Sample records for highly specific immunohistochemical

  1. Immunohistochemical identification of neuron-specific enolase, synaptophysin, chromogranin and endocrine granule constituent in neuroendocrine tumours

    DEFF Research Database (Denmark)

    Vyberg, M; Horn, T; Francis, D


    Immunohistochemical identification of neuroendocrine tumour markers in paraffin embedded material from 22 tumours (5 small cell carcinomas of the lung (SCCL), 12 carcinoids, 2 medullary thyroid carcinomas, 2 pheochromocytomas and one paraganglioma) with electron microscopically verified dense...

  2. Prostate adenocarcinoma with negative immunohistochemical stain of prostate-specific antigen presenting with cervical mass: A case report

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    Liming Zhu


    Full Text Available Cervical regional nodal involvement, as the first manifestation of prostatic cancer, has been rarely reported. Prostate cancer metastasis to the supraclavicular lymph nodes with negative immunohistochemical stain of PSA is even rarer. We report a case of prostate cancer with negative immunohistochemical stain of prostate-specific antigen presenting with left supraclavicular node enlargement. A 63-year-old man was referred to our hospital for a left supraclavicular mass. He had a family history of gastric cancer (two brothers had died of gastric cancer. Enhanced computed tomography of the abdomen revealed retroperitoneal lymph node enlargement. Gastroscopy revealed no evidence of any gastric tumor. Biopsy of the left-sided supraclavicular lymph nodes revealed metastatic adenocarcinoma with a negative prostate-specific antigen (PSA stain. The serum tumor markers were examined, revealing PSA levels of 21.820 ng/ml. Biopsy of the prostate disclosed poorly-to-moderately differentiated adenocarcinoma (Gleason 4 + 4 = 8. Although rare, prostate cancer should be considered in the differential diagnoses of elderly men with undetermined original adenocarcinoma, metastatic to the supraclavicular lymph nodes. Rectal examination, serum PSA and pelvic image can be helpful to lead the diagnosis. PSA stain may be weak or negative in some poorly differentiated patients, however, it is still a sensitive and specific marker of prostatic differentiation and must be routinely applied.

  3. Immunohistochemical identification of mast cells in formaldehyde-fixed tissue using monoclonal antibodies specific for tryptase. (United States)

    Walls, A F; Jones, D B; Williams, J H; Church, M K; Holgate, S T


    An avidin-biotin enhanced immunoperoxidase procedure using monoclonal antibodies (AA1, AA3, and AA5) prepared against human mast cell tryptase resulted in intense staining of mast cells in paraffin-embedded tissue. The distribution of mast cells observed was similar to that seen when adjacent serial sections were stained using a standard procedure with toluidine blue, though the immunoperoxidase technique permitted the identification of significantly more mast cells. With monoclonal antibody AA1, immunostaining was entirely specific for mast cell granules, and there was negligible background staining in a range of tissues including lung, tonsil, colon, gastric mucosa, skin, and pituitary. There was no staining of antibody on basophils or on any other normal blood leukocyte. The technique was effective with tissue fixed in either Carnoy's or neutral buffered formalin, though the internal mast cell structure was better preserved with formaldehyde fixation. The immunoperoxidase staining procedure with monoclonal antibody AA1 is a highly specific and sensitive means for the detection of mast cells in routinely processed tissues.

  4. High Ki-67 Immunohistochemical Reactivity Correlates With Poor Prognosis in Bladder Carcinoma (United States)

    Luo, Yihuan; Zhang, Xin; Mo, Meile; Tan, Zhong; Huang, Lanshan; Zhou, Hong; Wang, Chunqin; Wei, Fanglin; Qiu, Xiaohui; He, Rongquan; Chen, Gang


    Abstract Ki-67 is considered as one of prime biomarkers to reflect cell proliferation and immunohistochemical Ki-67 staining has been widely applied in clinical pathology. To solve the widespread controversy whether Ki-67 reactivity significantly predicts clinical prognosis of bladder carcinoma (BC), we performed a comprehensive meta-analysis by combining results from different literature. A comprehensive search was conducted in the Chinese databases of WanFang, China National Knowledge Infrastructure and Chinese VIP as well as English databases of PubMed, ISI web of science, EMBASE, Science Direct, and Wiley online library. Independent studies linking Ki-67 to cancer-specific survival (CSS), disease-free survival (DFS), overall survival (OS), progression-free survival (PFS), and recurrence-free survival (RFS) were included in our meta-analysis. With the cut-off values literature provided, hazard ratio (HR) values between the survival distributions were extracted and later combined with STATA 12.0. In total, 76 studies (n = 13,053 patients) were eligible for the meta-analysis. It was indicated in either univariate or multivariate analysis for survival that high Ki-67 reactivity significantly predicted poor prognosis. In the univariate analysis, the combined HR for CSS, DFS, OS, PFS, and RFS were 2.588 (95% confidence interval [CI]: 1.623–4.127, P < 0.001), 2.697 (95%CI: 1.874–3.883, P < 0.001), 2.649 (95%CI: 1.632–4.300, P < 0.001), 3.506 (95%CI: 2.231–5.508, P < 0.001), and 1.792 (95%CI: 1.409–2.279, P < 0.001), respectively. The pooled HR of multivariate analysis for CSS, DFS, OS, PFS, and RFS were 1.868 (95%CI: 1.343–2.597, P < 0.001), 2.626 (95%CI: 2.089–3.301, P < 0.001), 1.104 (95%CI: 1.008–1.209, P = 0.032), 1.518 (95%CI: 1.299–1.773, P < 0.001), and 1.294 (95%CI: 1.203–1.392, P < 0.001), respectively. Subgroup analysis of univariate analysis by origin showed that Ki-67 reactivity significantly

  5. Immunohistochemical detection of a very high density lipoprotein (VHDL) in ovarian follicles of Triatoma infestans. (United States)

    González, M S; Ronderos, J R; Rimoldi, O J; Brenner, R R


    The ability of Triatoma infestans ovarian follicles to synthesize a very high-density lipoprotein (VHDL) has been examined by immunohistochemical methods. This kind of lipoprotein can be envisaged as a storage hexameric protein present in the hemolymph of some insect species. VHDL immunoreactivity is observed in oocytes at different stages of maturation. The antigen is present in the oocyte cytoplasm as well as in the follicular epithelial cells. The immunopositive reaction in the apical surface of follicle cells suggests both a VHDL synthesis and a secretion process. Furthermore, VHDL seems to be stored into oocyte in yolk granules. On the contrary, no immunopositive reaction is observed in the intracellular spaces between follicle cells, suggesting that VHDL is not incorporated from hemolymph into the oocyte.

  6. Immunohistochemical visualization of neurons and specific glial cells for stereological application in the porcine neocortex

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    Lyck, Lise; Jelsing, Jacob; Jensen, Pia Søndergaard;


    The pig is becoming an increasingly used non-primate model in basic experimental studies of human neurological diseases. In spite of the widespread use of immunohistochemistry and cell type specific markers, the application of immunohistochemistry in the pig brain has not been systematically desc...

  7. Effects of fixation and tissue processing on immunohistochemical demonstration of specific antigens. (United States)

    Arnold, M M; Srivastava, S; Fredenburgh, J; Stockard, C R; Myers, R B; Grizzle, W E


    Identification of biomarkers in archival tissues using immunochemistry is becoming increasingly important for determining the diagnosis and prognosis of tumors, for characterizing preinvasive neoplastic changes in glandular tissues such as prostate, for evaluating the response of tumors and preinvasive neoplastic changes to certain therapies (i.e., as a surrogate intermediate end point), for selecting patients who are candidates for specific therapies (e.g., immunotherapy) and for retrospective studies. For detecting specific biomarkers it is important to understand the limitations imposed by the fixation methods and processing of the tissues. This study was designed to determine the effects of fixation on the detection in archival paraffin blocks of selected antigens postulated to be important in tumor biology. We evaluated the antigens TGF alpha, p185erbB-2, broad spectrum keratins, p53, and TAG-72 (B72.3). Fixatives evaluated included standard preparations of neutral buffered formalin, acid formalin, zinc formalin, alcoholic formalin, ethanol, methanol, and Bouin's fixative. We found that in general neutral buffered formalin is the poorest fixative for maintaining antigen recognition by immunohistochemistry and that no single fixative was best for all antigens. The dehydrating (coagulant) fixatives (e.g., ethanol and methanol) preserved immunorecognition of p53 and broad spectrum keratins best while the slow cross-linking fixatives (e.g., unbuffered zinc formalin) were best for demonstrating TGF alpha and p185erbB-2. Fixatives other than neutral buffered formalin produced equivalent recognition of the epitope of TAG-72 by B72.3. In formalin fixed archival tissues, only a portion of the antigen signal can be detected by routine immunohistologic methods.

  8. Muscle-specific integrins in masseter muscle fibers of chimpanzees: an immunohistochemical study.

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    Gianluigi Vaccarino


    Full Text Available Most notably, recent comparative genomic analyses strongly indicate that the marked differences between modern human and chimpanzees are likely due more to changes in gene regulation than to modifications of the genes. The most peculiar aspect of hominoid karyotypes is that human have 46 chromosomes whereas gorillas and chimpanzees have 48. Interestingly, human and chimpanzees do share identical inversions on chromosome 7 and 9 that are not evident in the gorilla karyotype. Thus, the general phylogeny suggests that humans and chimpanzees are sister taxa; based on this, it seems that human-chimpanzee sequence similarity is an astonishing 99%. At this purpose, of particular interest is the inactivation of the myosin heavy chain 16 (MYH16 gene, most prominently expressed in the masticatory muscle of mammals. It has been showed that the loss of this gene in humans may have resulted in smaller masticatory muscle and consequential changes to cranio-facial morphology and expansion of the human brain case. Powerful masticatory muscles are found in most primates; contrarily, in both modern and fossil member Homo, these muscles are considerably smaller. The evolving hominid masticatory apparatus shifted towards a pattern of gracilization nearly simultaneously with accelerated encephalization in early Homo. To better comprehend the real role of the MYH16 gene, we studied the primary proteins present in the muscle fibers of humans and non-humans, in order to understand if they really can be influenced by MYH16 gene. At this aim we examined the muscle-specific integrins, alpha 7B and beta 1D-integrins, and their relative fetal isoforms, alpha 7A and beta 1A-integrins, analyzing, by immunohistochemistry, muscle biopsies of two components of a chimpanzee's group in captivity, an alpha male and a non-alpha male subjects; all these integrins participate in vital biological processes such as maintenance of tissue integrity, embryonic development, cell

  9. Prognostic Significance of Immunohistochemical Phenotypes in Patients Treated for High-Grade Cervical Intraepithelial Neoplasia

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    Massimo Origoni


    Full Text Available Strong evidence exists that the host’s immune system plays a crucial role for the development of human papillomavirus-related cervical premalignant and malignant lesions. In particular, effective cell-mediated immunity (CMI promotes spontaneous infection clearance and cancer precursors regression in healthy subjects, while immunosuppressed individuals are more likely to experience infection persistence, cervical intraepithelial neoplasia (CIN lesions, and cervical cancer. In this study, the prognostic significance of immunohistochemical profiling of CD4+ T-cells, CD8+ T-cells, dendritic cells (CD11c+, T-bet+, and GATA-3+ transcription factors has been studied in surgical specimens of 34 consecutive women affected by high-grade cervical intraepithelial neoplasia (CIN2-3 submitted to cervical conization. Results have been correlated with the clinical outcomes at 24 months after treatment and statistically analyzed. Higher rates of CD4+ T-cells, CD11c+ dendritic cells, and T-bet+ transcription factor positivity showed a strong statistically significative correlation with favourable clinical outcomes (P≤0.0001. These data reinforce the evidence of the relevance of the host’s immune status in the natural history of HPV-related cervical disease and add a prognostic significance of the cervical immunological profile in terms of predicting significant lower recurrence rates.

  10. Sensitivity and specificity of monoclonal and polyclonal immunohistochemical staining for West Nile virus in various organs from American crows (Corvus brachyrhynchos

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    Wu Ping


    Full Text Available Abstract Background Based on results of earlier studies, brain, heart and kidney are most commonly used for West Nile virus (WNV detection in avian species. Both monoclonal and polyclonal antibodies have been used for the immunohistochemical diagnosis of WNV in these species. Thus far, no studies have been performed to compare the sensitivity and specificity of monoclonal and polyclonal antibodies in detecting WNV in American crows (Corvus brachyrhynchos. Our objectives were to determine 1 the comparative sensitivities of monoclonal and polyclonal antibodies for immunohistochemical (IHC diagnosis of WNV infection in free-ranging American crows, 2 which organ(s is/are most suitable for IHC-based diagnosis of WNV, and 3 how real-time RT-PCR on RNA extracted from formalin-fixed paraffin-embedded tissues compared to IHC for the diagnosis of WNV infection. Methods Various combinations, depending on tissue availability, of sections of heart, kidney, brain, liver, lung, spleen, and small intestine from 85 free-ranging American crows were stained using a rabbit-polyclonal anti-WNV antibody as well as a monoclonal antibody directed against an epitope on Domain III of the E protein of WNV. The staining intensity and the extent of staining were determined for each organ using both antibodies. Real-time RT-PCR on formalin-fixed paraffin-embedded tissues from all 85 crows was performed. Results Forty-three crows were IHC-positive in at least one of the examined organs with the polyclonal antibody, and of these, only 31 were positive when IHC was performed with the monoclonal antibody. Real-time RT-PCR amplified WNV-specific sequences from tissue extracts of the same 43 crows that were IHC-positive using the polyclonal antibody. All other 42 crows tested negative for WNV with real-time PCR and IHC staining. Both antibodies had a test specificity of 100% when compared to PCR results. The test sensitivity of monoclonal antibody-based IHC staining was only 72


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    Tika Fiona Sari


    Full Text Available Dengue virus are transmitted from human to human by the bites of infective female Aedesmosquitoes from subgenus Stegomyia. One of the way to detect Dengue virus antigen is by usingimmunohistochemical technique. This method was reported to detect dengue vims antigen in lowlevels. The aims of this study is to measure sensitivity and specificity of monoclonal antibodyDSSE10 using SBPC to detect antigen Dengue virus in head squash Toxorhynchites splendenswere infected with dengue patient serum and RT-PCR as gold standart. Artificially-infected Tx.splendens mosquitoes with serum positif dengue virus were used as infectious samples and noninfectedTx. splendens mosquitoes were used as control negative. The immunohistochemichalSBPC assay using monoclonal antibody DSSE10 then applied in mosquitoes head squash todetect Dengue vims antigen. RT-PCR as a gold standart was applied in each mosquito thorax.The result were analyzed by descriptive stasistic test and 2x2 diagnostic test table. Monoclonalantibody DSSE10 using immunohistochemical SBPC assay in head squash Tx. splendens wasgave sensitivity 87,09% and specificity 92,5%. Conclussion of this study is DSSE10 Monoclonalantibodies can be used as primary antibodies for the detection of dengue vims antigen inmosquito head squashKeywords: Dengue viruses, SBPC, antibodies DSSE10, head squash, Toxorhynchitessplendens' Virus Dengue ditularkan dari orang ke orang melalui gigitan nyamuk Aedes dari subgenusStegomyia. Salah satu cara untuk mendeteksi antigen vims Dengue adalah dengan menggunakanteknik imunohistokimia. Metode imunohistokimia dilaporkan dapat mendeteksi antigen vimsDengue dalam kadar yang rendah. Tujuan penelitian ini adalah melakukan evaluasi sensitivitasdan spesifitas antibodi monoklonal DSSE10 dengan metode imunohistokimia Streptavidin BiotinPeroxidase Complex (SBPC untuk mendeteksi antigen Dengue melalui scdiaan head squashnyamuk Toxorhynchites splendens yang diinfeksi dengan scrum penderita

  12. Neoadjuvant multidrug chemotherapy including High-Dose Methotrexate modifies VEGF expression in Osteosarcoma: an immunohistochemical analysis

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    Rosa Michele A


    Full Text Available Abstract Background Angiogenesis plays a role in the progression of osteosarcoma, as well as in other mesenchymal tumors and carcinomas, and it is most commonly assessed by vascular endothelial growth factor (VEGF expression or tumor CD31-positive microvessel density (MVD. Tumor VEGF expression is predictive of poor prognosis, and chemotherapy can affect the selection of angiogenic pattern. The aim of the study was to investigate the clinical and prognostic significance of VEGF and CD31 in osteosarcoma, both at diagnosis and after neoadjuvant chemotherapy, in order to identify a potential role of chemotherapy in angiogenic phenotype. Methods A retrospective analysis was performed on 16 patients with high grade osteosarcoma. In each case archival pre-treatment biopsy tissue and post-chemotherapy tumor specimens were immunohistochemically stained against CD31 and VEGF, as markers of angiogenic proliferation both in newly diagnosed primary osteosarcoma and after multidrug chemotherapy including high-dose methotrexate (HDMTX. The correlation between clinicopathological parameters and the degree of tumor VEGF and CD31 expression was statistically assessed using the χ2 test verified with Yates' test for comparison of two groups. Significance was set at p Results Expression of VEGF was positive in 11 cases/16 of cases at diagnosis. Moreover, 8 cases/16 untreated osteosarcomas were CD31-negative, but the other 8 showed an high expression of CD31. VEGF expression in viable tumor cells after neoadjuvant chemotherapy was observed in all cases; in particular, there was an increased VEGF expression (post-chemotherapy VEGF - biopsy VEGF in 11 cases/16. CD31 expression increased in 11 cases/16 and decreased in 3 cases after chemotherapy. The data relating to the change in staining following chemotherapy appear statistically significant for VEGF expression (p p > 0,05. Conclusions Even if the study included few patients, these results confirm that VEGF and CD

  13. NY-ESO-1 is a sensitive and specific immunohistochemical marker for myxoid and round cell liposarcomas among related mesenchymal myxoid neoplasms. (United States)

    Hemminger, Jessica A; Iwenofu, O Hans


    Myxoid and round cell liposarcomas constitute approximately one-third of all liposarcomas, a relatively common group of fat-derived soft tissue sarcomas. The histomorphology is a continuum between highly differentiated myxoid and poorly differentiated round cell components. The gold standard of diagnosis is dependent on histomorphology and/or identification of t(12;16)(q13;p11) translocation by cytogenetics or demonstration of DDIT3 rearrangements by fluorescence in situ hybridization. There are currently no diagnostic immunohistochemical stains available. The broad range of myxoid neoplasms in the differential diagnosis includes a variety of sarcomas. Given the notable differences in disease biology among myxoid neoplasms, which range from benign to aggressive, an accurate diagnosis is imperative for proper treatment and prognostication. Prompted by our recent study showing frequent expression of the cancer testis antigen NY-ESO-1 in myxoid and round cell liposarcomas, we sought to evaluate the utility of NY-ESO-1 as an immunohistochemical marker for myxoid and round cell liposarcoma among mesenchymal myxoid neoplasms within the differential diagnosis. Formalin-fixed, paraffin-embedded blocks were obtained for the following mesenchymal myxoid neoplasms (n=138): myxoid and round cell liposarcoma (n=38); well-differentiated liposarcoma (n=12); lipoma (n=20; 4 with myxoid change); extra-cardiac soft tissue myxoma (n=39); extraskeletal myxoid chondrosarcoma (n=12); myxofibrosarcoma (n=10: 5 low grade, 2 intermediate grade, 3 high grade); and low-grade fibromyxoid sarcoma (n=7). Utilizing standard immunohistochemistry protocols, full sections were stained with NY-ESO-1 (clone E978), and staining was assessed for intensity (1-2+), percentage of tumor positivity, and location. In all, 36/38 (95%) of the myxoid and round cell liposarcomas demonstrated NY-ESO-1 immunoreactivity. The majority of the positive cases (34/36; 94%) showed strong, homogenous staining (>50% tumor

  14. Evaluation of an immunohistochemical test with polyclonal antibodies raised against mycobacteria used in formalin-fixed tissue compared with mycobacterial specific culture. (United States)

    Carabias, E; Palenque, E; Serrano, R; Aguado, J M; Ballestín, C


    An immunohistochemical (IH) test (commercially available polyclonal antiserum rabbit anti-Myco-bacterium bovis; DAKO A/S) was used to detect the presence of mycobacteria in 65 formalin-fixed, paraffin-embedded tissue blocks from different organs, showing necrotizing caseous granuloma lesions on hematoxylin and eosin sections from 65 patients. These 65 samples were dyed using an acid-fast fluorescent technique and compared using the immunohistochemical method. Both results were also compared with the mycobacterial cultures. The IH test, compared with the culture, showed a sensitivity (S) of 52%, a specificity (Sp) of 76%, a positive predictive value (PV pos) of 61% and a negative predictive value (PV neg) of 69%. We analyze these data and discuss the possible causes of false-positive and -negative results of the IH test. This rapid test on paraffin embedded tissue seems valuable in the period when waiting for the culture results.

  15. Specific yield, High Plains aquifer (United States)

    U.S. Geological Survey, Department of the Interior — This raster data set represents specific-yield ranges in the High Plains aquifer of the United States. The High Plains aquifer underlies 112.6 million acres (176,000...

  16. MAGE-A is More Highly Expressed Than NY-ESO-1 in a Systematic Immunohistochemical Analysis of 3668 Cases. (United States)

    Kerkar, Sid P; Wang, Zeng-Feng; Lasota, Jerzy; Park, Tristen; Patel, Krishna; Groh, Eric; Rosenberg, Steven A; Miettinen, Markku M


    Two cancer testis antigens, the New York esophageal squamous cell carcinoma-1 (NY-ESO-1) and the melanoma-antigen family A (MAGE-A), represent promising immunotherapy targets due to the low expression of these antigens in nonmalignant tissue. To assess overexpression patterns in various cancers, we performed a systematic immunohistochemical analysis for NY-ESO-1 and MAGE-A on tissue array samples of 3668 common epithelial carcinomas (CA) and germ cell tumors of high prevalence and mortality. Here, we find significantly higher expression of MAGE-A (>50% on tumor cells) compared with NY-ESO-1 in several CAs including cutaneous squamous cell carcinomas (SCC) (52.8%/2.8%), esophageal SCC (50%/0%), head and neck SCC (41.1%/ESO-1, was seen in whole slide evaluations of an independent cohort of metastatic SCC (45.5%/3.6%) and metastatic CA (13.5%/0%) of various primaries with significantly higher expression of MAGE-A in metastatic SCC compared with other metastatic CA. MAGE-A is also more highly expressed in germ cell tumors, seminomas (69%/3.5%) and nonseminomas (40.1%/4.7%). In summary, MAGE-A is more highly expressed than NY-ESO-1 in a majority of human malignancies, and targeting MAGE-A may benefit a large number of patients.

  17. Protocadherin 11X/Y a human-specific gene pair: an immunohistochemical survey of fetal and adult brains. (United States)

    Priddle, Thomas H; Crow, Tim J


    Protocadherins 11X and 11Y are cell adhesion molecules of the δ1-protocadherin family. Pcdh11X is present throughout the mammalian radiation; however, 6 million years ago (MYA), a reduplicative translocation of the Xq21.3 block onto what is now human Yp11 created the Homo sapiens-specific PCDH11Y. Therefore, modern human females express PCDH11X whereas males express both PCDH11X and PCDH11Y. PCDH11X/Y has been subject to accelerated evolution resulting in human-specific changes to both proteins, most notably 2 cysteine substitutions in the PCDH11X ectodomain that may alter binding characteristics. The PCDH11X/Y gene pair is postulated to be critical to aspects of human brain evolution related to the neural correlates of language. Therefore, we raised antibodies to investigate the temporal and spatial expression of PCDH11X/Y in cortical and sub-cortical areas of the human fetal brain between 12 and 34 postconceptional weeks. We then used the antibodies to determine if this expression was consistent in a series of adult brains. PCDH11X/Y immunoreactivity was detectable at all developmental stages. Strong expression was detected in the fetal neocortex, ganglionic eminences, cerebellum, and inferior olive. In the adult brain, the cerebral cortex, hippocampal formation, and cerebellum were strongly immunoreactive, with expression also detectable in the brainstem.

  18. Immunohistochemical myofiber typing and high-resolution myofibrillar lesion detection in LR white embedded muscle (United States)

    Thompson, J. L.; Vijayan, K.; Riley, D. A.


    We have developed a method of fixing, embedding, sectioning, and staining that allows high-resolution detection of myofibrillar structure and myosin immunocytochemical muscle fiber typing in serial semithin sections of LR White plastic embedded muscle at the light microscopic level. Traditional approaches, such as cryostat sections, permit fiber typing, but small myofibrillar lesions (1-3 sarcomeres) are difficult to detect because of section thickness. Semithin sections of hydrophobic resins do not stain well either histochemically or immunocytochemically. Electron microscopy can resolve lesions and discriminate fiber types based on morphology, but the sampling area is small. Our goal was to develop a rapid method for defining both fiber type and high-resolution primary myofibrillar lesion damage. Mild fixation (1-4% paraformaldehyde, 0. 05-0.1% glutaraldehyde) and embedment in a hydrophilic resin (LR White) were used. Myofibrillar structure was extremely well preserved at the light microscopic (LM) level, and lesions could be readily resolved in Toluidine blue stained 500-nm sections. Fiber type was defined by LM immunomyosin staining of serial plastic semithin sections, which demonstrated reciprocal staining patterns for "fast (Sigma M4276) and "total" (skeletal muscle) myosins (Sigma M7523). Copyright 2000 Wiley-Liss, Inc.

  19. Immunohistochemical expression of neural tissue markers (neuron-specific enolase, glial fibrillary acidic protein, S100 protein) in ameloblastic fibrodentinoma: a comparative study with ameloblastic fibroma. (United States)

    Takeda, Y; Sato, H; Satoh, M; Nakamura, S I; Yamamoto, H


    Formalin-fixed paraffin-embedded sections of three cases of ameloblastic fibrodentinoma (AFD) were studied by the avidin-biotin-peroxidase complex method using antibodies against neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and S100 protein and the results were compared with those in ameloblastic fibroma (AF). A striking histopathological characteristic of AFD was the formation of abortive dentin with various degrees of maturation at the epithelial-mesenchymal tissue interface. Central cells of enamel organ-like epithelia with various stages of abortive dentin induction in AFD were generally positive for NSE. Dental lamina-like epithelial cells also showed positive staining in some areas. No cells were positive for NSE in AF. Positive staining for GFAP was observed in the juxta-epithelial mesenchymal tissue of the formation stage of immature dentin with various numbers of entrapped cells in AFD, but GFAP staining was negative in other mesenchymal and epithelial tissues at other stages. In AF, no GFAP-positive cells were found. There were a few S100 protein-positive cells found in the foci of epithelial components in both AFD and AF. Mesenchymal cells showing a dendritic or spindle shape were positive for S100 protein in some areas of AFD and AF. Although such cells in the mesenchymal component of pigmented AFD were more numerous than in non-pigmented AFD and AF, their distribution pattern in the former condition was basically similar to that in the latter. Although the present results, obtained from conventional immunohistochemical procedures, do not directly reflect the expression of neural crest-derived cells in the dentinogenesis of AFD, such results do not disprove the possibility of the expression of neural proteins probably related to neural crest-derived cells in dentinogenesis under certain pathologic conditions in odontogenic mixed tumors. Such a phenomenon may also occur during dentinogenesis in other odontogenic mixed tumors and

  20. Immunohistochemically detected p53 mutations in epithelial tumors and results of treatment with chemotherapy and radiotherapy. A treatment-specific overview of the clinical data

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    Thames, H.D. [University of Texas, Houston, TX (United States); Petersen, C.; Baumann, M. [University Hospital Dresden (Germany); Petersen, S. [Hospital Dresden-Friedrichstadt (Germany); Nieder, C. [Klinikum rechts der Isar, Munich (Germany)


    Background: The aim was to ascertain whether many hundreds of clinical reports over the last decade are consistent with the prediction of a poorer outcome in cancer patients with p53 abnormalities treated with cytotoxic drugs and radiation. Material and method: There are 301 studies on the influence of p53 overexpression published through summer 2000, in which chemotherapy or radiotherapy was used alone or in combination with surgery. From 45 reports meeting stringent selection rules, comparison groups are identified in whom the same measure of outcome was reported for the same treatment applied to the same tumor, with results corrected for important prognostic factors. Metaanalysis techniques are then applied to the comparison groups. Attention was limited to reports using immunohistochemical techniques, to form comparison groups of sufficient size. Results: Four comparison groups were identified by treatment and endpoint: (1) Stage-I-III breast cancer (surgery and chemotherapy, disease-free survival, seven studies); (2) stage I-III breast cancer (surgery and chemotherapy, overall survival, six studies); (3) stage II-IV head and neck cancer (radiotherapy and chemotherapy, overall survival, five studies); (4) FIGO I-IV ovarian cancer (surgery and chemotherapy, overall survival, six studies). In the breast (disease-free survival) and ovarian (overall survival) comparison groups, the hazard ratio for a deleterious effect of p53 overexpression was significant or marginally significant, depending on assumed ranges for unreported hazard ratios in non-significant studies. Conclusions: Despite the many caveats related to metaanalysis applied to retrospective data, high variability of immunohistochemical technique, etc., a nearly significant negative effect of p53 overexpression on outcome of treatment with cytotoxic drugs and radiation emerges in the few studies where heterogeneity can be sufficiently reduced or accounted for. (orig.) [German] Hintergrund: Es sollte

  1. Highly expressed genes in human high grade gliomas: immunohistochemical analysis of data from the Human Protein Atlas

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    Michael A. Meyer


    Full Text Available Gene expression within human glioblastomas were analyzed from data on 20,083 genes entered into the on-line Human Protein Atlas. In selecting genes that are strongly expressed within normal human brain tissue, 58 genes were identified from a search of the 20,083 entries that were rated as showing 90% or greater intensity of expression within normal brain tissues. Of these 58, a subset of 48 genes was identified that not only had expression data for human glioblastomas but also for the human glioblastoma cell line U-251. Four of these 48 selected genes were found to be strongly expressed within the cytoplasm when assessed by both histologic sampling of high grade glioma patient cases as well as U-251 glioblastoma cell line immunofluoresence analysis. These four human genes are: AGBL2 (ATP/GTP binding protein-like 2, BLOC1S6 (biogenesis of lysosomal organelles complex-1, subunit 6, MAP1A (microtubule-associated protein 1A and ZSWIM5 (zinc finger, SWIM-type containing 5, also known as KIAA1511. Further research is advocated to investigate the role of ZSWIM5 and AGBL2 in glioma cell biology.

  2. VHDL Specification Methodology from High-level Specification

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    M. Benmohammed


    Full Text Available Design complexity has been increasing exponentially this last decade. In order to cope with such an increase and to keep up designers' productivity, higher level specifications were required. Moreover new synthesis systems, starting with a high level specification, have been developed in order to automate and speed up processor design. This study presents a VHDL specification methodology aimed to extend structured design methodologies to the behavioral level. The goal is to develop VHDL modeling strategies in order to master the design and analysis of large and complex systems. Structured design methodologies are combined with a high-level synthesis system, a VHDL based behavioral synthesis tool, in order to allow hierarchical design and component re-use.

  3. The novel neuropeptide phoenixin is highly co-expressed with nesfatin-1 in the rat hypothalamus, an immunohistochemical study. (United States)

    Pałasz, Artur; Rojczyk, Ewa; Bogus, Katarzyna; Worthington, John J; Wiaderkiewicz, Ryszard


    The hypothalamus regulates a number of autonomic functions essential for homeostasis; therefore, investigations concerning hypothalamic neuropeptides and their functions and distribution are of great importance in contemporary neuroscience. Recently, novel regulatory factors expressed in the hypothalamus have been discovered, of which nesfatin-1 and phoenixin (PNX), show intriguing similarities in their brain distributions. There are currently few studies characterizing PNX expression, so it is imperative to accurately trace its localization, with particular attention to the hypothalamic nuclei and nesfatin-1 co-expression. Using fluorescence and classical immunohistochemical stainings on adult rat brain, we visualized the potential co-expression of nesfatin-1 and PNX immunoreactive cells. We have demonstrated a distinct PNX-immunoreactivity in 21-32% of cells in the arcuate nucleus, paraventricular nucleus, ventromedial and lateral hypothalamus. Nesfatin-1 expression reached 45-68% of all neurons in the same sites, while co-expression was strikingly seen in the vast majority (70-86%) of PNX-immunoreactive neurons in the rat hypothalamic nuclei. Our results demonstrate for the first time, a wide distribution of PNX in the hypothalamus which could implicate a potential functional relationship with nesfatin-1, possibly in the regulation of the hypothalamic-pituitary-gonadal axis or other autonomic functions, which require further study. Copyright © 2015. Published by Elsevier Ireland Ltd.

  4. Protein tyrosine nitration in chronic intramuscular parasitism: immunohistochemical evaluation of relationships between nitration, and fiber type-specific responses to infection

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    Ted H. Elsasser


    Full Text Available The present study was conducted to determine whether preferential muscle catabolism [psoas major (PM > rectus femoris (RF] observed during the chronic intramuscular stage of Sarcocystis cruzi infection could be associated with the pathological consequences of increased protein tyrosine nitration in fibers characteristically more metabolically active due to higher mitochondrial density. Holstein calves were assigned to control (C, or S. cruzi-infected (I groups, n=5/group. Calves were euthanized on day 63 of infection. Samples of RF and PM were prepared for metabolic fiber typing (MFT: slow oxidative, SO – Type I; fast oxidative glycolytic, FOG - Type IIa; fast glycolytic, FG – Type IIb, fiber area, and immunohistochemical localization of fast myosin heavy chain 2a and 2b, nitrotyrosine (NT, and mitochondrial Complex V ATP-synthase. MFT analysis documented that PM contained twice the number of SO fibers compared to RF (32 v 16%, P<0.002. SO and FOG fibers (Both higher in mitochondrial density than FG fibers in both PM and RF were significantly smaller in area in I calves with mean FG areas not different between C and I. Muscle NT content (Western blot of myofibrillar protein fraction increased with infection; NT was immunohistochemically localized into three distinct patterns in fibers: i sparse fiber staining, ii dense punctuate intrafiber staining, and iii pericystic staining. By image analysis, the greatest punctuate intrafiber pixel density of NT was associated with SO fibers from I calves with the NT colocalizing with mitochondrial Complex V – F1F0 ATP synthase. More fibers were positive for the colocalization in PM than RF (P<0.04. The data are consistent with the concept that fibers rich in mitochondria possessing more inherent oxidative energy capacity generate more nitrated proteins than glycolytic fibers and as such are more affected by the proinflammatory response to infections like Sarcocystosis.

  5. High Ki-67 Immunohistochemical Reactivity Correlates With Poor Prognosis in Bladder Carcinoma: A Comprehensive Meta-Analysis with 13,053 Patients Involved. (United States)

    Luo, Yihuan; Zhang, Xin; Mo, Meile; Tan, Zhong; Huang, Lanshan; Zhou, Hong; Wang, Chunqin; Wei, Fanglin; Qiu, Xiaohui; He, Rongquan; Chen, Gang


    Ki-67 is considered as one of prime biomarkers to reflect cell proliferation and immunohistochemical Ki-67 staining has been widely applied in clinical pathology. To solve the widespread controversy whether Ki-67 reactivity significantly predicts clinical prognosis of bladder carcinoma (BC), we performed a comprehensive meta-analysis by combining results from different literature. A comprehensive search was conducted in the Chinese databases of WanFang, China National Knowledge Infrastructure and Chinese VIP as well as English databases of PubMed, ISI web of science, EMBASE, Science Direct, and Wiley online library. Independent studies linking Ki-67 to cancer-specific survival (CSS), disease-free survival (DFS), overall survival (OS), progression-free survival (PFS), and recurrence-free survival (RFS) were included in our meta-analysis. With the cut-off values literature provided, hazard ratio (HR) values between the survival distributions were extracted and later combined with STATA 12.0. In total, 76 studies (n = 13,053 patients) were eligible for the meta-analysis. It was indicated in either univariate or multivariate analysis for survival that high Ki-67 reactivity significantly predicted poor prognosis. In the univariate analysis, the combined HR for CSS, DFS, OS, PFS, and RFS were 2.588 (95% confidence interval [CI]: 1.623-4.127, P < 0.001), 2.697 (95%CI: 1.874-3.883, P < 0.001), 2.649 (95%CI: 1.632-4.300, P < 0.001), 3.506 (95%CI: 2.231-5.508, P < 0.001), and 1.792 (95%CI: 1.409-2.279, P < 0.001), respectively. The pooled HR of multivariate analysis for CSS, DFS, OS, PFS, and RFS were 1.868 (95%CI: 1.343-2.597, P < 0.001), 2.626 (95%CI: 2.089-3.301, P < 0.001), 1.104 (95%CI: 1.008-1.209, P = 0.032), 1.518 (95%CI: 1.299-1.773, P < 0.001), and 1.294 (95%CI: 1.203-1.392, P < 0.001), respectively. Subgroup analysis of univariate analysis by origin showed that Ki-67 reactivity significantly correlated with all 5

  6. Primary breast cancer tumours contain high amounts of IgA1 immunoglobulin: an immunohistochemical analysis of a possible carrier of the tumour-associated Tn antigen.

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    Charlotte Welinder

    Full Text Available The Tn antigen (GalNAc alpha-O-Ser/Thr as defined by the binding of the lectin, helix pomatia agglutinin (HPA or anti-Tn monoclonal antibodies, is known to be exposed in a majority of cancers, and it has also been shown to correlate positively with the metastatic capacity in breast carcinoma. The short O-glycan that forms the antigen is carried by a number of different proteins. One potential carrier of the Tn antigen is immunoglobulin A1 (IgA1, which we surprisingly found in tumour cells of the invasive parts of primary breast carcinoma. Conventional immunohistochemical analysis of paraffin-embedded sections from primary breast cancers showed IgA1 to be present in the cytoplasm and plasma membrane of 35 out of 36 individual primary tumours. The immunohistochemical staining of HPA and anti-Tn antibody (GOD3-2C4 did to some extent overlap with the presence of IgA1 in the tumours, but differences were seen in the percentage of stained cells and in the staining pattern in the different breast cancers analysed. Anti-Tn antibody and HPA were also shown to specifically bind to a number of possible constellations of the Tn antigen in the hinge region of IgA1. Both reagents could also detect the presence of Tn positive IgA in serum. On average 51% of the tumour cells in the individual breast cancer tumour sections showed staining for IgA1. The overall amount of staining in the invasive part of the tumour with the anti Tn antibody was 67%, and 93% with HPA. The intra-expression or uptake of IgA1 in breast cancer makes it a new potential carrier of the tumour associated and immunogenic Tn antigen.

  7. Immunohistochemical expression of myoepithelial markers in adenomyoepithelioma of the breast: a unique paradoxical staining pattern of high-molecular weight cytokeratins. (United States)

    Moritani, Suzuko; Ichihara, Shu; Yatabe, Yasushi; Hasegawa, Masaki; Iwakoshi, Akari; Hosoda, Waki; Narita, Michihiko; Nagai, Yuichiro; Asai, Masami; Ujihira, Nobuko; Yuba, Yoshiaki; Jijiwa, Mayumi


    To determine which immunohistochemical markers are useful for the identification of neoplastic myoepithelial cells in adenomyoepithelioma of the breast, the expression of seven myoepithelial markers (α-smooth muscle actin (α-SMA), calponin, p63, CD10, cytokeratin 5/6, cytokeratin 14, and S-100) was examined in 19 lesions from 16 patients. The lesion consisted of seven spindle and 12 clear cell lesions. For normal myoepithelial cells, α-SMA, calponin, and p63 were significantly more sensitive than cytokeratin 5/6, cytokeratin 14, and S-100. There was no significant difference in the expression of α-SMA, calponin, p63, and CD10 in neoplastic myoepithelial cells of adenomyoepithelioma regardless of spindle or clear cell types. In spindle cell lesions, high-molecular weight cytokeratins (HMWCK; cytokeratin 5/6 and cytokeratin 14) tended to show higher staining scores and S-100 showed lower staining scores than other markers. In clear cell lesions, HMWCK showed significantly lower staining scores than the other five markers. There was no significant difference in staining scores among the other five markers. HMWCK showed a unique paradoxical staining pattern in clear cell lesions, with diffusely positive inner epithelial cells and completely negative outer myoepithelial cells. Although the sensitivity of HMWCK in clear cell lesions is low, with this unique paradoxical staining pattern and relatively high sensitivity in spindle cell lesions, HMWCK could be useful in diagnosing adenomyoepithelioma. In choosing immunohistochemical markers, any of the seven markers are useful, but combining HMWCK and any one of α-SMA, calponin, and p63 would be a good panel for the diagnosis of adenomyoepithelioma.

  8. Aldehyde dehydrogenase 3B1 (ALDH3B1): immunohistochemical tissue distribution and cellular-specific localization in normal and cancerous human tissues. (United States)

    Marchitti, Satori A; Orlicky, David J; Brocker, Chad; Vasiliou, Vasilis


    Aldehyde dehydrogenase (ALDH) enzymes are critical in the detoxification of endogenous and exogenous aldehydes. Our previous findings indicate that the ALDH3B1 enzyme is expressed in several mouse tissues and is catalytically active toward aldehydes derived from lipid peroxidation, suggesting a potential role against oxidative stress. The aim of this study was to elucidate by immunohistochemistry the tissue, cellular, and subcellular distribution of ALDH3B1 in normal human tissues and in tumors of human lung, colon, breast, and ovary. Our results indicate that ALDH3B1 is expressed in a tissue-specific manner and in a limited number of cell types, including hepatocytes, proximal convoluted tubule cells, cerebellar astrocytes, bronchiole ciliated cells, testis efferent ductule ciliated cells, and histiocytes. ALDH3B1 expression was upregulated in a high percentage of human tumors (lung > breast = ovarian > colon). Increased ALDH3B1 expression in tumor cells may confer a growth advantage or be the result of an induction mechanism mediated by increased oxidative stress. Subcellular localization of ALDH3B1 was predominantly cytosolic in tissues, with the exception of normal human lung and testis, in which localization appeared membrane-bound or membrane-associated. The specificity of ALDH3B1 distribution may prove to be directly related to the functional role of this enzyme in human tissues.

