, the optimization of differentiation protocols has mostly relied on the manipulation of extrinsic signals to mimic the in vivo environment. We review studies that have taken an alternative approach to modulate intrinsic signals by enforced expression of transcription factors. Single and combinations of multiple transcription factors have been used in a variety of contexts to enhance the production of haematopoietic cells from human pluripotent stem cells. This programming approach, together with the recent advances in the production and use of synthetic transcription factors, holds great promise for the production of fully functional HSCs in the future. PMID:26996518

  3. Mechanopathology of red blood cell diseases—Why mechanics matters

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    During the onset of a disease a cell may experience alterations in both the composition and organization of its cellular and molecular structures.These alterations may eventually lead to changes in its geometrical and mechanical properties such as cell size and shape,deformability and adhesion.As such,knowing how diseased cells respond to mechanical forces can reveal ways by which they differ from healthy ones.Here,we will present biomechanistic insights into red blood cell related diseases that manifest...

  4. Canine mesenchymal stem cells are effectively labeled with silica nanoparticles and unambiguously visualized in highly autofluorescent tissues

    OpenAIRE

    Han Sei-Myoung; Lee Hee-Woo; Bhang Dong-Ha; Seo Kyoung-Won; Youn Hwa-Young

    2012-01-01

    Abstract Background Development of a method for long-term labeling of cells is critical to elucidate transplanted cell fate and migration as well as the contribution to tissue regeneration. Silica nanoparticles have been recently developed and demonstrated to be biocompatible with a high labeling capacity. Thus, our study was designed to assess the suitability of silica nanoparticles for labeling canine mesenchymal stem cells (MSCs) and the fluorescence afficiency in highly autofluorescent ti...

  5. Multiscale modeling of red blood cell mechanics and blood flow in malaria.

    Directory of Open Access Journals (Sweden)

    Dmitry A Fedosov

    2011-12-01

    Full Text Available Red blood cells (RBCs infected by a Plasmodium parasite in malaria may lose their membrane deformability with a relative membrane stiffening more than ten-fold in comparison with healthy RBCs leading to potential capillary occlusions. Moreover, infected RBCs are able to adhere to other healthy and parasitized cells and to the vascular endothelium resulting in a substantial disruption of normal blood circulation. In the present work, we simulate infected RBCs in malaria using a multiscale RBC model based on the dissipative particle dynamics method, coupling scales at the sub-cellular level with scales at the vessel size. Our objective is to conduct a full validation of the RBC model with a diverse set of experimental data, including temperature dependence, and to identify the limitations of this purely mechanistic model. The simulated elastic deformations of parasitized RBCs match those obtained in optical-tweezers experiments for different stages of intra-erythrocytic parasite development. The rheological properties of RBCs in malaria are compared with those obtained by optical magnetic twisting cytometry and by monitoring membrane fluctuations at room, physiological, and febrile temperatures. We also study the dynamics of infected RBCs in Poiseuille flow in comparison with healthy cells and present validated bulk viscosity predictions of malaria-infected blood for a wide range of parasitemia levels (percentage of infected RBCs with respect to the total number of cells in a unit volume.

  6. Natural Antioxidants Improve Red Blood Cell “Survival” in Non-Leukoreduced Blood Samples

    Directory of Open Access Journals (Sweden)

    Yuliya V Kucherenko

    2015-03-01

    Full Text Available Background: Blood collected in an anticoagulant can be kept refrigerated in an unmodified state within 5 - 6 weeks. Oxidative damage is considered to be a one of the major factors contributing to the development of storage lesions. Lipid and membrane proteins oxidation results in changes in cation gradients that affect the cell survival. Aim: In the present study we used the natural antioxidants and ion channels blockers (L-carnosine, spermine, phloretin and their mixtures to prolong “survival” of red blood cells (RBCs, measured as the lack of PS exposure and cell hemolysis, in the Alsever's preservative solution upon hypothermic storage. Results: We show that the mixture of carnosine (20 mM, spermine (20 µM and phloretin (100 µM effectively blunted phosphatidylserine (PS exposure, Ca2+ accumulation and RBCs hemolysis in non-leukoreduced low (∼2% hematocrit samples after 36 days of storage as well as after 1 day of post-storage incubation of the stored cells in physiological saline solution. In addition, a slight but significant decrease in PS exposure was observed in non-leukoreduced high (∼20% hematocrit samples after 36 days of storage with the mixture of substances. Conclusion: We conclude that the use of the mixture of natural antioxidants (carnosine, spermine, and phloretin as an additive to blood preservative solution provides better RBCs storage and “survival”.