  9. Development of High Specific Strength Envelope Materials (United States)

    Komatsu, Keiji; Sano, Masa-Aki; Kakuta, Yoshiaki

    Progress in materials technology has produced a much more durable synthetic fabric envelope for the non-rigid airship. Flexible materials are required to form airship envelopes, ballonets, load curtains, gas bags and covering rigid structures. Polybenzoxazole fiber (Zylon) and polyalirate fiber (Vectran) show high specific tensile strength, so that we developed membrane using these high specific tensile strength fibers as a load carrier. The main material developed is a Zylon or Vectran load carrier sealed internally with a polyurethane bonded inner gas retention film (EVOH). The external surface provides weather protecting with, for instance, a titanium oxide integrated polyurethane or Tedlar film. The mechanical test results show that tensile strength 1,000 N/cm is attained with weight less than 230g/m2. In addition to the mechanical properties, temperature dependence of the joint strength and solar absorptivity and emissivity of the surface are measured. 

  10. Production of high specific activity silicon-32

    Energy Technology Data Exchange (ETDEWEB)

    Phillips, D.R. [Los Alamos National Lab., NM (United States); Brzezinski, M.A. [Univ. of California, Santa Barbara, CA (United States). Marine Biotechnology Center


    This is the final report of a three-year, Laboratory Directed Research and Development Project (LDRD) at Los Alamos National Laboratory (LANL). There were two primary objectives for the work performed under this project. The first was to take advantage of capabilities and facilities at Los Alamos to produce the radionuclide {sup 32}Si in unusually high specific activity. The second was to combine the radioanalytical expertise at Los Alamos with the expertise at the University of California to develop methods for the application of {sup 32}Si in biological oceanographic research related to global climate modeling. The first objective was met by developing targetry for proton spallation production of {sup 32}Si in KCl targets and chemistry for its recovery in very high specific activity. The second objective was met by developing a validated field-useable, radioanalytical technique, based upon gas-flow proportional counting, to measure the dynamics of silicon uptake by naturally occurring diatoms.

  11. Endogenous avidin biotin activity (EABA in thyroid pathology: immunohistochemical study

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    Nikiel Barbara


    Full Text Available Abstract Background Immunohistochemical methods based on the high affinity of avidin and biotin (e.g. ABC, LSAB are characterized by high sensitivity and are widely used for detection of immunologic reaction. However, a non-specific reaction, observed in frozen tissues and in paraffin-embedded material, increasing after heat induced epitope retrieval (HIER, and caused either by endogenous biotin or any another chemical compound with high affinity for avidin, may lead to diagnostic mistakes. The aim of our investigation is to study presence of endogenous avidin biotin activity (EABA in thyrocytes originating from various thyroid pathological lesions (neoplastic and non-neoplastic. Materials and methods The immunohistochemical study was performed on paraffin-embedded specimens of surgically resected thyroid tissue from 97 patients with thyroid diseases: 65 patients with papillary carcinoma (PTC, 11 patients with nodular goiter in whom features of benign papillary hyperplasia were found, 9 with lymphocytic thyroiditis (LT, 8 with follicular adenoma, and 4 patients with follicular carcinoma. In PTC immunohistochemical study was performed both in primary tumors and in lymph node metastases. After HIER, incubation with streptavidin from LSAB+ (DakoCytomation kit was done. Results Strong cytoplasmic EABA was observed in 56 of 65 (87.5% PTC and in oxyphilic cells in 8 of 9 cases of LT. Significant correlation between EABA in primary PTC tumor and EABA in lymph node metastases was stated. Normal surrounding thyroid tissues showed absence or weak EABA. Aberrant intranuclear localization of biotin was noted in morules of cribriform-morular variant of PTC. No statistically significant correlation between patient's age, sex, metastases presence and EABA was observed. Conclusion Among thyroid lesions, false positive reactions are highly probable in papillary thyroid carcinoma and in lymphocytic thyroiditis if immunohistochemical detection is used on systems

  12. Distinguishing medullary carcinoma of the breast from high-grade hormone receptor-negative invasive ductal carcinoma: an immunohistochemical approach.

    NARCIS (Netherlands)

    Flucke, U.E.; Flucke, M.T.; Hoy, L.; Breuer, E.; Goebbels, R.; Rhiem, K.; Schmutzler, R.; Winzenried, H.; Braun, M.; Steiner, S.; Buettner, R.; Gevensleben, H.


    AIMS: Medullary carcinomas (MCs) represent a rare breast cancer subtype associated with a rather favourable prognosis compared with invasive ductal carcinomas (IDCs). Due to histopathological overlap, MCs are frequently misclassified as high-grade IDCs, potentially leading to overtreatment of MCs. O

  13. Immunohistochemical expression of intrarenal renin angiotensin system components in response to tempol in rats fed a high salt diet (United States)

    Cao, Gabriel; Della Penna, Silvana Lorena; Kouyoumdzian, Nicolás Martín; Choi, Marcelo Roberto; Gorzalczany, Susana; Fernández, Belisario Enrique; Toblli, Jorge Eduardo; Rosón, María Inés


    AIM To determine the effect of tempol in normal rats fed high salt on arterial pressure and the balance between antagonist components of the renal renin-angiotensin system. METHODS Sprague-Dawley rats were fed with 8% NaCl high-salt (HS) or 0.4% NaCl (normal-salt, NS) diet for 3 wk, with or without tempol (T) (1 mmol/L, administered in drinking water). Mean arterial pressure (MAP), glomerular filtration rate (GFR), and urinary sodium excretion (UVNa) were measured. We evaluated angiotensin II (Ang II), angiotensin 1-7 (Ang 1-7), angiotensin converting enzyme 2 (ACE2), mas receptor (MasR), angiotensin type 1 receptor (AT1R) and angiotensin type 2 receptor (AT2R) in renal tissues by immunohistochemistry. RESULTS The intake of high sodium produced a slight but significant increase in MAP and differentially regulated components of the renal renin-angiotensin system (RAS). This included an increase in Ang II and AT1R, and decrease in ACE-2 staining intensity using immunohistochemistry. Antioxidant supplementation with tempol increased natriuresis and GFR, prevented changes in blood pressure and reversed the imbalance of renal RAS components. This includes a decrease in Ang II and AT1R, as increase in AT2, ACE2, Ang (1-7) and MasR staining intensity using immunohistochemistry. In addition, the natriuretic effects of tempol were observed in NS-T group, which showed an increased staining intensity of AT2, ACE2, Ang (1-7) and MasR. CONCLUSION These findings suggest that a high salt diet leads to changes in the homeostasis and balance between opposing components of the renal RAS in hypertension to favour an increase in Ang II. Chronic antioxidant supplementation can modulate the balance between the natriuretic and antinatriuretic components of the renal RAS. PMID:28101449

  14. Prostatic tissue in testicular teratoma. A clinicopathologic and immunohistochemical study. (United States)

    Roma, Andres A; Humphrey, Peter A


    The presence of prostatic differentiation as part of teratoma is very unusual and has been reported less than 20 times in the literature; however, all but 1 case were described in ovarian teratomas. We reviewed 45 specimens of germ cell tumors with teratoma component in postpuberal male patients. Original hematoxylin and eosin review failed to identify glands morphologically consistent with prostatic differentiation. Immunohistochemical stains performed on 10 specimens from 10 patients with small glandular and/or tubular structures revealed 1 case with glands positive for prostatic-specific antigen, prostatic-specific acid phosphatase, and prostein/P501S, whereas high-molecular-weight cytokeratin and p63 highlighted only basal cells. The glands were irregular in size and shape and contained mostly cuboidal to columnar luminal-type cells with occasional basal-type cells. Re-review of all the specimens revealed a second block from the same testis as well as 1 retroperitoneal lymph node with metastatic teratoma in the same patient, also immunohistochemically confirmed. These glands were seen in a smooth muscle stromal background, adjacent to classic gastrointestinal and tracheobronchial teratoma components. Our findings show immunohistochemically confirmed prostatic differentiation in 2 specimens from 1 patient with teratoma. This study raises the possibility that prostatic differentiation, difficult to recognize on morphology alone, might not be that unusual and that immunostains can help detect it over the several different epithelial components of teratoma.

  15. Immunohistochemical study on the expression of matrix metalloproteinase 2 and high-risk human papilloma virus in the malignant progression of papillomas. (United States)

    Lee, Ho-Jin; Kim, Jin-Wook


    Papilloma frequently develops as a benign tumor of the head and neck area, but its potential for malignant transformation has yet to be studied. This study aims to provide basic information for papillomas using the immunohistochemical staining of matrix metalloproteinase 2 (MMP-2) and human papilloma virus (HPV) 16 and 18. To evaluate the malignant transformation of papillomas, the selected tissue samples were serially diagnosed with pre-cancerous papilloma (with epithelial dysplasia, pseudo-epitheliomatous hyperplasia) or malignant lesion (squamous cell carcinoma, SCC) after the first diagnosis (squamous papilloma, inverted papilloma). The selected tissues were stained with an antibody to MMP-2 and HPV 16-E7, HPV 18-L1. A statistical analysis was performed according to each transformation step. The epithelial layer of papilloma and pre-cancerous papilloma lesions had a similar MMP-2 expression, but that of the malignant lesion had a significantly increased MMP-2 expression. HPV 16 and 18 infection rates were 28.6%, 33.3% and 63.6% in papillomas, pre-cancerous papilloma lesions, and SCC. A relatively high MMP-2 expression and HPV 16 or 18 infection of papillomas may be associated with early events in the multistep processes of malignant transformation of papillomas.

  16. Double-hit lymphomas: clinical, morphological, immunohistochemical and cytogenetic study in a series of Brazilian patients with high-grade non-Hodgkin lymphoma. (United States)

    Oliveira, Cristiano Claudino; Maciel-Guerra, Helena; Kucko, Luan; Hirama, Eric Jun; Brilhante, Américo Delgado; Quevedo, Francisco Carlos; da Cunha, Isabela Werneck; Soares, Fernando Augusto; Niero-Melo, Ligia; Dos Reis, Patrícia Pintor; Domingues, Maria Aparecida Custodio


    Double-hit lymphomas (DHL) are rare high-grade neoplasms characterized by two translocations: one involving the gene MYC and another involving genes BCL2 or BCL6, whose diagnosis depends on cytogenetic examination. This research studied DHL and morphological and/or immunophenotypic factors associated with the detection of these translocations in a group of high-grade non-Hodgkin lymphoma cases. Clinical and morphological reviews of 120 cases diagnosed with diffuse large B-cell lymphoma and Burkitt lymphoma were conducted. Immunohistochemistry (CD20, CD79a, PAX5, CD10, Bcl6, Bcl2, MUM1, TDT and Myc) and fluorescence in situ hybridization for detection of MYC, BCL2 and BCL6 gene translocations were performed in a tissue microarray platform. Three cases of DHL were detected: two with translocations of MYC and BCL2 and one with translocations of MYC and BCL6, all leading to death in less than six months. Among 90 cytogenetically evaluable biopsies, associations were determined between immunohistochemistry and fluorescence in situ hybridization for MYC (p = 0.036) and BCL2 (p = 0.001). However, these showed only regular agreement, indicated by Kappa values of 0.23 [0.0;0.49] and 0.35 [0.13;0.56], respectively. "Starry sky" morphology was strongly associated with MYC positivity (p = 0.01). The detection of three cases of DHL, all resulting in death, confirms the rarity and aggressiveness of this neoplasm. The "starry sky" morphological pattern and immunohistochemical expression of Myc and Bcl2 represent possible selection factors for additional cytogenetic diagnostic testing.

  17. Local and distant recurrences in rectal cancer patients are predicted by the nonspecific immune response; specific immune response has only a systemic effect - a histopathological and immunohistochemical study

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    Mulder-Stapel Adri


    Full Text Available Abstract Background Invasion and metastasis is a complex process governed by the interaction of genetically altered tumor cells and the immunological and inflammatory host reponse. Specific T-cells directed against tumor cells and the nonspecific inflammatory reaction due to tissue damage, cooperate against invasive tumor cells in order to prevent recurrences. Data concerning involvement of individual cell types are readily available but little is known about the coordinate interactions between both forms of immune response. Patients and methods The presence of inflammatory infiltrate and eosinophils was determined in 1530 patients with rectal adenocarcinoma from a multicenter trial. We selected 160 patients to analyze this inflammatory infiltrate in more detail using immunohistochemistry. The association with the development of local and distant relapses was determined using univariate and multivariate log rank testing. Results Patients with an extensive inflammatory infiltrate around the tumor had lower recurrence rates (3.4% versus 6.9%, p = 0.03, showing the importance of host response against tumor cells. In particular, peritumoral mast cells prevent local and distant recurrence (44% versus 15%, p = 0.007 and 86% versus 21%, p Conclusions We showed that next to properties of tumor cells, the amount and type of inflammation is also relevant in the control of rectal cancer. Knowledge of the factors involved may lead to new approaches in the management of rectal cancer.

  18. Measuring Specific Heats at High Temperatures (United States)

    Vandersande, Jan W.; Zoltan, Andrew; Wood, Charles


    Flash apparatus for measuring thermal diffusivities at temperatures from 300 to 1,000 degrees C modified; measures specific heats of samples to accuracy of 4 to 5 percent. Specific heat and thermal diffusivity of sample measured. Xenon flash emits pulse of radiation, absorbed by sputtered graphite coating on sample. Sample temperature measured with thermocouple, and temperature rise due to pulse measured by InSb detector.

  19. Sensitivity and specificity of real-time reverse transcription polymerase chain reaction, histopathology, and immunohistochemical labeling for the detection of Rift Valley fever virus in naturally infected cattle and sheep. (United States)

    Odendaal, Lieza; Fosgate, Geoffrey T; Romito, Marco; Coetzer, Jacobus A W; Clift, Sarah J


    Real-time reverse transcription polymerase chain reaction (real-time RT-PCR), histopathology, and immunohistochemical labeling (IHC) were performed on liver specimens from 380 naturally infected cattle and sheep necropsied during the 2010 Rift Valley fever (RVF) epidemic in South Africa. Sensitivity (Se) and specificity (Sp) of real-time RT-PCR, histopathology, and IHC were estimated in a latent-class model using a Bayesian framework. The Se and Sp of real-time RT-PCR were estimated as 97.4% (95% confidence interval [CI] = 95.2-98.8%) and 71.7% (95% CI = 65-77.9%) respectively. The Se and Sp of histopathology were estimated as 94.6% (95% CI = 91-97.2%) and 92.3% (95% CI = 87.6-95.8%), respectively. The Se and Sp of IHC were estimated as 97.6% (95% CI = 93.9-99.8%) and 99.4% (95% CI = 96.9-100%), respectively. Decreased Sp of real-time RT-PCR was ascribed to cross-contamination of samples. Stratified analysis of the data suggested variations in test accuracy with fetuses and severely autolyzed specimens. The Sp of histopathology in fetuses (83%) was 9.3% lower than the sample population (92.3%). The Se of IHC decreased from 97.6% to 81.5% in the presence of severe autolysis. The diagnostic Se and Sp of histopathology was higher than expected, confirming the value of routine postmortem examinations and histopathology of liver specimens. Aborted fetuses, however, should be screened using a variety of tests in areas endemic for RVF, and results from severely autolyzed specimens should be interpreted with caution. The most feasible testing option for countries lacking suitably equipped laboratories seems to be routine histology in combination with IHC.

  20. In vivo microdialysis and immunohistochemical analyses of tendon tissue demonstrated high amounts of free glutamate and glutamate NMDAR1 receptors, but no signs of inflammation, in Jumper's knee. (United States)

    Alfredson, H; Forsgren, S; Thorsen, K; Lorentzon, R


    This investigation describes, to our knowledge, the first experiment where the microdialysis technique was used to study certain metabolic events in human patellar tendons in combination with immunohistochemical analyses of tendon biopsies. In five patients (four men and one woman) with a long duration (range 12-36 months) of pain symptoms from Jumper's knee (localized tenderness in the patellar tendon verified as tendon changes with ultrasonography or MRI), and in five controls (four men and one woman) with normal patellar tendons, a standard microdialysis catheter was inserted into the patellar tendon under local anestesia. The local concentrations of glutamate (excitatory neurotransmitter) and prostaglandin E2 (PGE2) were registered under resting conditions. Samplings were done every 15 min during a 2 h period. In all individuals (patients and controls) biopsies were taken for immunohistochemical analyses. The results showed that it was possible to detect and measure the concentrations of glutamate and PGE2 in the patellar tendon with the use of microdialysis technique. There were significantly higher concentrations of free glutamate, but not PGE2, in tendons with tendinosis compared to normal tendons. In the biopsies, there were no inflammatory cell infiltrates, but, for the first time, it was shown that there was immunoreaction for the glutamate receptor NMDAR1 in association with nerve structures in human patellar tendons. These findings altogether indicate that glutamate might be involved in painful Jumper's knee, and further emphasizes that there is no chemical inflammation (normal PGE2 levels) in this chronic condition.

  1. Versatility of immunohistochemical reactions: comprehensive survey of detection systems. (United States)

    Mokrý, J


    The field of immunohistochemistry comprises histological methods enabling detection of tissue antigens via specific antibodies. Although all these techniques take an advantage of a large specificity of antibody to a particular tissue antigen there are many different approaches for enhancement and visualization of the signal. The aim of the present review article was to briefly outline the historical milestones that made the rapid progress of this discipline possible and give a comprehensive survey of immunohistochemical methods applicable to biomedical research. The survey starts with a description of the direct immunohistochemical method and then pays attention to a huge number of indirect methods. For better explanation of principles of individual techniques, the text is accompanied with graphical schemes. The highest attention is given to immunohistochemical methods that are most generally used, i.e. enzyme anti-enzyme complex methods (e.g. Peroxidase Anti-Peroxidase/PAP/ or Alkaline Phosphatase Anti-Alkaline Phosphatase /APAAP/) and methods based on avidin-biotin interactions (Bridged Avidin-Biotin/BRAB/, Avidin-Biotin Complex /ABC/, Labelled Avidin-Biotin/LAB/). Nevertheless, the principles of other immunohistochemical methods like two- or three-step indirect immunohistochemical methods, methods based on protein A-antibody interaction and Hapten Antibody Anti-Hapten method (HAAH), are also thoroughly characterized. Usefulness of each method for a specific utilization, its advantages and disadvantages are mentioned and compared with the latest immunohistochemical techniques, like Multi-Layered Peroxidase-Labelled Antibody (MLP) and water soluble polymer conjugates, e.g. Enhanced Polymer One-Step staining (EPOS) or EnVision. The last paragraphs are devoted to immunohistochemical amplification systems (Catalyzed Signal Amplification/CSA/ and label anti-label) that dramatically increase sensitivity of detection systems and enable to compare sensitivity of

  2. COTA (colon-ovarian tumor antigen). An immunohistochemical study. (United States)

    Pant, K D; Fenoglio-Preiser, C M; Berry, C O; Zamora, P O; Ram, M D; Fulks, R M; Rhodes, B A


    A goat anti-serum was prepared against mucinous ovarian cyst fluid and absorbed with normal colon and a variety of normal tissues until the only residual immunoreactivity was directed against colon cancer and ovarian tumor mucin. The set of antigenic determinants defined by this anti-serum has been called COTA, standing for colon-ovarian-tumor-antigen. This highly absorbed anti-serum (anti-COTA) was used for immunohistochemical staining of 42 different tissues in parallel with staining with a goat anti-CEA, which was also highly absorbed. The results suggest that COTA is a highly sensitive and specific antigen for colon carcinoma and may have potential for the early detection of malignant changes predictive of cancer of the colon.


    Institute of Scientific and Technical Information of China (English)

    信明军; 施诚仁; 张忠德; 李敏; 陈方; 潘伟华


    Objective To evaluate the sensitivity and specificity of regularly used immunohistochemical markers, including Vimentin (Vim), Desmin (Des), Myoglobin (MG), Myosin (MS), Smooth-muscle actin(SMA) and Sarcomeric actin ( Sr-A), in the diagnosis of rhabdomyosarcoma (RMS). Methods After resection, 24 RMSs and other childhood tumor specimens were fixed in 10% neutral-buffered formalin and embeded in paraffin. The immunohistochemical staining was performed by LSAB procedure. Heat-induced epitope retrieval of Des, MS, Sr-A was processed in order to enhence positive rate and positive strength. Results Vim, MG, MS,Des, Sr-A, SMA were arranged in the order of sensitivity from higher to lower. About specificity, Sr-A, Des, SMA,MG, Vim standed in a sequence from higher to lower (the data of MS is insufficient); Des, MG, Sr-A possessed higher experimental efficiency, followed by SMA, Vim in a succession. Conclusion Vim and MG are of the higher sensitivity but lower specificity. On the reverse, Sr-A and Des hold the better specificity but lower sensitivity.So the combination of multiple antibody reactions should be considered to improve the diagnostic ability in poorly differentiated RMS. According to the result of experimental efficiency, we suggest that the combination of Des, MG and Sr-A can make it possible to diagnose the majority of RMS clearly.

  4. Specifics of marketing strategy in the segment of high fashion


    Butigan, Ružica; Grilec Kaurić, Alica; UJEVIĆ, Darko


    The success of high fashion designers is not only in a specificity of the products but also in specific and very well executed marketing strategy. Emphasis is placed on the design of very specific marketing program and marketing strategies that must concider all the characteristics of the high fashion market. Therefore, a scientific research problem is defined as follows: although the market of high fashion at first glance does not imply a completely different marketing approach than other fa...

  5. High expression of testes-specific protease 50 is associated with poor prognosis in colorectal carcinoma.

    Directory of Open Access Journals (Sweden)

    Lei Zheng

    Full Text Available BACKGROUND: Testes-specific protease 50 (TSP50 is normally expressed in testes and abnormally expressed in breast cancer, but whether TSP50 is expressed in colorectal carcinoma (CRC and its clinical significance is unclear. We aimed to detect TSP50 expression in CRC, correlate it with clinicopathological factors, and assess its potential diagnostic and prognostic value. METHODOLOGY/PRINCIPAL FINDINGS: TSP50 mRNAs and proteins were detected in 7 CRC cell lines and 8 CRC specimens via RT-PCR and Western blot analysis. Immunohistochemical analysis of TSP50, p53 and carcinoembryonic antigen (CEA with tissue microarrays composed of 95 CRCs, 20 colorectal adenomas and 20 normal colorectal tissues were carried out and correlated with clinicopathological characteristics and disease-specific survival for CRC patients. There was no significant correlation between the expression levels of TSP50 and p53 (P = 0.751 or CEA (P = 0.663. Abundant expression of TSP50 protein was found in CRCs (68.4% while it was poorly expressed in colorectal adenomas and normal tissues (P<0.0001. Thus, CRCs can be distinguished from them with high specificity (92.5% and positive predictive value (PPV, 95.6%. The survival of CRC patients with high TSP50 expression was significantly shorter than that of the patients with low TSP50 expression (P = 0.010, specifically in patients who had early-stage tumors (stage I and II; P = 0.004. Multivariate Cox regression analysis indicated that high TSP50 expression was a statistically significant independent risk factor (hazard ratio  = 2.205, 95% CI = 1.214-4.004, P = 0.009. CONCLUSION: Our data demonstrate that TSP50 is a potential effective indicator of poor survival for CRC patients, especially for those with early-stage tumors.

  6. [Biochemical and immunohistochemical markers of brain injury]. (United States)

    Vajtr, D; Průsa, R; Houst'ava, L; Sámal, F; Kukacka, J; Pachl, J


    Proteins released to circulation from affected tissues during primary or secondary trauma brain injury might be used as serum markers of glial or ganglial cells damage (neuron specific enolasis and S100 B protein). Other markers of trauma can be proved as relatively specific of diffuse axonal injury by immunohistochemical detectoin (amyloid prekurzor protein, neuron specific enolasis, glial fibrilar acidic protein and superficial antigen receptor CD 68). Some markers are associated with blood brain barrier damage (matrix metaloproteinases (MMP-2, MMP-9) and synthase of nitric oxide (iNOS)). We aimed in our short communication on biomechanics of developed of trauma, primary or secondary kinds of trauma brain injury and use of trauma brain injury markers for clinical diagnostics and management of patients.

  7. Human vomeronasal epithelium development: An immunohistochemical overview. (United States)

    Dénes, Lóránd; Pap, Zsuzsanna; Szántó, Annamária; Gergely, István; Pop, Tudor Sorin


    The vomeronasal organ (VNO) is the receptor structure of the vomeronasal system (VNS) in vertebrates. It is found bilaterally in the submucosa of the inferior part of the nasal septum. There are ongoing controversies regarding the functionality of this organ in humans. In this study we propose the immunohistochemical evaluation of changes in components of the human vomeronasal epithelium during foetal development. We used 45 foetuses of different age, which were included in three age groups. After VNO identification immunohistochemical reactions were performed using primary antibodies against the following: neuron specific enolase, calretinin, neurofilament, chromogranin, synaptophysin, cytokeratin 7, pan-cytokeratin and S100 protein. Digital slides were obtained and following colorimetric segmentation, surface area measurements were performed. The VNO was found in less than half of the studied specimens (42.2%). Neuron specific enolase and calretinin immunoexpression showed a decreasing trend with foetal age, while the other neural/neuroendocrine markers were negative in all specimens. Cytokeratin 7 expression increased with age, while Pan-Ctk had no significant variations. S100 protein immunoexpression also decreased around the VNO. The results of the present work uphold the theory of regression of the neuroepithelium that is present during initial stages of foetal development.

  8. Immunohistochemical diagnosis of urinary bladder tuberculosis

    Directory of Open Access Journals (Sweden)

    S. A. Semenov


    Full Text Available Diagnostics of urinary bladder tuberculosis bases on pathological verification. Standard histological staining (hematoxylin–eosin reveals glaucomatous inflammation, but cannot estimate its etiology.Aim of our study was to evaluate the role of complex immunohistochemical method in diagnostic of tuberculosis infection in bladder. Our study included 21 histological specimen of the resected bladder in case of nephrotuberculosis. Standard histological examination revealed specific changes in bladder tissue only in 2 cases, while immunohistochemical method with antibodies to Mycobacterium tuberculosis (MBT demonstrated positive reaction at 5 patients. Investigation of lower urinary tract function in late postoperative period showed that patients with positive anti-MBT reaction had clinically significant chronic urinary retention, as well as their degree of urinary disorders assessed using a questionnaire IPSS-Qol was higher. Thus, the use of IHC method in combination with standard histological examination improves diagnostics of urinary bladder tuberculosis, and it may serve the predictor of long-term results of surgical treatment of microcystis.

  9. Tympanomastoid cholesterol granulomas: Immunohistochemical evaluation of angiogenesis. (United States)

    Iannella, Giannicola; Di Gioia, Cira; Carletti, Raffaella; Magliulo, Giuseppe


    This study investigates the immunohistochemical expression of vascular endothelial growth factor (VEGF) and CD34 in patients treated for middle ear and mastoid cholesterol granulomas to evaluate the angiogenesis and vascularization of this type of lesion. A correlation between the immunohistochemical data and the radiological and intraoperative evidence of temporal bone marrow invasion and blood source connection was performed to validate this hypothesis. Retrospective study. Immunohistochemical expression of VEGF and CD34 in a group of 16 patients surgically treated for cholesterol granuloma was examined. Middle ear cholesteatomas with normal middle ear mucosa and external auditory canal skin were used as the control groups. The radiological and intraoperative features of cholesterol granulomas were also examined. In endothelial cells, there was an increased expression of angiogenetic growth factor receptors in all the cholesterol granulomas in this study. The quantitative analysis of VEGF showed a mean value of 37.5, whereas the CD34 quantitative analysis gave a mean value of 6.8. Seven patients presented radiological or intraoperative evidence of bone marrow invasion, hematopoietic potentialities, or blood source connections that might support the bleeding theory. In all of these cases there was computed tomography or intraoperative evidence of bone erosion of the middle ear and/or temporal bone structures. The mean values of VEGF and CD34 were 41.1 and 7.7, respectively. High values of VEGF and CD34 are present in patients with cholesterol granulomas. Upregulation of VEGF and CD34 is indicative of a remarkable angiogenesis and a widespread vascular concentration in cholesterol granulomas. 3b. Laryngoscope, 127:E283-E290, 2017. © 2017 The American Laryngological, Rhinological and Otological Society, Inc.

  10. Phenotypic and immunohistochemical characterization of sarcoglycanopathies

    Directory of Open Access Journals (Sweden)

    Ana F. B. Ferreira


    Full Text Available INTRODUCTION: Limb-girdle muscular dystrophy presents with heterogeneous clinical and molecular features. The primary characteristic of this disorder is proximal muscular weakness with variable age of onset, speed of progression, and intensity of symptoms. Sarcoglycanopathies, which are a subgroup of the limb-girdle muscular dystrophies, are caused by mutations in sarcoglycan genes. Mutations in these genes cause secondary deficiencies in other proteins, due to the instability of the dystrophin-glycoprotein complex. Therefore, determining the etiology of a given sarcoglycanopathy requires costly and occasionally inaccessible molecular methods. OBJECTIVE: The aim of this study was to identify phenotypic differences among limb-girdle muscular dystrophy patients who were grouped according to the immunohistochemical phenotypes for the four sarcoglycans. METHODS: To identify phenotypic differences among patients with different types of sarcoglycanopathies, a questionnaire was used and the muscle strength and range of motion of nine joints in 45 patients recruited from the Department of Neurology - HC-FMUSP (Clinics Hospital of the Faculty of Medicine of the University of São Paulo were evaluated. The findings obtained from these analyses were compared with the results of the immunohistochemical findings. RESULTS: The patients were divided into the following groups based on the immunohistochemical findings: a-sarcoglycanopathies (16 patients, b-sarcoglycanopathies (1 patient, y-sarcoglycanopathies (5 patients, and nonsarcoglycanopathies (23 patients. The muscle strength analysis revealed significant differences for both upper and lower limb muscles, particularly the shoulder and hip muscles, as expected. No pattern of joint contractures was found among the four groups analyzed, even within the same family. However, a high frequency of tiptoe gait was observed in patients with a-sarcoglycanopathies, while calf pseudo-hypertrophy was most common in

  11. Immunohistochemical differentiation of inflammatory myopathies

    Directory of Open Access Journals (Sweden)

    Jayalakshmi B Panicker


    Full Text Available Background: Idiopathic inflammatory myopathies are a heterogeneous group of acquired muscle disorders with considerable overlap in the histological features, making histological diagnosis difficult at times. Aims: To determine the immunohistochemical profile of clinically suspected cases of inflammatory myopathies, using monoclonal antibodies to HLA-1 and membrane attack complex (MAC, and to correlate the clinical, serological, and electromyographic profile and the histopathological picture, with the immunohistochemical profile. Settings and Design: This was a retrospective study analyzing the clinical and histopathological features in muscle of clinically suspected cases of inflammatory myopathy and correlating it to their HLA-1 and MAC immunostaining profiles. Material and Methods: The study subjects included 33 cases with suspected inflammatory myopathy and 59 with non-inflammatory muscle disease, as controls. Clinical data, electromyographic findings, serological profile, and details of therapy were obtained from patient records. Statistical Analysis: Student ′T′ test, Pearson′s Chi square test, and Kappa statistics were used appropriately. Results: Although HLA-1 and MAC immunostaining did not help to differentiate the individual subtypes of inflammatory myopathy, when either HLA-1 or MAC was positive, inflammatory myopathy could be ruled in with 86.5% certainty and when both HLA-1 and MAC were negative, it could be ruled out with 95% certainty. Conclusions: A combination of clinical presentation, serological profile, electromyographic and histopathological features, together with the immunoprofile for HLA-1 and MAC, contribute toward making a diagnosis of inflammatory myopathy.

  12. Immunohistochemical diagnosis of fusariosis with monoclonal antibodies

    DEFF Research Database (Denmark)

    Jensen, H.E.; Aalbæk, B.; Jungersen, Gregers

    for fusariosis. A panel of newly developed Mabs for immunohistochemical diagnosis of fusariosis was screened for specificity on experimentally infected laboratory animal tissue and on skin tissue biopsies from two neutropenic patients with Fusarium sepsis. Methods: Somatic antigens were made from F. solani (CBS......Objectives: Fusariosis is an emerging fungal infection, especially in neutropenic patients. Proper identification of Fusarium spp. is important because the choice of antifungal treatment of fusariosis differs from that of aspergillosis, candidosis or scedosporidiosis (pseudalleceriosis). Cultural...... isolation attempts from fusarium lesions often fail, and because the tissue forms of Fusarium spp. are histologically indistinguishable from fungal elements of aspergillosis, true hyphae containing candidosis and scedosporidiosis (pseudalleceriosis), alternative diagnostic techniques are often necessary...

  13. An Efficient Role Specification Management Model for Highly Distributed Environments

    Directory of Open Access Journals (Sweden)

    Soomi Yang


    Full Text Available Highly distributed environments such as pervasive computing environments not having global or broad control, need another attribute certificate management technique. For an efficient role based access control using attribute certificate, we use a technique of structuring role specification certificates. It can provide more flexible and secure collaborating environments. The roles are grouped and made them into the relation tree. It can reduce management cost and overhead incurred when changing the specification of the role. Further we use caching of frequently used role specification certificate for better performance in case applying the role. Tree structured role specification results secure and efficient role renewing and distribution. Caching of role specification helps an application of role. In order to be scalable distribution of the role specification certificate, we use multicasting packets. Also, performance enhancement of structuring role specification certificates is quantified in the sense of taking into account of the packet loss. In the experimental section, it is shown that role updating and distribution are secured and efficient.

  14. Intense neutron source: high-voltage power supply specifications

    Energy Technology Data Exchange (ETDEWEB)

    Riedel, A.A.


    This report explains the need for and sets forth the electrical, mechanical and safety specifications for a high-voltage power supply to be used with the intense neutron source. It contains sufficient information for a supplier to bid on such a power supply.

  15. Immunohistochemical detection of human intestinal spirochetosis. (United States)

    Ogata, Sho; Shimizu, Ken; Oda, Tomohiro; Tominaga, Susumu; Nakanishi, Kuniaki


    Human intestinal spirochetosis (HIS) is a colorectal infection by Brachyspira species of spiral bacteria. Immunohistochemical cross-reaction to an antibody for Treponema pallidum aids its histologic diagnosis. This study's aim was to analyze the immunohistochemical characteristics of HIS. In this analysis, on 223 specimens from 83 HIS cases, we focused on so-called fringe formation (a histologic hallmark of HIS), spiral organisms within mucus or within crypts, and strong immunopositive materials in the mucosa, together with their location and the types of lesions. Fringe formation was found in 81.6% of all specimens and spiral organisms within mucus or within crypts in 97.3% and 57.0%, respectively. Strong immunopositive materials were observed in the surface epithelial layer in 87.9%, in the subepithelial layer in 94.6%, and in deeper mucosa in 2.2% of all specimens. The positive rates in conventional adenomas (24.0%, n = 146) and hyperplastic nodules (100%, n = 17) were each different from that found in inflammation (70.8%, n = 24), and spiral organisms were seen more frequently in the right-side large intestine than in the left (within mucus, 100%, n = 104 versus 95.0%, n = 119; within crypts, 65.4%, n = 104 versus 49.6%, n = 119). Thus, immunohistochemistry was effective not only in supporting the diagnosis of HIS but also in highlighting spiral organisms within mucus or crypts that were invisible in routine histology. Possibly, these spiral organisms may spread throughout the entire large intestine, although there is a potential problem with antibody specificity.

  16. Development of a recombinant antibody towards PAPP-A for immunohistochemical use in multiple animal species

    DEFF Research Database (Denmark)

    Mikkelsen, Jakob H; Steffensen, Lasse B; Oxvig, Claus


    normal physiology and under different pathological conditions. However, antibodies for the detection of PAPP-A in non-human tissues have been lacking, although needed for use with several animal models which are currently being developed. To develop a monoclonal antibody suitable...... for the immunohistochemical detection of PAPP-A, we therefore selected a phage-derived scFv antibody, PAC1, specifically recognizing an epitope of PAPP-A, which is highly conserved between multiple animal species. We first converted this antibody into bivalent IgG, and verified its ability to recognize PAPP-A in sections...

  17. High specificity in plant leaf metabolic responses to arbuscular mycorrhiza. (United States)

    Schweiger, Rabea; Baier, Markus C; Persicke, Marcus; Müller, Caroline


    The chemical composition of plants (phytometabolome) is dynamic and modified by environmental factors. Understanding its modulation allows to improve crop quality and decode mechanisms underlying plant-pest interactions. Many studies that investigate metabolic responses to the environment focus on single model species and/or few target metabolites. However, comparative studies using environmental metabolomics are needed to evaluate commonalities of chemical responses to certain challenges. We assessed the specificity of foliar metabolic responses of five plant species to the widespread, ancient symbiosis with a generalist arbuscular mycorrhizal fungus. Here we show that plant species share a large 'core metabolome' but nevertheless the phytometabolomes are modulated highly species/taxon-specifically. Such a low conservation of responses across species highlights the importance to consider plant metabolic prerequisites and the long time of specific plant-fungus coevolution. Thus, the transferability of findings regarding phytometabolome modulation by an identical AM symbiont is severely limited even between closely related species.

  18. Effective synaptome analysis of itch-mediating neurons in the spinal cord: A novel immunohistochemical methodology using high-voltage electron microscopy. (United States)

    Satoh, Keita; Takanami, Keiko; Murata, Kazuyoshi; Kawata, Mitsuhiro; Sakamoto, Tatsuya; Sakamoto, Hirotaka


    Transmission electron microscopy (TEM) is used for three-dimensional (3-D) analysis of synaptic connections in neuroscience research. However, 3-D reconstruction of the synapses using serial ultrathin sections is a powerful but tedious approach requiring advanced technical skills. High-voltage electron microscopy (HVEM) allows examination of thicker sections of biological specimens due to the increased penetration of the more accelerated electrons, which is useful to analyze the 3-D structure of biological specimens. However, it is still difficult to visualize the neural networks and synaptic connections in 3-D using HVEM because of insufficient and non uniform heavy metal staining in the membranous structures in semi-thin sections. Here, we present the successful chemical 3-D neuroanatomy of the rat spinal dorsal horn at the ultrastructural level as a first step for effective synaptome analysis by applying a high-contrast en bloc staining method to immune-HVEM tomography. Our new approach made it possible to examine many itch-mediating synaptic connections and neural networks in the spinal cord simultaneously using HVEM tomography. This novel 3-D electron microscopy is very useful for the analysis of synaptic structure and the chemical neuroanatomy at the 3-D ultrastructural level. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  19. Immunohistochemical expression of p16 in lipoblastomas. (United States)

    Cappellesso, Rocco; d'Amore, Emanuele S G; Dall'Igna, Patrizia; Guzzardo, Vincenza; Vassarotto, Elisa; Rugge, Massimo; Alaggio, Rita


    Lipoblastoma (LB) is a rare benign adipocytic tumor of childhood occasionally showing histological similarities to myxoid liposarcoma (ML) or well-differentiated liposarcoma (WDL). p16 immunohistochemistry has proved to be useful in distinguishing various types of liposarcomas, in particular WDL from lipoma, with higher sensitivity and specificity than MDM2 and CDK4 immunohistochemistry. In this study, we reported the histologic features of a series of 30 LB with emphasis on the potential diagnostic pitfalls and investigated the immunohistochemical expression of p16. Moreover, p16 immunostaining was performed in 16 liposarcomas (11 WDL and 5 ML), 16 lipomas, and 16 cases of liponecrosis in order to evaluate its usefulness in the differential diagnosis of challenging lesions occurring in older children. Overall, p16 immunostaining was positive in 3 LBs and in 12 out of 16 liposarcomas (10 WDL and 2 ML), with a sensitivity of 75%, a specificity of 90%, a positive predictive value of 80%, and a negative predictive value of 87%. All lipomas were p16 negative, whereas 5 liponecroses were positive. Accounting altogether the benign lesions versus liposarcomas, p16 showed a sensitivity of 75%, a specificity of 87%, a positive predictive value of 60%, and a negative predictive value of 93%. Our data suggest that a negative p16 immunostaining may be helpful in excluding a liposarcoma when occurring in unusual clinical contexts, such as in adolescence or late recurrence. However, such finding should be interpreted with caution since also some liposarcomas lack p16 and occasional LBs are positive.

  20. Nano-porous ultra-high specific surface ultrafine fibers

    Institute of Scientific and Technical Information of China (English)

    LI Xinsong; NIE Guangyu


    Nano-porous ultra-high specific surface ultrafine fibers are created by the method of "electrospinning-phase separation-leaching" (EPL) for the first time. First of all, polymer solutions of polyacrylonitrile (PAN) and polyvinylpyrrolidone (PVP) blends dissolved in co-solvent are electrospun to make ultrafine fibers when charged to high voltages. The incompatibility of PAN and PVP induces phase separation to form microdomains of PVP in the polymer blend ultrafine fibers. Then, PVP microdomains in the blend fibers are leached out in water, and porous PAN ultrafine fibers are obtained. Lastly, the surface and cross-section of the porous ultrafine fibers are observed in detail by field emission scanning electron microscope (FESEM), and the specific surface of the ultrafine fibers is measured by means of nitrogen absorption. With increasing the content of PVP, the specific surface area of the ultrafine fibers increases apparently. The specific surface area of the porous ultrafine fibers with the diameter of 2130 nm is more than 70 m2·g-1. The cross-section of the PAN porous ultrafine fibers after leaching of PVP microdomains from polymer blend fibers with the feed ratio of PAN/PVP of 10/20 shows the characteristic of porous structure with pore diameter of ca 30 nm according to FESEM photo.

  1. Bilateral synchronous high-grade serous carcinoma and clear cell carcinoma in right and left ovaries with immunohistochemical confirmation: An exceptional finding

    Directory of Open Access Journals (Sweden)

    Agarwal Preeti


    Full Text Available Synchronous epithelial or mixed epithelial and germ cells tumors in the same ovary is a recognized event, however, having two different surface epithelial tumors in contra lateral ovaries is a rare occurrence; prognosis and pathogenesis of which is still not clear. We came across similar finding in a 60-year-old female with different types of surface epithelial neoplasm in right and left ovaries at the same time; both of which were malignant. Clinicoradiologically only the left ovary revealed tumor, right ovary was atrophic. To our surprise, left ovary revealed high grade serous carcinoma and the right ovary displayed clear cell carcinoma. We performed immunohistochemistry to rule out the possibility of clear cell variant of serous papillary carcinoma. On literature search, we found; only single case with synchronous presentation of two different surface epithelial ovarian tumors in the same patient, both of which were benign.

  2. The possible prognostic role of histone deacetylase and transforming growth factor β/Smad signaling in high grade gliomas treated by radio-chemotherapy: a preliminary immunohistochemical study

    Directory of Open Access Journals (Sweden)

    Roberta Sferra


    Full Text Available Glioblastoma (GBM is the most common and aggressive tumor of the central nervous system. Unfortunately, patients affected by this disease have a very poor prognosis, due to high level of invasiveness and resistance to standard therapies. Although the molecular profile of GBM has been extensively investigated, the events responsible for its pathogenesis and progression remain largely unknown. Histone Deacetylases (HDAC dependent epigenetic modifications and transforming growth factor (TGF-β/Smad pathway seem to play an important role in GBM tumorigenesis, resistance to common therapies and poor clinical outcome.  The aim of this study was to evaluate the involvement and the possible interaction between these two molecular cascades in the pathogenesis and prognosis of GBM. Immunohistochemistry (IHC was performed on microdissected GBM samples, collected from 14 patients (6 men and 8 women ranging in age from 43 to 74 years. The patients were previously divided, on the basis of their overall survival (OS, into two groups: short and long OS. Patients with poor prognosis showed hyperexpression of HDAC4 and HDAC6, an activation of the TGF-β/Smad pathway, with high levels of IL-13, Smad2, PDGF and MMP3 expression, compared to the long survivors. The short OS group exhibits a decrease in Smad 7 expression and also low levels of p21 immunostaining, which represents a common target of the two pathways. The IHC data was confirmed by quantitative analysis and Immunoblotting. Our preliminary results suggest that both HDAC4 and HDAC6 together with the TGF-β/Smad pathway may be involved in progression of GBM and this cross talking could be a useful prognostic marker in this deadly disease.

  3. Accelerator Production and Separations for High Specific Activity Rhenium-186

    Energy Technology Data Exchange (ETDEWEB)

    Jurisson, Silvia S. [Univ. of Missouri, Columbia, MO (United States); Wilbur, D. Scott [Univ. of Washington, Seattle, WA (United States)


    Tungsten and osmium targets were evaluated for the production of high specific activity rhenium-186. Rhenium-186 has potential applications in radiotherapy for the treatment of a variety of diseases, including targeting with monoclonal antibodies and peptides. Methods were evaluated using tungsten metal, tungsten dioxide, tungsten disulfide and osmium disulfide. Separation of the rhenium-186 produced and recycling of the enriched tungsten-186 and osmium-189 enriched targets were developed.

  4. Benchmarking procedures for high-throughput context specific reconstruction algorithms

    Directory of Open Access Journals (Sweden)

    Maria ePires Pacheco


    Full Text Available Recent progress in high-throughput data acquisition has shifted the focus from data generation to processing and understanding of how to integrate collected information. Context specific reconstruction based on generic genome scale models like ReconX (Duarte et al., 2007; Thiele et al., 2013 or HMR (Agren et al., 2013 has the potential to become a diagnostic and treatment tool tailored to the analysis of specific individuals. The respective computational algorithms require a high level of predictive power, robustness and sensitivity. Although multiple context specific reconstruction algorithms were published in the last ten years, only a fraction of them is suitable for model building based on human high-throughput data. Beside other reasons, this might be due to problems arising from the limitation to only one metabolic target function or arbitrary thresholding.This review describes and analyses common validation methods used for testing model building algorithms. Two major methods can be distinguished, consistency testing and comparison based testing. The former includes methods like cross validation or testing with artificial networks. The latter covers methods comparing sets of functionalities, comparison with existing networks or additional databases. We test those methods on several available algorithms and deduce properties of these algorithms, that can be compared with future developments. The set of tests performed, can therefore serve as a benchmarking procedure for future algorithms

  5. P53 expression is significantly correlated with high risk of malignancy and epithelioid differentiation in GISTs. An immunohistochemical study of 104 cases

    Directory of Open Access Journals (Sweden)

    Klöppel Günter


    Full Text Available Abstract Background Molecular analyses of the c-kit and PDGFRα genes have contributed greatly to our understanding of the development of gastrointestinal stromal tumors (GISTs, but little is known about their malignant potential. The aim of our study was to evaluate cell cycle regulators as potential prognostic markers in GISTs. Methods We investigated 104 KIT positive GISTs from various tumor sites in immunoassays on CD34, Ki67 and particularly on P53, BCL-2 and Cyclin D1. The results were compared with tumor size, mitotic rate, proliferative activity, histological subtype, nuclear atypia and risk assessment according to Fletcher and Miettinen. Occurrence of metastases and survival were also taken into account. Results The expression of P53 was significantly correlated with high risk criteria towards malignancy and epithelioid differentiation in GISTs. Likewise P53 label correlated significantly with the established prognostic indicators: tumor size, mitotic rate, nuclear atypia and proliferative activity. Regarding the site of tumor presentation, P53 was not a decisive factor. BCL-2 and Cyclin D1 expression was not related to any of the prognostic indicators. Conclusion The present data identified P53 being a recommendable marker for predicting the risk of malignancy in GISTs. In addition, we found P53 significantly correlated with epithelioid tumor differentiation, independent of tumor site. BCL-2 and Cyclin D1, however, did not prove to be deciding markers for diagnosis and prognosis.

  6. [Immuno-histochemical study of medullary thyroid carcinoma]. (United States)

    Bessho, T


    Five peptide hormones including calcitonin (CT) and gastrin-releasing peptide (GRP), serotonin (5HT), CEA, nervous tissue specific proteins and monoclonal antibody Leu-7 were immuno-histochemically studied on 60 cases of medullary thyroid carcinoma (MTC). In addition, localization of varied products in the tumor cells and its relations with the clinical features in some cases were evaluated. MTC contains a variety of products in many cases, and CT and CEA were positive in all cases. In 50 of the 57 cases (87.7%), GRP was positive, which suggested that GRP could be a novel tumor marker for this tumor. Furthermore, in tumor cells and C-cell hyperplastic foci, identical cells were sometimes revealed to possess both CT and GRP. Existence of somatostatin (SS), substance-P (SP), beta-MSH, 5 HT, Leu-7 and NSE in the tumor cells were confirmed. NSE was positive in 32 of the 47 cases (61.8%) which could confirm that MTC possesses neuroendocrine nature. In two cases of autopsy in which the tumors were highly malignant in clinical course and undifferentiated in histology, most tumor cells showed poor stainability for peptide hormones, suggesting that specific qualities as neuroendocrine tumor had been lost. In familial cases, the tumor tended to contain multiple substances.

  7. Preparation of a novel antiserum to aromatase with high affinity and specificity: Its clinicopathological significance on breast cancer tissue. (United States)

    Kanomata, Naoki; Matsuura, Shiro; Nomura, Tsunehisa; Kurebayashi, Junichi; Mori, Taisuke; Kitawaki, Jo; Moriya, Takuya


    Aromatase inhibitors have been widely used for the endocrine treatment of estrogen-dependent breast cancer in postmenopausal patients. However, clinicopathological studies of aromatase have been limited due to unsatisfactory specificity and/or restricted availability of anti-aromatase antibodies. Here, we have generated a polyclonal antiserum with high affinity and specificity for human aromatase using a monoclonal antibody tagged immunoaffinity chromatography on an industrial production scale. Our preliminary immunohistochemical analysis of 221 invasive breast cancer cases indicated that 87.3% (193/221) had at least 5% aromatase positive cells. The histoscore for aromatase was inversely correlated with pT (p = 0.019), pN (p = 0.001), stage (p cancer aromatase expression was independent of estrogen receptor (ER), progesterone receptor (PgR), and human epidermal growth factor receptor 2 statuses. This antiserum will be applicable to clinicopathological examination of aromatase in addition to ER and PgR for an appropriate use of aromatase inhibitor on the treatment of breast cancer. Further studies on the relationship between Aromatase inhibitors have been widely used for the endocrine treatment of estrogen-dependent breast cancer in postmenopausal patients. However, clinicopathological studies of aromatase have been limited due to unsatisfactory specificity and/or restricted availability of anti-aromatase antibodies. Here, we have generated a polyclonal antiserum with high affinity and specificity for human aromatase using a monoclonal antibody tagged immunoaffinity chromatography on an industrial production scale. Our preliminary immunohistochemical analysis of 221 invasive breast cancer cases indicated that 87.3% (193/221) had at least 5% aromatase positive cells. The histoscore for aromatase was inversely correlated with pT (p = 0.019), pN (p = 0.001), stage (p cancer aromatase expression was independent of estrogen receptor (ER), progesterone receptor (PgR), and

  8. High efficiency cell-specific targeting of cytokine activity (United States)

    Garcin, Geneviève; Paul, Franciane; Staufenbiel, Markus; Bordat, Yann; van der Heyden, José; Wilmes, Stephan; Cartron, Guillaume; Apparailly, Florence; de Koker, Stefaan; Piehler, Jacob; Tavernier, Jan; Uzé, Gilles


    Systemic toxicity currently prevents exploiting the huge potential of many cytokines for medical applications. Here we present a novel strategy to engineer immunocytokines with very high targeting efficacies. The method lies in the use of mutants of toxic cytokines that markedly reduce their receptor-binding affinities, and that are thus rendered essentially inactive. Upon fusion to nanobodies specifically binding to marker proteins, activity of these cytokines is selectively restored for cell populations expressing this marker. This ‘activity-by-targeting’ concept was validated for type I interferons and leptin. In the case of interferon, activity can be directed to target cells in vitro and to selected cell populations in mice, with up to 1,000-fold increased specific activity. This targeting strategy holds promise to revitalize the clinical potential of many cytokines.

  9. Cellulose antibody films for highly specific evanescent wave immunosensors (United States)

    Hartmann, Andreas; Bock, Daniel; Jaworek, Thomas; Kaul, Sepp; Schulze, Matthais; Tebbe, H.; Wegner, Gerhard; Seeger, Stefan


    For the production of recognition elements for evanescent wave immunosensors optical waveguides have to be coated with ultrathin stable antibody films. In the present work non amphiphilic alkylated cellulose and copolyglutamate films are tested as monolayer matrices for the antibody immobilization using the Langmuir-Blodgett technique. These films are transferred onto optical waveguides and serve as excellent matrices for the immobilization of antibodies in high density and specificity. In addition to the multi-step immobilization of immunoglobulin G(IgG) on photochemically crosslinked and oxidized polymer films, the direct one-step transfer of mixed antibody-polymer films is performed. Both planar waveguides and optical fibers are suitable substrates for the immobilization. The activity and specificity of immobilized antibodies is controlled by the enzyme-linked immunosorbent assay (ELISA) technique. As a result reduced non-specific interactions between antigens and the substrate surface are observed if cinnamoylbutyether-cellulose is used as the film matrix for the antibody immobilization. Using the evanescent wave senor (EWS) technology immunosensor assays are performed in order to determine both the non-specific adsorption of different coated polymethylmethacrylat (PMMA) fibers and the long-term stability of the antibody films. Specificities of one-step transferred IgG-cellulose films are drastically enhanced compared to IgG-copolyglutamate films. Cellulose IgG films are used in enzymatic sandwich assays using mucine as a clinical relevant antigen that is recognized by the antibodies BM2 and BM7. A mucine calibration measurement is recorded. So far the observed detection limit for mucine is about 8 ng/ml.

  10. Procurement specification high vacuum test chamber and pumping system

    Energy Technology Data Exchange (ETDEWEB)

    Cormick, J. E.


    The specification establishes requirements for a high-vacuum test chamber, associated vacuum pumps, valves, controls, and instrumentation that shall be designed and fabricated for use as a test chamber for testing a closed loop Brayton Isotope Power System (BIPS) Ground Demonstration System (GDS). The vacuum system shall include all instrumentation required for pressure measurement and control of the vacuum pumping system. A general outline of the BIPS-GDS in the vacuum chamber and the preliminary piping and instrumentation interface to the vacuum chamber are shown.

  11. Production of N-13 labeled compounds with high specific activity

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, Kazutoshi; Sasaki, Motoji; Yoshida, Yuichiro; Haradahira, Terushi; Inoue, Osamu [National Inst. of Radiological Sciences, Chiba (Japan)


    Nitrogen-13 was produced by irradiating ultra pure water saturated with a pure gas (N2, O2, He, H2) with 18 MeV protons. Ion species generated by irradiation were analyzed with radio ion chromatography systems. An automated equipment was developed to synthesize anhydrous (13N)NH3 as a synthetic precursor and (13N)p-nitrophenyl carbamate ((13N)NPC) as a model compound, using the (13N)NH3. The radiochemical yield and specific activity of (13N)NPC was high enough to carry out the receptor study with PET. (author)

  12. Highly specific urine-based marker of bladder cancer. (United States)

    Van Le, Thu-Suong; Miller, Raymond; Barder, Timothy; Babjuk, Marko; Potter, Douglas M; Getzenberg, Robert H


    Bladder cancer represents a major health problem throughout the world, but advances in tumor biomarker development are revolutionizing how physicians diagnose the disease. We previously used an indirect immunoassay to demonstrate that the bladder cancer specific biomarker, BLCA-4, is present in urine samples from patients with bladder cancer, but not in samples from healthy individuals. In this study, a sandwich immunoassay was used to measure BLCA-4 in urine samples from patient populations with various urologic conditions and healthy individuals. Urine was collected from healthy individuals and from patients with bladder cancer, benign urologic conditions, or prostate cancer. BLCA-4 levels were evaluated by a sandwich immunoassay using two antibodies directed against distinct epitopes on BLCA-4. Using a prospectively determined cutoff of an absorbance unit (OD) of 0.04, 67 of the 75 samples from patients with bladder cancer were positive for BLCA-4, resulting in an assay sensitivity of 89%. Also, 62 of the 65 samples from individuals without bladder cancer were negative for BLCA-4, resulting in an assay specificity of 95%. The high sensitivity and specificity of the sandwich BLCA-4 immunoassay may allow for earlier detection and treatment of disease, thus greatly improving patient care.

  13. Flexible and Lightweight Fuel Cell with High Specific Power Density. (United States)

    Ning, Fandi; He, Xudong; Shen, Yangbin; Jin, Hehua; Li, Qingwen; Li, Da; Li, Shuping; Zhan, Yulu; Du, Ying; Jiang, Jingjing; Yang, Hui; Zhou, Xiaochun


    Flexible devices have been attracting great attention recently due to their numerous advantages. But the energy densities of current energy sources are still not high enough to support flexible devices for a satisfactory length of time. Although proton exchange membrane fuel cells (PEMFCs) do have a high-energy density, traditional PEMFCs are usually too heavy, rigid, and bulky to be used in flexible devices. In this research, we successfully invented a light and flexible air-breathing PEMFC by using a new design of PEMFC and a flexible composite electrode. The flexible air-breathing PEMFC with 1 × 1 cm(2) working area can be as light as 0.065 g and as thin as 0.22 mm. This new PEMFC exhibits an amazing specific volume power density as high as 5190 W L(-1), which is much higher than traditional (air-breathing) PEMFCs. Also outstanding is that the flexible PEMFC retains 89.1% of its original performance after being bent 600 times, and it retains its original performance after being dropped five times from a height of 30 m. Moreover, the research has demonstrated that when stacked, the flexible PEMFCs are also useful in mobile applications such as mobile phones. Therefore, our research shows that PEMFCs can be made light, flexible, and suitable for applications in flexible devices. These innovative flexible PEMFCs may also notably advance the progress in the PEMFC field, because flexible PEMFCs can achieve high specific power density with small size, small volume, low weight, and much lower cost; they are also much easier to mass produce.

  14. Immunohistochemical detection of Mycoplasma mycoides mycoides ...

    African Journals Online (AJOL)

    Immunohistochemical detection of Mycoplasma mycoides mycoides small colony type in lungs of slaughtered cattle at ... Tanzania Veterinary Journal ... healthy slaughtered animals to confirm presence of the disease in these animals. A total of ...

  15. Features of immunohistochemical diagnostic of melanocytic tumours

    Directory of Open Access Journals (Sweden)

    Shpon’ka I.S.


    Full Text Available Background. The malignant melanomas are the most important group of skin cancers. Although less common than the familiar basal and squamous cell tumours of the skin, they are much more frequently fatal, due to their intrinsic tendency to lymphatic and haematogenic metastasis. Objective. The article is devoted to parsing cases melanocytic tumours that were established through immunohistochemical study. Methods. In the study analyzed 236 patient material (150 women and 86 men aged 28 to 77 years during 2010-2013 turned out to clarify the histological diagnosis of skin tumors or metastases to lymph nodes (rare at other sites. The primary monoclonal antibodies used Сytokeratin, Рan Ab1 (clone AE1/AE3, S100 (clone 4C4.9, Ki-67 (clone SP6, Vimentin (clone V9, Melanoma gp100 (clone HMB-45. Results. Naevus proliferation rate showed a statistically significant difference with respect to proliferation rate of malignant melanomas (p<0,05. All samples (100% showed positive expression of high-intensity staining (+++ or moderate (++ intensity on the marker S100; 98,30% of samples (232 of 236 showed positive expression of marker HMB-45 at least in terms of tumor cells with intensity color from the high (+++ to weak (+ and 83.89% of the samples (198 of 236 were negative (– Сytokeratin, Рan Ab1 (other 38 cases showed weakly positive expression (+/– of tumor cells. Conclusions. 1. In the differential diagnosis of melanoma and naevus, we must bear in mind the uniformity immunophenotype of these tumors and consider only the cytological features of the tumor, changes in the structure of the epidermis and dermis (contour, symmetry, depth, inflammatory infiltration and proliferation rate. 2. Patients whose lymph nodes were the first clinical signs of cancer are always in need for additional immunohistochemical studies to avoid diagnostic errors. 3. The most common phenotype of melanocytic tumours responsible Сytokeratin, Рan–, Vimintin+, S100+, HMB-45

  16. Histopathological and immunohistochemical profile in anaplastic gangliogliomas. (United States)

    Romero-Rojas, Alfredo E; Diaz-Perez, Julio A; Chinchilla-Olaya, Sandra I; Amaro, Deirdre; Lozano-Castillo, Alfonso; Restrepo-Escobar, Ligia I


    The anaplastic ganglioglioma (AG) is the high-grade counterpart of ganglioglioma, a rare mixed tumor composed of neuronal/ganglion and glial cells. We describe the histopathology and immunohistochemistry in 7 cases of AG and correlate them with the clinical and radiological features. Our AG patients correspond to 2.5% of the central nervous system tumor patients evaluated in our institution. The mean age at presentation was 25.7 years, with a male predominance. The most common clinical presentation was generalized tonic-clonic seizures (3/7 cases), in correlation with frequent cortical/subcortical location (6/7 cases). Histopathologically, all our cases showed high-grade features in glial (glial fibrillary acid protein-positive) and neuron-ganglion cells (synaptophysin, PGP-9.5, neurofilament, NSE and CD56-positive), as well as moderate cellularity, frequent mitotic figures and a Ki-67 labeling index >5%. All our patients had poor survival. We found that a typical histopathological and immunohistochemical profile is constant and can be useful in early diagnosis of these aggressive neoplasms. Copyright © 2013 Sociedad Española de Neurocirugía. Published by Elsevier España. All rights reserved.

  17. A highly specific coding system for structural chromosomal alterations. (United States)

    Martínez-Frías, M L; Martínez-Fernández, M L


    The Spanish Collaborative Study of Congenital Malformations (ECEMC, from the name in Spanish) has developed a very simple and highly specific coding system for structural chromosomal alterations. Such a coding system would be of value at present due to the dramatic increase in the diagnosis of submicroscopic chromosomal deletions and duplications through molecular techniques. In summary, our new coding system allows the characterization of: (a) the type of structural anomaly; (b) the chromosome affected; (c) if the alteration affects the short or/and the long arm, and (d) if it is a non-pure dicentric, a non-pure isochromosome, or if it affects several chromosomes. We show the distribution of 276 newborn patients with these types of chromosomal alterations using their corresponding codes according to our system. We consider that our approach may be useful not only for other registries, but also for laboratories performing these studies to store their results on case series. Therefore, the aim of this article is to describe this coding system and to offer the opportunity for this coding to be applied by others. Moreover, as this is a SYSTEM, rather than a fixed code, it can be implemented with the necessary modifications to include the specific objectives of each program. Copyright © 2013 Wiley Periodicals, Inc.

  18. Nanoporous ultra-high specific surface inorganic fibres (United States)

    Kanehata, Masaki; Ding, Bin; Shiratori, Seimei


    Nanoporous inorganic (silica) nanofibres with ultra-high specific surface have been fabricated by electrospinning the blend solutions of poly(vinyl alcohol) (PVA) and colloidal silica nanoparticles, followed by selective removal of the PVA component. The configurations of the composite and inorganic nanofibres were investigated by changing the average silica particle diameters and the concentrations of colloidal silica particles in polymer solutions. After the removal of PVA by calcination, the fibre shape of pure silica particle assembly was maintained. The nanoporous silica fibres were assembled as a porous membrane with a high surface roughness. From the results of Brunauer-Emmett-Teller (BET) measurements, the BET surface area of inorganic silica nanofibrous membranes was increased with the decrease of the particle diameters. The membrane composed of silica particles with diameters of 15 nm showed the largest BET surface area of 270.3 m2 g-1 and total pore volume of 0.66 cm3 g-1. The physical absorption of methylene blue dye molecules by nanoporous silica membranes was examined using UV-vis spectrometry. Additionally, the porous silica membranes modified with fluoroalkylsilane showed super-hydrophobicity due to their porous structures.

  19. Nanoporous ultra-high specific surface inorganic fibres

    Energy Technology Data Exchange (ETDEWEB)

    Kanehata, Masaki [Faculty of Science and Technology, Keio University, Yokohama 223-8522 (Japan); Ding Bin [Fiber and Polymer Science, University of California, Davis, CA 95616 (United States); Shiratori, Seimei [Faculty of Science and Technology, Keio University, Yokohama 223-8522 (Japan)


    Nanoporous inorganic (silica) nanofibres with ultra-high specific surface have been fabricated by electrospinning the blend solutions of poly(vinyl alcohol) (PVA) and colloidal silica nanoparticles, followed by selective removal of the PVA component. The configurations of the composite and inorganic nanofibres were investigated by changing the average silica particle diameters and the concentrations of colloidal silica particles in polymer solutions. After the removal of PVA by calcination, the fibre shape of pure silica particle assembly was maintained. The nanoporous silica fibres were assembled as a porous membrane with a high surface roughness. From the results of Brunauer-Emmett-Teller (BET) measurements, the BET surface area of inorganic silica nanofibrous membranes was increased with the decrease of the particle diameters. The membrane composed of silica particles with diameters of 15 nm showed the largest BET surface area of 270.3 m{sup 2} g{sup -1} and total pore volume of 0.66 cm{sup 3} g{sup -1}. The physical absorption of methylene blue dye molecules by nanoporous silica membranes was examined using UV-vis spectrometry. Additionally, the porous silica membranes modified with fluoroalkylsilane showed super-hydrophobicity due to their porous structures.

  20. Highly efficient site-specific transgenesis in cancer cell lines

    Directory of Open Access Journals (Sweden)

    Michael Iacovos P


    Full Text Available Abstract Background Transgenes introduced into cancer cell lines serve as powerful tools for identification of genes involved in cancer. However, the random nature of genomic integration site of a transgene highly influences the fidelity, reliability and level of its expression. In order to alleviate this bottleneck, we characterized the potential utility of a novel PhiC31 integrase-mediated site-specific insertion system (PhiC31-IMSI for introduction of transgenes into a pre-inserted docking site in the genome of cancer cells. Methods According to this system, a “docking-site” was first randomly inserted into human cancer cell lines and clones with a single copy were selected. Subsequently, an “incoming” vector containing the gene of interest was specifically inserted in the docking-site using PhiC31. Results Using the Pc-3 and SKOV-3 cancer cell lines, we showed that transgene insertion is reproducible and reliable. Furthermore, the selection system ensured that all surviving stable transgenic lines harbored the correct integration site. We demonstrated that the expression levels of reporter genes, such as green fluorescent protein and luciferase, from the same locus were comparable among sister, isogenic clones. Using in vivo xenograft studies, we showed that the genetically altered cancer cell lines retain the properties of the parental line. To achieve temporal control of transgene expression, we coupled our insertion strategy with the doxycycline inducible system and demonstrated tight regulation of the expression of the antiangiogenic molecule sFlt-1-Fc in Pc-3 cells. Furthermore, we introduced the luciferase gene into the insertion cassette allowing for possible live imaging of cancer cells in transplantation assays. We also generated a series of Gateway cloning-compatible intermediate cassettes ready for high-throughput cloning of transgenes and demonstrated that PhiC31-IMSI can be achieved in a high throughput 96-well plate

  1. Dendritic cells in melanoma - immunohistochemical study and research trends. (United States)

    Nedelcu, Roxana Ioana; Ion, Daniela Adriana; Holeab, Cosmin Adrian; Cioplea, Mirela Daniela; Brînzea, Alice; Zurac, Sabina Andrada


    Cutaneous dendritic cells play multiple physiological roles and are involved in various pathophysiological processes. Research studies of dendritic cells abound in the medical literature. Nevertheless, the role of dendritic cells in melanoma regression phenomenon is not completely understood. We conducted a scientometric analysis in order to highlight the current state on research regarding dendritic cells and melanoma. We also performed an immunohistochemical study, using specific markers for dendritic cells (CD1a, langerin). We evaluated the frequency and distribution of dendritic cells in areas of tumor regression compared to the areas of inflammatory infiltrate of melanoma without regression. The immunohistochemical study we performed revealed that dendritic cells are more frequent in the regressed areas, comparing with non-regressed ones. In regressed areas, dendritic cells have a predominant nodular pattern (19 cases), followed by diffuse isolate pattern (eight cases) and mixed pattern (diffuse and nodular) (three cases). In melanoma without regression, most cases presented a diffuse pattern (27 cases) of dendritic cells distribution. In conclusion, our immunohistochemical study stressed differences between frequency and distribution of dendritic cells located in the melanoma with regression and melanoma without regression. These data suggest that dendritic cells are involved in the regression phenomenon. Following the literature analysis we obtained, we observed that dendritic cells profile in melanoma with regression was poorly studied. Insights into antitumor immune response and dendritic cells may be essential for the understanding of the potential prognostic role of dendritic cells in melanoma and for the development of new promising therapeutic strategies for melanoma.

  2. High epidermal growth factor receptor immunohistochemical expression in urothelial carcinoma of the bladder is not associated with EGFR mutations in exons 19 and 21: a study using formalin-fixed, paraffin-embedded archival tissues. (United States)

    Chaux, Alcides; Cohen, Julie S; Schultz, Luciana; Albadine, Roula; Jadallah, Sana; Murphy, Kathleen M; Sharma, Rajni; Schoenberg, Mark P; Netto, George J


    Epidermal growth factor receptor (EGFR) is a member of the erbB tyrosine kinase family reported to be overexpressed in a variety of solid malignancies. Mutations in exons 19 to 21 of the tyrosine kinase domain have been detected in a subset of these tumors and its presence associated with a better response to EGFR inhibitors. Several clinical trials are currently underway to evaluate the performance of such drugs in patients with bladder cancer, but data on EGFR mutation status are limited. The current study assesses EGFR immunohistochemical expression and the presence of mutations in exons 19 and 21 by polymerase chain reaction in 19 bladder urothelial carcinomas from formalin-fixed, paraffin-embedded tissues. Representative paraffin sections were microdissected for DNA extraction using a pinpoint isolation system. Parallel sections were immunostained using a monoclonal anti-EGFR antibody. No mutations in exons 19 and 21 of EGFR were identified in any of the cases. Immunohistochemical EGFR positivity was observed in 14 of 19 cases. In summary, we found EGFR protein expression in 74% of urothelial carcinomas, but we failed to detect EGFR mutations at exons 19 to 21, suggesting that EGFR overexpression is not related to the presence of mutations in the tyrosine kinase domain of the gene. Mutation analysis of EGFR exons 19 and 21 is feasible in microdissected paraffin sections from archival tissues. Immunohistochemical expression of EGFR may not be useful to predict therapeutic response to EGFR inhibitors in patients with urothelial carcinomas. To explain EGFR immunohistochemical overexpression, other mechanisms besides mutations in the EGFR kinase domain should be investigated in future studies.

  3. High voltage and high specific capacity dual intercalating electrode Li-ion batteries (United States)

    West, William C. (Inventor); Blanco, Mario (Inventor)


    The present invention provides high capacity and high voltage Li-ion batteries that have a carbonaceous cathode and a nonaqueous electrolyte solution comprising LiF salt and an anion receptor that binds the fluoride ion. The batteries can comprise dual intercalating electrode Li ion batteries. Methods of the present invention use a cathode and electrode pair, wherein each of the electrodes reversibly intercalate ions provided by a LiF salt to make a high voltage and high specific capacity dual intercalating electrode Li-ion battery. The present methods and systems provide high-capacity batteries particularly useful in powering devices where minimizing battery mass is important.

  4. Indirect defense in a highly specific ant-plant mutualism. (United States)

    Grangier, Julien; Dejean, Alain; Malé, Pierre-Jean G; Orivel, Jérôme


    Although associations between myrmecophytes and their plant ants are recognized as a particularly effective form of protective mutualism, their functioning remains incompletely understood. This field study examined the ant-plant Hirtella physophora and its obligate ant associate Allomerus decemarticulatus. We formulated two hypotheses on the highly specific nature of this association: (1) Ant presence should be correlated with a marked reduction in the amount of herbivory on the plant foliage; (2) ant activity should be consistent with the "optimal defense" theory predicting that the most vulnerable and valuable parts of the plant are the best defended. We validated the first hypothesis by demonstrating that for ant-excluded plants, expanding leaves, but also newly matured ones in the long term, suffered significantly more herbivore damage than ant-inhabited plants. We showed that A. decemarticulatus workers represent both constitutive and inducible defenses for their host, by patrolling its foliage and rapidly recruiting nestmates to foliar wounds. On examining how these activities change according to the leaves' developmental stage, we found that the number of patrolling ants dramatically decreased as the leaves matured, while leaf wounds induced ant recruitment regardless of the leaf's age. The resulting level of these indirect defenses was roughly proportional to leaf vulnerability and value during its development, thus validating our second hypothesis predicting optimal protection. This led us to discuss the factors influencing ant activity on the plant's surface. Our study emphasizes the importance of studying both the constitutive and inducible components of indirect defense when evaluating its efficacy and optimality.

  5. High Performance Organ-Specific Nuclear Medicine Imagers. (United States)

    Majewski, Stan


    One of the exciting applications of nuclear science is nuclear medicine. Well-known diagnostic imaging tools such as PET and SPECT (as well as MRI) were developed as spin-offs of basic scientific research in atomic and nuclear physics. Development of modern instrumentation for applications in particle physics experiments offers an opportunity to contribute to development of improved nuclear medicine (gamma and positron) imagers, complementing the present set of standard imaging tools (PET, SPECT, MRI, ultrasound, fMRI, MEG, etc). Several examples of new high performance imagers developed in national laboratories in collaboration with academia will be given to demonstrate this spin-off activity. These imagers are designed to specifically image organs such as breast, heart, head (brain), or prostate. The remaining and potentially most important challenging application field for dedicated nuclear medicine imagers is to assist with cancer radiation treatments. Better control of radiation dose delivery requires development of new compact in-situ imagers becoming integral parts of the radiation delivery systems using either external beams or based on radiation delivery by inserting or injecting radioactive sources (gamma, beta or alpha emitters) into tumors.

  6. Specification and analysis of a high speed transport protocol (United States)

    Tipici, Huseyin A.


    While networks have been getting faster, perceived throughput at the application has not always increased accordingly and the bottleneck has moved to the communications processing part of the system. The issues that cause the performance bottlenecks in the current transport protocols are discussed in this thesis, and a further study on a high speed transport protocol which tries to overcome these difficulties with some unique features is presented. By using the Systems of Communicating Machines (SCM) model as a framework, a refined and improved version of the formal protocol specification is built over the previous work, and it is analyzed to verify that the protocol is free from logical errors such as deadlock, unspecified reception, unexecuted transitions and blocking loops. The analysis is conducted in two phases which consists of the application of the associated system state analysis and the simulation of the protocol using the programming language ADA. The thesis also presents the difficulties encountered during the course of the analysis, and suggests possible solutions to some of the problems.

  7. Highly specific spectroscopic photoacoustic molecular imaging of dynamic optical absorption shifts of an antibody-ICG contrast agent (Conference Presentation) (United States)

    Wilson, Katheryne E.; Bachawal, Sunitha; Abou-Elkacem, Lotfi; Jensen, Kristen C.; Machtaler, Steven; Tian, Lu; Willmann, Juergen K.


    Improved techniques for breast cancer screening are critically needed as current methods lack diagnostic accuracy. Using spectroscopic photoacoustic (sPA) molecular imaging with a priori knowledge of optical absorption spectra allows suppression of endogenous background signal, increasing the overall sensitivity and specificity of the modality to exogenous contrast agents. Here, sPA imaging was used to monitor antibody-indocyanine green (ICG) conjugates as they undergo optical absorption spectrum shifts after cellular endocytosis and degradation to allow differentiation between normal murine mammary glands from breast cancer by enhancing molecular imaging signal from target (B7-H3)-bound antibody-ICG. First, B7-H3 was shown to have highly specific (AUC of 0.93) expression on both vascular endothelium and tumor stroma in malignant lesions through quantitative immunohistochemical staining of B7-H3 on 279 human samples (normal (n=53), benign lesions (11 subtypes, n=182), breast cancers (4 subtypes, n=97)), making B7-H3 a promising target for sPA imaging. Second, absorption spectra of intracellular and degraded B7-H3-ICG and Isotype control (Iso-ICG) were characterized through in vitro and in vivo experiments. Finally, a transgenic murine breast cancer model (FVB/N-Tg(MMTVPyMT)634Mul) was imaged, and sPA imaging in found a 3.01 (IQR 2.63, 3.38, Panimals (n=60), Iso-ICG (n=30), blocking B7-H3+B7-H3-ICG (n=20), and free ICG (n=20)) despite significant tumor accumulation of Iso-ICG, confirmed through ex vivo histology. Overall, leveraging anti-B7-H3 antibody-ICG contrast agents, which have dynamic optical absorption spectra representative of molecular interactions, allows for highly specific sPA imaging of murine breast cancer.

  8. Immunohistochemical identification and quantitative analysis of cytoplasmic Cu/Zn superoxide dismutase in mouse organogenesis (United States)

    Yon, Jung-Min; Baek, In-Jeoung; Lee, Se-Ra; Kim, Mi-Ra; Lee, Beom Jun; Yun, Young Won


    Cytoplasmic Cu/Zn superoxide dismutase (SOD1) is an antioxidant enzyme that converts superoxide to hydrogen peroxide in cells. Its spatial distribution matches that of superoxide production, allowing it to protect cells from oxidative stress. SOD1 deficiencies result in embryonic lethality and a wide range of pathologies in mice, but little is known about normal SOD1 protein expression in developing embryos. In this study, the expression pattern of SOD1 was investigated in post-implantation mouse embryos and extraembryonic tissues, including placenta, using Western blotting and immunohistochemical analyses. SOD1 was detected in embryos and extraembryonic tissues from embryonic day (ED) 8.5 to 18.5. The signal in embryos was observed at the lowest level on ED 9.5-11.5, and the highest level on ED 17.5-18.5, while levels remained constant in the surrounding extraembryonic tissues during all developmental stages examined. Immunohistochemical analysis of SOD1 expression on ED 13.5-18.5 revealed its ubiquitous distribution throughout developing organs. In particular, high levels of SOD1 expression were observed in the ependymal epithelium of the choroid plexus, ganglia, sensory cells of the olfactory and vestibulocochlear epithelia, blood cells and vessels, hepatocytes and hematopoietic cells of the liver, lymph nodes, osteogenic tissues, and skin. Thus, SOD1 is highly expressed at late stages of embryonic development in a cell- and tissue-specific manner, and can function as an important antioxidant enzyme during organogenesis in mouse embryos. PMID:18716442

  9. Indirect defense in a highly specific ant-plant mutualism (United States)

    Grangier, Julien; Dejean, Alain; Malé, Pierre-Jean G.; Orivel, Jérôme


    Although associations between myrmecophytes and their plant ants are recognized as a particularly effective form of protective mutualism, their functioning remains incompletely understood. This field study examined the ant-plant Hirtella physophora and its obligate ant associate Allomerus decemarticulatus. We formulated two hypotheses on the highly specific nature of this association: (1) Ant presence should be correlated with a marked reduction in the amount of herbivory on the plant foliage; (2) ant activity should be consistent with the "optimal defense" theory predicting that the most vulnerable and valuable parts of the plant are the best defended. We validated the first hypothesis by demonstrating that for ant-excluded plants, expanding leaves, but also newly matured ones in the long term, suffered significantly more herbivore damage than ant-inhabited plants. We showed that A. decemarticulatus workers represent both constitutive and inducible defenses for their host, by patrolling its foliage and rapidly recruiting nestmates to foliar wounds. On examining how these activities change according to the leaves’ developmental stage, we found that the number of patrolling ants dramatically decreased as the leaves matured, while leaf wounds induced ant recruitment regardless of the leaf’s age. The resulting level of these indirect defenses was roughly proportional to leaf vulnerability and value during its development, thus validating our second hypothesis predicting optimal protection. This led us to discuss the factors influencing ant activity on the plant’s surface. Our study emphasizes the importance of studying both the constitutive and inducible components of indirect defense when evaluating its efficacy and optimality.

  10. Collagenous gastritis: a morphologic and immunohistochemical study of 40 patients. (United States)

    Arnason, Thomas; Brown, Ian S; Goldsmith, Jeffrey D; Anderson, William; O'Brien, Blake H; Wilson, Claire; Winter, Harland; Lauwers, Gregory Y


    Collagenous gastritis is a rare condition defined histologically by a superficial subepithelial collagen layer. This study further characterizes the morphologic spectrum of collagenous gastritis by evaluating a multi-institutional series of 40 patients (26 female and 14 male). The median age at onset was 16 years (range 3-89 years), including 24 patients (60%) under age 18. Twelve patients (30%) had associated celiac disease, collagenous sprue, or collagenous colitis. Hematoxylin and eosin slides were reviewed in biopsies from all patients and tenascin, gastrin, eotaxin, and IgG4/IgG immunohistochemical stains were applied to a subset. The distribution of subepithelial collagen favored the body/fundus in pediatric patients and the antrum in adults. There were increased surface intraepithelial lymphocytes (>25 lymphocytes/100 epithelial cells) in five patients. Three of these patients had associated celiac and/or collagenous sprue/colitis, while the remaining two had increased duodenal lymphocytosis without specific etiology. An eosinophil-rich pattern (>30 eosinophils/high power field) was seen in 21/40 (52%) patients. Seven patients' biopsies demonstrated atrophy of the gastric corpus mucosa. Tenascin immunohistochemistry highlighted the subepithelial collagen in all 21 specimens evaluated and was a more sensitive method of collagen detection in biopsies from two patients with subtle subepithelial collagen. No increased eotaxin expression was identified in 16 specimens evaluated. One of the twenty-three biopsies tested had increased IgG4-positive cells (100/high power field) with an IgG4/IgG ratio of 55%. In summary, collagenous gastritis presents three distinct histologic patterns including a lymphocytic gastritis-like pattern, an eosinophil-rich pattern, and an atrophic pattern. Eotaxin and IgG4 were not elevated enough to implicate these pathways in the pathogenesis. Tenascin immunohistochemistry can be used as a sensitive method of collagen detection.

  11. Probucol suppresses oxidant stress in hypertensive arteries. Immunohistochemical evidence. (United States)

    Sharma, R C; Hodis, H N; Mack, W J; Sevanian, A; Kramsch, D M


    Oxy-free radicals may be involved in the pathogenesis of accelerated atherosclerosis in hypertension. We evaluated the direct antioxidant potential of probucol in hypertensive arteries by studying the spatial immunohistochemical distribution of three primary antioxidant enzymes (AEs). Nineteen normocholesterolemic New Zealand White rabbits were divided into two groups: normotensive controls (NT; n = 6) and 13 animals rendered hypertensive by surgical coarctation of abdominal aorta. The hypertensive group was subdivided into hypertensive alone (HT; n = 8) and hypertensive treated with 1% probucol (PO) for 9 weeks (HT-P; n = 5). Blood pressure rose significantly in both hypertensive groups (P < .005). At autopsy, both hypertensive groups showed similarly significant increases in mean arterial intima-media thickness (IMT) whether or not they were treated with probucol. However, only HT rabbits revealed significant increases in the intima-media depth penetration of glutathione peroxidase, superoxide dismutase, and catalase AEs. By contrast, in HT-P animals probucol produced significant reductions of immunostaining of all three AEs compared to the HT group (P < .05). Additionally, specific macrophage immunostaining revealed that the arterial wall of HT rabbits had numerous (10 to 12 per high power field) subintimal and medial macrophages as compared to the HT-P animals (1 to 2 per high power field). The blood pressure level correlated significantly with IMT in all three groups, but with depth penetration of the three AEs only in the NT and HT groups. Probucol, therefore, appears to act in concert with the native arterial antioxidant enzymes as a potent free radical scavenger to reduce oxidative stress and thus attenuate the macrophage invasive response in hypertensive arteries.