  7. 78 FR 23571 - Advisory Council on Blood Stem Cell Transplantation; Notice of Meeting

    Science.gov (United States)

    2013-04-19

    ... HUMAN SERVICES Health Resources and Services Administration Advisory Council on Blood Stem Cell... amended), the Advisory Council on Blood Stem Cell Transplantation (ACBSCT) advises the Secretary of the... Hematopoietic Stem Cell Transplantation for Hemoglobinopathies. The Council will also hear presentations...

  8. The role of red blood cells in inflammation and remodeling

    OpenAIRE

    Fredriksson, Karin

    2004-01-01

    Besides being carriers of oxygen and carbon dioxide, red blood cells (RBCs) also have a scavenger function, binding inflammatory mediators to surface receptors. Animal and experimental models have suggested a role for RBCs in inflammatory and fibrotic responses and patients with idiopathic pulmonary hemosiderosis, a disease characterized by lung hemorrhage, frequently develop fibrosis. Fibroblasts, the resident cell in the connective tissue, play an active role in tissue rem...

  9. Shear stress-induced improvement of red blood cell deformability

    OpenAIRE

    Meram, Ece; Yılmaz, Bahar D.; Bas, Ceren; Atac, Nazlı; Yalçın, Ö.; Başkurt, Oguz K.; Meiselman, Herbert J.

    2013-01-01

    Classically, it is known that red blood cell (RBC) deformability is determined by the geometric and material properties of these cells. Experimental evidence accumulated during the last decade has introduced the concept of active regulation of RBC deformability. This regulation is mainly related to altered associations between membrane skeletal proteins and integral proteins, with the latter serving to anchor the skeleton to the lipid matrix. It has been hypothesized that shear stress induces...

  10. Automated counting of white blood cells in synovial fluid.

    NARCIS (Netherlands)

    R. de Jonge (Robert); R.W. Brouwer (Reinoud); M. Smit (Marij); M. de Frankrijker-Merkestijn; R.J. Dolhain; J.M.W. Hazes (Mieke); A.W. van Toorenenbergen (Albert); J. Lindemans (Jan)

    2004-01-01

    textabstractOBJECTIVES: To evaluate the performance of automated leucocyte (white blood cell; WBC) counting by comparison with manual counting. METHODS: The number of WBC was determined in heparinized synovial fluid samples by the use of (i) a standard urine cytometer (Kova) and a

  11. Red blood cell antibodies in pregnancy and their clinical consequences

    DEFF Research Database (Denmark)

    Nordvall, Maria; Dziegiel, Morten Hanefeld; Hegaard, Hanne Kristine;

    2009-01-01

    The objective was to determine clinical consequences of various specificities for the infant/fetus. The population was patients referred between 1998 and 2005 to the tertiary center because of detected red blood cell (RBC) alloimmunization. Altogether 455 infants were delivered by 390 alloimmunized...

  12. Red Blood Cell Spectrin Skeleton in the Spotlight.

    Science.gov (United States)

    Braun-Breton, Catherine; Abkarian, Manouk

    2016-02-01

    Das et al. recently reported a role for the major merozoite surface protein MSP1 in malarial parasite egress from the red blood cell (RBC). On the basis of these new data and physical considerations, we propose an updated model for the main steps of this essential process for parasite proliferation. PMID:26652974

  13. Viscoelastic properties of whole blood. Influence of fast sedimenting red blood cell aggregates.

    Science.gov (United States)