  12. The immunohistochemical characterization of sarcomatoid malignant mesothelioma of the pleura. (United States)

    Chirieac, Lucian R; Pinkus, Geraldine S; Pinkus, Jack L; Godleski, John; Sugarbaker, David J; Corson, Joseph M


    The immunohistochemical characteristics of epithelioid malignant mesothelioma are well described. However, immunohistochemical analyses of sarcomatoid mesothelioma, the less common type, are limited and its distinction from other tumors of the chest wall, lung and pleura is often problematic. We evaluated 24 patients with pleural sarcomatoid mesothelioma who had surgery (12 extrapleural pneumonectomies, 9 pleurectomies and 3 large biopsies) between 1989 and 2005. Clinicopathologic features and demographic data were recorded. We describe immunohistochemical results for 10 antibodies: AE1/AE3, CAM5.2 and MNF-116 keratins, calretinin, WT-1 protein, bcl-2, CD34, desmin, D2-40 and podoplanin. The patients were 23 men and one woman with a median age at diagnosis of 64.7 years (range 47 to 76). Tumor cells were positive for the keratin proteins AE1/AE3 in 18/24 cases, CAM 5.2 in 23/24 cases and MNF-116 in 21/21 cases. Calretinin was positive in 6/24 cases, WT-1 (nuclear) in 8/24 cases, bcl-2 in 0/24 cases, CD34 in 0/24 cases, desmin in 0/24 cases, D2-40 in 24/24 cases and podoplanin in 24/24 cases. This panel of antibodies may be helpful in establishing a pathologic diagnosis of sarcomatoid mesothelioma. In our study, D2-40 and podoplanin are highly sensitive immunohistochemical markers for sarcomatoid mesothelioma. Additional studies are required to define their role in the differential diagnosis of other spindle cell tumors.

  13. Massive retinal gliosis: An unusual case with immunohistochemical study

    Directory of Open Access Journals (Sweden)

    Sanjay D Deshmukh


    Full Text Available Massive retinal gliosis (MRG is a rare, benign intraocular condition that results from the proliferation of well-differentiated glial cells. Immunohistochemically, these cells show positivity for glial fibrillary acid protein (GFAP, neuron specific enolase (NSE, and S-100 protein. We encountered a case of a 45-year-old female with loss of vision in the left eye. She had a history of trauma to that eye two years ago. Enucleation was carried out, because malignancy was suspected due to retinal calcification. On the basis of light microscopy and immunohistochemistry (IHC performed on the enucleated eye, it was diagnosed as massive retinal gliosis.

  14. Ingrown toenail: histopathologic and immunohistochemical study. (United States)

    Fernandez-Flores, Angel; Martínez-Nova, Alfonso; Salgado-Fernandez, Sara


    Although ingrown toenail (IT) is a common problem, the morphologic alterations presented in this condition have rarely been presented in literature. We have studied 14 cases from 12 different patients and studied the biopsies of all of them, not only with routine techniques but also with histochemical stains (Masson trichrome, periodic acid-Schiff, and Gram) and with immunohistochemical techniques mindbomb homolog-1 (MIB-1), p63, D2-40, and AE1-AE3). The patients were clinically classified into 4 stages (1, 2a, 2b, and 3). The main changes evidenced were ingrowing of the squamous epithelium (either from the matrix or mostly from the hyponichium) and chronic plasma-cell rich inflammation with prominent vascularization. The intensity of these changes varied depending on the stage: whereas they were mild in stages 1 and 2a, they were prominent in stages 2b and 3. In advanced stages, the epithelial nests showed certain dyskeratosis. The expression of MIB-1 was, nevertheless, only basal and parabasal. p63 was intensively expressed by the epithelial nests. We therefore conclude that (a) IT shows epithelial ingrowing with accompanying granulation tissue; (b) according to the clinical stage of the disease, this pattern can be more or less complex; (c) in advanced stages of IT, the pattern can mimic a well-differentiated squamous cell carcinoma; (d) MIB-1 is not helpful in distinguishing IT from squamous cell carcinoma; (e) the high expression of p63 by the ingrowing epithelium should not be the source of a misdiagnosis of malignancy.

  15. An Immunohistochemical Algorithm for Ovarian Carcinoma Typing (United States)

    Rahimi, Kurosh; Rambau, Peter F.; Naugler, Christopher; Le Page, Cécile; Meunier, Liliane; de Ladurantaye, Manon; Lee, Sandra; Leung, Samuel; Goode, Ellen L.; Ramus, Susan J.; Carlson, Joseph W.; Li, Xiaodong; Ewanowich, Carol A.; Kelemen, Linda E.; Vanderhyden, Barbara; Provencher, Diane; Huntsman, David; Lee, Cheng-Han; Gilks, C. Blake; Mes Masson, Anne-Marie


    There are 5 major histotypes of ovarian carcinomas. Diagnostic typing criteria have evolved over time, and past cohorts may be misclassified by current standards. Our objective was to reclassify the recently assembled Canadian Ovarian Experimental Unified Resource and the Alberta Ovarian Tumor Type cohorts using immunohistochemical (IHC) biomarkers and to develop an IHC algorithm for ovarian carcinoma histotyping. A total of 1626 ovarian carcinoma samples from the Canadian Ovarian Experimental Unified Resource and the Alberta Ovarian Tumor Type were subjected to a reclassification by comparing the original with the predicted histotype. Histotype prediction was derived from a nominal logistic regression modeling using a previously reclassified cohort (N=784) with the binary input of 8 IHC markers. Cases with discordant original or predicted histotypes were subjected to arbitration. After reclassification, 1762 cases from all cohorts were subjected to prediction models (χ2 Automatic Interaction Detection, recursive partitioning, and nominal logistic regression) with a variable IHC marker input. The histologic type was confirmed in 1521/1626 (93.5%) cases of the Canadian Ovarian Experimental Unified Resource and the Alberta Ovarian Tumor Type cohorts. The highest misclassification occurred in the endometrioid type, where most of the changes involved reclassification from endometrioid to high-grade serous carcinoma, which was additionally supported by mutational data and outcome. Using the reclassified histotype as the endpoint, a 4-marker prediction model correctly classified 88%, a 6-marker 91%, and an 8-marker 93% of the 1762 cases. This study provides statistically validated, inexpensive IHC algorithms, which have versatile applications in research, clinical practice, and clinical trials. PMID:26974996

  16. Immunohistochemical diagnosis of myocarditis on (infantile) autopsy material: Does it improve the diagnosis? (United States)

    Grasmeyer, Sarah; Madea, Burkhard


    The standard for the histopathologic diagnosis of myocarditis has been the Dallas criteria. Recently, immunohistochemical studies that include the specification and quantification of interstitial inflammatory cells have been proposed as the diagnostic approach for myocarditis. Cut-off limits regarding inflammatory cell numbers for the positive diagnosis of myocarditis have been recommended. However, it is unclear whether these can be applied to postmortem tissues or to infants, as they were established from endomyocardial biopsies and for adults. Nevertheless, cut-off limits for the postmortem diagnosis of myocarditis in the first year of life have been proposed. Studies using these cut-off limits identified myocarditis in a high percentage of presumed sudden infant death syndrome (SIDS) cases. These results were re-evaluated by the present study, which examined heart specimens from infants less than 1 year of age. The study had a test group of 92 SIDS cases and a control group of 15. Myocardial tissue was examined from eight standardized locations, stained with hematoxylin-eosin and for three different immunohistochemical reagents (LCA for leukocytes, CD68 for macrophages, CD45-RO for T-lymphocytes). Histopathological assessment of the number of inflammatory cells was carried out on an aggregate of 80 mm(2) of myocardial tissue per case. Myocarditis, based on the Dallas criteria, was histologically diagnosed in only two cases. Immunohistochemical quantification revealed elevated cell counts in the SIDS group for LCA and CD45-RO. However, those differences were neither statistically significant nor clinically relevant as the mean cell counts per mm(2) were low. The density of inflammatory cells differed considerably from section to section and even within single sections. Therefore the commonly used arithmetic mean value was not diagnostically relevant, suggesting cut-off values based on the arithmetic mean value as recommended in the literature, cannot be regarded

  17. High Specificity of Quantitative Methylation-Specific PCR Analysis for MGMT Promoter Hypermethylation Detection in Gliomas

    Directory of Open Access Journals (Sweden)

    Paola Parrella


    Full Text Available Normal brain tissue from 28 individuals and 50 glioma samples were analyzed by real-time Quantitative Methylation-Specific PCR (QMSP. Data from this analysis were compared with results obtained on the same samples by MSP. QMSP analysis demonstrated a statistically significant difference in both methylation level (P=.000009 Mann Whitney Test and frequencies (P=.0000007, Z-test in tumour samples as compared with normal brain tissues. Although QMSP and MSP showed similar sensitivity, the specificity of QMSP analysis was significantly higher (93%; CI95%: 84%–100% as compared with MSP (64%; 95%CI: 46%–82%. Our results suggest that QMSP analysis may represent a powerful tool to identify glioma patients that will benefit from alkylating agents chemotherapy.

  18. System Specification for Immobilized High-Level Waste Interim Storage

    Energy Technology Data Exchange (ETDEWEB)

    CALMUS, R.B.


    This specification establishes the system-level functional, performance, design, interface, and test requirements for Phase 1 of the IHLW Interim Storage System, located at the Hanford Site in Washington State. The IHLW canisters will be produced at the Hanford Site by a Selected DOE contractor. Subsequent to storage the canisters will be shipped to a federal geologic repository.

  19. Specification of High Activity Gamma-Ray Sources. (United States)

    International Commission on Radiation Units and Measurements, Washington, DC.

    The report is concerned with making recommendations for the specifications of gamma ray sources, which relate to the quantity of radioactive material and the radiation emitted. Primary consideration is given to sources in teletherapy and to a lesser extent those used in industrial radiography and in irradiation units used in industry and research.…

  20. Immunohistochemical visualization of mouse interneuron subtypes

    DEFF Research Database (Denmark)

    Jensen, Simon Mølgaard; Ulrichsen, Maj; Boggild, Simon;


    The activity of excitatory neurons is controlled by a small, but highly diverse population of inhibitory interneurons. These cells show a high level of physiological, morphological and neurochemical heterogeneity, and play highly specific roles in neuronal circuits. In the mammalian hippocampus...

  1. Modified rapid urease test for Helicobacter pylori detection in relation to an immunohistochemical stain. (United States)

    Tokunaga, Y; Shirahase, H; Yamamoto, E; Inao, R; Hamaguchi, S; Kanaji, K; Kitaoka, A; Yagi, T; Tokuka, A; Ohsumi, K


    The rapid urease test and touch cytology have been used for the rapid detection of Helicobacter pylori infection. Recently, a modified rapid urease (MRU) test, which provides results in 20 min has been available on a commercial basis. To date, few reports have evaluated the accuracy of this test. This study evaluated the sensitivity, specificity, and accuracy of the MRU test and touch cytology to detect H. pylori in relation to the density of H. pylori infection determined semi-quantitatively by using immunohistochemical stains. Biopsy specimens obtained from a total of 60 patients who underwent endoscopy for evaluation of gastroduodenal diseases were studied by using the MRU test, Giemsa stain for touch smear tissue and histological methods. An immunohistochemical stain was used as a standard, and the density of H. pylori infection was graded according to the number of individual bacteria seen as follows: grade 0 = 0; grade 1+ = 1-9; grade 2+ = 10-29; grade 3+ = 30-99; grade 4+ > or = 100. The severity of gastritis was evaluated histologically in each specimen and compared with the density of H. pylori infection. The MRU test had an overall sensitivity of 73%, specificity of 100% and accuracy of 85%. The Giemsa stain had a sensitivity of 91%, specificity of 100% and accuracy of 95%.The sensitivities of the MRU test and Giemsa stain decreased in mild H. pylori infection. In the MRU test, the sensitivity was 47% when the density of H. pylori infection was 1+, while 80-100% sensitivities were obtained when the densities of infection were > or = 2+. With the Giemsa stain, the sensitivity was 80% when the density was 1+, while the sensitivity increased to 100% when the densities were > or = 2+. The severity of gastritis determined by the Rauws scores showed a positive correlation with the density of H. pylori infection as evaluated by immunohistochemical staining. The MRU test had high sensitivity and specificity for moderate to severe H. pylori infection, but it may

  2. High specific power flexible integrated IMM photovoltaic blanket Project (United States)

    National Aeronautics and Space Administration — Originally designed for space applications, multi-junction solar cells have a high overall power conversion efficiency (>30%) which compares favorably to...

  3. Dermatophytosis caused by Trichophyton spp. in a Tenerife Lizard (Gallotia galloti): an immunohistochemical study. (United States)

    Orós, J; Hernández, J D; Gallardo, J; Lupiola, P; Jensen, H E


    Reports of dermatophytosis in reptiles are rare. This report describes the microscopical and immunohistochemical findings in a case of dermatophytosis caused by Trichophyton spp. in a 2-year-old Tenerife lizard (Gallotia galloti) with ulcerative and pustular skin lesions. Microscopically, the lesions were characterized by superficial epidermal pustules containing heterophils with numerous fungal hyphae that stained by periodic acid-Schiff and Grocott's stain. Fungal culture was not performed, but a panel of polyclonal antibodies specific for different fungal genera was applied to tissue sections. These immunohistochemical studies demonstrated reactivity of the hyphae only with antiserum specific for Trichophyton spp.

  4. Highly Specific Pose Estimation with a Catadioptric Omnidirectional Camera



    International audience; This article presents a new method for estimating the pose of para-catadioptric vision systems. It is based on the estimation of vanishing points associated with vertical edges of the environment. However, unlike classical approaches no feature (line, circle) extraction and/or identification is needed. A sampled domain of possible vanishing points is tested and histograms are build to characterize the soundness of these points. A specificity index allows to find the mo...

  5. Coaxial plasma thrusters for high specific impulse propulsion (United States)

    Schoenberg, Kurt F.; Gerwin, Richard A.; Barnes, Cris W.; Henins, Ivars; Mayo, Robert; Moses, Ronald, Jr.; Scarberry, Richard; Wurden, Glen


    A fundamental basis for coaxial plasma thruster performance is presented and the steady-state, ideal MHD properties of a coaxial thruster using an annular magnetic nozzle are discussed. Formulas for power usage, thrust, mass flow rate, and specific impulse are acquired and employed to assess thruster performance. The performance estimates are compared with the observed properties of an unoptimized coaxial plasma gun. These comparisons support the hypothesis that ideal MHD has an important role in coaxial plasma thruster dynamics.

  6. High tumour cannabinoid CB1 receptor immunoreactivity negatively impacts disease-specific survival in stage II microsatellite stable colorectal cancer.

    Directory of Open Access Journals (Sweden)

    Sofia B Gustafsson

    Full Text Available BACKGROUND: There is good evidence in the literature that the cannabinoid system is disturbed in colorectal cancer. In the present study, we have investigated whether CB(1 receptor immunoreactive intensity (CB(1IR intensity is associated with disease severity and outcome. METHODOLOGY/PRINCIPAL FINDINGS: CB(1IR was assessed in formalin-fixed, paraffin-embedded specimens collected with a consecutive intent during primary tumour surgical resection from a series of cases diagnosed with colorectal cancer. Tumour centre (n = 483 and invasive front (n = 486 CB(1IR was scored from 0 (absent to 3 (intense staining and the data was analysed as a median split i.e. CB(1IR <2 and ≥2. In microsatellite stable, but not microsatellite instable tumours (as adjudged on the basis of immunohistochemical determination of four mismatch repair proteins, there was a significant positive association of the tumour grade with the CB(1IR intensity. The difference between the microsatellite stable and instable tumours for this association of CB(1IR was related to the CpG island methylation status of the cases. Cox proportional hazards regression analyses indicated a significant contribution of CB(1IR to disease-specific survival in the microsatellite stable tumours when adjusting for tumour stage. For the cases with stage II microsatellite stable tumours, there was a significant effect of both tumour centre and front CB(1IR upon disease specific survival. The 5 year probabilities of event-free survival were: 85±5 and 66±8%; tumour interior, 86±4% and 63±8% for the CB(1IR<2 and CB(1IR≥2 groups, respectively. CONCLUSIONS/SIGNIFICANCE: The level of CB(1 receptor expression in colorectal cancer is associated with the tumour grade in a manner dependent upon the degree of CpG hypermethylation. A high CB(1IR is indicative of a poorer prognosis in stage II microsatellite stable tumour patients.

  7. Immunohistochemical localization of TGF-B1 during morphogenetic movements of the developing mouse palate. (United States)

    Williams, J M; Robinson, R A; Solursh, M


    Transforming growth factor-beta (TGF-B1) may play an important role in developmentally active tissues in which it is found in high concentrations. We localized TGF-B1 in the developing fetal mouse palate immunohistochemically using a polyclonal antibody. Mouse fetal palates at 12-17 days (inclusive) of gestation were examined and specific focal concentrations of TGF-B1 identified regions undergoing active morphogenesis. The association of TGF-B1 with aggregates of mesenchymal cells in the palate and chondroblasts, rhabdomyocytes, and epithelia of the craniofacial complex strongly implicates its role in proliferation and differentiation in the developing mouse palate. We believe these findings have important bearing on the normal development of the palate as well as cleft anomalies.

  8. A Method for Sectioning and Immunohistochemical Analysis of Stem Cell-Derived 3-D Organoids. (United States)

    Wiley, Luke A; Beebe, David C; Mullins, Robert F; Stone, Edwin M; Tucker, Budd A


    This unit describes a protocol for embedding, sectioning, and immunocytochemical analysis of pluripotent stem cell-derived 3-D organoids. Specifically, we describe a method to embed iPSC-derived retinal cups in low-melt agarose, acquire thick sections using a vibratome tissue slicer, and perform immunohistochemical analysis. This method includes an approach for antibody labeling that minimizes the amount of antibody needed for individual experiments and that utilizes large-volume washing to increase the signal-to-noise ratio, allowing for clean, high-resolution imaging of developing cell types. The universal methods described can be employed regardless of the type of pluripotent stem cell used and 3-D organoid generated. © 2016 by John Wiley & Sons, Inc.

  9. Plantar fibromatosis: an immunohistochemical and ultrastructural study. (United States)

    de Palma, L; Santucci, A; Gigante, A; Di Giulio, A; Carloni, S


    The analogies between plantar fibromatosis and Dupuytren's disease (palmar fibromatosis) are well known. The latter is clinically more frequent and has been the object of extensive immunohistochemical and ultrastructural studies, with a view to investigating its pathogenesis. By contrast, such data on plantar fibromatosis are quite scarce. A histochemical, immunohistochemical, and ultrastructural study was performed on nodule tissue from six patients who were subjected to total fasciectomy for plantar fibromatosis. The study of myofibroblasts revealed features suggestive of their fibroblastic origin and evidenced a cytoskeleton and an extracellular filamentous system that could enable myofibroblasts to generate and exert the intracellular forces that contribute to the contraction of the aponeurosis. These aspects are similar to those observed in Dupuytren's disease and seem to lend support to the theory that the two diseases are expressions of the same disorder.

  10. Advanced Space Power Systems (ASPS): High Specific Energy Li-ion Battery Cells Project (United States)

    National Aeronautics and Space Administration — The objective of the High Specific Energy Battery project element is to develop high specific energy battery technologies that enable new capabilities for future...

  11. SOX11 expression is highly specific for mantle cell lymphoma and identifies the cyclin D1-negative subtype (United States)

    Mozos, Ana; Royo, Cristina; Hartmann, Elena; De Jong, Daphne; Baró, Cristina; Valera, Alexandra; Fu, Kai; Weisenburger, Dennis D.; Delabie, Jan; Chuang, Shih-Sung; Jaffe, Elaine S.; Ruiz-Marcellan, Carmen; Dave, Sandeep; Rimsza, Lisa; Braziel, Rita; Gascoyne, Randy D.; Solé, Francisco; López-Guillermo, Armando; Colomer, Dolors; Staudt, Louis M.; Rosenwald, Andreas; Ott, German; Jares, Pedro; Campo, Elias


    Background Cyclin D1-negative mantle cell lymphoma is difficult to distinguish from other small B-cell lymphomas. The clinical and pathological characteristics of patients with this form of lymphoma have not been well defined. Overexpression of the transcription factor SOX11 has been observed in conventional mantle cell lymphoma. The aim of this study was to determine whether this gene is expressed in cyclin D1-negative mantle cell lymphoma and whether its detection may be useful to identify these tumors. Design and Methods The microarray database of 238 mature B-cell neoplasms was re-examined. SOX11 protein expression was investigated immunohistochemically in 12 cases of cyclin D1-negative mantle cell lymphoma, 54 cases of conventional mantle cell lymphoma, and 209 additional lymphoid neoplasms. Results SOX11 mRNA was highly expressed in conventional and cyclin D1-negative mantle cell lymphoma and in 33% of the cases of Burkitt’s lymphoma but not in any other mature lymphoid neoplasm. SOX11 nuclear protein was detected in 50 cases (93%) of conventional mantle cell lymphoma and also in the 12 cyclin D1-negative cases of mantle cell lymphoma, the six cases of lymphoblastic lymphomas, in two of eight cases of Burkitt’s lymphoma, and in two of three T-prolymphocytic leukemias but was negative in the remaining lymphoid neoplasms. Cyclin D2 and D3 mRNA levels were significantly higher in cyclin D1-negative mantle cell lymphoma than in conventional mantle cell lymphoma but the protein expression was not discriminative. The clinico-pathological features and outcomes of the patients with cyclin D1-negative mantle cell lymphoma identified by SOX11 expression were similar to those of patients with conventional mantle cell lymphoma. Conclusions SOX11 mRNA and nuclear protein expression is a highly specific marker for both cyclin D1-positive and negative mantle cell lymphoma. PMID:19880778

  12. High intensity training may reverse the fiber type specific decline in myogenic stem cells in multiple sclerosis patients

    Directory of Open Access Journals (Sweden)

    Jean eFarup


    Full Text Available Multiple sclerosis (MS is associated with loss of skeletal muscle mass and function. The myogenic stem cells (satellite cells – SCs are instrumental to accretion of myonuclei, but remain to be investigated in MS. The present study aimed to compare the SC and myonuclei content between MS patients (n=23 and age matched healthy controls (HC, n=18. Furthermore, the effects of 12 weeks of high intensity training on SC and myonuclei content were explored in MS. Muscle biopsies were obtained from m. Vastus Lateralis at baseline (MS+HC and following 12 weeks of training (MS only. Frozen biopsies were sectioned followed by immunohistochemical analysis for fiber type specific SCs (Pax7+, myonuclei (MN and central nuclei content and fiber cross-sectional area (fCSA using ATPase histochemistry. At baseline the SCs per fiber was lower in type II compared to type I fiber in both MS (119%, p<0.01 and HC (69%, p<0.05, whereas the SCs per fCSA was lower in type II fibers compared to type I only in MS (72%, p<0.05. No differences were observed in MN or central nuclei between MS and HC. Following training the type II fiber SCs per fiber and fCSA in MS patients increased by 165% (p<0.05 and 135% (p<0.05, respectively. Furthermore, the type II fiber MN content increased by 35% (p<0.05 following training. In conclusion, the SC content is lower in type II compared to type I fibers in both MS and HC. Furthermore, high intensity training was observed to selectively increase the SC and myonuclei content in type II fibers in MS patients.

  13. HPV immunohistochemical testing and cervical dysplasia




    Background and aim HPV (Human Papilloma Virus) infection represents a necessary condition for cervical carcinogenesis. The purpose of this study was to evaluate the efficiency of HPV testing using an immunohistochemical staining kit with implications upon both diagnosis and treatment of cervical intraepithelial neoplasia (CIN). Methods Seventy-nine patients and eighty-six controls were enrolled in the study. Each patient had completed a physical examination, gynecological examination with cer...

  14. Surprisingly high substrate specificities observed in complex biofilms

    DEFF Research Database (Denmark)

    Nierychlo, Marta; Kindaichi, Tomonori; Kragelund, Caroline;

    The behavior of microorganisms in natural ecosystems (e.g. biofilms) differs significantly from laboratory studies. In nature microorganisms experience alternating periods of surplus nutrients, nutrient-limitation, and starvation. Literature data suggests that to survive and compete successfully......, microorganisms can regulate their metabolism expressing wide range of uptake and catabolic systems. However, ecophysiological studies of natural biofilms indicate that bacteria are very specialized in their choice of substrate, so even minor changes in substrate composition can affect the community composition...... by selection for different specialized species. We hypothesized that bacteria growing in natural environment express strongly conserved substrate specificity which is independent on short-term (few hours) variations in growth conditions. In this study, biofilm from Aalborg wastewater treatment plant was used...

  15. Differentiation in cutaneous adnexal tumors: Immunohistochemical study

    Directory of Open Access Journals (Sweden)

    Figen BARUT


    Full Text Available Cutaneous adnexal neoplasms are rare tumors that display differentiation in different ways. The aim of this study is, to present embryonic developmental properties and to determine the way of differentiation of adnexal neoplasms by evaluating the immunohistochemical expression of various markers.Forty-seven cases with adnexal tumors enrolled in this study. Histopathologic groups of these 47 cases were: 15 (32% hair follicle tumors, 11 (23.4% sebaceous tumors, 8 (17% apocrine tumors, and 13 (27.6% eccrine tumors. CK5-6, CK6, CK7, CK8, CK10, CK19, GCDFP-15, carcinoembryonic antigen, epithelial membrane antigen and S100 protein expressions were examined by immunohistochemical methods.As a result of this immunohistochemical study it was concluded that the expression of CK5-6 and CK8 carries more importance than other markers in determining certain types of differentiation of hair follicle tumors. It was also determined that, epithelial membrane antigen expression is important for the diagnosis of sebaceous tumors and the markers like CK8, CK10 and carcinoembryonic antigen may aid for the same purpose as well. It was found that, GCDFP-15 as well as CK5-6 expressions are significant for apocrine tumors, and carcinoembryonic antigen reaction as well as CK8 positivity will aid in determining differentiation of eccrine tumors. The presence of similar CK6 expression in all kinds of adnexal tumors has demonstrated that this marker is useless in differential diagnosis.

  16. Histopathologic and immunohistochemical features of Hashimoto thyroiditis

    Directory of Open Access Journals (Sweden)

    H Kazem Amani


    Full Text Available Background: Intrathyroid lymphoid tissue is accrued in Hashimoto thyroiditis (HT. Histologically, this acquired lymphoid tissue bears a close resemblance to mucosa-associated lymphoid tissue (MALT and can evolve to lymphoma. Aim: To demonstrate the morphological, and immunohistochemical profiles of Hashimoto thyroiditis and to ascertain the importance of light chain restriction in distinguishing HT with extensive lymphoplasmacytoid infiltrate from MALT lymphoma. Materials and Methods: We studied histopathologically and immunohistochemically (CD20, CD3, Igk, Igl and cytokeratin 30 cases of HT for evaluation of the lymphoid infiltrate and the presence of lymphoepithelial lesions (LELs. Distinguishing between early thyroid lymphoma and HT was evaluated by light chain restriction. These findings were compared with two cases of primary thyroid lymphoma. Results: The histopathological findings were characteristic of HT. Immunohistochemistry confirmed inconspicuous, rare B-cell LELs as well as a prominent T-lymphocyte population. Testing for light chain restriction showed polyclonal population of plasma cells. The cases of MALT lymphoma had distinct destructive lymphoepithelial lesions, B-cell immunophenotyping and showed kappa light chain restriction in the plasmacytoid population. Conclusions: Hashimoto thyroiditis differs both histopathologically and immunohistochemically from thyroid lymphoma. In suspicious cases, immunohistochemistry could be helpful in reaching a definitive diagnosis.


    Directory of Open Access Journals (Sweden)

    Katarina Bevizova


    Full Text Available Malignant cancers of urinary bladder are the second most common malignancy of the urinary tract and the fourth most common malignancy in general, especially in men. The aim of this study was a retrospective analysis of selected markers (p53, Ki-67 and E-cadherin of urinary bladder cancers from the Department of Urology in Bratislava, Slovak Republic between years 2007 and 2009. We analysed 244 patients (202 males, 42 females with diagnosed bladder cancer via cystoscopy and subsequent transurethral resection. Patients’ age varied from 36 to 98 years. Obtained samples were fixed by 10% buffered formalin for 24 to 48 h. Subsequently, they were dehydrated in ascending ethanol series and embedded in paraffin. The parafin sections of 5 µm were prepared by microtome and they were stained by haematoxylin and eosin. The antibodies against to p53, Ki-67 and E-cadherin were used in immunohistochemical analysis. Statistical evaluation was performed via SPSS using non-parametric Kruskal-Wallis test and p values<0.05 were considered statistically significant. No significant differences in the expression of selected markers were found between genders. Expression of p53 and Ki-67, in G1 and G2 of low grade tumours was lower in comparison to their expression in G3 tumors. Expression of E-cadherin was the opposite in this case. The expression of p53 and Ki-67 positively correlated with tumor’s depth of invasion, while the expression of E-cadherin significantly decreased. In case of T4 tumors, the expression of all markers exhibited consistently high values. When analysing tumor multiplicity, the expression of p53 and Ki-67 significantly decreased, while the expression of E-cadherin significantly increased. Based on the obtained results it can be concluded that the analysis of p53, Ki-67 and E-cadherin expression is essential for diagnostics and prognostics of bladder cancer and should be routinely used in daily practise together with

  18. High Specific Energy Pulsed Electric Discharge Laser Research. (United States)


    drop out excess water, filtered, dried, filtered again, and then pumped up to the storage bottle pressure (Fig. 47). At the exit of the high...pressure pump, an oil filter was used to remove any oil that may have been introduced by the compressor. Bottles were pumped up to 2000 psig...Lowder, R. S. , "Air-Combustion Product N2-C02 Electric Laser, " J. Appl. Phys. Lett. 26, 373 (1975). 5. Miller, D. J. and Millikan , R. C

  19. Range-Specific High-resolution Mesoscale Model Setup (United States)

    Watson, Leela R.


    This report summarizes the findings from an AMU task to determine the best model configuration for operational use at the ER and WFF to best predict winds, precipitation, and temperature. The AMU ran test cases in the warm and cool seasons at the ER and for the spring and fall seasons at WFF. For both the ER and WFF, the ARW core outperformed the NMM core. Results for the ER indicate that the Lin microphysical scheme and the YSU PBL scheme is the optimal model configuration for the ER. It consistently produced the best surface and upper air forecasts, while performing fairly well for the precipitation forecasts. Both the Ferrier and Lin microphysical schemes in combination with the YSU PBL scheme performed well for WFF in the spring and fall seasons. The AMU has been tasked with a follow-on modeling effort to recommended local DA and numerical forecast model design optimized for both the ER and WFF to support space launch activities. The AMU will determine the best software and type of assimilation to use, as well as determine the best grid resolution for the initialization based on spatial and temporal availability of data and the wall clock run-time of the initialization. The AMU will transition from the WRF EMS to NU-WRF, a NASA-specific version of the WRF that takes advantage of unique NASA software and datasets. 37

  20. Ductal adenocarcinoma of the prostate: immunohistochemical findings and clinical significance

    Directory of Open Access Journals (Sweden)

    Sha JJ


    Full Text Available Jianjun Sha,1,2 Juanjie Bo,1 Jiahua Pan,1 Lianhua Zhang,1 Hanqing Xuan,1 Wei Chen,1 Dong Li,1 Zhaoliang Wang,1 Dongming Liu,1 Yiran Huang1,2 1Department of Urology, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, 2School of Biomedical Engineering, Shanghai Jiaotong University, Shanghai, People's Republic of China Introduction: To investigate the clinical features, diagnosis, treatment, and prognosis of ductal adenocarcinoma of the prostate. Methods: The clinicopathological and immunohistochemical data of seven patients with ductal adenocarcinoma of the prostate were retrospectively analyzed. All patients underwent physical examination, magnetic resonance imaging (MRI, bone scan, cystoscopy, and computed tomography (CT scan. The level of prostate-specific antigen (PSA before and after surgery was assessed. Different prostate cancer markers were used for immunohistochemical staining. Results: The mean age of the seven patients diagnosed with prostatic ductal adenocarcinoma in this study was 76.2 years (range 57–88. Five patients presented with intermittent and painless gross hematuria, one patient with progressive dysuria, and one patient with elevated serum PSA on routine health examination. The level of PSA before surgery ranged from 1.3 to 45.0 ng/mL. Immunohistochemical staining results of the prostatic ductal adenocarcinoma confirmed positivity for PSA, prostatic acid phosphatase, androgen receptor, and alpha-methyacyl co-enzyme A (CoA-reductase markers. Two of the patients underwent bilateral orchiectomy combined with anti-androgen therapy, three underwent transurethral resection of prostate, one received radical prostatectomy, and one received medical castration therapy. The clinical outcomes of all patients were satisfactory, based on follow-up data. The symptoms of hematuria and dysuria were ameliorated well, and the postoperative PSA level decreased below 4.0 ng/mL. Recurrence or metastasis of disease was

  1. Manganese Dioxide with High Specific Surface Area for Alkaline Battery

    Institute of Scientific and Technical Information of China (English)

    HUANG You-ju; LIN Yu-li; LI Wei-shan


    The authors reported a facile method for the synthesis of manganese dioxide without any template and catalyst at a low-temperature.The prepared sample was characterized with X-ray diffraction(XRD),scanning electron microscopy(SEM),Brunauer-Emmett-Teller(BET) surface analysis,Fourier transform infrared(FTIR) spectrometry,cyclic voltammetry,altemative current(AC) impedance test and battery discharge test.It is found that the prepared sample belongs to α-MnO2 and has a microsphere morphology and a large BET surface area.The electrochemical characterization indicates that the prepared sample displays a larger electrochemical capacitance than the commercial electrolytic manganese dioxides(EMD) in Na2SO4 solution,and exhibits larger discharge capacity than EMD,especially at a high rate discharge condition when it is used as cathode of alkaline Zn/MnO2 battery.

  2. Reprogrammable Controller Design From High-Level Specification

    Directory of Open Access Journals (Sweden)

    M. Benmohammed


    Full Text Available Existing techniques in high-level synthesis mostly assume a simple controller architecture model in the form of a single FSM. However, in reality more complex controller architectures are often used. On the other hand, in the case of programmable processors, the controller architecture is largely defined by the available control-flow instructions in the instruction set. With the wider acceptance of behavioral synthesis, the application of these methods for the design of programmable controllers is of fundamental importance in embedded system technology. This paper describes an important extension of an existing architectural synthesis system targeting the generation of ASIP reprogrammable architectures. The designer can then generate both style of architecture, hardwired and programmable, using the same synthesis system and can quickly evaluate the trade-offs of hardware decisions.

  3. High Performance Computing - Power Application Programming Interface Specification.

    Energy Technology Data Exchange (ETDEWEB)

    Laros, James H.,; Kelly, Suzanne M.; Pedretti, Kevin; Grant, Ryan; Olivier, Stephen Lecler; Levenhagen, Michael J.; DeBonis, David


    Measuring and controlling the power and energy consumption of high performance computing systems by various components in the software stack is an active research area [13, 3, 5, 10, 4, 21, 19, 16, 7, 17, 20, 18, 11, 1, 6, 14, 12]. Implementations in lower level software layers are beginning to emerge in some production systems, which is very welcome. To be most effective, a portable interface to measurement and control features would significantly facilitate participation by all levels of the software stack. We present a proposal for a standard power Application Programming Interface (API) that endeavors to cover the entire software space, from generic hardware interfaces to the input from the computer facility manager.

  4. Immunohistochemical study of doublecortin and nucleostemin in canine brain

    Directory of Open Access Journals (Sweden)

    E. De Nevi


    Full Text Available Finding a marker of neural stem cells remains a medical research priority. It was reported that the proteins doublecortin and nucleostemin were related with stem/progenitor cells in central nervous system. The aim of the present immunohistochemical study was to evaluate the expression of these proteins and their pattern of distribution in canine brain, including age-related changes, and in non-nervous tissues. We found that doublecortin had a more specific expression pattern, related with neurogenesis and neuronal migration, while nucleostemin was expressed in most cells of almost every tissue studied. The immunolabeling of both proteins decreased with age. We may conclude that nucleostemin is not a specific marker of stem/progenitor cells in the dog. Doublecortin, however, is not an exclusive marker of neural stem cells, but also of neuronal precursors.

  5. Immunohistochemical demonstration of airway epithelial cell markers of guinea pig. (United States)

    Li, Yong; Wang, Jing; He, Hai Yan; Ma, Ling Jie; Zeng, Jin; Deng, Guang Cun; Liu, Xiaoming; Engelhardt, John F; Wang, Yujiong


    The guinea pig (Cavea porcellus) is a mammalian non-rodent species in the Caviidae family. The sensitivity of the respiratory system and the susceptibility to infectious diseases allows the guinea pig to be a useful model for both infectious and non-infectious lung diseases such as asthma and tuberculosis. In this report, we demonstrated for the first time, the major cell types and composition in the guinea pig airway epithelium, using cell type-specific markers by immunohistochemical staining using the commercial available immunological reagents that cross-react with guinea pig. Our results revealed the availability of antibodies cross-reacting with airway epithelial cell types of basal, non-ciliated columnar, ciliated, Clara, goblet and alveolar type II cells, as well as those cells expressing Mucin 5AC, Mucin 2, Aquaporin 4 and Calcitonin Gene Related Peptide. The distribution of these various cell types were quantified in the guinea pig airway by immunohistochemical staining and were comparable with morphometric studies using an electron microscopy assay. Moreover, this study also demonstrated that goblet cells are the main secretory cell type in the guinea pig's airway, distinguishing this species from rats and mice. These results provide useful information for the understanding of airway epithelial cell biology and mechanisms of epithelial-immune integration in guinea pig models.

  6. C4d staining as immunohistochemical marker in inflammatory myopathies. (United States)

    Pytel, Peter


    The diagnosis of an inflammatory myopathy is often established based on basic histologic studies. Additional immunohistochemical studies are sometimes required to support the diagnosis and the classification of inflammatory myopathies. Staining for major histocompatibility complex 1 (MHC1) often shows increased sarcolemmal labeling in inflammatory myopathies. Endomysial capillary staining C5b-9 (membrane attack complex) is a feature that is reported as frequently associated with dermatomyositis. Immunohistochemical staining for C4d is widely used for various applications including the assessment of antibody-mediated rejection after solid organ transplantation. In the context of dermatomyositis, C4d staining has been described in skin biopsies but not in muscle biopsies. A total of 32 muscle biopsy specimens were examined. The hematoxylin and eosin-stained slides were reviewed, and immunohistochemical studies for MHC1, C5b-9, and C4d were conducted. The staining observed for C5b-9 and C4d was compared. Overall, the staining pattern for C4d mirrored the one observed for C5b-9 in the examined muscle biopsy specimens. There was high and statistically significant (P<0.0001) correlation between the staining seen with these 2 antibodies. Both antibodies labeled the cytoplasm of degenerating necrotic myofibers. In addition, both antibodies showed distinct endomysial capillary labeling in a subset of dermatomyositis. Areas with perifascicular atrophy often exhibited the most prominent vascular labeling for C4d and C5b-9. In conclusion, C4d and C5b-9 show similar expression patterns in muscle biopsies of patients with inflammatory myopathies and both highlight the presence of vascular labeling associated with dermatomyositis. C4d antibodies are widely used and may offer an alternative for C5b-9 staining.

  7. Myeloid sarcomas: a histologic, immunohistochemical, and cytogenetic study

    Directory of Open Access Journals (Sweden)

    Rodgers William H


    Full Text Available Abstract Context. - Myeloid sarcoma (MS is a neoplasm of immature granulocytes, monocytes, or both involving any extramedullary site. The correct diagnosis of MS is important for adequate therapy, which is often delayed because of a high misdiagnosis rate. Objective. - To evaluate the lineage differentiation of neoplastic cells in MS by immunohistochemistry, and to correlate the results with clinicopathologic findings and cytogenetic studies. Design. - Histologic and immunohistochemical examinations were performed on formalin-fixed paraffin-embedded tissue samples from 13 cases of MS. They were classified according to the World Health Organization criteria. Chromosomal analysis data were available in 11 cases. Clinical, pathological, and cytogenetic findings were analyzed. Results. - The study included six male and seven female patients with an age range of 25 to 72 years (mean, 49.3 years and a male to female ratio of 1:1.2. MS de novo occurred in 4/13 (31% of cases examined. The most sensitive immunohistochemical markers were CD43 and lysozyme present in all cases with MS (13/13, 100%. All de novo MS showed a normal karyotype, monoblastic differentiation, and lack of CD34. The most common chromosomal abnormalities in MS associated with a hematopoietic disorder were trisomy 8 and inv(16 (2/11, 18%. Conclusion. - An immunohistochemical panel including CD43, lysozyme, myeloperoxidase (MPO, CD68 (or CD163, CD117, CD3 and CD20 can successfully identify the vast majority of MS variants in formalin-fixed paraffin-embedded tissue sections. The present report expands the spectrum of our knowledge showing that de novo MS has frequent monoblastic differentiation and frequently carries a normal karyotype.

  8. Unallocated Off-Specification Highly Enriched Uranium: Recommendations for Disposition

    Energy Technology Data Exchange (ETDEWEB)

    Bridges, D. N.; Boeke, S. G.; Tousley, D. R.; Bickford, W.; Goergen, C.; Williams, W.; Hassler, M.; Nelson, T.; Keck, R.; Arbital, J.


    The U.S. Department of Energy (DOE) has made significant progress with regard to disposition planning for 174 metric tons (MTU) of surplus Highly Enriched Uranium (HEU). Approximately 55 MTU of this 174 MTU are ''offspec'' HEU. (''Off-spec'' signifies that the isotopic or chemical content of the material does not meet the American Society for Testing and Materials standards for commercial nuclear reactor fuel.) Approximately 33 of the 55 MTU have been allocated to off-spec commercial reactor fuel per an Interagency Agreement between DOE and the Tennessee Valley Authority (1). To determine disposition plans for the remaining {approx}22 MTU, the DOE National Nuclear Security Administration (NNSA) Office of Fissile Materials Disposition (OFMD) and the DOE Office of Environmental Management (EM) co-sponsored this technical study. This paper represents a synopsis of the formal technical report (NNSA/NN-0014). The {approx} 22 MTU of off-spec HEU inventory in this study were divided into two main groupings: one grouping with plutonium (Pu) contamination and one grouping without plutonium. This study identified and evaluated 26 potential paths for the disposition of this HEU using proven decision analysis tools. This selection process resulted in recommended and alternative disposition paths for each group of HEU. The evaluation and selection of these paths considered criteria such as technical maturity, programmatic issues, cost, schedule, and environment, safety and health compliance. The primary recommendations from the analysis are comprised of 7 different disposition paths. The study recommendations will serve as a technical basis for subsequent programmatic decisions as disposition of this HEU moves into the implementation phase.

  9. American Tegumentary Leishmaniasis: Effectiveness of an Immunohistochemical Protocol for the Detection of Leishmania in Skin (United States)

    Alves, Cibele Fontes; Alves, Cintia Fontes; Figueiredo, Maria Marta; Souza, Carolina Carvalho; Machado-Coelho, George Luiz Lins; Melo, Maria Norma; Tafuri, Washington Luiz; Raso, Pedro; Soares, Rodrigo Pedro; Tafuri, Wagner Luiz


    Background American tegumentary leishmaniasis (ATL) is endemic in Latin America, where Brazil has over 27 thousand cases per year. The aim of the present study was to develop an immunohistochemical method (IHC) for ATL diagnosis. For this purpose, we used serum from a dog naturally infected with Leishmania (L) infantum (canine hyperimmune serum) as the primary antibody, followed by a detection system with a secondary biotinylated antibody. Methodology Skin samples were obtained from 73 patients in an endemic area of Caratinga, Minas Gerais (MG) State, Brazil all testing positive for ATL with the Montenegro skin test, microscopy, and PCR. Canine hyperimmune serum of a dog naturally infected with Leishmania (L.) infantum was employed as a primary antibody in an immunohistochemical diagnostic method using streptavidin-biotin peroxidase. To assess the specificity of this reaction, IHC assays employing two monoclonal antibodies were carried out. As the polymer-based technology is less time-consuming and labor intensive than the IHC labeled streptavidin-biotin peroxidase method, we compared the two methods for all samples. Results The IHC method detected ATL in 67 of the 73 cases (91.8%). Immunolabeled parasites were primarily detected inside macrophages either in the superficial or the deep dermis. Detection was facilitated by the high contrast staining of amastigotes (dark brown) against the light blue background. A lower detection rate (71.2%) was observed with the both of the monoclonal Leishmania antibodies compared to the canine hyperimmune serum. This may have been due to a non-specific background staining observed in all histological samples rendering positive detection more difficult. The higher efficacy of the canine hyperimmune serum in the IHC method was confirmed by the method using streptavidin-biotin peroxidase as well as that with the polymer-based technology (biotin-avidin-free system). Conclusions The data are encouraging with regard to validating IHC

  10. Identification of potential prognostic markers for vulvar cancer using immunohistochemical staining of tissue microarrays.

    NARCIS (Netherlands)

    Fons, G.; Burger, M.P.; Kate, F.J. ten; Velden, J. van der


    The aim of this study is to determine immunohistochemical markers with prognostic significance for disease-specific survival in patients with squamous cell cancer of the vulva. The study material consisted of slides and paraffin blocks of 50 vulvectomy specimens. A tissue microarray was constructed

  11. Immunohistochemical Analysis of the Structure of Injured Peripheral Nerve Neuroma after Electrosurgical Welding Intervention. (United States)

    Korsak, A V; Chaikovskii, Yu B


    Immunohistochemical analysis of changes in neuroma after surgical treatment of damaged peripheral nerve with the use of high frequency electrosurgical device for high frequency current welding of soft tissues was carried out. No adverse effects of this technology and the bipolar instrument on degeneration and regeneration of damaged nerve stem were detected.

  12. Automated quantification of nuclear immunohistochemical markers with different complexity. (United States)

    López, Carlos; Lejeune, Marylène; Salvadó, María Teresa; Escrivà, Patricia; Bosch, Ramón; Pons, Lluis E; Alvaro, Tomás; Roig, Jordi; Cugat, Xavier; Baucells, Jordi; Jaén, Joaquín


    Manual quantification of immunohistochemically stained nuclear markers is still laborious and subjective and the use of computerized systems for digital image analysis have not yet resolved the problems of nuclear clustering. In this study, we designed a new automatic procedure for quantifying various immunohistochemical nuclear markers with variable clustering complexity. This procedure consisted of two combined macros. The first, developed with a commercial software, enabled the analysis of the digital images using color and morphological segmentation including a masking process. All information extracted with this first macro was automatically exported to an Excel datasheet, where a second macro composed of four different algorithms analyzed all the information and calculated the definitive number of positive nuclei for each image. One hundred and eighteen images with different levels of clustering complexity was analyzed and compared with the manual quantification obtained by a trained observer. Statistical analysis indicated a great reliability (intra-class correlation coefficient > 0.950) and no significant differences between the two methods. Bland-Altman plot and Kaplan-Meier curves indicated that the results of both methods were concordant around 90% of analyzed images. In conclusion, this new automated procedure is an objective, faster and reproducible method that has an excellent level of accuracy, even with digital images with a high complexity.

  13. Hepatocellular Tumors: Immunohistochemical Analyses for Classification and Prognostication

    Institute of Scientific and Technical Information of China (English)

    Regina Cheuk-Lam Lo; Irene Oi-Lin Ng


    Following the classification of hepatocellular nodules by the International Working Party in 1995 and further elaboration by the International Consensus Group for Hepatocellular Neoplasia in 2009,entities under the spectrum of hepatocellular nodules have been better characterized.Research work hence has been done to answer questions such as distinguishing high-grade dysplastic nodules from early hepatocellular carcinoma (HCC),delineating the tumor cell origin of HCC,identifying its prognostic markers,and subtyping hepatocellular adenomas.As a result,a copious amount of data at immunohistochemical and molecular levels has emerged.A panel of immunohistochemical markers including glypican-3,heat shock protein 70 and glutamine synthetase has been found to be of use in the diagnosis of small,well differentiated hepatocellular tumors and particularly of HCC.The use of liver fatty acid binding protein (L-FABP),β-catenin,glutamine synthetase,serum amyloid protein and C-reactive protein is found to be helpful in the subtyping of hepatocellular adenomas.The role of tissue biomarkers for prognostication in HCC and the use of biomarkers in subclassifying HCC based on tumor cell origin are also discussed.

  14. Synthesis of a high specific activity methyl sulfone tritium isotopologue of fevipiprant (NVP-QAW039). (United States)

    Luu, Van T; Goujon, Jean-Yves; Meisterhans, Christian; Frommherz, Matthias; Bauer, Carsten


    The synthesis of a triple tritiated isotopologue of the CRTh2 antagonist NVP-QAW039 (fevipiprant) with a specific activity >3 TBq/mmol is described. Key to the high specific activity is the methylation of a bench-stable dimeric disulfide precursor that is in situ reduced to the corresponding thiol monomer and methylated with [(3)H3]MeONos having per se a high specific activity. The high specific activity of the tritiated active pharmaceutical ingredient obtained by a build-up approach is discussed in the light of the specific activity usually to be expected if hydrogen tritium exchange methods were applied.

  15. An immunohistochemical study of verrucous hemangiomas. (United States)

    Trindade, Felicidade; Torrelo, Antonio; Requena, Luis; Tellechea, Oscar; Del Pozo, Jesus; Sacristán, Felipe; Esteve-Martínez, Altea; de Unamuno, Blanca; de Miquel, Víctor Alegre; Colmenero, Isabel


    According to the International Society for the study of vascular Anomalies, vascular anomalies are classified as vascular neoplasms and vascular malformations. In some vascular lesions, categorization as a neoplasm or malformation has not been established with confidence so far. In order to further clarify the nosology of verrucous hemangioma, we studied 13 cases. To analyze immunohistochemical characteristics of verrucous hemangiomas in order to gain further insight in its histogenesis. We carried out a retrospective review. Immunohistochemical expression for Wilms tumor 1 (WT1), Glut-1 and D2-40 was performed in 13 cases. Immunohistochemistry performed with Glut-1 and WT1 showed positive staining in all lesions. All verrucous hemangiomas lacked D2-40 immunostaining. This is the first report in the literature investigating WT1 in verrucous hemangioma in order to further clarify the nosology of this vascular anomaly. Despite the clinical features of verrucous hemangioma, which are similar to those seen in vascular malformations, verrucous hemangioma exhibited an immunoprofile similar to vascular neoplasms, according to WT1 and Glut-1 positivity. © 2013 John Wiley & Sons A/S. Published by Blackwell Publishing Ltd.

  16. Immunohistochemical detection of brain tissue in heated meat products. (United States)

    Tersteeg, M H G; Koolmees, P A; van Knapen, F


    Immunohistochemical methods were used to determine whether brain tissue could be detected in test batches of meat products prepared with known levels of this tissue (0, 1, 5, 10, or 20% bovine brain tissue or 5% porcine brain tissue). Four different, commercially-available antibodies were examined: anti-Neurofilament (anti-NF), anti-MyelinBasicProtein (anti-MBP), anti-NeuronSpecificEnolase (anti-NSE) and anti-GlialFibrillaryAcidicProtein (anti-GFAP). Results obtained with the four antibodies differed with the heat treatment applied to the products (pasteurisation or sterilisation). The amount of immunoreaction product in the raw meat product varied with the antibody, even when the sample contained the same amount of brain tissue. The staining pattern also varied with the antibody. Overall, the anti-MBP antibody proved to be most useful in detecting brain tissue in finely comminuted heated meat products.

  17. Immunohistochemical investigations of genital ulcers caused by Haemophilus ducreyi. (United States)

    Abeck, D; Freinkel, A L; Korting, H C; Szeimis, R M; Ballard, R C


    To gain information on the specific composition of the inflammatory infiltrate of genital ulcers caused by Haemophilus ducreyi, biopsies of 6 genital ulcers which were diagnosed as chancroid on clinical and microbiological grounds were subjected to immunohistochemical investigations after conventional haematoxylineosin staining. A variety of antibodies reactive against B- and T-cells, plasma cells and granulocytes were used with each tissue sections. The lymphocytic infiltrate of chancroid ulcers consisted of both B- and T-lymphocytes and showed a cluster-like formation. B-lymphocytes were preferentially localized perivascularly in the middle layer, T-lymphocytes mainly in the deep layer of the inflamed oedematous tissue. Results stress the importance of both B- and T-cell mediated immune responses in Haemophilus ducreyi infection.

  18. Immunohistochemical detection of hTERT in urothelial lesions: a potential adjunct to urine cytology

    Directory of Open Access Journals (Sweden)

    Khalbuss Walid


    Full Text Available Abstract Background Urine cytology has a critical role in evaluation for bladder carcinoma. Due to the low sensitivity of this technique, ancillary modalities such as the detection of markers of malignancy by immunochemistry are desirable. Promising factors in this context are components of the human telomerase enzyme complex. Telomerase repairs and extend telomeres, which when eroded beyond a critical limit trigger a senescence checkpoint. Accordingly, while absent in normal somatic cells, telomerase activity has been detected in the great majority of malignant tumor specimens tested, and so has potential value for the recognition of malignant cells in clinical specimens. Methods In this study, we investigated whether the immunohistochemical detection of the catalytic subunit of telomerase (hTERT can aid cytology in the diagnosis of bladder lesions. Findings from the retrospective evaluation of over 100 cell blocks, including urine sediments from confirmed malignant and benign conditions, were compared with routine urine cytology data. Results The presence of hTERT protein was indicative of the transformation of urothelia to a malignant phenotype. Nucleolar hTERT was expressed in 27 (93% of 29 samples obtained from patients with confirmed primary bladder cancer. Conversely, hTERT was detectable in only 3 (0.8% of 39 samples from benign conditions. The hTERT assay showed higher diagnostic sensitivity (84.8% than published urine cytology data (~65% for confirmed bladder carcinoma, however, the hTERT assay was less specific than cytology (65.2% vs. ~95% respectively. Conclusion As a highly sensitive marker, immunohistochemical hTERT detection in urine sediments represents a reliable adjunct to cytology in the accurate diagnosis of urothelial neoplasms.

  19. Immunohistochemical evaluation of tissue factor, fibrin/fibrinogen and D-dimers in canine gliomas. (United States)

    de la Fuente, Cristian; Pumarola, Martí; Blasco, Ester; Fernández, Francisco; Viu, Judit; Añor, Sònia


    In human gliomas, tissue factor (TF) is overexpressed, associated with the grade of malignancy and influences tumour biology. Intra-tumoural fibrin/fibrinogen deposition and activation of the fibrinolytic system also play a role in tumour cell proliferation and angiogenesis. The first aim of the present study was to investigate TF expression and the presence of fibrin/fibrinogen and D-dimers in canine glioma biopsies, graded according to the World Health Organization (WHO) classification of tumours of the central nervous system. The second aim was to investigate the occurrence of intravascular thrombosis (IVT) in canine gliomas, as a potential histological marker of glioma type or grade of malignancy. An immunohistochemical study using antibodies against TF, fibrin/fibrinogen and D-dimers was performed with 24 glioma samples, including 15 oligodendrogliomas, 6 astrocytomas and 3 mixed gliomas. Immunohistochemical data were statistically analysed to determine whether there was any relationship between glioma type and grade of malignancy. All gliomas were moderate to strongly positive for TF and the staining score was significantly higher (P = 0.04) in high-grade (III or IV) than in low-grade (II) gliomas. Intra-tumoural fibrin/fibrinogen deposition was detected in all tumour biopsies assessed, and D-dimers were detected in 17/24 gliomas. IVT was a frequent finding, but was not linked to a specific glioma type or malignancy grade. TF expression, fibrin/fibrinogen deposition, extravascular fibrinolytic system activation and IVT occur in canine gliomas. Canine glioma might be a suitable model for studying coagulation and fibrinolysis as potential therapeutic targets for human gliomas.

  20. Successful Treatment of Granuloma Faciale with Topical Tacrolimus: A Case Report and Immunohistochemical Study

    Directory of Open Access Journals (Sweden)

    Gen-ichi Tojo


    Full Text Available We report the case of a 55-year-old Japanese patient with granuloma faciale (GF successfully treated with topical tacrolimus and describe the immunohistochemical study. Immunohistochemical staining revealed that the patient’s granuloma contained CD3+, CD4+, CD8+, CD68+ and CD163+ cells. Interestingly, these cells contained granulysin+ T cells and lacked Foxp3high+ regulatory T cells. In addition, the macrophages were mainly CD163+, which suggested that the alternatively activated macrophage is one of the main components of GF. In summary, the present data shed light on the granuloma-composing cells and possible mechanisms in the treatment of GF with topical tacrolimus.

  1. Pneumatic Performance Study of a High Pressure Ejection Device Based on Real Specific Energy and Specific Enthalpy

    Directory of Open Access Journals (Sweden)

    Jie Ren


    Full Text Available In high-pressure dynamic thermodynamic processes, the pressure is much higher than the air critical pressure, and the temperature can deviate significantly from the Boyle temperature. In such situations, the thermo-physical properties and pneumatic performance can’t be described accurately by the ideal gas law. This paper proposes an approach to evaluate the pneumatic performance of a high-pressure air catapult launch system, in which esidual functions are used to compensate the thermal physical property uncertainties of caused by real gas effects. Compared with the Nelson-Obert generalized compressibility charts, the precision of the improved virial equation of state is better than Soave-Redlich-Kwong (S-R-K and Peng-Robinson (P-R equations for high pressure air. In this paper, the improved virial equation of state is further used to establish a compressibility factor database which is applied to evaluate real gas effects. The specific residual thermodynamic energy and specific residual enthalpy of the high-pressure air are also derived using the modified corresponding state equation and improved virial equation of state which are truncated to the third virial coefficient. The pneumatic equations are established on the basis of the derived residual functions. The comparison of the numerical results shows that the real gas effects are strong, and the pneumatic performance analysis indicates that the real dynamic thermodynamic process is obviously different from the ideal one.

  2. Nerve sheath myxoma (neurothekeoma) of the skin: light microscopic and immunohistochemical reappraisal of the cellular variant. (United States)

    Argenyi, Z B; LeBoit, P E; Santa Cruz, D; Swanson, P E; Kutzner, H


    Nerve sheath myxoma (NSM) is a rare cutaneous neoplasm, the histogenesis of which is controversial. Fifteen cases of NSM were studied by routine light microscopy and with a broad panel of immunohistochemical stains. NSM were classified into three groups based on cellularity, mucin content and growth pattern. 1) The hypocellular (myxoid) type (5/15 cases) showed frequent encapsulation or sharp circumscription. Immunohistochemically this type was strongly positive for S-100 protein and collagen type IV and variably positive for epithelial membrane antigen. 2) The cellular type (4/15 cases) had scant mucin and ill-defined nodular or infiltrating growth. Immunostaining showed positive reaction for neuron specific enolase (2/4), Leu-7 (1/4) and smooth muscle specific actin (2/4), and was negative with the other antibodies. 3) The "mixed type" (6/15 cases) had variable cellularity and mucin content with poor demarcation and variable immunolabeling. We conclude that: 1) there are major light microscopic and immunohistochemical differences between the classical hypocellular (myxoid) and the cellular forms of NSM (neurothekeoma); 2) while the immunohistochemical results support the presence of nerve sheath differentiation in the classical forms of NSM, and to some extent in the mixed forms, there is an absence of convincing evidence of neural differentiation in the cellular variant by either light microscopy or immunohistochemistry; 3) the variable immunophenotypes suggest that differentiation other than neural may take place in CNT.

  3. High Efficiency, High Mass Specific Power Two-Terminal Solar Cells Project (United States)

    National Aeronautics and Space Administration — Future NASA missions will require significant improvements in photovoltaic energy conversion efficiency (>30%) and mass specific power (>600 W/kg), and...

  4. Scrub typhus hepatitis confirmed by immunohistochemical staining

    Institute of Scientific and Technical Information of China (English)

    Jong-Hoon Chung; Sung-Chul Lim; Na-Ra Yun; Sung-Heui Shin; Choon-Mee Kim; Dong-Min Kim


    Scrub typhus is an acute febrile disease caused by Orientia tsutsugamushi (O.tsutsugamushi).We report herein the case of a woman who presented with fever and elevated serum levels of liver enzymes and who was definitively diagnosed with scrub typhus by histopathological examination of liver biopsy specimens,serological tests and nested polymerase chain reaction.Immunohistochemical staining using a monoclonal anti-O.tsutsugamushi antibody showed focally scattered positive immunoreactions in the cytoplasm of some hepatocytes.This case suggests that scrub typhus hepatitis causes mild focal inflammation due to direct liver damage without causing piecemeal necrosis or interface hepatitis.Thus,scrub typhus hepatitis differs from acute viral hepatitis secondary to liver damage due to host immune responses,which causes severe Iobular disarray with diffuse hepatocytic degeneration,necrosis and apoptosis as well as findings indicative of hepatic cholestasis,such as hepatic bile plugs or brown pigmentation of hepatocytes.

  5. Immunohistochemical demonstration of glial markers in retinoblastomas

    DEFF Research Database (Denmark)

    Schrøder, H D


    Twenty retinoblastomas were studied immunohistochemically in order to visualize glial cells. In the retina, the glial cells in the ganglion cell layer and the Müller cells were GFAP positive, while only the glial cells of the ganglion cell layer expressed S-100 reactivity. In the tumours S-100/GFAP...... positive glial cells were found in areas near the retina and along many tumour vessels. Some S-100 reactive cells previously interpreted as tumour cells were refound in a few tumours. In areas with Flexner-Winterstein rosettes and in areas with light cells showing photoreceptor-like differentiation, glial...... cells reactive for both S-100 and GFAP were demonstrated. The latter findings may represent differentiation in a glial direction in the more mature parts of retinoblastoma....

  6. Fine specificities of two lectins from Cymbosema roseum seeds: a lectin specific for high-mannose oligosaccharides and a lectin specific for blood group H type II trisaccharide. (United States)

    Dam, Tarun K; Cavada, Benildo S; Nagano, Celso S; Rocha, Bruno Am; Benevides, Raquel G; Nascimento, Kyria S; de Sousa, Luiz Ag; Oscarson, Stefan; Brewer, C Fred


    The legume species of Cymbosema roseum of Diocleinae subtribe produce at least two different seed lectins. The present study demonstrates that C. roseum lectin I (CRL I) binds with high affinity to the "core" trimannoside of N-linked oligosaccharides. Cymbosema roseum lectin II (CRL II), on the other hand, binds with high affinity to the blood group H trisaccharide (Fucα1,2Galα1-4GlcNAc-). Thermodynamic and hemagglutination inhibition studies reveal the fine binding specificities of the two lectins. Data obtained with a complete set of monodeoxy analogs of the core trimannoside indicate that CRL I recognizes the 3-, 4- and 6-hydroxyl groups of the α(1,6) Man residue, the 3- and 4-hydroxyl group of the α(1,3) Man residue and the 2- and 4-hydroxyl groups of the central Man residue of the trimannoside. CRL I possesses enhanced affinities for the Man5 oligomannose glycan and a biantennary complex glycan as well as glycoproteins containing high-mannose glycans. On the other hand, CRL II distinguishes the blood group H type II epitope from the Lewis(x), Lewis(y), Lewis(a) and Lewis(b) epitopes. CRL II also distinguishes between blood group H type II and type I trisaccharides. CRL I and CRL II, respectively, possess differences in fine specificities when compared with other reported mannose and fucose recognizing lectins. This is the first report of a mannose-specific lectin (CRL I) and a blood group H type II-specific lectin (CRL II) from seeds of a member of the Diocleinae subtribe.

  7. Merkel cell carcinoma with an unusual immunohistochemical profile

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    L. Pilloni


    Full Text Available The clinical and morphological picture of Merkel cell carcinoma (MCC may be rather challenging; therefore, the immunohistochemical profile plays a relevant role in confirming the microscopic diagnosis. A panel of antibodies including cytokeratins 20, 7 and epithelial membrane antigen, and neuronspecific enolase is used in confirming the morphological diagnosis of MCC. The majority of MCCs express CK20 and are CK7-negative. Herein, we present a case of primary cutaneous neuroendocrine carcinoma with an atypical immunohistochemical pattern. A 83-years old woman presented with a painless plaque, red to violaceous in colour, located in the leg. The skin tumor was excided, formalin-fixed and paraffinembedded. Tissue sections were immunostained with a panel of antibodies routinely utilized in complex primary skin tumors for evidencing epithelial and neuroendocrine differentiation of tumor cells. The neuroendocrine differentiation of tumor cells was evidenced by their immunoreactivity for synaptophysin, chromograninA and neuron-specific enolase. Tumor cells also showed diffuse cytoplasmic staining for CK7. No immunoreactivity was detected for CK20 and thyroid transcription factor-1. Our data, together with previous rare reports of CK20-/CK7+ MCCs, lay stress on the importance of routinely utilizing a panel of antibodies in the differential diagnosis of complex primary carcinomas of the skin and may have important implications in expanding the differential diagnosis of skin tumors. In particular, caution should be taken in excluding the diagnosis of MCC only on the basis of the absence of reactivity of tumor cells for CK20, favouring the wrong diagnosis of less aggressive skin tumors.

  8. Cyclin D1 is a useful marker for soft tissue Ewing's sarcoma/peripheral Primitive Neuroectodermal Tumor in children and adolescents: A comparative immunohistochemical study with rhabdomyosarcoma. (United States)

    Magro, Gaetano; Brancato, Franca; Musumeci, Giuseppe; Alaggio, Rita; Parenti, Rosalba; Salvatorelli, Lucia


    Cyclin D1 amplification and/or overexpression contribute to the loss of the regulatory circuits that govern G1-S transition phase of the cell cycle, playing pivotal roles in different human malignant tumors, including breast, colon, prostate cancer, lymphoma, melanoma and neuroblastoma. In vitro studies have shown that cyclin D1 is overexpressed in Ewing's sarcoma (EWS)/peripheral Primitive Neuroectodermal Tumor (pPNET), but not in rhabdomyosarcoma cell lines. Only a few immunohistochemical studies are available on cyclin D1 expression in EWS/pPNET, which confirmed its expression only in a limited number of cases. The aim of the present study was a comparative immunohistochemical analysis of the expression and distribution of cyclin D1 in a large series of pediatric/adolescent soft tissue EWS/pPNETs and rhabdomyosarcomas (both embryonal and alveolar subtypes) to assess its potential usefulness in their differential diagnosis. Notably cyclin D1 was strongly and diffusely expressed in all cases (20/20) of EWS/pPNET, while it was lacked in all cases (15/15) of rhabdomyosarcomas. Immunohistochemical overexpression of cyclin D1 in EWS/pPNET is a novel finding which could be exploitable as a diagnostic immunomarker for this tumor. Although highly sensitive, cyclin D1 is not specific for EWS/pPNET, and thus it should not be evaluated alone but in the context of a wide immunohistochemical panel. Accordingly, we first emphasize that when pathologists are dealing with a small round blue cell tumor of soft tissues in pediatric/adolescent patients, a strong and diffuse nuclear expression of cyclin D1 is of complementary diagnostic value to CD99 and FLI-1 in confirming diagnosis of EWS/pPNET and in ruling out rhabdomyosarcoma.

  9.  Immunohistochemical Expression of ki-67 and p53 in Colorectal Adenomas: A Clinicopathological Study

    Directory of Open Access Journals (Sweden)

    Hussam Hasson Ali


    Full Text Available  Objectives: To evaluate the significance of P53 and Ki-67 expression as immunohistochemical markers in early detection of premalignant changes in different types of colorectal adenomas. Also, to correlate immunohistochemical expression of the two markers with different clinicopathological parameters including; age, and sex of the patient, type, site, size and grade of dysplasia of colorectal adenomas.Methods: Forty-seven polypectomy specimens of colorectal adenomas were retrieved from the archival materials of the Gastrointestinal and Hepatic Diseases Teaching Hospital in Baghdad from 2009 - 2010. Four µm section specimens were stained by immunohistochemical technique with Ki-67 and P53 tumor markers. P-values <0.05 were considered statistically significant.Results: Immunohistochemical expressions of Ki-67 and P53 had a significant correlation with the size and grade of dysplasia in colorectal adenomas. However, there was no significant correlation among the immunohistochemical expression of Ki-67 and P53 with the age and gender of the patient, and the type and site of colorectal adenomas. There was no significant correlation between Ki-67 and P53 expressions in colorectal adenomas. Villous adenomas of colorectum showed a significant correlation with the grade of dysplasia, while there was no significant correlation between size and site of colorectal adenoma with the grade of dysplasia.Conclusion: High grade dysplasia with significant positive immunohistochemical markers of Ki-67 and P53 could be valuable parameters for selecting from the total colorectal adenoma population, those most deserving of close surveillance in follow-up cancer prevention programs. It is closely linked with increasing age particularly in patients with a large size adenoma of villous component in their histology.

  10. [Prospects for using immunohistochemical methods in forensic medical thanatology]. (United States)

    Bogomolov, D V; Bogomolova, I N; Karavaeva, I E


    This review of Russian and foreign literature is focused on the use of immunohistochemical methods in forensic medical practice. It shows that forensic medical specialists not infrequently underestimate the value of these techniques. Recommendations are proposed for a more extensive application of immunohistochemical methods in practical and fundamental medico-legal thanatology.

  11. Pancreatic neuroendocrine tumor and solid-pseudopapillary neoplasm: Key immunohistochemical profiles for differential diagnosis (United States)

    Ohara, Yusuke; Oda, Tatsuya; Hashimoto, Shinji; Akashi, Yoshimasa; Miyamoto, Ryoichi; Enomoto, Tsuyoshi; Satomi, Kaishi; Morishita, Yukio; Ohkohchi, Nobuhiro


    AIM To reveal better diagnostic markers for differentiating neuroendocrine tumor (NET) from solid-pseudopapillary neoplasm (SPN), focusing primarily on immunohistochemical analysis. METHODS We reviewed 30 pancreatic surgical specimens of NET (24 cases) and SPN (6 cases). We carried out comprehensive immunohistochemical profiling using 9 markers: Synaptophysin, chromogranin A, pan-cytokeratin, E-cadherin, progesterone receptor, vimentin, α-1-antitrypsin, CD10, and β-catenin. RESULTS E-cadherin staining in NETs, and nuclear labeling of β-catenin in SPNs were the most sensitive and specific markers. Dot-like staining of chromogranin A might indicate the possibility of SPNs rather than NETs. The other six markers were not useful because their expression overlapped widely between NETs and SPNs. Moreover, two cases that had been initially diagnosed as NETs on the basis of their morphological features, demonstrated SPN-like immunohistochemical profiles. Careful diagnosis is crucial as we actually found two confusing cases showing disagreement between the tumor morphology and immunohistochemical profiles. CONCLUSION E-cadherin, chromogranin A, and β-catenin were the most useful markers which should be employed for differentiating between NET and SPN.

  12. Morphological and immunohistochemical characterisation of seminomas in Norwegian dogs

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    Thorvaldsen Tor


    Full Text Available Abstract Background Seminomas in the dog have traditionally been assumed to resemble human spermatocytic seminomas, based on their low malignancy and high occurrence in old individuals. However, recently published studies indicate that canine seminomas can be classified as classical and spermatocytic seminomas in a similar way as in man, and that classical seminomas comprise a substantial proportion of seminomas in the dog. These two factors both contribute to increasing the potential of canine seminoma as a relevant model for human testicular cancer. The aim of the present study was to characterise seminoma in Norwegian dogs using morphology and immunohistochemistry, and determine whether these tumours are comparable with human classical seminoma. Methods By applying diagnostic criteria from human pathology, 45 seminomas from the Norwegian Canine Cancer Register were examined histologically with hematoxylin and eosin (HE and periodic acid-Schiff (PAS stains. All sections were stained immunohistochemically with antibodies against human placental alkaline phosphatase (PLAP and the transmembrane receptor c-KIT. Results Although two of the seminomas showed immunohistochemical staining characteristics indicative of classical seminoma (PLAP+/c-KIT+, all 45 examined seminomas were morphologically consistent with spermatocytic seminoma. Conclusions The value of canine seminoma as a model for SE in man remains unclear. Among the 45 investigated tumours from Norwegian dogs, none were classified as classical seminoma based on morphological criteria consistent with human seminomas. Regional or breed differences in the occurrence of classical seminoma in the dog, as well as the lack of uniform diagnostic criteria, might explain the discrepancy between the findings in the current study and the results presented by other authors.

  13. High frequency, high temperature specific core loss and dynamic B-H hysteresis loop characteristics of soft magnetic alloys (United States)

    Wieserman, W. R.; Schwarze, G. E.; Niedra, J. M.


    Limited experimental data exists for the specific core loss and dynamic B-H loops for soft magnetic materials for the combined conditions of high frequency and high temperature. This experimental study investigates the specific core loss and dynamic B-H loop characteristics of Supermalloy and Metglas 2605SC over the frequency range of 1 to 50 kHz and temperature range of 23 to 300 C under sinusoidal voltage excitation. The experimental setup used to conduct the investigation is described. The effects of the maximum magnetic flux density, frequency, and temperature on the specific core loss and on the size and shape of the B-H loops are examined.

  14. 78 FR 36685 - Atlantic Highly Migratory Species; 2013 Atlantic Bluefin Tuna Quota Specifications (United States)


    ... discard estimate used to set the specifications is wrong, especially if it is too high and as a result the... the Federal Register, emailing individuals who have subscribed to the Atlantic HMS News...

  15. Immunohistochemical localization of collagen VI in arthrofibrosis. (United States)

    Zeichen, J; van Griensven, M; Albers, I; Lobenhoffer, P; Bosch, U


    Arthrofibrosis is a disabling complication after knee trauma and surgery. Clinically, it is characterized by pain and joint stiffness due to massive connective tissue proliferation. In similar pathological conditions with fibrotic transformation such as lung fibrosis or superficial fibromatoses, an increased expression of collagen type VI has been reported. Collagen VI, which forms a filamentous network, is thought to serve as an anchoring element between collagen I/III fibrils and basement membranes and as a cell binding structure. Collagen VI may also play a contributing role in the pathogenesis of arthrofibrosis. The aim of the present study was therefore to demonstrate the localization and distribution of type VI collagen in arthrofibrotic tissue. Tissue samples from the infrapatellar fat pad and intercondylar synovia of 13 patients suffering from arthrofibrosis were taken at surgery. The expression of type VI collagen was studied immunohistochemically using an immunoperoxidase method for light microscopic visualization. Histologic analysis showed a synovial hyperplasia with inflammatory cell infiltration and vascular proliferation. Compared with normal synovial tissue, type VI collagen was widely distributed as a network subsynovially and around the capillary walls. The results of the present study suggest that dysregulation of collagen VI synthesis could be an important contributing factor in the complex mechanisms of disordered matrix protein deposition leading to arthrofibrosis.

  16. Cytohistopathological and immunohistochemical correlation of soft tissue tumors

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    J. Chandralekha


    Full Text Available Background: Fine needle aspiration cytology has become an established tool in the diagnostic armamentarium of many clinical practices. The initial diagnosis of many mass lesions, both superficial and deep-seated, can often be readily and safely assessed by fine needle aspiration cytology. In our study, we assessed 361 cases of soft tissue tumors by fine needle aspiration cytology during a period of three years. We tried to follow up as many cases as possible to obtain corresponding excision biopsies for histopathological examination. Immunohistochemical studies were also performed on biopsy sections in some cases for confirmation of diagnoses. Aims and objectives: 1 To study the age, sex and site-wise distribution of soft tissue tumors. 2 To assess the utility of fine needle aspiration cytology in diagnosing various types of soft tissue tumors. 3 To assess the sensitivity, specificity, positive and negative predictive values, and overall histological correlation percentage of fine needle aspiration cytology in diagnosing soft tissue tumors. Methods: Aspirations were carried out using a 22 gauge disposable needle and a 10c.c disposable syringe for suction. Wet-fixed smears were stained with hematoxylin and eosin and pap stain. Dry-fixed smears were stained with May-Grunwald Giemsa stain. Periodic Acid Schiff stain was used in some cases of extraskeletal Ewing's sarcoma. Corresponding biopsy sections were stained with hematoxylin and eosin. Immunohistochemical stains were also used in some of the cases for confirmation of diagnosis. Results: Of the 361 cases recorded in our study, 320 patients could be successfully followed up and excision biopsies were obtained. The remaining 41 patients were excluded from the study due to inability to obtain biopsy. Of the 320 cases, 200 were diagnosed as benign soft tissue tumors, while 120 were diagnosed as malignant on cytological examination. The median age of occurrence of benign soft tissue tumors was 34

  17. Preparation of Zirconia-Ceria Powders with High Specific Surface Area

    Institute of Scientific and Technical Information of China (English)

    Wang Enguo; Mei Fang


    Zirconia-ceria mixed oxide powders were prepared by high temperature aging method.The effects of the temperature and the time of aging, cerium content and calcination on powder performance were studied.The result shows that high temperature aging is an efficient way of preparation of ZrO2-CeO2 mixed oxide powders with high specific surface area and good thermal stability, and that addition of a small amount of cerium to hydrous zirconia can promote the preparation of high specific surface area powders.

  18. Canine classical seminoma: a specific malignant type with human classifications is highly correlated with tumor angiogenesis

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    Kim Jong-Hyuk


    Full Text Available Abstract Background Human seminoma is classified as classical seminoma (SE and spermatocytic seminoma (SS. Human SE is known to be more malignant and metastasizing more frequently than SS. Tumor angiogenesis is highly related with tumor progression and metastasis, with microvessel density (MVD being an important parameter of metastatic potential. Canine seminoma is not yet well-established as SE or SS type including correlation with angiogenesis. We classified canine SE and SS, and then compared them to tumor associated vessels. Methods Twenty-three cases of canine seminomas (2 intratubular, 9 diffuse, and 12 intratubular/diffuse seminomas showing both intratubular and diffuse patterns were classified as SE or SS by immunohistochemistry (IHC using monoclonal antibody against PLAP and by PAS stain. The histopathological data were then compared to see if there was a correlation with SE or SS. Angiogenesis of seminomas were evaluated by immunohistochemical assay using polyclonal antibody against Von Willebrand factor (vWF and by calculating the means of MVD, vessels area and perimeters using computerized image analysis. Statistical Package for Social Sciences (SPSS program was used for various statistical analyses. Results The numbers of PLAP+/PAS+ canine SEs were 8/23 (34.8% and PLAP-/PAS- SSs were 15/23 (61.2%. All SE cases (8/8, 100% were intratubular/diffuse types. SS types included 2 intratubular (2/15, 13.3%, 9 diffuse (9/15, 60%, and 4 intratubular/diffuse (4/15, 26.7% types. MVD and vascular parameters in SEs were significantly higher than in SSs, showing the highest value in the intratubular/diffuse type. Seminomas observed with neoplastic cells invasion of vessels presented higher perimeter and area values than seminomas without conformed neoplastic cells invasion. Conclusion In this study, we demonstrated a positive relationship between canine SE and tumor angiogenesis. Furthermore, we also showed that a tumor cells invasion of vessels

  19. Canine classical seminoma: a specific malignant type with human classifications is highly correlated with tumor angiogenesis (United States)


    Background Human seminoma is classified as classical seminoma (SE) and spermatocytic seminoma (SS). Human SE is known to be more malignant and metastasizing more frequently than SS. Tumor angiogenesis is highly related with tumor progression and metastasis, with microvessel density (MVD) being an important parameter of metastatic potential. Canine seminoma is not yet well-established as SE or SS type including correlation with angiogenesis. We classified canine SE and SS, and then compared them to tumor associated vessels. Methods Twenty-three cases of canine seminomas (2 intratubular, 9 diffuse, and 12 intratubular/diffuse seminomas showing both intratubular and diffuse patterns) were classified as SE or SS by immunohistochemistry (IHC) using monoclonal antibody against PLAP and by PAS stain. The histopathological data were then compared to see if there was a correlation with SE or SS. Angiogenesis of seminomas were evaluated by immunohistochemical assay using polyclonal antibody against Von Willebrand factor (vWF) and by calculating the means of MVD, vessels area and perimeters using computerized image analysis. Statistical Package for Social Sciences (SPSS) program was used for various statistical analyses. Results The numbers of PLAP+/PAS+ canine SEs were 8/23 (34.8%) and PLAP-/PAS- SSs were 15/23 (61.2%). All SE cases (8/8, 100%) were intratubular/diffuse types. SS types included 2 intratubular (2/15, 13.3%), 9 diffuse (9/15, 60%), and 4 intratubular/diffuse (4/15, 26.7%) types. MVD and vascular parameters in SEs were significantly higher than in SSs, showing the highest value in the intratubular/diffuse type. Seminomas observed with neoplastic cells invasion of vessels presented higher perimeter and area values than seminomas without conformed neoplastic cells invasion. Conclusion In this study, we demonstrated a positive relationship between canine SE and tumor angiogenesis. Furthermore, we also showed that a tumor cells invasion of vessels were a correlated

  20. Immunohistochemical expression of IDH1 in gliomas: A tissue microarray-based approach

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    Varuna Sipayya


    Conclusion : Monoclonal antibody to IDH1 (R132 is a useful and less-labor-intensive method to detect mutations in gliomas. IDH1 is a useful immunohistochemical marker to differentiate reactive gliosis from low-grade astrocytoma, has potential as an independent prognostic marker and also helps in distinguishing primary from secondary GBM. Its sensitivity and specificity need to be assessed by simultaneous sequencing and its validation on clinically annotated samples.

  1. The diagnostic utility of p16 FISH and GLUT-1 immunohistochemical analysis in mesothelial proliferations. (United States)

    Monaco, Sara E; Shuai, Yongli; Bansal, Mona; Krasinskas, Alyssa M; Dacic, Sanja


    Two promising ancillary tests used in the diagnosis of mesothelioma include GLUT-1 immunohistochemical analysis and fluorescence in situ hybridization (FISH) testing for the p16 deletion. This study compared the diagnostic usefulness of p16 FISH and GLUT-1 immunohistochemical analysis in the diagnosis of mesothelial proliferations in 158 cases with a diagnosis of benign (45.4%), atypical (10.4%), or malignant/mesothelioma (44.2%). Of the 70 benign cases, none had a deletion of p16 and 5 cases (7%) were positive for GLUT-1. Of the 68 mesotheliomas, 40 (59%) had a deletion of p16 (sensitivity, 59%; specificity, 100%) and 27 (40%) were positive for GLUT-1 (sensitivity, 40%; specificity, 93%). GLUT-1 showed lower sensitivity in pleural (56% vs 70%) and peritoneal (29% vs 51%) mesotheliomas (P = .004). Our results demonstrate that p16 FISH is a more sensitive and specific test than GLUT-1 immunohistochemical analysis and can be a more reliable ancillary tool to support the diagnosis of mesothelioma.

  2. Correlation between PET/CT results and histological and immunohistochemical findings in breast carcinomas

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    Almir Galvão Vieira Bitencourt


    Full Text Available Objective To correlate the results of 18F-fluoro-2-deoxy-D-glucose (18F-FDG positron emission tomography/computed tomography (PET/CT performed with a specific protocol for assessment of breasts with histological/immunohistochemical findings in breast carcinoma patients. Materials and Methods Cross-sectional study with prospective data collection, where patients with biopsy-confirmed breast carcinomas were studied. The patients underwent PET/CT examination in prone position, with a specific protocol for assessment of breasts. PET/CT findings were compared with histological and immunohistochemical data. Results The authors identified 59 malignant breast lesions in 50 patients. The maximum diameter of the lesions ranged from 6 to 80 mm (mean: 32.2 mm. Invasive ductal carcinoma was the most common histological type (n = 47; 79.7%. At PET/CT, 53 (89.8% of the lesions demonstrated anomalous concentrations of 18F-FDG, with maximum SUV ranging from 0.8 to 23.1 (mean: 5.5. A statistically significant association was observed between higher values of maximum SUV and histological type, histological grade, molecular subtype, tumor diameter, mitotic index and Ki-67 expression. Conclusion PET/CT performed with specific protocol for assessment of breasts has demonstrated good sensitivity and was associated with relevant histological/immunohistochemical factors related to aggressiveness and prognosis of breast carcinomas.

  3. Immunohistochemical studies in equine recurrent uveitis (ERU). (United States)

    Romeike, A; Brügmann, M; Drommer, W


    Despite extensive clinical research, the etiology of equine recurrent uveitis (ERU) is still unknown. After an immunologic pathogenesis was established in recurrent uveitis in humans, a similar pathogenic mechanism was assumed to exist in ERU. To investigate whether immunopathologic mechanisms are involved in ERU, 20 eyes of 15 horses with ERU were examined immunohistochemically with a T cell marker, B cell marker, and anti-major histocompatibility complex (MHC) class II antibodies. Twenty-six eyes of 20 horses were used for investigation of MHC class II antigen expression in normal equine eyes. In 18 eyes of 14 horses, the number of T cells in the inflammatory cell population within the uvea was assessed. In 16/18 eyes (89%), the T lymphocyte fraction was > 70%. This cell population was distributed mostly in a diffuse manner throughout the uvea and also within the mantle zone of follicular lymphocytic aggregates. Foci of B lymphocytes could be found within the center of follicular aggregates in three eyes. The expression of MHC class II antigen on resident ocular cells was evaluated in 10 eyes of six horses with ERU. An increase of MHC class II antigen expression in the trabecular meshwork and on the nonpigmented ciliary epithelium was noted as was a deviant expression on proliferating Müller cells and retinal pigment epithelial cells. The predominance of T cells in the inflammatory infiltrates supports the central role of a cell-mediated immune response. Furthermore, the observation of a deviant MHC class II expression on resident ocular cells suggests that aberrant immune regulation may play a role in the pathogenesis of ERU.