    Schneditz, D; Rainer, F; Kenner, T

    1987-01-01

    Red blood cell (RBC) aggregation is known to be of deciding influence on erythrocyte sedimentation-rate (ESR) and on whole blood viscoelastic properties. The rheological behaviour of blood collected from a control-group with normal ESR is compared to the viscoelastic behaviour of blood collected from two groups with high to very high ESR, whose individuals are suffering from chronical polyarthritis and Morbus Bechterew, respectively. The rheological properties are evaluated by means of an oscillating-flow capillary-rheometer where the viscous (eta') and elastic (eta") component of the complex viscosity (eta) is measured at a constant frequency of 2 Hz. Correcting for the varying hematocrit of the different blood samples according to an exponential equation, the viscoelastic data are found to be elevated in the groups with high ESR. For the viscous properties this is only due to the increase of the plasma viscosity. A correction for the plasma viscosity, however, shows that the viscous properties at low shear- rates (2s-1) are significantly reduced, whereas elastic properties in a range of medium shear-rates (10s-1 to 50s-1) are significantly increased (P less than 0.001, t-test of Student). This result is discussed to be due to the high packing density of the RBC in fast sedimenting aggregates. High packing density reduces the effective volume of the RBC but increases the stiffness of the aggregates. PMID:3651579

  14. A novel in vivo cell-wall labeling approach sheds new light on peptidoglycan synthesis in Escherichia coli

    NARCIS (Netherlands)

    N.K. Olrichs; M.E.G. Aarsman; J. Verheul; C.J. Arnusch; N.I. Martin; M. Hervé; W. Vollmer; B. de Kruijff; E. Breukink; T. den Blaauwen

    2011-01-01

    Peptidoglycan synthesis and turnover in relation to cell growth and division has been studied by using a new labeling method. This method involves the incorporation of fluorescently labeled peptidoglycan precursors into the cell wall by means of the cell-wall recycling pathway. We show that Escheric

  15. Gene Expression Profiles from Peripheral Blood Mononuclear Cells Are Sensitive to Short Processing Delays.

    Science.gov (United States)

    Barnes, Michael G; Grom, Alexei A; Griffin, Thomas A; Colbert, Robert A; Thompson, Susan D

    2010-09-29

    In the analysis of peripheral blood gene expression, timely processing of samples is essential to ensure that measurements reflect in vivo biology, rather than ex vivo sample processing variables. The effect of processing delays on global gene expression patterns in peripheral blood mononuclear cells (PBMCs) was assessed by isolating and stabilizing PBMC-derived RNA from 3 individuals either immediately after phlebotomy or after a 4 h delay. RNA was labeled using NuGEN Ovation labeling and probed using the Affymetrix HG U133 Plus 2.0 GeneChip(®). Comparison of gene expression levels (≥2-fold expression change and P trends in expression patterns associated with delayed processing were also apparent in an independent set of 276 arrays of RNA from human PBMC samples with varying processing times. These data indicate that the time between sample acquisition, initiation of processing, and when the RNA is stabilized should be a prime consideration when designing protocols for translational studies involving PBMC gene expression analysis. PMID:21743826

  16. A novel technique for in-vitro labelling of colonic cancer cells

    International Nuclear Information System (INIS)

    Full text: The aim of this study was to determine a reliable method for labelling human colorectal tumour cells in order to study the movement of tumour cells in real-time during laparoscopic assisted colectomies in an animal model. The radiopharmaceutical technetium-99m hexamethylpropylene amine oxime (HMPAO) was used to radiolabel human colorectal tumour cells. The effects of temperature, incubation time and cell number were examined. Increasing incubation time and temperature showed that 99mTc-HMPAO accumulated more rapidly in cells, reaching a plateau in 15-30 minutes. Accumulation of 99mTc-HMPAO in tumour cells was lower at room temperature than at 37 deg C. 99mTc-HMPAO had the highest labelling efficiency (greater than 85%) with the largest number of cells (7x106) so that the maximum number of cells need to be used. As accumulation of 99mTc-HMPAO within the tumour cells did not greatly increase when incubation time increased from 15-30 minutes, it was concluded that optimal labelling of human colorectal tumour cells with 99mTc-HMPAO occurs at 15 minutes with incubation at 37 deg C. This cell labelling technique has been utilised in a scintigraphic model to determine the real-time movement of human colorectal tumour cells in a porcine model in vivo. This is to aid the evaluation of possible correlation between tumour recurrence at the site of the surgical scar of where the trocars were placed during the procedure via contamination of instruments used by 'spilled' tumour cells. Copyright (2000) The Australian and New Zealand Society of Nuclear Medicine Inc

  17. Red blood cell aggregation, aggregate strength and oxygen transport potential of blood are abnormal in both homozygous sickle cell anemia and sickle-hemoglobin C disease

    OpenAIRE

    Tripette, Julien; Alexy, Tamas; Hardy-Dessources, Marie-Dominique; Mougenel, Daniele; Beltan, Eric; Chalabi, Tawfik; Chout, Roger; Etienne-Julan, Maryse; Hue, Olivier; Meiselman, Herbert J.; Connes, Philippe

    2009-01-01

    Recent evidence suggests that red cell aggregation and the ratio of hematocrit to blood viscosity, an index of the oxygen transport potential of blood, might considerably modulate blood flow dynamics in the microcirculation. The findings of this study indicate that patients with sickle cell disease and those with sickle cell hemoglobin C disease have low ratios of hematocrit to blood viscosity as compared to normal controls. This may play a role in tissue hypoxia and clinical status of these ...

  18. Measuring density and compressibility of white blood cells and prostate cancer cells by microchannel acoustophoresis

    DEFF Research Database (Denmark)

    Barnkob, Rune; Augustsson, Per; Magnusson, Cecilia; Lilja, Hans; Laurell, Thomas; Bruus, Henrik

    determine the density and compressibility of individual cells enables the prediction and alteration of the separation outcome for a given cell mixture. We apply the method on white blood cells (WBCs) and DU145 prostate cancer cells (DUCs) aiming to improve isolation of circulating tumor cells from blood, an......We present a novel method for the determination of density and compressibility of individual particles and cells undergoing microchannel acoustophoresis in an arbitrary 2D acoustic field. Our method is a critical advancement within acoustophoretic separation of biological cells, as the ability to...... emerging tool in the monitoring and characterizing of metastatic cancer....

  19. Identification of Progenitor Cell Survival in Peripheral Blood

    International Nuclear Information System (INIS)

    The myeloid progenitors can not survive properly under the usual conditions of blood banking.The aim of work is to assay the survival of myeloid progenitors during varying periods of blood storage, under the usual condition of blood banking. It is an attempt to detect whether or not ,these circulating myeloid progenitors could be stored under the blood banking condition to be used in clinical transplantation protocols to treat a wide variety of refractory diseases.Individual blood samples from forty healthy adults were examined clinically, laboratory and ultrasonography. Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque gradients . Serial dilutions of human placental conditioned medium were made, and tested for optimal activity by In vitro cultured technique.This study estimated that the mean levels of absolute number of myeloid progenitors per c.mm. at zero time was 137.7±68.3 (Range 54-297),at day 3 was 71.0±40.2 (Range 54-297), at day 7 was 94.8±45.7 (Range 30 -232) and at day 14 was 45.5±22.7). There was statistically significant decrease in the number of colonies from zero time to day 14. There was statistically significant decrease in the number of myeloid progenitors from zero time to day 14

  20. Indolic uremic solutes enhance procoagulant activity of red blood cells through phosphatidylserine exposure and microparticle release.

    Science.gov (United States)

    Gao, Chunyan; Ji, Shuting; Dong, Weijun; Qi, Yushan; Song, Wen; Cui, Debin; Shi, Jialan

    2015-11-01

    Increased accumulation of indolic uremic solutes in the blood of uremic patients contributes to the risk of thrombotic events. Red blood cells (RBCs), the most abundant blood cells in circulation, may be a privileged target of these solutes. However, the effect of uremic solutes indoxyl sulfate (IS) and indole-3-acetic acid (IAA) on procoagulant activity (PCA) of erythrocyte is unclear. Here, RBCs from healthy adults were treated with IS and IAA (mean and maximal concentrations reported in uremic patients). Phosphatidylserine (PS) exposure of RBCs and their microparticles (MPs) release were labeled with Alexa Fluor 488-lactadherin and detected by flow cytometer. Cytosolic Ca(2+) ([Ca(2+)]) with Fluo 3/AM was analyzed by flow cytometer. PCA was assessed by clotting time and purified coagulation complex assays. We found that PS exposure, MPs generation, and consequent PCA of RBCs at mean concentrations of IS and IAA enhanced and peaked in maximal uremic concentrations. Moreover, 128 nM lactadherin, a PS inhibitor, inhibited over 90% PCA of RBCs and RMPs. Eryptosis or damage, by indolic uremic solutes was due to, at least partially, the increase of cytosolic [Ca(2+)]. Our results suggest that RBC eryptosis in uremic solutes IS and IAA plays an important role in thrombus formation through releasing RMPs and exposing PS. Lactadherin acts as an efficient anticoagulant in this process. PMID:26516916