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    Yu. G. KOLENKO


    Full Text Available In recent years, substantial changes have occurred in the structure of oral mucosa diseases, in particular an increased ratio of precancerous diseases, so that an effective non-invasive detection of any sign of malignancy appears as an urgent and most actual task of dentistry. Aim: To study the proliferative activity of epithelial cells in Ki-67 antigenin patients with leukoplakia of the oral mucosa. Materials and method: A complex clinical and laboratory examination was performed on 155 patients with oral leukoplakia, who addressed the Operative Dentistry Department of the “A.A.Bogomolets” National Medical University of Kiev between 2010 and 2014. All patients have been subjected to a careful clinical examination, which included: dental anamnesis, visual inspection, oral examination and digital palpation of oral mucosa and tongue mucosa, biopsy of leukoplakia lesions for cytological and histological examination. Results: Histological evaluation of the material has been performed according to the WHO (2005 classification of leukoplakia. 10 (14% sites of unaltered mucosa, 10 (14% samples of hyperkeratosis without atypia, 14 (19% biopsy specimens of hyperkeratosis SIN1, 15 (21% – hyperkeratosis SIN2, 10 (14% - SIN3 and 13 (18% cases of squamous cell carcinoma were evidenced. Immunohistochemical investigation evidenced the presence of protein Ki-67 in the nuclei of epithelial cells. In the unmodified epithelium of the oral mucosa, all epithelial cells with stained nuclei are virtually located in the basal layer. Conclusion: Against the general increase of the proliferative activity of epithelial cells with increasing SIN, a characteristic distribution of proliferating cells in the thickness of the epithelium was revealed for each studied group, as follows: in the control group and in leukoplakia without atypia, immunopositive cells are located in the basal layer, in leukoplakia (SIN1, SIN2 and SIN3 – in parabasal position while, in squamous

  5. Electrolytes with Improved Safety Characteristics for High Voltage, High Specific Energy Li-ion Cells (United States)

    Smart, M. C.; Krause, F. C.; Hwang, C.; West, W. C.; Soler, J.; Whitcanack, L. W.; Prakash, G. K. S.; Ratnakumar, B. V.


    (1) NASA is actively pursuing the development of advanced electrochemical energy storage and conversion devices for future lunar and Mars missions; (2) The Exploration Technology Development Program, Energy Storage Project is sponsoring the development of advanced Li-ion batteries and PEM fuel cell and regenerative fuel cell systems for the Altair Lunar Lander, Extravehicular Activities (EVA), and rovers and as the primary energy storage system for Lunar Surface Systems; (3) At JPL, in collaboration with NASA-GRC, NASA-JSC and industry, we are actively developing advanced Li-ion batteries with improved specific energy, energy density and safety. One effort is focused upon developing Li-ion battery electrolyte with enhanced safety characteristics (i.e., low flammability); and (4) A number of commercial applications also require Li-ion batteries with enhanced safety, especially for automotive applications.

  6. Immunohistochemical Study of Glypican-3 and HepPar-1 in Differentiating Hepatocellular Carcinoma from Metastatic Carcinomas in FNA of the Liver. (United States)

    Ibrahim, Taiseer R; Abdel-Raouf, Samar M


    Hepatocellular carcinoma (HCC) remains a common malignant cancer worldwide, it is considered the fifth most common malignant cancer. On the other hand, metastatic tumors are widespread in the liver , with metastatic adenocarcinoma (MA) constituting the greatest part, therefore differentiation of HCC from MA is a frequent problem facing the pathologist especially in liver fine-needle aspiration biopsies. Evaluating the diagnostic value of glypican-3 (GPC-3) and HepPar-1 immunostaining in differentiating hepatocellular carcinoma from metastatic tumors in liver cell block material. Fourty eight cell blocks prepared from FNA from the liver ( 30 cases HCC, 18 cases metastatic carcinoma in liver) stained by Glypican -3 and HepPar-1 immunohistochemical markers. Glypican-3 was immunoexpressed in 97% of cases of HCC while all cases of metastatic carcinoma were negative. HepPar-1 was expressed in 93% of cases of HCC and 11% of metastatic carcinoma of the liver. In this study the sensitivity of GPC3 in the diagnosis of HCC in cytological material was 96.7% and the specificity was 100% while the sensitivity and specificity of HepPar-1 was 93.3% and 88.9% respectively. Immunohistochemical staining for GPC-3 in cell block material of the liver is highly sensitive and specific and it is a valuable tool capable of differentiating HCC from most of metastatic tumors of the liver.


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    Kanwardeep Singh


    Full Text Available BACKGROUND Primary central nervous system lymphoma (PCNSL is a rare form of extranodal non-Hodgkin lymphoma (NHL confined to the brain, spinal cord and/or eye, occurring in immunocompetent individuals. Histologically, they are diffuse large B-cell lymphomas. Over the last few decades there has been a gradual increase in their incidence. AIM To study the clinical, histopathological and immunohistochemical profile of primary central nervous system lymphoma. SETTING AND DESIGN Retrospective audit of seven cases of PCNSL diagnosed over a period of five years in a tertiary referral hospital of North India. MATERIAL AND METHODS The clinical, radiological and laboratory findings were retrieved from the hospital records. Histopathology slides were reviewed, studied in detail and a panel of immunohistochemical markers comprising of CD3, CD5, CD20, CD10, BCL6, BCL2, MUM1, CD30, EBV (LMP1, Ki-67 and p53 was done on all cases. RESULTS The male to female ratio was 3:4 with a median age of 60 years. The most common form of presentation was neurological deficits and altered sensorium. Imaging showed contrast enhancing, single or multiple, deep seated lesions within the cerebral hemispheres. Histologically, all were high-grade diffuse large B-cell lymphomas showing typical angiocentricity and a median Ki-67 proliferative index of 80%. Based on immunohistochemistry (Hans classifier three cases had germinal centre B-cell (GCB and four had non-germinal centre B-cell (non-GCB phenotype. p53 was expressed in all cases with strong expression in four of them. Four patients died before treatment could be initiated, one received palliative chemo-radiotherapy and two did not follow up after diagnosis. CONCLUSIONS Primary CNS lymphomas are high-grade diffuse large B-cell lymphomas which show high Ki-67 proliferative indices and frequent overexpression of p53. Irrespective of histological subtype, GCB or non-GCB, outcome is uniformly poor. Early and prompt diagnosis is

  8. Immunohistochemical LRIG3 expression in cervical intraepithelial neoplasia and invasive squamous cell cervical cancer: association with expression of tumor markers, hormones, high-risk HPV-infection, smoking and patient outcome

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    A.K. Lindström


    Full Text Available The novel biomarker LRIG3 is a member of the LRIG family (LRIG1-3. While LRIG1 has been associated with favorable prognosis and LRIG2 with poor prognosis in invasive cervical cancer, little is known about the role of LRIG3. The aim of this study was to investigate the expression of LRIG3 in invasive cancer and cervical intraepithelial neoplasia (CIN for possible correlation with other tumor markers, to hormones and smoking, as a diagnostic adjunct in CIN, and prognostic value in invasive cancer. Cervical biopsies from 129 patients with invasive squamous cell carcinoma and 170 biopsies showing low grade and high grade CIN, or normal epithelium were stained for LRIG3 and 17 additional tumor markers. Among other variables the following were included: smoking habits, hormonal contraceptive use, serum progesterone, serum estradiol, high-risk HPV-infection, menopausal status and ten-year survival. In CIN, high expression of the tumor suppressors retinoblastoma protein, p53, and p16, and E-cadherin (cell-cell interaction, or low expression of CK10, correlated to LRIG3 expression. In addition, progestogenic contraceptive use correlated to high expression of LRIG3. In invasive cancer there was a correlation between expression of the major tumor promoter c-myc and high LRIG3 expression. High LRIG3 expression correlated significantly to presence of high-risk HPV infection in patients with normal epithelium and CIN. There was no correlation between LRIG3 expression and 10-year survival in patients with invasive cell cervical cancer. LRIG3 expression is associated with a number of molecular events in CIN. Expression also correlates to hormonal contraceptive use. The results on expression of other tumor markers suggest that LRIG3 is influenced by or influences a pattern of tumor markers in cancer and precancerous cells. Further studies are needed to elucidate if LRIG3 expression might be clinically useful.

  9. Osteoclastic giant cell tumor of the pancreas: an immunohistochemical study

    DEFF Research Database (Denmark)

    Dizon, M A; Multhaupt, H A; Paskin, D L


    A case of an osteoclastic giant cell tumor of the pancreas is presented. Immunohistochemical studies were performed, which showed keratin (CAM, AE1) and epithelial membrane antigen positivity in the tumor cells. The findings support an epithelial origin for this tumor.......A case of an osteoclastic giant cell tumor of the pancreas is presented. Immunohistochemical studies were performed, which showed keratin (CAM, AE1) and epithelial membrane antigen positivity in the tumor cells. The findings support an epithelial origin for this tumor....

  10. Hybrid-PIC Modeling of a High-Voltage, High-Specific-Impulse Hall Thruster (United States)

    Smith, Brandon D.; Boyd, Iain D.; Kamhawi, Hani; Huang, Wensheng


    The primary life-limiting mechanism of Hall thrusters is the sputter erosion of the discharge channel walls by high-energy propellant ions. Because of the difficulty involved in characterizing this erosion experimentally, many past efforts have focused on numerical modeling to predict erosion rates and thruster lifespan, but those analyses were limited to Hall thrusters operating in the 200-400V discharge voltage range. Thrusters operating at higher discharge voltages (V(sub d) >= 500 V) present an erosion environment that may differ greatly from that of the lower-voltage thrusters modeled in the past. In this work, HPHall, a well-established hybrid-PIC code, is used to simulate NASA's High-Voltage Hall Accelerator (HiVHAc) at discharge voltages of 300, 400, and 500V as a first step towards modeling the discharge channel erosion. It is found that the model accurately predicts the thruster performance at all operating conditions to within 6%. The model predicts a normalized plasma potential profile that is consistent between all three operating points, with the acceleration zone appearing in the same approximate location. The expected trend of increasing electron temperature with increasing discharge voltage is observed. An analysis of the discharge current oscillations shows that the model predicts oscillations that are much greater in amplitude than those measured experimentally at all operating points, suggesting that the differences in oscillation amplitude are not strongly associated with discharge voltage.

  11. Immunohistochemical characterization of basal cell adenomas of the salivary gland. (United States)

    Takahashi, H; Fujita, S; Okabe, H; Tsuda, N; Tezuka, F


    Seven cases of basal cell adenomas of the salivary gland were analyzed by immunohistochemical methods with a broad panel of routinely used antibodies. Histologically the epithelial elements were classified as tubuloglandular, trabecular and solid patterns. The authors' results indicated the following: 1) The duct lining cells of tubuloglandular and trabecular patterns have distinct epithelial features with cytokeratins (KL 1, PKK 1, *PKK 2 and PKK 3), alpha-one-antichymotrypsin (alpha 1-ACT), carcinoembryonic antigen (CEA) and S-100 alpha subunit positivity. 2) The basaloid cells in the trabecular and solid patterns expressed two immunophenotypes: one had actin, neuron-specific enolase (NSE), S-100 protein and S-100 beta subunit patterns typical of myoepithelial cells in normal glands. The other basaloid cells had vimentin and S-100 protein patterns. The former cell type could be found in 4 of 7 cases and the latter was found in 7 cases. This represents a minor participation of the myoepithelial cells in the basal cell adenoma. 3) The basement membrane and stromal connective tissue around the neoplastic cells were positive for alpha-one-antitrypsin (alpha 1-AT). This antibody is a good marker in identifying the basement membrane-like material.

  12. Novel immunohistochemical monoclonal antibody against rat B cell receptor Associated Protein 31 (BAP31). (United States)

    Song, Chaojun; Yan, Binyuan; Chen, Lihua; Li, Yongming; Wei, Yuying; Sun, Yuanjie; Yang, Angang; Yang, Kun; Jin, Boquan


    BAP31 is an evolutionarily conserved polytopic integral protein of the endoplasmic reticulum (ER) membrane implicated in regulating the export of selected membrane proteins from the ER to downstream compartments of the secretory pathway. BAP31 interacts with mIgD, cellubrevin, major histocompatibility complex class I, and BCL-2/BCL-X(L) and plays an important role in regulating the egress of these proteins and in apoptosis. Although BAP31 RNA is ubiquitous, the protein's anatomic localization in rat tissues has not been determined. This is partially because production of high affinity antibodies, especially monoclonal antibodies (MAbs) suitable for immunohistochemical staining, has lagged. To gain further insight into its possible functions, we generated a novel MAb specific for rat BAP31 in immunocytochemistry and immunohistochemistry and localized BAP31 in some rat tissues. Immunoreactivity of BAP31 was prominent in fundic glands, colon, pancreatic acinuses, and liver but not in skeleton muscle and lung. Thus, successful production of rat BAP31 monoclonal antibodies provides a new powerful tool for investigation of BAP31 function in the rat model.

  13. Denatured G-protein coupled receptors as immunogens to generate highly specific antibodies. (United States)

    Talmont, Franck; Moulédous, Lionel; Boué, Jérôme; Mollereau, Catherine; Dietrich, Gilles


    G-protein coupled receptors (GPCRs) play a major role in a number of physiological and pathological processes. Thus, GPCRs have become the most frequent targets for development of new therapeutic drugs. In this context, the availability of highly specific antibodies may be decisive to obtain reliable findings on localization, function and medical relevance of GPCRs. However, the rapid and easy generation of highly selective anti-GPCR antibodies is still a challenge. Herein, we report that highly specific antibodies suitable for detection of GPCRs in native and unfolded forms can be elicited by immunizing animals against purified full length denatured recombinant GPCRs. Contrasting with the currently admitted postulate, our study shows that an active and well-folded GPCR is not required for the production of specific anti-GPCR antibodies. This new immunizing strategy validated with three different human GPCR (μ-opioid, κ-opioid, neuropeptide FF2 receptors) might be generalized to other members of the GPCR family.


    Directory of Open Access Journals (Sweden)

    Arockiasamy Babiya Infant


    Full Text Available BACKGROUND Primary adenocarcinoma of cervix constitutes 10-15% of all cases of carcinoma of cervix, which is the second most common carcinoma next to squamous cell carcinoma. Endocervical adenocarcinoma have a considerable morphological overlap with endometrial adenocarcinoma though they differ in their aetiologies, behaviour, and treatments. This makes their diagnosis very difficult particularly in biopsy or curetting specimens or when a fractional dilation and curettage specimens show adenocarcinoma in both components of it. This study was done in the aim to suggest the possible origin of the tumour with the help of immunohistochemistry. AIMS AND OBJECTIVES To identify the incidence, distribution, clinicopathological, histomorphological features of endocervical adenocarcinomas and to determine the immunohistochemical expression of CEA, Vimentin, ER and PR in endometrioid type of adenocarcinoma detected in endocervical biopsies, fractional dilation and curettage specimens (Both the components showing similar morphology, and in hysterectomy specimens to suggest the site of origin of tumour. MATERIALS AND METHODS It is a retrospective descriptive study of cervical adenocarcinomas conducted in the Institute of Obstetrics and Gynaecology, Madras Medical College, Chennai for a period of 4 years during the period between 2009 November to 2013 October. The statistical analysis was performed using statistical package for social science software version 11.5 the clinicopathological profile of the tumour were calculated using Student t-test and chi-square test. OBSERVATION AND RESULTS Among the total 13499 cases received during the study period, 2489 were cervical malignancies comprising 148 adenocarcinoma. It includes 101 mucinous (Endocervical type, 44 endometrioid type, 2 serous type, and 1 clear cell type. Among the 30 cases of endometrioid type, 16 cases showed definite immunophenotype of cervical origin, 9 cases of endometrial origin and in the

  15. Localization of West Nile Virus in monkey brain: double staining antigens immunohistochemically of neurons, neuroglia cells and West Nile Virus. (United States)

    He, Xianli; Ren, Junping; Xu, Fangling; Ferguson, Monique R; Li, Guangyu


    West Nile virus (WNV) can cause encephalitis or meningitis that affects brain tissue, which can also lead to permanent neurological damage that can be fatal. To our knowledge, no consistent double immunohistochemical staining of neurons, neuroglia cells, and WNV has yet been reported. To establish a method for performing double-label immunohistochemical detection of neurons, neuroglia cells and WNV, examining the pathological characteristics of WNV-infected neurons, neuroglia cells, and investigating distribution of WNV in monkey brain, paraffin-embedded monkey brain tissue were retrospectively studied by immunohistochemical staining of neurons, neuroglia cells and WNV. Antibodies against neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and WNV were used to develop the method of double-label immunohistochemical staining, which allowed independent assessment of neuron status and WNV distribution. A range of immunohistochemical WNV infection in monkey brain was observed in both neurons and neuroglia cells in terms of the thickness of lesion staining, and the WNV staining was slightly higher in neuroglia cells than in neurons. All these findings suggest that WNV invasion in the brain plays a crucial role in neurological damage by inducing central nervous system (CNS) cell dysfunction or cell death directly.

  16. Hollow Carbon Nanofiber-Encapsulated Sulfur Cathodes for High Specific Capacity Rechargeable Lithium Batteries

    KAUST Repository

    Zheng, Guangyuan


    Sulfur has a high specific capacity of 1673 mAh/g as lithium battery cathodes, but its rapid capacity fading due to polysulfides dissolution presents a significant challenge for practical applications. Here we report a hollow carbon nanofiber-encapsulated sulfur cathode for effective trapping of polysulfides and demonstrate experimentally high specific capacity and excellent electrochemical cycling of the cells. The hollow carbon nanofiber arrays were fabricated using anodic aluminum oxide (AAO) templates, through thermal carbonization of polystyrene. The AAO template also facilitates sulfur infusion into the hollow fibers and prevents sulfur from coating onto the exterior carbon wall. The high aspect ratio of the carbon nanofibers provides an ideal structure for trapping polysulfides, and the thin carbon wall allows rapid transport of lithium ions. The small dimension of these nanofibers provides a large surface area per unit mass for Li2S deposition during cycling and reduces pulverization of electrode materials due to volumetric expansion. A high specific capacity of about 730 mAh/g was observed at C/5 rate after 150 cycles of charge/discharge. The introduction of LiNO3 additive to the electrolyte was shown to improve the Coulombic efficiency to over 99% at C/5. The results show that the hollow carbon nanofiber-encapsulated sulfur structure could be a promising cathode design for rechargeable Li/S batteries with high specific energy. © 2011 American Chemical Society.

  17. R1: Immunohistochemical study of mucins in human intestinal spirochetosis. (United States)

    Ogata, Sho; Shimizu, Ken; Tominaga, Susumu; Nakanishi, Kuniaki


    Most patients with human intestinal spirochetosis (HIS; a colorectal bacterial infection caused by Brachyspira species) seem asymptomatic, and its pathogenicity remains unclear. Recently, alterations in mucin expression were reported in animal Brachyspira infection. The present question was "Is mucin expression altered in HIS?". Using antibodies for MUCs 1, 2, 4, 5 AC, and 6, we immunohistochemically compared 215 specimens from 83 histology-confirmed HIS cases with 106 specimens from 26 non-HIS cases. Positive staining (which included even focal positive staining) was rated "high (+)" or "low (+)". Results were analysed for four categories of lesions, and associations between MUC expression and spirochetal presence were also analysed. In the "specimens without polyps or adenocarcinoma" category: high (+) MUC2-positivity was more frequent in HIS than in control. In the hyperplasia/serrated polyp category: in HIS (vs. control), the MUC5AC-positivity rate was lower, while high (+) MUC4-positivity was more frequent. In the conventional adenoma category: in HIS (vs. control), the MUC1-positivity rate was lower, while both high (+) MUC2-positivity and high (+) MUC5AC-positivity were less frequent. In the adenocarcinoma category: high (+) MUC2-positivity was more frequent in HIS than in control. Among the above mucins, only MUC1-positivity was significantly associated with an absence of the so-called fringe formation, an absence of spiral organisms within mucus, and an absence of strong immunopositive materials within the epithelial layer and within the subepithelial layer. The results suggest that Brachyspira infection or a related change in the microbiome may alter the large intestine mucin-expression profile in humans.

  18. Novel method for the high-throughput production of phosphorylation site-specific monoclonal antibodies (United States)

    Kurosawa, Nobuyuki; Wakata, Yuka; Inobe, Tomonao; Kitamura, Haruki; Yoshioka, Megumi; Matsuzawa, Shun; Kishi, Yoshihiro; Isobe, Masaharu


    Threonine phosphorylation accounts for 10% of all phosphorylation sites compared with 0.05% for tyrosine and 90% for serine. Although monoclonal antibody generation for phospho-serine and -tyrosine proteins is progressing, there has been limited success regarding the production of monoclonal antibodies against phospho-threonine proteins. We developed a novel strategy for generating phosphorylation site-specific monoclonal antibodies by cloning immunoglobulin genes from single plasma cells that were fixed, intracellularly stained with fluorescently labeled peptides and sorted without causing RNA degradation. Our high-throughput fluorescence activated cell sorting-based strategy, which targets abundant intracellular immunoglobulin as a tag for fluorescently labeled antigens, greatly increases the sensitivity and specificity of antigen-specific plasma cell isolation, enabling the high-efficiency production of monoclonal antibodies with desired antigen specificity. This approach yielded yet-undescribed guinea pig monoclonal antibodies against threonine 18-phosphorylated p53 and threonine 68-phosphorylated CHK2 with high affinity and specificity. Our method has the potential to allow the generation of monoclonal antibodies against a variety of phosphorylated proteins. PMID:27125496

  19. Specification Improvement Through Analysis of Proof Structure (SITAPS): High Assurance Software Development (United States)


    SPECIFICATION IMPROVEMENT THROUGH ANALYSIS OF PROOF STRUCTURE (SITAPS): HIGH ASSURANCE SOFTWARE DEVELOPMENT BAE SYSTEMS FEBRUARY...ANALYSIS OF PROOF STRUCTURE (SITAPS): HIGH ASSURANCE SOFTWARE DEVELOPMENT 5a. CONTRACT NUMBER FA8750-13-C-0240 5b. GRANT NUMBER N/A 5c. PROGRAM...General adoption of these techniques has had limited penetration in the software development community. Two interrelated causes may account for

  20. Quantifying domain-ligand affinities and specificities by high-throughput holdup assay. (United States)

    Vincentelli, Renaud; Luck, Katja; Poirson, Juline; Polanowska, Jolanta; Abdat, Julie; Blémont, Marilyne; Turchetto, Jeremy; Iv, François; Ricquier, Kevin; Straub, Marie-Laure; Forster, Anne; Cassonnet, Patricia; Borg, Jean-Paul; Jacob, Yves; Masson, Murielle; Nominé, Yves; Reboul, Jérôme; Wolff, Nicolas; Charbonnier, Sebastian; Travé, Gilles


    Many protein interactions are mediated by small linear motifs interacting specifically with defined families of globular domains. Quantifying the specificity of a motif requires measuring and comparing its binding affinities to all its putative target domains. To this end, we developed the high-throughput holdup assay, a chromatographic approach that can measure up to 1,000 domain-motif equilibrium binding affinities per day. After benchmarking the approach on 210 PDZ-peptide pairs with known affinities, we determined the affinities of two viral PDZ-binding motifs derived from human papillomavirus E6 oncoproteins for 209 PDZ domains covering 79% of the human 'PDZome'. We obtained sharply sequence-dependent binding profiles that quantitatively describe the PDZome recognition specificity of each motif. This approach, applicable to many categories of domain-ligand interactions, has wide potential for quantifying the specificities of interactomes.

  1. Highly specific and sensitive electrochemical genotyping via gap ligation reaction and surface hybridization detection. (United States)

    Huang, Yong; Zhang, Yan-Li; Xu, Xiangmin; Jiang, Jian-Hui; Shen, Guo-Li; Yu, Ru-Qin


    This paper developed a novel electrochemical genotyping strategy based on gap ligation reaction with surface hybridization detection. This strategy utilized homogeneous enzymatic reactions to generate molecular beacon-structured allele-specific products that could be cooperatively annealed to capture probes stably immobilized on the surface via disulfide anchors, thus allowing ultrasensitive surface hybridization detection of the allele-specific products through redox tags in close proximity to the electrode. Such a unique biphasic architecture provided a universal methodology for incorporating enzymatic discrimination reactions in electrochemical genotyping with desirable reproducibility, high efficiency and no interferences from interficial steric hindrance. The developed technique was demonstrated to show intrinsic high sensitivity for direct genomic analysis, and excellent specificity with discriminativity of single nucleotide variations.

  2. Feasibility of High-Repetition, Task-Specific Training for Individuals With Upper-Extremity Paresis (United States)

    Waddell, Kimberly J.; Birkenmeier, Rebecca L.; Moore, Jennifer L.; Hornby, T. George


    OBJECTIVE. We investigated the feasibility of delivering an individualized, progressive, high-repetition upper-extremity (UE) task-specific training protocol for people with stroke in the inpatient rehabilitation setting. METHOD. Fifteen patients with UE paresis participated in this study. Task-specific UE training was scheduled for 60 min/day, 4 days/wk, during occupational therapy for the duration of a participant’s inpatient stay. During each session, participants were challenged to complete ≥300 repetitions of various tasks. RESULTS. Participants averaged 289 repetitions/session, spending 47 of 60 min in active training. Participants improved on impairment and activity level outcome measures. CONCLUSION. People with stroke in an inpatient setting can achieve hundreds of repetitions of task-specific training in 1-hr sessions. As expected, all participants improved on functional outcome measures. Future studies are needed to determine whether this high-repetition training program results in better outcomes than current UE interventions. PMID:25005508

  3. Effects of Collaborative Preteaching on Science Performance of High School Students with Specific Learning Disabilities (United States)

    Thornton, Amanda; McKissick, Bethany R.; Spooner, Fred; Lo, Ya-yu; Anderson, Adrienne L.


    Investigating the effectiveness of inclusive practices in science instruction and determining how to best support high school students with specific learning disabilities (SLD) in the general education classroom is a topic of increasing research attention in the field. In this study, the researchers conducted a single-subject multiple probe across…

  4. Highly specific protease-based approach for detection of Porphyromonas gingivalis in diagnosis of periodontitis

    NARCIS (Netherlands)

    Kaman, W.E.; Galassi, F.; de Soet, J.J.; Bizzarro, S.; Loos, B.G.; Veerman, E.C.I.; van Belkum, A.; Hays, J.P.; Bikker, F.J.


    Porphyromonas gingivalis is associated with the development of periodontitis. Here we describe the development of a highly specific protease-based diagnostic method for the detection of P. gingivalis in gingival crevicular fluid. Screening of a proteolytic peptide substrate library, including fluoro

  5. Highly specific protease-based approach for detection of porphyromonas gingivalis in diagnosis of periodontitis

    NARCIS (Netherlands)

    J.P. Hays (John); W.E. Kaman (Wendy); F. Galassi (Fabiano); J.J. de Soet (Johannes); S. Bizzarro (Sergio); B.G. Loos (Bruno G.); E.C.I. Veerman (Enno); A.F. van Belkum (Alex); F.J. Bikkerk


    textabstractPorphyromonas gingivalis is associated with the development of periodontitis. Here we describe the development of a highly specific protease-based diagnostic method for the detection of P. gingivalis in gingival crevicular fluid. Screening of a proteolytic peptide substrate library, incl

  6. Synthesis of tritium-labelled imipramine and desipramine with high specific activities

    Energy Technology Data Exchange (ETDEWEB)

    Yakovleva, L.A.; Kaminski, Ju.L.; Sosnova, L.P.; Kudelin, B.K. (Khlopin Radium Inst., St. Petersburg (Russian Federation))


    [G-[sup 3]H]imipramine and [G-[sup 3]H] desipramine with specific activities 110-160 and 80-100 kCi/mol, respectively were synthesized by high temperature solid-phase catalytic isotope exchange with gaseous tritium. (author).

  7. Using the SCR Specification Technique in a High School Programming Course. (United States)

    Rosen, Edward; McKim, James C., Jr.


    Presents the underlying ideas of the Software Cost Reduction (SCR) approach to requirements specifications. Results of applying this approach to the teaching of programing to high school students indicate that students perform better in writing programs. An appendix provides two examples of how the method is applied to problem solving. (MDH)

  8. Sonic hedgehog in oral squamous cell carcinoma: An immunohistochemical study (United States)

    Srinath, Sahana; Iyengar, Asha R; Mysorekar, Vijaya


    Background: Recent studies have revealed the involvement of hedgehog (Hh) signaling component in proliferation and invasive behavior of many carcinomas. Aim: This study aims to identify the expression of sonic Hh (SHH) protein of SHH pathway in oral epithelial dysplasia and oral squamous cell carcinoma (OSCC) using SHH (H-160) (Santa Cruz, sc-9042) which could have therapeutic implication in future. Materials and Methods: A total of 250 cases comprising 50 normal oral mucosa, 50 cases of oral epithelial dysplasia, 50 well, 50 moderate and 50 poorly differentiated OSCCs were included in the study. Immunohistochemical evaluation of SHH protein expression was conducted using monoclonal antibody. Interpretation of the expression was done by immunoreactive score of Remmele and Stegner (IRS) scoring method. Statistical Analysis: Chi-Square test was used to analyze the results. Results: The study showed that SHH signaling molecules are highly expressed in OSCC, and their expression was mainly in the cytoplasm of epithelial cells. Conclusion: The SHH signaling component is associated with the pathological parameter in OSCC and oral epithelial dysplasia. PMID:27721600

  9. Immunohistochemical staining of radixin and moesin in prostatic adenocarcinoma

    Directory of Open Access Journals (Sweden)

    Becich Michael J


    Full Text Available Abstract Background Some members of the Protein 4.1 superfamily are believed to be involved in cell proliferation and growth, or in the regulation of these processes. While the expression levels of two members of this family, radixin and moesin, have been studied in many tumor types, to our knowledge they have not been investigated in prostate cancer. Methods Tissue microarrays were immunohistochemically stained for either radixin or moesin, with the staining intensities subsequently quantified and statistically analyzed using One-Way ANOVA or nonparametric equivalent with subsequent Student-Newman-Keuls tests for multiple comparisons. There were 11 cases of normal donor prostates (NDP, 14 cases of benign prostatic hyperplasia (BPH, 23 cases of high-grade prostatic intraepithelial neoplasia (HGPIN, 88 cases of prostatic adenocarcinoma (PCa, and 25 cases of normal tissue adjacent to adenocarcinoma (NAC analyzed in the microarrays. Results NDP, BPH, and HGPIN had higher absolute staining scores for radixin than PCa and NAC, but with a significant difference observed between only HGPIN and PCa (p = Conclusions To our knowledge, these studies represent the first reports on the expression profiles of radixin and moesin in prostatic adenocarcinoma. The current study has shown that there were statistically significant differences observed between HGPIN and PCa and HGPIN and NAC in terms of radixin expression. The differences in the moesin profiles by tissue type were not statistically significant. Additional larger studies with these markers may further elucidate their potential roles in prostatic neoplasia progression.

  10. Mechanism of benign biliary stricture: A morphological and immunohistochemical study

    Institute of Scientific and Technical Information of China (English)

    Zhi-Min Geng; Ying-Min Yao; Qing-Guang Liu; Xin-Jie Niu; Xiao-Gong Liu


    AIM: To explore the mechanism of benign biliary stricture.METHODS: A model of trauma of bile duct was established in 28 dogs. The anastomosed tissues were resected and examined by light and electron microscopes on day 3, in wk 1, 3 and mo 3, 6 after operation. CD68, TGF-β1 and α-SMA were examined by immunohistochemical staining, respectively.RESULTS: The mucosal epithelium of the bile duct was slowly recovered, chronic inflammation lasted for a long time,fibroblasts proliferated actively, extracellular matrix was over-deposited. Myofibroblasts functioned actively and lasted through the whole process. The expression of macrophages in lamina propria under mucosa, TGF-β1 in granulation tissue,fibroblasts and endothelial cells of blood vessels, α-SMA in myofiroblasts were rather strong from the 1st wk to the 6th mo after operation.CONCLUSION: The type of healing occurring in bile duct belongs to overhealing. Myofibroblasts are the main cause for scar contracture and stricture of bile duct. High expressions of CD68, TGF-β1 and α-SMA are closely related to the active proliferation of fibroblasts, extracellular matrix overdeposition and scar contracture of bile duct.

  11. Temporal Processing Capacity in High-Level Visual Cortex Is Domain Specific. (United States)

    Stigliani, Anthony; Weiner, Kevin S; Grill-Spector, Kalanit


    Prevailing hierarchical models propose that temporal processing capacity--the amount of information that a brain region processes in a unit time--decreases at higher stages in the ventral stream regardless of domain. However, it is unknown if temporal processing capacities are domain general or domain specific in human high-level visual cortex. Using a novel fMRI paradigm, we measured temporal capacities of functional regions in high-level visual cortex. Contrary to hierarchical models, our data reveal domain-specific processing capacities as follows: (1) regions processing information from different domains have differential temporal capacities within each stage of the visual hierarchy and (2) domain-specific regions display the same temporal capacity regardless of their position in the processing hierarchy. In general, character-selective regions have the lowest capacity, face- and place-selective regions have an intermediate capacity, and body-selective regions have the highest capacity. Notably, domain-specific temporal processing capacities are not apparent in V1 and have perceptual implications. Behavioral testing revealed that the encoding capacity of body images is higher than that of characters, faces, and places, and there is a correspondence between peak encoding rates and cortical capacities for characters and bodies. The present evidence supports a model in which the natural statistics of temporal information in the visual world may affect domain-specific temporal processing and encoding capacities. These findings suggest that the functional organization of high-level visual cortex may be constrained by temporal characteristics of stimuli in the natural world, and this temporal capacity is a characteristic of domain-specific networks in high-level visual cortex. Significance statement: Visual stimuli bombard us at different rates every day. For example, words and scenes are typically stationary and vary at slow rates. In contrast, bodies are dynamic

  12. Preparation and characterization of boron nitride/carbon fiber composite with high specific surface area

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Yan; Fan, Mingwen [Wuhan Univ. (China). Key Laboratory for Oral Biomedical Engineering; Yuan, Songdong; Xiong, Kun; Hu, Kunpeng; Luo, Yi [Hubei Univ. of Technology, Wuhan (China). School of Chemistry and Chemical Engineering; Li, Dong [Hubei Univ. of Technology, Wuhan (China). School of Chemistry and Chemical Engineering; Oxford Univ. (United Kingdom). Chemistry Research Lab.


    Boron nitride can be used as a good catalyst carrier because of its high thermal conductivity and chemical stability. However, a high specific surface area of boron nitride is still desirable. In this work, a carbon fiber composite coated with boron nitride villous nano-film was prepared, and was also characterized by means of scanning electron microscopy, high resolution transmission electron microscopy, Fourier transform infrared spectroscopy, and Brunauer-Emmett-Teller analysis. The results indicated that the carbon fibers were covered by uniform villous boron nitride films whose thickness was about 150 - 200 nm. The specific surface area of the boron nitride/carbon fiber composite material was 96 m{sup 2} g{sup -1}, which was markedly improved compared with conventional boron nitride materials. (orig.)

  13. A novel marker of ameloblastoma and systematic review of immunohistochemical findings. (United States)

    Khalele, Bacem A E O; Al-Shiaty, Rami A


    This study aims at investigating the pathogenesis and oncogenesis of ameloblastoma. Being the commonest odontogenic tumor with idiopathic nature, ameloblastoma poses a fierce controversy about its oncogenesis. Immunohistochemical markers, over years, have highlighted specific pathways which are inherently undertaken in the tumorigenic process of ameloblastoma. Besides the recently pronounced clue of BRAF V600E mutant gene, this study introduces a new marker with its outstanding impact on our contemporary knowledge about ameloblastoma. Extrapolating from the systematic review of medical literature and recruiting a novel immunohistochemical marker, ameloblastoma enacts a new scenario supporting the approved involvement of MAPK by overexpressing WT1 a total of 37 archival cases, regardless of the histological variant in study. There evinces a significant contribution of Wilm's tumor gene, as an oncogene rather than a suppressor gene, to the pathogenesis of the ameloblastomatous tumorigenesis. Moreover, no ameloblastomatous histological phenotype has established, given the literature underpinned, a concrete impact on the clinical behavior. Immunohistochemical research papers which investigated tumorigenesis - although they do not quantitatively measure much- had the most significant impact on the diagnostic and prognostic levels. WT1 may play, therefore, a remarkable role in the oncogenesis of ameloblastoma. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Effective Inhibition of Bone Morphogenetic Protein Function by Highly Specific Llama-Derived Antibodies. (United States)

    Calpe, Silvia; Wagner, Koen; El Khattabi, Mohamed; Rutten, Lucy; Zimberlin, Cheryl; Dolk, Edward; Verrips, C Theo; Medema, Jan Paul; Spits, Hergen; Krishnadath, Kausilia K


    Bone morphogenetic proteins (BMP) have important but distinct roles in tissue homeostasis and disease, including carcinogenesis and tumor progression. A large number of BMP inhibitors are available to study BMP function; however, as most of these antagonists are promiscuous, evaluating specific effects of individual BMPs is not feasible. Because the oncogenic role of the different BMPs varies for each neoplasm, highly selective BMP inhibitors are required. Here, we describe the generation of three types of llama-derived heavy chain variable domains (VHH) that selectively bind to either BMP4, to BMP2 and 4, or to BMP2, 4, 5, and 6. These generated VHHs have high affinity to their targets and are able to inhibit BMP signaling. Epitope binning and docking modeling have shed light into the basis for their BMP specificity. As opposed to the wide structural reach of natural inhibitors, these small molecules target the grooves and pockets of BMPs involved in receptor binding. In organoid experiments, specific inhibition of BMP4 does not affect the activation of normal stem cells. Furthermore, in vitro inhibition of cancer-derived BMP4 noncanonical signals results in an increase of chemosensitivity in a colorectal cancer cell line. Therefore, because of their high specificity and low off-target effects, these VHHs could represent a therapeutic alternative for BMP4(+) malignancies.

  15. Specific heat of pristine and brominated graphite fibers, composites and HOPG. [Highly Oriented Pyrolytic Graphite (United States)

    Hung, Ching-Chen; Maciag, Carolyn


    Differential scanning calorimetry was used to obtain specific heat values of pristine and brominated P-100 graphite fibers and brominated P-100/epoxy composite as well as pristine and brominated highly oriented pyrolytic graphite (HOPG) for comparison. Based on the experimental results obtained, specific heat values are calculated for several different temperatures, with a standard deviation estimated at 1.4 percent of the average values. The data presented here are useful in designing heat transfer devices (such as airplane de-icing heaters) from bromine fibers.

  16. Immunohistochemical expression of Skp2 protein in oral nevi and melanoma (United States)

    León, Jorge E.; Carlos, Román; Delgado-Azañero, Wilson; Mosqueda-Taylor, Adalberto; Paes-de-Almeida, Oslei


    Objective: The aim of this study was to analyze the immunohistochemical expression of Skp2 protein in 38 oral nevi and 11 primary oral melanomas. Study Design: Expression of this ubiquitin protein was evaluated by immunohistochemistry in 49 oral melanocytic lesions, including 38 intramucosal nevi and 11 primary oral melanomas. The labeling index (LI) was assessed considering the percentage of cells expressing nuclear positivity out of the total number of cells, counting 1000 cells per slide. Results: Skp2 protein was rarely expressed in intramucosal nevi, in contrast to oral melanomas, which showed high levels of this protein. Conclusion: These results indicate that Skp2 protein may play a role in the development and progression of oral melanomas, and it also could be useful as an immunohistochemical marker for differential diagnosis of oral benign and malignant melanocytic lesions. Key words:Oral melanoma, oral nevi, Skp2, cell cycle, immunohistochemistry. PMID:23385514

  17. Hydrazide functionalized core-shell magnetic nanocomposites for highly specific enrichment of N-glycopeptides. (United States)

    Liu, Liting; Yu, Meng; Zhang, Ying; Wang, Changchun; Lu, Haojie


    In view of the biological significance of glycosylation for human health, profiling of glycoproteome from complex biological samples is highly inclined toward the discovery of disease biomarkers and clinical diagnosis. Nevertheless, because of the existence of glycopeptides at relatively low abundances compared with nonglycosylated peptides and glycan microheterogeneity, glycopeptides need to be highly selectively enriched from complex biological samples for mass spectrometry analysis. Herein, a new type of hydrazide functionalized core-shell magnetic nanocomposite has been synthesized for highly specific enrichment of N-glycopeptides. The nanocomposites with both the magnetic core and the polymer shell hanging high density of hydrazide groups were prepared by first functionalization of the magnetic core with polymethacrylic acid by reflux precipitation polymerization to obtain the Fe3O4@poly(methacrylic acid) (Fe3O4@PMAA) and then modification of the surface of Fe3O4@PMAA with adipic acid dihydrazide (ADH) to obtain Fe3O4@poly(methacrylic hydrazide) (Fe3O4@PMAH). The abundant hydrazide groups toward highly specific enrichment of glycopeptides and the magnetic core make it suitable for large-scale, high-throughput, and automated sample processing. In addition, the hydrophilic polymer surface can provide low nonspecific adsorption of other peptides. Compared to commercially available hydrazide resin, Fe3O4@PMAH improved more than 5 times the signal-to-noise ratio of standard glycopeptides. Finally, this nanocomposite was applied in the profiling of N-glycoproteome from the colorectal cancer patient serum. In total, 175 unique glycopeptides and 181 glycosylation sites corresponding to 63 unique glycoproteins were identified in three repeated experiments, with the specificities of the enriched glycopeptides and corresponding glycoproteins of 69.6% and 80.9%, respectively. Because of all these attractive features, we believe that this novel hydrazide functionalized

  18. Preparation and Characterization of Porous Yttrium Oxide Powders with High Specific Surface Area

    Institute of Scientific and Technical Information of China (English)


    The porous cubic yttrium oxides with high specific surface area were prepared by the explosive decomposition of yttrium nitrate and its complex formed with methyl salicylate. The specific surface area and properties of powders synthesized at various temperatures were characterized using BET, X-ray diffraction (XRD), infrared spectra (IR), and scanning electron microscopy (SEM). The results indicate that the highest specific surface area is found to be 65.37 m2*g-1 at the calcination temperature of 600 ℃, and then decreases to 20.33 m2*g-1 with the calcination temperature rising from 600 to 900 ℃. The powders show strong surface activity for adsorping water and carbon dioxide in air, which also decreases with the rising calcination temperature. The drop both on the surface area and surface activity of samples at higher temperatures may be due to pore-narrowing(sintering) effects.

  19. Synthesis of high specific activity (1- sup 3 H) farnesyl pyrophosphate

    Energy Technology Data Exchange (ETDEWEB)

    Saljoughian, M.; Morimoto, H.; Williams, P.G.


    The synthesis of tritiated farnesyl pyrophosphate with high specific activity is reported. trans-trans Farnesol was oxidized to the corresponding aldehyde followed by reduction with lithium aluminium tritide (5%-{sup 3}H) to give trans-trans (1-{sup 3}H)farnesol. The specific radioactivity of the alcohol was determined from its triphenylsilane derivative, prepared under very mild conditions. The tritiated alcohol was phosphorylated by initial conversion to an allylic halide, and subsequent treatment of the halide with tris-tetra-n-butylammonium hydrogen pyrophosphate. The hydride procedure followed in this work has advantages over existing methods for the synthesis of tritiated farnesyl pyrophosphate, with the possibility of higher specific activity and a much higher yield obtained. 10 refs., 3 figs.

  20. A Rapid In-Clinic Test Detects Acute Leptospirosis in Dogs with High Sensitivity and Specificity. (United States)

    Kodjo, Angeli; Calleja, Christophe; Loenser, Michael; Lin, Dan; Lizer, Joshua


    A rapid IgM-detection immunochromatographic test (WITNESS® Lepto, Zoetis) has recently become available to identify acute canine leptospirosis at the point of care. Diagnostic sensitivity and specificity of the test were evaluated by comparison with the microscopic agglutination assay (MAT), using a positive cut-off titer of ≥800. Banked serum samples from dogs exhibiting clinical signs and suspected leptospirosis were selected to form three groups based on MAT titer: (1) positive (n = 50); (2) borderline (n = 35); and (3) negative (n = 50). Using an analysis to weight group sizes to reflect French prevalence, the sensitivity and specificity were 98% and 93.5% (88.2% unweighted), respectively. This test rapidly identifies cases of acute canine leptospirosis with high levels of sensitivity and specificity with no interference from previous vaccination.

  1. Differences in tumor thickness between hematoxylin and eosin and Melan-A immunohistochemically stained primary cutaneous melanomas. (United States)

    Drabeni, Marina; Lopez-Vilaró, Laura; Barranco, Carlos; Trevisan, Giusto; Gallardo, Fernando; Pujol, Ramon M


    Tumor thickness (Breslow thickness) represents the main prognostic factor in primary melanoma. Potential differences in melanoma tumor thickness measurements between conventional hematoxylin and eosin (H&E) and Melan-A immunohistochemical staining were evaluated. Ninety-nine excisional biopsies were included in the study. From each sample, 2 consecutive histological sections were stained with H&E and Melan-A, respectively. Tumor thickness was measured from both sections by 2 independent observers. In 59 biopsy specimens (59.6%), higher tumor thickness measurements were recorded in Melan-A-stained than in H&E-stained sections. In 42.4% of such cases (25 biopsies), the observed differences were ≥0.2 mm. After Melan-A evaluation, 33% of in situ melanoma cases were reclassified as invasive melanoma, with thickness measurements ranging from 0.15 to 0.35 mm. In 23 biopsies, identical values were recorded with both techniques, whereas in 17 cases, measurements obtained with H&E staining were slightly higher (from 0.01 to 0.18 mm) than those obtained with Melan-A staining. A high rate of interobserver agreement was noted, and significant intertechnique measurement differences were detected. Significant discrepancies (≥0.2 mm) in thickness measurements between the 2 techniques were mainly attributed to the presence of individual or small clusters of melanocytic cells in the papillary dermis. These melanocytic cells could be easily overlooked in H&E-stained sections, especially in sections showing dense lymphohistiocytic inflammatory infiltrates, numerous melanin-containing histiocytic cells in the upper dermis, or extensive fibrotic changes or regression phenomena. This study confirms the practical interest of immunohistochemical staining with Melan-A in evaluating primary melanoma and, specifically, in situ melanoma cases.

  2. A highly sensitive and specific system for large-scale gene expression profiling

    Directory of Open Access Journals (Sweden)

    Wang Hui-Yun


    Full Text Available Abstract Background Rapid progress in the field of gene expression-based molecular network integration has generated strong demand on enhancing the sensitivity and data accuracy of experimental systems. To meet the need, a high-throughput gene profiling system of high specificity and sensitivity has been developed. Results By using specially designed primers, the new system amplifies sequences in neighboring exons separated by big introns so that mRNA sequences may be effectively discriminated from other highly related sequences including their genes, unprocessed transcripts, pseudogenes and pseudogene transcripts. Probes used for microarray detection consist of sequences in the two neighboring exons amplified by the primers. In conjunction with a newly developed high-throughput multiplex amplification system and highly simplified experimental procedures, the system can be used to analyze >1,000 mRNA species in a single assay. It may also be used for gene expression profiling of very few (n = 100 or single cells. Highly reproducible results were obtained from duplicate samples with the same number of cells, and from those with a small number (100 and a large number (10,000 of cells. The specificity of the system was demonstrated by comparing results from a breast cancer cell line, MCF-7, and an ovarian cancer cell line, NCI/ADR-RES, and by using genomic DNA as starting material. Conclusion Our approach may greatly facilitate the analysis of combinatorial expression of known genes in many important applications, especially when the amount of RNA is limited.

  3. Production of high quality distillate to meet a fit-for-purpose boiler feedwater specification

    Energy Technology Data Exchange (ETDEWEB)

    Minnich, K. [Veolia Water Solutions Oil and Gas, Calgary, AB (Canada); Neu, D. [Veolia Water Solutions and Technologies/HPD, Pewaukee, WI (United States); Drone, J.L. [Veolia Water Solutions and Technologies/HPD, Plainfield, IL (United States)


    Veolia Water Solutions and Technologies has significant experience managing boiler water chemistry and is the world's largest manufacturer of evaporation systems. The company has conducted extensive testing and analysis for produced water evaporation distillate from multiple facilities. In order to produce boiler feed water, evaporation of produced water is used at several steam assisted gravity drainage (SAGD) facilities. There are no official guidelines for the required quality of evaporator distillate to feed a once through steam generator (OTSG) or high pressure industrial watertube boiler (IWT) that will produce injection steam. This paper presented a basis for a fit-for-purpose specification for IWT boilers as well as data on the performance of a Vapor Washer, which produces high quality distilled water that meets fit-for-purpose specifications even during normal variations in feed conditions. Specifically, the paper discussed boiler water requirements for steam injection IWTs; the quality of distillate from a produced water evaporator; the benefits of vapour washing to maintain distillate quality; and suggested boiler chemistry limits for a fit-for-purpose specification. Oxygen, iron, and copper were discussed as being implicated with corrosion and reaction in boilers. Hardness contaminants such as calcium and magnesium were also presented. Suggested limits for boiler water in the fit-for-purpose specification were also presented for silica, total alkalinity, free OH alkalinity, and total dissolved solids in steam. It was concluded that foaming episodes can occur in produced water evaporators due to normal variations, and the distillate can fail to meet the fit-for-purpose specification during foam upsets. 3 refs., 9 figs.

  4. A novel and highly specific phage endolysin cell wall binding domain for detection of Bacillus cereus. (United States)

    Kong, Minsuk; Sim, Jieun; Kang, Taejoon; Nguyen, Hoang Hiep; Park, Hyun Kyu; Chung, Bong Hyun; Ryu, Sangryeol


    Rapid, specific and sensitive detection of pathogenic bacteria is crucial for public health and safety. Bacillus cereus is harmful as it causes foodborne illness and a number of systemic and local infections. We report a novel phage endolysin cell wall-binding domain (CBD) for B. cereus and the development of a highly specific and sensitive surface plasmon resonance (SPR)-based B. cereus detection method using the CBD. The newly discovered CBD from endolysin of PBC1, a B. cereus-specific bacteriophage, provides high specificity and binding capacity to B. cereus. By using the CBD-modified SPR chips, B. cereus can be detected at the range of 10(5)-10(8) CFU/ml. More importantly, the detection limit can be improved to 10(2) CFU/ml by using a subtractive inhibition assay based on the pre-incubation of B. cereus and CBDs, removal of CBD-bound B. cereus, and SPR detection of the unbound CBDs. The present study suggests that the small and genetically engineered CBDs can be promising biological probes for B. cereus. We anticipate that the CBD-based SPR-sensing methods will be useful for the sensitive, selective, and rapid detection of B. cereus.

  5. High Transferability of Homoeolog-Specific Markers between Bread Wheat and Newly Synthesized Hexaploid Wheat Lines (United States)

    Zeng, Deying; Luo, Jiangtao; Li, Zenglin; Chen, Gang; Zhang, Lianquan; Ning, Shunzong; Yuan, Zhongwei; Zheng, Youliang; Hao, Ming; Liu, Dengcai


    Bread wheat (Triticum aestivum, 2n = 6x = 42, AABBDD) has a complex allohexaploid genome, which makes it difficult to differentiate between the homoeologous sequences and assign them to the chromosome A, B, or D subgenomes. The chromosome-based draft genome sequence of the ‘Chinese Spring’ common wheat cultivar enables the large-scale development of polymerase chain reaction (PCR)-based markers specific for homoeologs. Based on high-confidence ‘Chinese Spring’ genes with known functions, we developed 183 putative homoeolog-specific markers for chromosomes 4B and 7B. These markers were used in PCR assays for the 4B and 7B nullisomes and their euploid synthetic hexaploid wheat (SHW) line that was newly generated from a hybridization between Triticum turgidum (AABB) and the wild diploid species Aegilops tauschii (DD). Up to 64% of the markers for chromosomes 4B or 7B in the SHW background were confirmed to be homoeolog-specific. Thus, these markers were highly transferable between the ‘Chinese Spring’ bread wheat and SHW lines. Homoeolog-specific markers designed using genes with known functions may be useful for genetic investigations involving homoeologous chromosome tracking and homoeolog expression and interaction analyses. PMID:27611704

  6. Functional differences in the specific B-cell compartment in high or low antibody responder mice. (United States)

    de Franco, M; Vidard, L; Mouton, D; Decreusefond, C; Gille Perramant, M F; Couderc, J


    The role of antigen-presenting cells (APC) in quantitative antibody synthesis regulation was studied in mice genetically selected for high (HI) or low (LI) antibody response. Irradiated spleen cells and enriched specific B cells from HI and LI mice co-isogenic at H-2s locus, were compared for their capacity to present chicken ovalbumin (OVA) to specific T-cell hybridomas. Minor differences were observed between HI and LI mice when three distinct hybridomas were stimulated in the presence of OVA and splenic macrophages APC. These differences were totally abolished when APC were pulsed with OVAxAb complexes. Looking at the B-cell compartment, hybridoma IL-2 responses were similar when TNP primed B cells were pulsed with OVA. However, when OVA was targeted on TNP-specific enriched B cells by pulsing with TNP-OVA, the IL-2 production by the T-cell hybridomas was stronger in the presence of HI B cells than in the presence of LI B cells. These results strongly suggest that an efficient Ag handling/processing by specific B cells is a major component of the high Ab responder status in Biozzi mice.

  7. Production and characterization of highly specific monoclonal antibodies to D-glutamic acid. (United States)

    Sakamoto, Seiichi; Matsuura, Yurino; Yonenaga, Yayoi; Tsuneura, Yumi; Aso, Mariko; Kurose, Hitoshi; Tanaka, Hiroyuki; Morimoto, Satoshi


    Most of the functions of D-amino acids (D-AA) remain unclear because of little analytic methods for specific detection/determination. In this study, a highly specific monoclonal antibody to D-glutamic acid (D-Glu-MAb) was produced using a hybridoma method. Characterization of D-Glu-MAb by indirect enzyme-linked immunosorbent assay (ELISA) revealed that it has high selectivity against D-Glu-glutaraldehyde (GA) conjugates, while no cross-reaction was observed when 38 other kinds of AA-GA conjugates were used. Moreover, subsequent indirect competitive ELISA disclosed that an epitope of D-Glu-MAb is a D-Glu-GA molecule in the conjugates, suggesting that D-Glu-MAb could be a useful tool to investigate the functional analysis of D-Glu in immunostaining.

  8. Human PHOSPHO1 exhibits high specific phosphoethanolamine and phosphocholine phosphatase activities (United States)


    Human PHOSPHO1 is a phosphatase enzyme for which expression is upregulated in mineralizing cells. This enzyme has been implicated in the generation of Pi for matrix mineralization, a process central to skeletal development. PHOSPHO1 is a member of the haloacid dehalogenase (HAD) superfamily of Mg2+-dependent hydrolases. However, substrates for PHOSPHO1 are, as yet, unidentified and little is known about its activity. We show here that PHOSPHO1 exhibits high specific activities toward phosphoethanolamine (PEA) and phosphocholine (PCho). Optimal enzymic activity was observed at approx. pH 6.7. The enzyme shows a high specific Mg2+-dependence, with apparent Km values of 3.0 μM for PEA and 11.4 μM for PCho. These results provide a novel mechanism for the generation of Pi in mineralizing cells from PEA and PCho. PMID:15175005

  9. Selective culling of high avidity antigen-specific CD4+ T cells after virulent Salmonella infection. (United States)

    Ertelt, James M; Johanns, Tanner M; Mysz, Margaret A; Nanton, Minelva R; Rowe, Jared H; Aguilera, Marijo N; Way, Sing Sing


    Typhoid fever is a persistent infection caused by host-adapted Salmonella strains adept at circumventing immune-mediated host defences. Given the importance of T cells in protection, the culling of activated CD4+ T cells after primary infection has been proposed as a potential immune evasion strategy used by this pathogen. We demonstrate that the purging of activated antigen-specific CD4+ T cells after virulent Salmonella infection requires SPI-2 encoded virulence determinants, and is not restricted only to cells with specificity to Salmonella-expressed antigens, but extends to CD4+ T cells primed to expand by co-infection with recombinant Listeria monocytogenes. Unexpectedly, however, the loss of activated CD4+ T cells during Salmonella infection demonstrated using a monoclonal population of adoptively transferred CD4+ T cells was not reproduced among the endogenous repertoire of antigen-specific CD4+ T cells identified with MHC class II tetramer. Analysis of T-cell receptor variable segment usage revealed the selective loss and reciprocal enrichment of defined CD4+ T-cell subsets after Salmonella co-infection that is associated with the purging of antigen-specific cells with the highest intensity of tetramer staining. Hence, virulent Salmonella triggers the selective culling of high avidity activated CD4+ T-cell subsets, which re-shapes the repertoire of antigen-specific T cells that persist later after infection.

  10. Specific binding of eukaryotic ORC to DNA replication origins depends on highly conserved basic residues. (United States)

    Kawakami, Hironori; Ohashi, Eiji; Kanamoto, Shota; Tsurimoto, Toshiki; Katayama, Tsutomu


    In eukaryotes, the origin recognition complex (ORC) heterohexamer preferentially binds replication origins to trigger initiation of DNA replication. Crystallographic studies using eubacterial and archaeal ORC orthologs suggested that eukaryotic ORC may bind to origin DNA via putative winged-helix DNA-binding domains and AAA+ ATPase domains. However, the mechanisms how eukaryotic ORC recognizes origin DNA remain elusive. Here, we show in budding yeast that Lys-362 and Arg-367 residues of the largest subunit (Orc1), both outside the aforementioned domains, are crucial for specific binding of ORC to origin DNA. These basic residues, which reside in a putative disordered domain, were dispensable for interaction with ATP and non-specific DNA sequences, suggesting a specific role in recognition. Consistent with this, both residues were required for origin binding of Orc1 in vivo. A truncated Orc1 polypeptide containing these residues solely recognizes ARS sequence with low affinity and Arg-367 residue stimulates sequence specific binding mode of the polypeptide. Lys-362 and Arg-367 residues of Orc1 are highly conserved among eukaryotic ORCs, but not in eubacterial and archaeal orthologs, suggesting a eukaryote-specific mechanism underlying recognition of replication origins by ORC.

  11. Immunohistochemical localization and quantification of the 3-(cystein-S-yl)-acetaminophen protein adduct in acetaminophen hepatotoxicity. (United States)

    Roberts, D W; Bucci, T J; Benson, R W; Warbritton, A R; McRae, T A; Pumford, N R; Hinson, J A


    Acetaminophen overdose causes severe hepatotoxicity in humans and laboratory animals, presumably by metabolism to N-acetyl-p-benzoquinone imine: and binding to cysteine groups as 3-(cystein-S-yl)acetaminophen-protein adduct. Antiserum specific for the adduct was used immunohistochemically to demonstrate the formation, distribution, and concentration of this specific adduct in livers of treated mice and was correlated with cell injury as a function of dose and time. Within the liver lobule, immunohistochemically demonstrable adduct occurred in a temporally progressive, central-to-peripheral pattern. There was concordance between immunohistochemical staining and quantification of the adduct in hepatic 10,000g supernate, using a quantitative particle concentration fluorescence immunoassay. Findings include: 1) immunochemically detectable adduct before the appearance of centrilobular necrosis, 2) distinctive lobular zones of adduct localization with subsequent depletion during the progression of toxicity, 3) drug-protein binding in hepatocytes at subhepatotoxic doses and before depletion of total hepatic glutathione, 4) immunohistochemical evidence of drug binding in the nucleus, and 5) adduct in metabolically active and dividing hepatocytes and in macrophagelike cells in the regenerating liver.

  12. Histological and immunohistochemical evaluation of sentinel lymph ...

    African Journals Online (AJOL)

    [6,7] The technique is sensitive and specific to ... of sentinel lymph nodes in breast cancer at a tertiary hospital in ... tumour excision or simple mastectomy as well as SLN biopsies. All the ... [6,7] Significant resource implications of the increasing ...

  13. Status of JENDL High Energy File. Evaluation method, tools, specification, release procedure, etc

    Energy Technology Data Exchange (ETDEWEB)

    Fukahori, Tokio [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment


    The ENDF-6 format file should be kept as a standard distribution file and it is not difficult to convert into some other form for code`s libraries. From this point of view, status of JENDL High Energy File is introduced in this report as well as evaluation strategy, recommended specification, stored nuclides and quantities, a format structure, evaluation methods and tools, and release plan. (author)

  14. The use of Element-Specific Detectors Coupled with High-Performance Liquid Chromatographs. (United States)


    approach to determining the amount of various chelating agents in solution, Jones and Manahan reacted the indicator metal, A 20 copper(II) with a...latter two cases, GFAA was employed as the element specific detector. Jones and Manahan employing a high performance absorption column directly...Chromatogr. Sci., 17: 395 (1979). 35. D. R. Jones, IV and S. E. Manahan , Anal. Chem., 48: 1897 (1976). 36. D. R. Jones, IV and S. E. Manahan , Anal. Chem

  15. High specificity of a novel Zika virus ELISA in European patients after exposure to different flaviviruses. (United States)

    Huzly, Daniela; Hanselmann, Ingeborg; Schmidt-Chanasit, Jonas; Panning, Marcus


    The current Zika virus (ZIKV) epidemic in the Americas caused an increase in diagnostic requests in European countries. Here we demonstrate high specificity of the Euroimmun anti-ZIKV IgG and IgM ELISA tests using putative cross-reacting sera of European patients with antibodies against tick-borne encephalitis virus, dengue virus, yellow fever virus and hepatitis C virus. This test may aid in counselling European travellers returning from regions where ZIKV is endemic.

  16. Immunohistochemical localization of epidermal growth factor in rat and man

    DEFF Research Database (Denmark)

    Poulsen, Steen Seier; Nexø, Ebba


    Epidermal growth factor (EGF) is a peptide which stimulates cell mitotic activity and differentiation, has a cytoprotective effect on the gastroduodenal mucosa, and inhibits gastric acid secretion. The immunohistochemical localization of EGF in the Brunner's glands and the submandibular glands...... is well documented. The localization of EGF in other tissues is still unclarified. In the present study, the immunohistochemical localization of EGF in tissues from rat, man and a 20 week human fetus were investigated. In man and rat, immunoreaction was found in the submandibular glands, the serous glands...

  17. Villoglandular papillary adenocarcinoma of the uterine cervix with immunohistochemical characteristics. (United States)

    Polat, A; Düsmez, D; Pata, O; Aydin, O; Egilmez, R


    Villoglandular papillary adenocarcinoma of uterine cervix has been recently described and to date fewer than a hundred cases have been reported in the world literature. Here we present a 38-year-old woman who underwent radical hysterectomy combined bilateral pelvic lymphoadenectomy and after 28 months postoperatively no lymph node metastasis and no evidence of recurrent disease ocurred. Immunohistochemically Ki-67 overexpression was detected in the tumour, with no immunoreactivity with p53, estrogen and progesteron receptors and broadly-reactive human papilloma virus including types 6, 11, 16, 18, 31, 33, 42, 51, 52, 56, and 58. In this paper, clinical, macroscopical, microscopical and immunohistochemical characteristics of this tumour are reviewed.

  18. Immunohistochemical study of muscle biopsy in children with cerebral palsy. (United States)

    Marbini, Adriana; Ferrari, A; Cioni, G; Bellanova, M F; Fusco, C; Gemignani, F


    Muscle biopsy was examined in 20 children with cerebral palsy, using immunohistochemical methods for marker of denervation neural cell adhesion molecules (N-CAM) in addition to standard techniques. Histological and histochemical study showed mild myopathic changes, type 1 predominance, and type 1 and type 2 hypotrophy, in accord with previous observations. Immunohistochemical study showed N-CAM expression in most biopsies (15/20), usually in scattered fibers, whereas in four patients aged less than 6 years it was expressed in grouped fibers. Our study supports the hypothesis of motor unit remodeling as a consequence of spasticity, especially in early phases of the disease.

  19. Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing


    Tsai, Shengdar Q.; Wyvekens, Nicolas; Khayter, Cyd; Foden, Jennifer A.; Thapar, Vishal; Reyon, Deepak; Goodwin, Mathew J.; Aryee, Martin J.; Joung, J. Keith


    Monomeric CRISPR-Cas9 nucleases are widely used for targeted genome editing but can induce unwanted off-target mutations with high frequencies. Here we describe dimeric RNA-guided FokI Nucleases (RFNs) that recognize extended sequences and can edit endogenous genes with high efficiencies in human cells. The cleavage activity of an RFN depends strictly on the binding of two guide RNAs (gRNAs) to DNA with a defined spacing and orientation and therefore show improved specificities relative to wi...

  20. Theory of specific heat of vortex liquid of high T c superconductors (United States)

    Bai, Chen; Chi, Cheng; Wang, Jiangfan


    Superconducting thermal fluctuation (STF) plays an important role in both thermodynamic and transport properties in the vortex liquid phase of high T c superconductors. It was widely observed in the vicinity of the critical transition temperature. In the framework of Ginzburg-Landau-Lawrence-Doniach theory in magnetic field, a self-consistent analysis of STF including all Landau levels is given. Besides that, we calculate the contribution of STF to specific heat in vortex liquid phase for high T c cuprate superconductors, and the fitting results are in good agreement with experimental data. Project supported by the National Natural Science Foundation of China (Grant No. 11274018).

  1. Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing


    Tsai, Shengdar Q.; Wyvekens, Nicolas; Khayter, Cyd; Foden, Jennifer A.; Thapar, Vishal; Reyon, Deepak; Goodwin, Mathew J.; Aryee, Martin J; Joung, J. Keith


    Monomeric CRISPR-Cas9 nucleases are widely used for targeted genome editing but can induce unwanted off-target mutations with high frequencies. Here we describe dimeric RNA-guided FokI Nucleases (RFNs) that recognize extended sequences and can edit endogenous genes with high efficiencies in human cells. The cleavage activity of an RFN depends strictly on the binding of two guide RNAs (gRNAs) to DNA with a defined spacing and orientation and therefore show improved specificities relative to wi...

  2. Fast and highly specific DNA-based multiplex detection on a solid support. (United States)

    Barišić, Ivan; Kamleithner, Verena; Schönthaler, Silvia; Wiesinger-Mayr, Herbert


    Highly specific and fast multiplex detection methods are essential to conduct reasonable DNA-based diagnostics and are especially important to characterise infectious diseases. More than 1000 genetic targets such as antibiotic resistance genes, virulence factors and phylogenetic markers have to be identified as fast as possible to facilitate the correct treatment of a patient. In the present work, we developed a novel ligation-based DNA probe concept that was combined with the microarray technology and used it for the detection of bacterial pathogens. The novel linear chain (LNC) probes identified all tested species correctly within 1 h based on their 16S rRNA gene in a 25-multiplex reaction. Genomic DNA was used directly as template in the ligation reaction identifying as little as 10(7) cells without any pre-amplification. The high specificity was further demonstrated characterising a single nucleotide polymorphism leading to no false positive fluorescence signals of the untargeted single nucleotide polymorphism (SNP) variants. In comparison to conventional microarray probes, the sensitivity of the novel LNC3 probes was higher by a factor of 10 or more. In summary, we present a fast, simple, highly specific and sensitive multiplex detection method adaptable for a wide range of applications.

  3. Highly specific SNP detection using 2D graphene electronics and DNA strand displacement. (United States)

    Hwang, Michael T; Landon, Preston B; Lee, Joon; Choi, Duyoung; Mo, Alexander H; Glinsky, Gennadi; Lal, Ratnesh


    Single-nucleotide polymorphisms (SNPs) in a gene sequence are markers for a variety of human diseases. Detection of SNPs with high specificity and sensitivity is essential for effective practical implementation of personalized medicine. Current DNA sequencing, including SNP detection, primarily uses enzyme-based methods or fluorophore-labeled assays that are time-consuming, need laboratory-scale settings, and are expensive. Previously reported electrical charge-based SNP detectors have insufficient specificity and accuracy, limiting their effectiveness. Here, we demonstrate the use of a DNA strand displacement-based probe on a graphene field effect transistor (FET) for high-specificity, single-nucleotide mismatch detection. The single mismatch was detected by measuring strand displacement-induced resistance (and hence current) change and Dirac point shift in a graphene FET. SNP detection in large double-helix DNA strands (e.g., 47 nt) minimize false-positive results. Our electrical sensor-based SNP detection technology, without labeling and without apparent cross-hybridization artifacts, would allow fast, sensitive, and portable SNP detection with single-nucleotide resolution. The technology will have a wide range of applications in digital and implantable biosensors and high-throughput DNA genotyping, with transformative implications for personalized medicine.

  4. [High prevalence of specific language impairment in Robinson Crusoe Island. A possible founder effect]. (United States)

    Villanueva, Pía; de Barbieri, Zulema; Palomino, Hernán M; Palomino, Hernán


    Specific language impairment (SLI) occurs in 2% to 8% of preschool children. Major and candidate genes are probably involved. Genetic drift is a cause for the presence of high frequencies of deleterious alíeles of a specific disease and the founder effect is one of its forms. Robinson Crusoe Island has 633 inhabitants and its actual population began with 8 families that repopulated the island at the end of XIXth century. To assess the frequency of specific language impairment among children living in Robinson Crusoe Island. All 66 children aged between 3 and 9 years living in the island, were studied. Parents were interviewed and in children, non verbal intelligence, audiometric parameters, comprehension and expression of oral language were assessed. Extended genealogies were also performed. Forty children had at least one parent that was descending of founder families. Among these, 35% had SLI. Eighth five percent of SLI affected children came from the same colonizer family. The prevalence of SLI in Robinson Crusoe Island is higher than that reported in mainland Chile and abroad. This high prevalence, associated to a high frequency of consanguinity, supports the influence of genetic mechanisms in SLI transmission, based on a founder effect.

  5. Starvation, together with the SOS response, mediates high biofilm-specific tolerance to the fluoroquinolone ofloxacin.

    Directory of Open Access Journals (Sweden)

    Steve P Bernier

    Full Text Available High levels of antibiotic tolerance are a hallmark of bacterial biofilms. In contrast to well-characterized inherited antibiotic resistance, molecular mechanisms leading to reversible and transient antibiotic tolerance displayed by biofilm bacteria are still poorly understood. The physiological heterogeneity of biofilms influences the formation of transient specialized subpopulations that may be more tolerant to antibiotics. In this study, we used random transposon mutagenesis to identify biofilm-specific tolerant mutants normally exhibited by subpopulations located in specialized niches of heterogeneous biofilms. Using Escherichia coli as a model organism, we demonstrated, through identification of amino acid auxotroph mutants, that starved biofilms exhibited significantly greater tolerance towards fluoroquinolone ofloxacin than their planktonic counterparts. We demonstrated that the biofilm-associated tolerance to ofloxacin was fully dependent on a functional SOS response upon starvation to both amino acids and carbon source and partially dependent on the stringent response upon leucine starvation. However, the biofilm-specific ofloxacin increased tolerance did not involve any of the SOS-induced toxin-antitoxin systems previously associated with formation of highly tolerant persisters. We further demonstrated that ofloxacin tolerance was induced as a function of biofilm age, which was dependent on the SOS response. Our results therefore show that the SOS stress response induced in heterogeneous and nutrient-deprived biofilm microenvironments is a molecular mechanism leading to biofilm-specific high tolerance to the fluoroquinolone ofloxacin.

  6. Molecular inversion probe: a new tool for highly specific detection of plant pathogens.

    Directory of Open Access Journals (Sweden)

    Han Yih Lau

    Full Text Available Highly specific detection methods, capable of reliably identifying plant pathogens are crucial in plant disease management strategies to reduce losses in agriculture by preventing the spread of diseases. We describe a novel molecular inversion probe (MIP assay that can be potentially developed into a robust multiplex platform to detect and identify plant pathogens. A MIP has been designed for the plant pathogenic fungus Fusarium oxysporum f.sp. conglutinans and the proof of concept for the efficiency of this technology is provided. We demonstrate that this methodology can detect as little as 2.5 ng of pathogen DNA and is highly specific, being able to accurately differentiate Fusarium oxysporum f.sp. conglutinans from other fungal pathogens such as Botrytis cinerea and even pathogens of the same species such as Fusarium oxysporum f.sp. lycopersici. The MIP assay was able to detect the presence of the pathogen in infected Arabidopsis thaliana plants as soon as the tissues contained minimal amounts of pathogen. MIP methods are intrinsically highly multiplexable and future development of specific MIPs could lead to the establishment of a diagnostic method that could potentially screen infected plants for hundreds of pathogens in a single assay.

  7. Identification of fluorescent compounds with non-specific binding property via high throughput live cell microscopy.

    Directory of Open Access Journals (Sweden)

    Sangeeta Nath

    Full Text Available INTRODUCTION: Compounds exhibiting low non-specific intracellular binding or non-stickiness are concomitant with rapid clearing and in high demand for live-cell imaging assays because they allow for intracellular receptor localization with a high signal/noise ratio. The non-stickiness property is particularly important for imaging intracellular receptors due to the equilibria involved. METHOD: Three mammalian cell lines with diverse genetic backgrounds were used to screen a combinatorial fluorescence library via high throughput live cell microscopy for potential ligands with high in- and out-flux properties. The binding properties of ligands identified from the first screen were subsequently validated on plant root hair. A correlative analysis was then performed between each ligand and its corresponding physiochemical and structural properties. RESULTS: The non-stickiness property of each ligand was quantified as a function of the temporal uptake and retention on a cell-by-cell basis. Our data shows that (i mammalian systems can serve as a pre-screening tool for complex plant species that are not amenable to high-throughput imaging; (ii retention and spatial localization of chemical compounds vary within and between each cell line; and (iii the structural similarities of compounds can infer their non-specific binding properties. CONCLUSION: We have validated a protocol for identifying chemical compounds with non-specific binding properties that is testable across diverse species. Further analysis reveals an overlap between the non-stickiness property and the structural similarity of compounds. The net result is a more robust screening assay for identifying desirable ligands that can be used to monitor intracellular localization. Several new applications of the screening protocol and results are also presented.

  8. Immunohistochemical consistency between primary tumors and lymph node metastases of gastric neuroendocrine carcinoma

    Directory of Open Access Journals (Sweden)

    Uchiyama Chieko


    Full Text Available Abstract Background Gastric neuroendocrine carcinoma (G-NEC is a rare, highly malignant tumor that exhibits aggressive growth leading to vascular invasion, distant metastasis and extremely poor prognosis. We studied the clinicopathological findings of seven patients at our institute to better under this disease. Methods Seven cases of G-NEC were identified among 1,027 cases of gastric carcinoma that underwent gastrectomy at Kansai Rousai Hospital between 2002 and 2010. We studied the pathological and immunohistochemical features of gastric neuroendocrine carcinomas at both the primary site and metastatic lymph nodes. Results The mean patient age was 73 years (range 63 to 86 years. There were no females in this series. The final staging was Stage I in one case, Stage II in two, Stage III in two and Stage IV in two. A total of 31 metastatic lymph nodes were found in these patients. This study revealed that the ratio of neuroendocrine cells was similar between the primary and metastatic sites, which tended to show the same expression patterns of neuroendocrine markers. Conclusions Metastatic lymph nodes showed heterogeneous immunohistochemical expression patterns similar to the primary sites. G-NEC is far advanced at diagnosis and rapidly reaches the lymph nodes retaining its heterogeneity, carrying a worse prognosis than common gastric cancer. Mini abstract G-NEC grows rapidly and metastasizes to the lymph nodes, retaining its pathological and immunohistochemical heterogeneity even at the metastatic sites.

  9. Immunohistochemical detection of CCR2 and CX3CR1 in sepsis-induced lung injury. (United States)

    An, Jun-Ling; Ishida, Yuko; Kimura, Akihiko; Tsokos, Michael; Kondo, Toshikazu


    Sepsis is a systemic inflammatory disease with high mortality. In the present study, we immunohistochemically examined CCR2 and CX3CR1 expression in sepsis-induced lung injury, and discussed its availability for the postmortem diagnosis of sepsis. Lung samples were obtained from different lung lobes of nine sepsis and eight control cases with postmortem intervals between 12 and 48h. Immunohistochemically, mononuclear cells recruited into the lungs expressed CCR2 and CX3CR1 in both sepsis and non-septic groups. In double-color immunofluorescence analysis, CCR2- or CX3CR1-positive cells could be identified as CD68-positive macrophages. Moreover, most of CD68-positive macrophages expressed both CCR2 and CX3CR1. Morphometrically, the average of CCR2- and CX3CR1-positive macrophages was significantly increased in sepsis group, compared with control group (sepsis vs. control: 41.6+/-1.3% vs. 8.0+/-0.4% in CCR2; 36.2+/-1.3% vs. 9.2+/-0.4% in CX3CR1). These observations implied that CCR2- or CX3CR1-positive macrophages were recruited into the lungs under several pathological conditions. In particular, their recruitment might be more evident in sepsis. Moreover, from the forensic aspects, immunohistochemical detection of CCR2 and CX3CR1 expression in the lungs can be considered as valuable diagnostic tools for the postmortem diagnosis of sepsis.

  10. Identification of HIV-1 specific T lymphocyte responses in highly exposed persistently seronegative Chinese

    Institute of Scientific and Technical Information of China (English)

    LIU Hong-wei; SHAO Yi-ming; HONG Kun-xue; MA Jun; YUAN Lin; LIU Sha; CHEN Jian-ping; ZHANG Yuan-zhi; RUAN Yu-hua; XU Jian-qing


    Background Studies of highly exposed persistently seronegative (HEPS) individuals may provide valuable information on mechanisms of protection and on vaccine design. Cellular immune responses play a critical role in containing human immunodeficiency virus. However, the cellular immune responses in HEPS individuals have not been thoroughly assessed at the entire viral genome level.Methods Ten HEPS Chinese with a history of frequent penetrative vaginal intercourse (mean frequency, at least once a week), with some unprotected sexual contact occurring in the weeks or days immediately before enrollment, 25 HIV-1 seropositive individuals, 10 HIV-1-seronegative healthy individuals with low-risk sexual behavior and no history suggestive of exposure to HIV-1 infection were enrolled. HIV-1-specific T cell responses were comprehensively analyzed by an interferon- γ Elispot assay against 770 overlapping peptides spanning all HIV-1 proteins.Results HIV-1-specific T-cell responses of interferon- γ secretion were identified in 3 (30%) out of 10 HEPS individuals; the specific cytotoxic T lymphocytes were targeted at Pol (2/10), Env (2/10), and Tat (1/10).HIV-1-specific T-cell responses of interferon- γ secretion were identified in 20 (80%) out of 25 seropositive intravenous drug users (IDUs), revealing that all HIV-1 proteins and protein subunits could serve as targets for HIV-1-specific CD8+ T cell responses with 85% recognizing Gag, 80% recognizing Nef, 75% recognizing Pol,60% recognizing Env, 55% recognizing Vpu, 45% recognizing Vpr, 20% recognizing Vif, 20% recognizing Tat and 15% recognizing Rev in these seropositive individuals. None of the seronegative healthy individuals gave the positive T-cell responses.Conclusions About 30% of HEPS Chinese mounted HIV-1 specific T cell immune responses. Cell-mediated immunity against HIV-1 may be developed through non-productive infections.

  11. A Highly Photostable Hyperbranched Polyglycerol-Based NIR Fluorescence Nanoplatform for Mitochondria-Specific Cell Imaging. (United States)

    Dong, Chunhong; Liu, Zhongyun; Liu, Junqing; Wu, Changzhu; Neumann, Falko; Wang, Hanjie; Schäfer-Korting, Monika; Kleuser, Burkhard; Chang, Jin; Li, Wenzhong; Ma, Nan; Haag, Rainer


    Considering the critical role of mitochondria in the life and death of cells, non-invasive long-term tracking of mitochondria has attracted considerable interest. However, a high-performance mitochondria-specific labeling probe with high photostability is still lacking. Herein a highly photostable hyperbranched polyglycerol (hPG)-based near-infrared (NIR) quantum dots (QDs) nanoplatform is reported for mitochondria-specific cell imaging. Comprising NIR Zn-Cu-In-S/ZnS QDs as extremely photostable fluorescent labels and alkyl chain (C12 )/triphenylphosphonium (TPP)-functionalized hPG derivatives as protective shell, the tailored QDs@hPG-C12 /TPP nanoprobe with a hydrodynamic diameter of about 65 nm exhibits NIR fluorescence, excellent biocompatibility, good stability, and mitochondria-targeted ability. Cell uptake experiments demonstrate that QDs@hPG-C12 /TPP displays a significantly enhanced uptake in HeLa cells compared to nontargeted QDs@hPG-C12 . Further co-localization study indicates that the probe selectively targets mitochondria. Importantly, compared with commercial deep-red mitochondria dyes, QDs@hPG-C12 /TPP possesses superior photostability under continuous laser irradiation, indicating great potential for long-term mitochondria labeling and tracking. Moreover, drug-loaded QDs@hPG-C12 /TPP display an enhanced tumor cell killing efficacy compared to nontargeted drugs. This work could open the door to the construction of organelle-targeted multifunctional nanoplatforms for precise diagnosis and high-efficient tumor therapy.

  12. An RNA-based signature enables high specificity detection of circulating tumor cells in hepatocellular carcinoma. (United States)

    Kalinich, Mark; Bhan, Irun; Kwan, Tanya T; Miyamoto, David T; Javaid, Sarah; LiCausi, Joseph A; Milner, John D; Hong, Xin; Goyal, Lipika; Sil, Srinjoy; Choz, Melissa; Ho, Uyen; Kapur, Ravi; Muzikansky, Alona; Zhang, Huidan; Weitz, David A; Sequist, Lecia V; Ryan, David P; Chung, Raymond T; Zhu, Andrew X; Isselbacher, Kurt J; Ting, David T; Toner, Mehmet; Maheswaran, Shyamala; Haber, Daniel A


    Circulating tumor cells (CTCs) are shed into the bloodstream by invasive cancers, but the difficulty inherent in identifying these rare cells by microscopy has precluded their routine use in monitoring or screening for cancer. We recently described a high-throughput microfluidic CTC-iChip, which efficiently depletes hematopoietic cells from blood specimens and enriches for CTCs with well-preserved RNA. Application of RNA-based digital PCR to detect CTC-derived signatures may thus enable highly accurate tissue lineage-based cancer detection in blood specimens. As proof of principle, we examined hepatocellular carcinoma (HCC), a cancer that is derived from liver cells bearing a unique gene expression profile. After identifying a digital signature of 10 liver-specific transcripts, we used a cross-validated logistic regression model to identify the presence of HCC-derived CTCs in nine of 16 (56%) untreated patients with HCC versus one of 31 (3%) patients with nonmalignant liver disease at risk for developing HCC (P digital CTC scoring was not correlated with the standard HCC serum protein marker alpha fetoprotein (P = 0.57). Modeling the sequential use of these two orthogonal markers for liver cancer screening in patients with high-risk cirrhosis generates positive and negative predictive values of 80% and 86%, respectively. Thus, digital RNA quantitation constitutes a sensitive and specific CTC readout, enabling high-throughput clinical applications, such as noninvasive screening for HCC in populations where viral hepatitis and cirrhosis are prevalent.

  13. Maltodextrin-based imaging probes detect bacteria in vivo with high sensitivity and specificity (United States)

    Ning, Xinghai; Lee, Seungjun; Wang, Zhirui; Kim, Dongin; Stubblefield, Bryan; Gilbert, Eric; Murthy, Niren


    The diagnosis of bacterial infections remains a major challenge in medicine. Although numerous contrast agents have been developed to image bacteria, their clinical impact has been minimal because they are unable to detect small numbers of bacteria in vivo, and cannot distinguish infections from other pathologies such as cancer and inflammation. Here, we present a family of contrast agents, termed maltodextrin-based imaging probes (MDPs), which can detect bacteria in vivo with a sensitivity two orders of magnitude higher than previously reported, and can detect bacteria using a bacteria-specific mechanism that is independent of host response and secondary pathologies. MDPs are composed of a fluorescent dye conjugated to maltohexaose, and are rapidly internalized through the bacteria-specific maltodextrin transport pathway, endowing the MDPs with a unique combination of high sensitivity and specificity for bacteria. Here, we show that MDPs selectively accumulate within bacteria at millimolar concentrations, and are a thousand-fold more specific for bacteria than mammalian cells. Furthermore, we demonstrate that MDPs can image as few as 105 colony-forming units in vivo and can discriminate between active bacteria and inflammation induced by either lipopolysaccharides or metabolically inactive bacteria.

  14. Two strains of Crocosphaera watsonii with highly conserved genomes are distinguished by strain-specific features

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    Shellie Roxanne Bench


    Full Text Available Unicellular nitrogen-fixing cyanobacteria are important components of marine phytoplankton. Although non-nitrogen-fixing marine phytoplankton generally exhibit high gene sequence and genomic diversity, gene sequences of natural populations and isolated strains of Crocosphaera watsonii, one of two most abundant open ocean unicellular cyanobacteria groups, have been shown to be 98-100% identical.. The low sequence diversity in Crocosphaera is a dramatic contrast to sympatric species of Prochlorococcus and Synechococcus, and raises the question of how genome differences can explain observed phenotypic diversity among Crocosphaera strains. Here we show, through whole genome comparisons of two phenotypically different strains, that there are strain-specific sequences in each genome, and numerous genome rearrangements, despite exceptionally low sequence diversity in shared genomic regions. Some of the strain-specific sequences encode functions that explain observed phenotypic differences, such as exopolysaccharide biosynthesis. The pattern of strain-specific sequences distributed throughout the genomes, along with rearrangements in shared sequences is evidence of significant genetic mobility that may be attributed to the hundreds of transposase genes found in both strains. Furthermore, such genetic mobility appears to be the main mechanism of strain divergence in Crocosphaera which do not accumulate DNA microheterogeneity over the vast majority of their genomes. The strain-specific sequences found in this study provide tools for future physiological studies, as well as genetic markers to help determine the relative abundance of phenotypes in natural populations.

  15. Highly Specific Detection of Five Exotic Quarantine Plant Viruses using RT-PCR

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    Hoseong Choi


    Full Text Available To detect five plant viruses (Beet black scorch virus, Beet necrotic yellow vein virus, Eggplant mottled dwarf virus, Pelargonium zonate spot virus, and Rice yellow mottle virus for quarantine purposes, we designed 15 RT-PCR primer sets. Primer design was based on the nucleotide sequence of the coat protein gene, which is highly conserved within species. All but one primer set successfully amplified the targets, and gradient PCRs indicated that the optimal temperature for the 14 useful primer sets was 51.9°C. Some primer sets worked well regardless of annealing temperature while others required a very specific annealing temperature. A primer specificity test using plant total RNAs and cDNAs of other plant virus-infected samples demonstrated that the designed primer sets were highly specific and generated reproducible results. The newly developed RT-PCR primer sets would be useful for quarantine inspections aimed at preventing the entry of exotic plant viruses into Korea.

  16. Neurogenic bladder: Highly selective rhizotomy of specific dorsal rootlets maybe a better choice. (United States)

    Zhu, Genying; Zhou, Mouwang; Wang, Wenting; Zeng, Fanshuo


    Spinal cord injury results not only in motor and sensory dysfunctions, but also in loss of normal urinary bladder functions. A number of clinical studies were focused on the strategies for improvement of functions of the bladder. Completely dorsal root rhizotomy or selective specific S2-4 dorsal root rhizotomy suppress autonomic hyper-reflexia but have the same defects: it could cause detrusor and sphincter over-relaxation and loss of reflexive erection in males. So precise operation needs to be considered. We designed an experimental trail to test the possibility on the basis of previous study. We found that different dorsal rootlets which conduct impulses from the detrusor or sphincter can be distinguished by electro-stimulation in SD rats. Highly selective rhizotomy of specific dorsal rootlets could change the intravesical pressure and urethral perfusion pressure respectively. We hypothese that for neurogenic bladder following spinal cord injury, highly selective rhizotomy of specific dorsal rootlets maybe improve the bladder capacity and the detrusor sphincter dyssynergia, and at the same time, the function of other pelvic organ could be maximize retainment.

  17. Experiences with the hydraulic design of the high specific speed Francis turbine (United States)

    Obrovsky, J.; Zouhar, J.


    The high specific speed Francis turbine is still suitable alternative for refurbishment of older hydro power plants with lower heads and worse cavitation conditions. In the paper the design process of such kind of turbine together with the results comparison of homological model tests performed in hydraulic laboratory of ČKD Blansko Engineering is introduced. The turbine runner was designed using the optimization algorithm and considering the high specific speed hydraulic profile. It means that hydraulic profiles of the spiral case, the distributor and the draft tube were used from a Kaplan turbine. The optimization was done as the automatic cycle and was based on a simplex optimization method as well as on a genetic algorithm. The number of blades is shown as the parameter which changes the resulting specific speed of the turbine between ns=425 to 455 together with the cavitation characteristics. Minimizing of cavitation on the blade surface as well as on the inlet edge of the runner blade was taken into account during the design process. The results of CFD analyses as well as the model tests are mentioned in the paper.

  18. Identification of an epigenetic biomarker panel with high sensitivity and specificity for colorectal cancer and adenomas

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    Lind Guro E


    Full Text Available Abstract Background The presence of cancer-specific DNA methylation patterns in epithelial colorectal cells in human feces provides the prospect of a simple, non-invasive screening test for colorectal cancer and its precursor, the adenoma. This study investigates a panel of epigenetic markers for the detection of colorectal cancer and adenomas. Methods Candidate biomarkers were subjected to quantitative methylation analysis in test sets of tissue samples from colorectal cancers, adenomas, and normal colonic mucosa. All findings were verified in independent clinical validation series. A total of 523 human samples were included in the study. Receiver operating characteristic (ROC curve analysis was used to evaluate the performance of the biomarker panel. Results Promoter hypermethylation of the genes CNRIP1, FBN1, INA, MAL, SNCA, and SPG20 was frequent in both colorectal cancers (65-94% and adenomas (35-91%, whereas normal mucosa samples were rarely (0-5% methylated. The combined sensitivity of at least two positives among the six markers was 94% for colorectal cancers and 93% for adenoma samples, with a specificity of 98%. The resulting areas under the ROC curve were 0.984 for cancers and 0.968 for adenomas versus normal mucosa. Conclusions The novel epigenetic marker panel shows very high sensitivity and specificity for both colorectal cancers and adenomas. Our findings suggest this biomarker panel to be highly suitable for early tumor detection.

  19. The effects of varying resistance-training loads on intermediate- and high-velocity-specific adaptations. (United States)

    Jones, K; Bishop, P; Hunter, G; Fleisig, G


    The purpose of this study was to compare changes in velocity-specific adaptations in moderately resistance-trained athletes who trained with either low or high resistances. The study used tests of sport-specific skills across an intermediate- to high-velocity spectrum. Thirty NCAA Division I baseball players were randomly assigned to either a low-resistance (40-60% 1 repetition maximum [1RM]) training group or a high-resistance (70-90% 1RM) training group. Both of the training groups intended to maximallv accelerate each repetition during the concentric phase (IMCA). The 10 weeks of training consisted of 4 training sessions a week using basic core exercises. Peak force, velocity, and power were evaluated during set angle and depth jumps as well as weighted jumps using 30 and 50% 1RM. Squat 1RMs were also tested. Although no interactions for any of the jump tests were found, trends supported the hypothesis of velocity-specific training. Percentage gains suggest that the combined use of heavier training loads (70-90% 1RM) and IMCA tend to increase peak force in the lower-body leg and hip extensors. Trends also show that the combined use of lighter training loads (40-60% 1RM) and IMCA tend to increase peak power and peak velocity in the lower-body leg and hip extensors. The high-resistance group improved squats more than the low-resistance group (p IMCA to increase 1RM strength in the lower bodies of resistance-trained athletes.

  20. Record-high specific conductance and water temperature in San Francisco Bay during water year 2015 (United States)

    Work, Paul; Downing-Kunz, Maureen; Livsey, Daniel


    The San Francisco estuary is commonly defined to include San Francisco Bay (bay) and the adjacent Sacramento–San Joaquin River Delta (delta). The U.S. Geological Survey (USGS) has operated a high-frequency (15-minute sampling interval) water-quality monitoring network in San Francisco Bay since the late 1980s (Buchanan and others, 2014). This network includes 19 stations at which sustained measurements have been made in the bay; currently, 8 stations are in operation (fig. 1). All eight stations are equipped with specific conductance (which can be related to salinity) and water-temperature sensors. Water quality in the bay constantly changes as ocean tides force seawater in and out of the bay, and river inflows—the most significant coming from the delta—vary on time scales ranging from those associated with storms to multiyear droughts. This monitoring network was designed to observe and characterize some of these changes in the bay across space and over time. The data demonstrate a high degree of variability in both specific conductance and temperature at time scales from tidal to annual and also reveal longer-term changes that are likely to influence overall environmental health in the bay.In water year (WY) 2015 (October 1, 2014, through September 30, 2015), as in the preceding water year (Downing-Kunz and others, 2015), the high-frequency measurements revealed record-high values of specific conductance and water temperature at several stations during a period of reduced freshwater inflow from the delta and other tributaries because of persistent, severe drought conditions in California. This report briefly summarizes observations for WY 2015 and compares them to previous years that had different levels of freshwater inflow.

  1. Endophytic Fungal Communities Associated with Vascular Plants in the High Arctic Zone Are Highly Diverse and Host-Plant Specific.

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    Tao Zhang

    Full Text Available This study assessed the diversity and distribution of endophytic fungal communities associated with the leaves and stems of four vascular plant species in the High Arctic using 454 pyrosequencing with fungal-specific primers targeting the ITS region. Endophytic fungal communities showed high diversity. The 76,691 sequences obtained belonged to 250 operational taxonomic units (OTUs. Of these OTUs, 190 belonged to Ascomycota, 50 to Basidiomycota, 1 to Chytridiomycota, and 9 to unknown fungi. The dominant orders were Helotiales, Pleosporales, Capnodiales, and Tremellales, whereas the common known fungal genera were Cryptococcus, Rhizosphaera, Mycopappus, Melampsora, Tetracladium, Phaeosphaeria, Mrakia, Venturia, and Leptosphaeria. Both the climate and host-related factors might shape the fungal communities associated with the four Arctic plant species in this region. These results suggested the presence of an interesting endophytic fungal community and could improve our understanding of fungal evolution and ecology in the Arctic terrestrial ecosystems.

  2. Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity

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    Wang, Deng-Liang [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China); Song, Yan-Ling; Zhu, Zhi; Li, Xi-Lan; Zou, Yuan [State Key Laboratory for Physical Chemistry of Solid Surfaces, Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Chemistry, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005 (China); Yang, Hai-Tao; Wang, Jiang-Jie [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Yao, Pei-Sen [Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China); Pan, Ru-Jun [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Yang, Chaoyong James, E-mail: [State Key Laboratory for Physical Chemistry of Solid Surfaces, Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Chemistry, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005 (China); Kang, De-Zhi, E-mail: [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China)


    Highlights: • This is the first report of DNA aptamer against EGFR in vitro. • Aptamer can bind targets with high affinity and selectivity. • DNA aptamers are more stable, cheap and efficient than RNA aptamers. • Our selected DNA aptamer against EGFR has high affinity with K{sub d} 56 ± 7.3 nM. • Our selected DNA aptamer against EGFR has high selectivity. - Abstract: Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher’s attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with K{sub d} 56 ± 7.3 nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy.

  3. Immunohistochemical detections of EGFR status in NSCLC

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    Jie WANG


    Full Text Available Background and objective Patients with non-small cell lung cancer (NSCLC harboring mutations of the epidermal growth factor receptor (EGFR respond well to EGFR-tyrosine kinase inhibitor therapy. Immunohistochemistry (IHC is a simple and widely used technique in clinical pathology laboratories. IHC also features cost effectiveness and rapid detection of EGFR mutations compared with molecular methods. This study aims to determine the accuracy of IHC for EGFR mutation detection in NSCLC. Methods Specimens (obtained from surgery or biopsy from 97 NSCLC cases were stained through IHC with mutation-specific antibodies. The clinicopathological features of patients with positive immunostaining results were analyzed. Positive specimens were subjected to liquid chip technology to detect the actual EGFR status. Forty NSCLC specimens obtained from surgery and confirmed to have EGFR mutations through liquid chip technology were collected. These specimens were then subjected to IHC analyses with mutation-specific antibodies. The sensitivity of IHC in detecting EGFR mutations was calculated. Results Seventeen of the 97 NSCLC specimens were stained positive, and positive results were mostly observed in females, patients with adenocarcinoma, and non-smokers. About 76.9% of specimens with positive IHC results harbored mutations. The sensitivity of IHC was 40% among the 40 cases identified as containing EGFR mutations through liquid chip technology. Conclusion The strong positive immunostaining result is accurate, but the sensitivity of the method may not be optimal and significantly varies in different studies. The widespread application of IHC in clinics must be further investigated.

  4. Detection of pork adulteration by highly-specific PCR assay of mitochondrial D-loop. (United States)

    Karabasanavar, Nagappa S; Singh, S P; Kumar, Deepak; Shebannavar, Sunil N


    We describe a highly specific PCR assay for the authentic identification of pork. Accurate detection of tissues derived from pig (Sus scrofa) was accomplished by using newly designed primers targeting porcine mitochondrial displacement (D-loop) region that yielded an unique amplicon of 712 base pairs (bp). Possibility of cross-amplification was precluded by testing as many as 24 animal species (mammals, birds, rodent and fish). Suitability of PCR assay was confirmed in raw (n = 20), cooked (60, 80 and 100 °C), autoclaved (121 °C) and micro-oven processed pork. Sensitivity of detection of pork in other species meat using unique pig-specific PCR was established to be at 0.1%; limit of detection (LOD) of pig DNA was 10 pg (pico grams). The technique can be used for the authentication of raw, processed and adulterated pork and products under the circumstances of food adulteration related disputes or forensic detection of origin of pig species.

  5. A new ethylene glycol-silane monolayer for highly-specific DNA detection on Silicon Chips (United States)

    Carrara, Sandro; Cavallini, Andrea; Maruyama, Yuki; Charbon, Edoardo; De Micheli, Giovanni


    Monolayer thin films with ethylene-glycol function onto gold surfaces by using thiols have been extensively investigated. They have been proposed as precursors for applications to bio-detection, where their hydrophilic character improves both specificity and sensitivity. The aim of this letter is to characterize ethylene-glycol monolayer precursors formed onto silicon chips by using silanes. The importance of the ethylene-glycol function is demonstrated by comparing with the well known 3-Aminopropyltriethoxysilane. The different nano-scale structures of the two precursor monolayers are investigated by using atomic force microscopy (AFM). Longer, wider, and deeper grooves were measured in the images acquired on 3-Aminopropyltriethoxysilane. Fluorescence investigation demonstrates that the presence of ethylene-glycol function improves target hybridization onto silicon chips, assuring highly-specific detection of DNA.

  6. Polarity-specific high-level information propagation in neural networks. (United States)

    Lin, Yen-Nan; Chang, Po-Yen; Hsiao, Pao-Yueh; Lo, Chung-Chuan


    Analyzing the connectome of a nervous system provides valuable information about the functions of its subsystems. Although much has been learned about the architectures of neural networks in various organisms by applying analytical tools developed for general networks, two distinct and functionally important properties of neural networks are often overlooked. First, neural networks are endowed with polarity at the circuit level: Information enters a neural network at input neurons, propagates through interneurons, and leaves via output neurons. Second, many functions of nervous systems are implemented by signal propagation through high-level pathways involving multiple and often recurrent connections rather than by the shortest paths between nodes. In the present study, we analyzed two neural networks: the somatic nervous system of Caenorhabditis elegans (C. elegans) and the partial central complex network of Drosophila, in light of these properties. Specifically, we quantified high-level propagation in the vertical and horizontal directions: the former characterizes how signals propagate from specific input nodes to specific output nodes and the latter characterizes how a signal from a specific input node is shared by all output nodes. We found that the two neural networks are characterized by very efficient vertical and horizontal propagation. In comparison, classic small-world networks show a trade-off between vertical and horizontal propagation; increasing the rewiring probability improves the efficiency of horizontal propagation but worsens the efficiency of vertical propagation. Our result provides insights into how the complex functions of natural neural networks may arise from a design that allows them to efficiently transform and combine input signals.

  7. Clinical implications for biochemical diagnostic thresholds of adrenal sufficiency using a highly specific cortisol immunoassay. (United States)

    Kline, G A; Buse, J; Krause, R D


    Recent guidelines recommend a diagnosis of adrenal insufficiency when a stimulated peak cortisol level falls below 500nmol/L. This may not be valid using a highly specific cortisol immunoassay or liquid chromatography-mass spectroscopy (LCMS/MS). We sought to determine the diagnostic threshold for adrenal insufficiency using a new and widely available, highly specific cortisol immunoassay. All patients having a dynamic test of adrenal reserve had results measured using the historical cortisol assay (Roche Cortisol) and the newer assay (Roche Cortisol II). Subjects were categorized according to the traditional assay (normal>500nmol/L) with clinical case adjudication where necessary. Results from Cortisol II assay were concomitantly measured along with cortisol levels measured by LCMS/MS. ROC curve analysis was performed to generate new diagnostic thresholds. The Roche Cortisol II compared favourably with measures by LCMS/MS, generating cortisol levels approximately 30% lower than the older immunoassay. Many normal subjects had peak cortisols as low as 300nmol/L with Cortisol II. The optimized diagnostic threshold for adrenal insufficiency was 350nmol/L with a sensitivity of 91% and specificity 97%. Use of the old diagnostic threshold with the Cortisol II assay would have inappropriately doubled the rate of patient-classification as adrenal insufficient. Transition to a more specific cortisol assay requires revision of diagnostic thresholds for dynamic tests of adrenal insufficiency. With the Roche Cortisol II assay, a cut-off of 350nmol/L should replace the traditional 500nmol/L although some healthy subjects may be very close to this level. Copyright © 2017 The Canadian Society of Clinical Chemists. All rights reserved.

  8. Whole cell-SELEX aptamers for highly specific fluorescence molecular imaging of carcinomas in vivo.

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    Hui Shi

    Full Text Available BACKGROUND: Carcinomas make up the majority of cancers. Their accurate and specific diagnoses are of great significance for the improvement of patients' curability. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we report an effectual example of the in vivo fluorescence molecular imaging of carcinomas with extremely high specificity based on whole cell-SELEX aptamers. Firstly, S6, an aptamer against A549 lung carcinoma cells, was adopted and labeled with Cy5 to serve as a molecular imaging probe. Flow cytometry assays revealed that Cy5-S6 could not only specifically label in vitro cultured A549 cells in buffer, but also successfully achieve the detection of ex vivo cultured target cells in serum. When applied to in vivo imaging, Cy5-S6 was demonstrated to possess high specificity in identifying A549 carcinoma through a systematic comparison investigation. Particularly, after Cy5-S6 was intravenously injected into nude mice which were simultaneously grafted with A549 lung carcinoma and Tca8113 tongue carcinoma, a much longer retention time of Cy5-S6 in A549 tumor was observed and a clear targeted cancer imaging result was presented. On this basis, to further promote the application to imaging other carcinomas, LS2 and ZY8, which are two aptamers selected by our group against Bel-7404 and SMMC-7721 liver carcinoma cells respectively, were tested in a similar way, both in vitro and in vivo. Results showed that these aptamers were even effective in differentiating liver carcinomas of different subtypes in the same body. CONCLUSIONS/SIGNIFICANCE: This work might greatly advance the application of whole cell-SELEX aptamers to carcinomas-related in vivo researches.

  9. Common and specific brain regions in high- versus low-confidence recognition memory (United States)

    Kim, Hongkeun; Cabeza, Roberto


    The goal of the present functional magnetic resonance imaging (fMRI) study was to investigate whether and to what extent brain regions involved in high-confidence recognition (HCR) versus low-confidence recognition (LCR) overlap or separate from each other. To this end, we performed conjunction analyses involving activations elicited during high-confidence hit, low-confidence hit, and high-confidence correct-rejection responses. The analyses yielded 3 main findings. First, sensory/perceptual and associated posterior regions were common to HCR and LCR, indicating contribution of these regions to both HCR and LCR activity. This finding may help explain why these regions are among the most common in functional neuroimaging studies of episodic retrieval. Second, medial temporal lobe (MTL) and associated midline regions were associated with HCR, possibly reflecting recollection-related processes, whereas specific prefrontal cortex (PFC) regions were associated with LCR, possibly reflecting executive control processes. This finding is consistent with the notion that the MTL and PFC networks play complementary roles during episodic retrieval. Finally, within posterior parietal cortex, a dorsal region was associated with LCR, possibly reflecting top-down attentional processes, whereas a ventral region was associated with HCR, possibly reflecting bottom-up attentional processes. This finding may help explain why functional neuroimaging studies have found diverse parietal effects during episodic retrieval. Taken together, our findings provide strong evidence that HCR versus LCR, and by implication, recollection versus familiarity processes, are represented in common as well as specific brain regions. PMID:19501072

  10. Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity. (United States)

    Wang, Deng-Liang; Song, Yan-Ling; Zhu, Zhi; Li, Xi-Lan; Zou, Yuan; Yang, Hai-Tao; Wang, Jiang-Jie; Yao, Pei-Sen; Pan, Ru-Jun; Yang, Chaoyong James; Kang, De-Zhi


    Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher's attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with Kd 56±7.3nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy.

  11. Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. (United States)

    Tsai, Shengdar Q; Wyvekens, Nicolas; Khayter, Cyd; Foden, Jennifer A; Thapar, Vishal; Reyon, Deepak; Goodwin, Mathew J; Aryee, Martin J; Joung, J Keith


    Monomeric CRISPR-Cas9 nucleases are widely used for targeted genome editing but can induce unwanted off-target mutations with high frequencies. Here we describe dimeric RNA-guided FokI nucleases (RFNs) that can recognize extended sequences and edit endogenous genes with high efficiencies in human cells. RFN cleavage activity depends strictly on the binding of two guide RNAs (gRNAs) to DNA with a defined spacing and orientation substantially reducing the likelihood that a suitable target site will occur more than once in the genome and therefore improving specificities relative to wild-type Cas9 monomers. RFNs guided by a single gRNA generally induce lower levels of unwanted mutations than matched monomeric Cas9 nickases. In addition, we describe a simple method for expressing multiple gRNAs bearing any 5' end nucleotide, which gives dimeric RFNs a broad targeting range. RFNs combine the ease of RNA-based targeting with the specificity enhancement inherent to dimerization and are likely to be useful in applications that require highly precise genome editing.

  12. Pyrosequencing reveals highly diverse and species-specific microbial communities in sponges from the Red Sea

    KAUST Repository

    Lee, Onon


    Marine sponges are associated with a remarkable array of microorganisms. Using a tag pyrosequencing technology, this study was the first to investigate in depth the microbial communities associated with three Red Sea sponges, Hyrtios erectus, Stylissa carteri and Xestospongia testudinaria. We revealed highly diverse sponge-associated bacterial communities with up to 1000 microbial operational taxonomic units (OTUs) and richness estimates of up to 2000 species. Altogether, 26 bacterial phyla were detected from the Red Sea sponges, 11 of which were absent from the surrounding sea water and 4 were recorded in sponges for the first time. Up to 100 OTUs with richness estimates of up to 300 archaeal species were revealed from a single sponge species. This is by far the highest archaeal diversity ever recorded for sponges. A non-negligible proportion of unclassified reads was observed in sponges. Our results demonstrated that the sponge-associated microbial communities remained highly consistent in the same sponge species from different locations, although they varied at different degrees among different sponge species. A significant proportion of the tag sequences from the sponges could be assigned to one of the sponge-specific clusters previously defined. In addition, the sponge-associated microbial communities were consistently divergent from those present in the surrounding sea water. Our results suggest that the Red Sea sponges possess highly sponge-specific or even sponge-species-specific microbial communities that are resistant to environmental disturbance, and much of their microbial diversity remains to be explored. © 2011 International Society for Microbial Ecology All rights reserved.

  13. The triazine-based porous organic polymer: Novel synthetic strategy for high specific surface area

    Energy Technology Data Exchange (ETDEWEB)

    Park, Jin Kuen [Dept. of Chemistry, Hankuk University of Foreign Studies, Yongin (Korea, Republic of)


    A new type of microporous polymer has been successively synthesized via a simple polycondensation reaction with the 2,4-diaminotriazine moiety and dianhydride monomer. Diaminotriazine moieties in M1 especially can provide effective branching sites, resulting in high surface areas up to 1150 m{sup 2} /g. In addition, the specific pore structure of the polyimide POP in its solid state can be modified by the surface activation method. Therefore, it can be expected that the resulting material will be a promising candidate for gas storage, and with this synthetic strategy, various type of derivatives will also be optimized.

  14. Facile syntheses of 3-dimension graphene aerogel and nanowalls with high specific surface areas (United States)

    Zhou, Lina; Yang, Zhongbo; Yang, Jun; Wu, Yonggang; Wei, Dongshan


    We propose facile synthesis methods to prepare two three-dimension (3D) multi-porous graphene structures including graphene aerogel and graphene nanowalls. The graphene aerogel was prepared via the hydrothermal reduction and tert-butanol freeze-drying. The graphene nanowalls were prepared by growing the obtained graphene aerogel via a plasma enhanced chemical vapor deposition (PECVD). High specific surface areas of these two structures up to 795 m2/g were obtained by BET analyses. The crystallized, multi-porous, and thermal stable features of these two 3D graphene structures were verified via the X-ray diffraction, SEM, Raman spectroscopy and thermal gravity analysis characterizations.

  15. Production study of high specific activity NCA Re-186g by proton and deuteron cyclotron irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Bonardi, M.L., E-mail: mauro.bonardi@mi.infn.i [L.A.S.A., Radiochemistry Laboratory, Universita degli Studi di Milano, UNIMI and Istituto Nazionale di Fisica Nucleare, INFN, Via F.lli Cervi 201, I-20090 Segrate (Italy); Groppi, F.; Manenti, S.; Persico, E.; Gini, L. [L.A.S.A., Radiochemistry Laboratory, Universita degli Studi di Milano, UNIMI and Istituto Nazionale di Fisica Nucleare, INFN, Via F.lli Cervi 201, I-20090 Segrate (Italy)


    Very high specific activity (A{sub S}) {sup 186g}Re could be produced by either proton or deuteron cyclotron irradiation on highly enriched {sup 186}W target in no-carrier-added (NCA) form, leading to a A{sub S} very close to the theoretical carrier free (CF) value of 6.88 GBq {mu}g{sup -1}. Thick target yields (TTYs), obtained irradiating both thick metal W targets of natural isotopic composition and highly enriched pressed powdered {sup 186}W targets, were measured at different particles energies taking into account high accuracy and precision on both yield and beam energy. The measurement of radionuclidic purity of {sup 186g}Re obtained activating highly enriched {sup 186}W by both p and d beams were also carried out and accurately compared. The excitation function as thin-target yields (tty, i.e. proportional to the reaction cross-sections) and the integrated TTYs for all Re (A=181, 182, 183, 184, 186 and their metastable levels), W and Ta co-produced radionuclides will be presented elsewhere in deep details.

  16. Leukocyte chemotactic factor 2 amyloidosis cannot be reliably diagnosed by immunohistochemical staining. (United States)

    Paueksakon, Paisit; Fogo, Agnes B; Sethi, Sanjeev


    We investigated the role of leukocyte chemotactic factor (LECT2) immunohistochemical staining in the diagnosis of type of renal amyloidosis. Fifty renal amyloidosis cases with available paraffin blocks in our 2002 to 2012 renal biopsy files were reviewed. Patients were designated as a defined amyloid, including amyloid light chain (AL) and amyloid-associated amyloid (AA), or a non-AL/non-AA amyloid group. LECT2-specific antibody immunohistochemistry was performed in all 50 cases. Laser microdissection and mass spectrometry (LMD/MS) were performed in 10 cases. Forty-five patients had amyloid classified as either AL (44) or AA (1), and 5 had undetermined amyloid. Three of the five non-AL/non-AA group patient biopsies showed positive LECT2 immunohistochemical staining, and of these, LECT2 was also identified by LMD/MS in 1 patient, fibrinogen-α was identified in 1 patient, and apolipoprotein IV was identified in 1 patient. Two of these non-AL/non-AA patients showed negative LECT2 staining, and LMD/MS showed apolipoprotein IV as a major protein component. Five of the 44 AL amyloid patients showed weakly positive LECT2 staining. However, LECT2 was not identified by LMD/MS in any of these 5 cases. The single patient with AA amyloid was negative for LECT2 by immunohistochemical staining. Among 5 non-AL and non-AA amyloidosis patients in our study, 1 had LECT2, 1 had fibrinogen-α, and 3 had apolipoprotein IV as a major protein component. The data from this study show that weak LECT2 staining should be regarded as indeterminate or a negative result and does not per se allow diagnosis of specific amyloid type. The diagnosis of LECT2 renal amyloidosis may require LMD/MS confirmation.

  17. An improved quantitative mass spectrometry analysis of tumor specific mutant proteins at high sensitivity. (United States)

    Ruppen-Cañás, Isabel; López-Casas, Pedro P; García, Fernando; Ximénez-Embún, Pilar; Muñoz, Manuel; Morelli, M Pia; Real, Francisco X; Serna, Antonio; Hidalgo, Manuel; Ashman, Keith


    New disease specific biomarkers, especially for cancer, are urgently needed to improve individual diagnosis, prognosis, and treatment selection, that is, for personalized medicine. Genetic mutations that affect protein function drive cancer. Therefore, the detection of such mutations represents a source of cancer specific biomarkers. Here we confirm the implementation of the mutant protein specific immuno-SRM (where SRM is selective reaction monitoring) mass spectrometry method of RAS proteins reported by Wang et al. [Proc. Natl. Acad. Sci. USA 2011, 108, 2444-2449], which exploits an antibody to simultaneously capture the different forms of the target protein and the resolving power and sensitivity of LC-MS/MS and improve the technique by using a more sensitive mass spectrometer. The mutant form G12D was quantified by SRM on a QTRAP 5500 mass spectrometer and the MIDAS workflow was used to confirm the sequence of the targeted peptides. This assay has been applied to quantify wild type and mutant RAS proteins in patient tumors, xenografted human tissue, and benign human epidermal tumors at high sensitivity. The limit of detection for the target proteins was as low as 12 amol (0.25 pg). It requires low starting amounts of tissue (ca.15 mg) that could be obtained from a needle aspiration biopsy. The described strategy could find application in the clinical arena and be applied to the study of expression of protein variants in disease.

  18. Knee and hip joint biomechanics are gender-specific in runners with high running mileage. (United States)

    Gehring, D; Mornieux, G; Fleischmann, J; Gollhofer, A


    Female runners are reported to be more prone to develop specific knee joint injuries than males. It has been suggested that increased frontal plane joint loading might be related to the incidence of these knee injuries in running. The purpose of this study was to evaluate if frontal plane knee and hip joint kinematics and kinetics are gender-specific in runners with high mileage. 3D-kinematics and kinetics were recorded from 16 female and 16 male runners at a speed of 3 m/s, 4 m/s, and 5 m/s. Frontal plane joint angles and joint moments were ascertained and compared between genders among speed conditions. Across all speed conditions, females showed increased hip adduction and reduced knee adduction angles compared to males (p running speed (p = 0.07). Hip and knee frontal plane joint kinematics are gender-specific. In addition, there are indications that frontal plane joint loading is increased in female runners. Future research should focus on the relationship of these observations regarding overuse running injuries.

  19. Highly specific epigenome editing by CRISPR-Cas9 repressors for silencing of distal regulatory elements. (United States)

    Thakore, Pratiksha I; D'Ippolito, Anthony M; Song, Lingyun; Safi, Alexias; Shivakumar, Nishkala K; Kabadi, Ami M; Reddy, Timothy E; Crawford, Gregory E; Gersbach, Charles A


    Epigenome editing with the CRISPR (clustered, regularly interspaced, short palindromic repeats)-Cas9 platform is a promising technology for modulating gene expression to direct cell phenotype and to dissect the causal epigenetic mechanisms of gene regulation. Fusions of nuclease-inactive dCas9 to the Krüppel-associated box (KRAB) repressor (dCas9-KRAB) can silence target gene expression, but the genome-wide specificity and the extent of heterochromatin formation catalyzed by dCas9-KRAB are not known. We targeted dCas9-KRAB to the HS2 enhancer, a distal regulatory element that orchestrates the expression of multiple globin genes, and observed highly specific induction of H3K9 trimethylation (H3K9me3) at the enhancer and decreased chromatin accessibility of both the enhancer and its promoter targets. Targeted epigenetic modification of HS2 silenced the expression of multiple globin genes, with minimal off-target changes in global gene expression. These results demonstrate that repression mediated by dCas9-KRAB is sufficiently specific to disrupt the activity of individual enhancers via local modification of the epigenome.

  20. A highly specific gold nanoprobe for live-cell single-molecule imaging

    CERN Document Server

    Leduc, Cecile; Gautier, Jérémie; Soto-Ribeiro, Martinho; Wehrle-Haller, B; Gautreau, Alexis; Giannone, Gregory; Cognet, Laurent; Lounis, Brahim


    Single molecule tracking in live cells is the ultimate tool to study subcellular protein dynamics, but it is often limited by the probe size and photostability. Due to these issues, long-term tracking of proteins in confined and crowded environments, such as intracellular spaces, remains challenging. We have developed a novel optical probe consisting of 5-nm gold nanoparticles functionalized with a small fragment of camelid antibodies that recognize widely used GFPs with a very high affinity, which we call GFP-nanobodies. These small gold nanoparticles can be detected and tracked using photothermal imaging for arbitrarily long periods of time. Surface and intracellular GFP-proteins were effectively labeled even in very crowded environments such as adhesion sites and cytoskeletal structures both in vitro and in live cell cultures. These nanobody-coated gold nanoparticles are probes with unparalleled capabilities; small size, perfect photostability, high specificity, and versatility afforded by combination with...

  1. Specific-heat measurement of single metallic, carbon, and ceramic fibers at very high temperature (United States)

    Pradère, C.; Goyhénèche, J. M.; Batsale, J. C.; Dilhaire, S.; Pailler, R.


    The main objective of this work is to present a method for measuring the specific heat of single metallic, carbon, and ceramic fibers at very high temperature. The difficulty of the measurement is due to the microscale of the fiber (≈10μm) and the important range of temperature (700-2700K). An experimental device, a modelization of the thermal behavior, and an analytic model have been developed. A discussion on the measurement accuracy yields a global uncertainty lower than 10%. The characterization of a tungsten filament with thermal properties identical to those of the bulk allows the validation of the device and the thermal estimation method. Finally, measurements on carbon and ceramic fibers have been done at very high temperature.

  2. Establishing Specifications for Low Enriched Uranium Fuel Operations Conducted Outside the High Flux Isotope Reactor Site

    Energy Technology Data Exchange (ETDEWEB)

    Pinkston, Daniel [ORNL; Primm, Trent [ORNL; Renfro, David G [ORNL; Sease, John D [ORNL


    The National Nuclear Security Administration (NNSA) has funded staff at Oak Ridge National Laboratory (ORNL) to study the conversion of the High Flux Isotope Reactor (HFIR) from the current, high enriched uranium fuel to low enriched uranium fuel. The LEU fuel form is a metal alloy that has never been used in HFIR or any HFIR-like reactor. This report provides documentation of a process for the creation of a fuel specification that will meet all applicable regulations and guidelines to which UT-Battelle, LLC (UTB) the operating contractor for ORNL - must adhere. This process will allow UTB to purchase LEU fuel for HFIR and be assured of the quality of the fuel being procured.

  3. Standardized methods for the production of high specific-activity zirconium-89

    Energy Technology Data Exchange (ETDEWEB)

    Holland, Jason P. [Memorial Sloan-Kettering Cancer Center, Department of Radiology, New York, NY 10065 (United States)], E-mail:; Sheh Yiauchung [Memorial Sloan-Kettering Cancer Center, Department of Radiology, New York, NY 10065 (United States)], E-mail:; Lewis, Jason S. [Memorial Sloan-Kettering Cancer Center, Department of Radiology, New York, NY 10065 (United States)], E-mail:


    Zirconium-89 is an attractive metallo-radionuclide for use in immuno-PET due to favorable decay characteristics. Standardized methods for the routine production and isolation of high-purity and high-specific-activity {sup 89}Zr using a small cyclotron are reported. Optimized cyclotron conditions reveal high average yields of 1.52{+-}0.11 mCi/{mu}A.h at a proton beam energy of 15 MeV and current of 15 {mu}A using a solid, commercially available {sup 89}Y-foil target (0.1 mm, 100% natural abundance). {sup 89}Zr was isolated in high radionuclidic and radiochemical purity (>99.99%) as [{sup 89}Zr]Zr-oxalate by using a solid-phase hydroxamate resin with >99.5% recovery of the radioactivity. The effective specific-activity of {sup 89}Zr was found to be in the range 5.28-13.43 mCi/{mu}g (470-1195 Ci/mmol) of zirconium. New methods for the facile production of [{sup 89}Zr]Zr-chloride are reported. Radiolabeling studies using the trihydroxamate ligand desferrioxamine B (DFO) gave 100% radiochemical yields in <15 min at room temperature, and in vitro stability measurements confirmed that [{sup 89}Zr]Zr-DFO is stable with respect to ligand dissociation in human serum for >7 days. Small-animal positron emission tomography (PET) imaging studies have demonstrated that free {sup 89}Zr(IV) ions administered as [{sup 89}Zr]Zr-chloride accumulate in the liver, whilst [{sup 89}Zr]Zr-DFO is excreted rapidly via the kidneys within <20 min. These results have important implication for the analysis of immuno-PET imaging of {sup 89}Zr-labeled monoclonal antibodies. The detailed methods described can be easily translated to other radiochemistry facilities and will facilitate the use of {sup 89}Zr in both basic science and clinical investigations.

  4. Highly selective nanocomposite sorbents for the specific recognition of S-ibuprofen from structurally related compounds (United States)

    Sooraj, M. P.; Mathew, Beena


    The aim of the present work was to synthesize highly homogeneous synthetic recognition units for the selective and specific separation of S-ibuprofen from its closely related structural analogues using molecular imprinting technology. The molecular imprinted polymer wrapped on functionalized multiwalled carbon nanotubes (MWCNT-MIP) was synthesized using S-ibuprofen as the template in the imprinting process. The characterization of the products and intermediates were done by FT-IR spectroscopy, PXRD, TGA, SEM and TEM techniques. The high regression coefficient value for Langmuir adsorption isotherm ( R 2 = 0.999) showed the homogeneous imprint sites and surface adsorption nature of the prepared polymer sorbent. The nano-MIP followed a second-order kinetics ( R 2 = 0.999) with a rapid adsorption rate which also suggested the formation of recognition sites on the surface of MWCNT-MIP. MWCNT-MIP showed 83.6 % higher rebinding capacity than its non-imprinted counterpart. The higher relative selectivity coefficient ( k') of the imprinted sorbent towards S-ibuprofen than that for its structural analogues evidenced the capability of the nano-MIP to selectively and specifically rebind the template rather than its analogues.

  5. Boechera species exhibit species-specific responses to combined heat and high light stress. (United States)

    Gallas, Genna; Waters, Elizabeth R


    As sessile organisms, plants must be able to complete their life cycle in place and therefore tolerance to abiotic stress has had a major role in shaping biogeographical patterns. However, much of what we know about plant tolerance to abiotic stresses is based on studies of just a few plant species, most notably the model species Arabidopsis thaliana. In this study we examine natural variation in the stress responses of five diverse Boechera (Brassicaceae) species. Boechera plants were exposed to basal and acquired combined heat and high light stress. Plant response to these stresses was evaluated based on chlorophyll fluorescence measurements, induction of leaf chlorosis, and gene expression. Many of the Boechera species were more tolerant to heat and high light stress than A. thaliana. Gene expression data indicates that two important marker genes for stress responses: APX2 (Ascorbate peroxidase 2) and HsfA2 (Heat shock transcription factor A2) have distinct species-specific expression patterns. The findings of species-specific responses and tolerance to stress indicate that stress pathways are evolutionarily labile even among closely related species.

  6. Module-assisted preparation of {sup 64}Cu with high specific activity

    Energy Technology Data Exchange (ETDEWEB)

    Thieme, S. [Institute of Radiopharmacy, Helmholtz-Zentrum Dresden-Rossendorf, P.O. Box 510 119, 01314 Dresden (Germany); Walther, M., E-mail: [Institute of Radiopharmacy, Helmholtz-Zentrum Dresden-Rossendorf, P.O. Box 510 119, 01314 Dresden (Germany); Pietzsch, H.-J. [Institute of Radiopharmacy, Helmholtz-Zentrum Dresden-Rossendorf, P.O. Box 510 119, 01314 Dresden (Germany); Henniger, J. [Institute of Nuclear and Particle Physics, University of Technology Dresden, 01062 Dresden (Germany); Preusche, S.; Maeding, P.; Steinbach, J. [Institute of Radiopharmacy, Helmholtz-Zentrum Dresden-Rossendorf, P.O. Box 510 119, 01314 Dresden (Germany)


    In this work the production of {sup 64}Cu via the {sup 64}Ni(p,n){sup 64}Cu reaction with optimized conditions for low current irradiation is presented. Different target setups and cleaning steps for lowering metal contaminations in the product were applied. {sup 64}Cu with high specific activities up to 1685 GBq/{mu}mol was produced despite low overall activity ({approx}4.2 GBq per run). The module processing leads to a highly reproducible, reliable product quality (<1 {mu}g Cu and <7 {mu}g Ni). Besides its diagnostic value {sup 64}Cu may be of interest even for therapeutic purposes due to its decay characteristics. - Highlights: Black-Right-Pointing-Pointer Theoretical comparison of {sup 64}Cu and {sup 67}Cu for therapeutical use. Black-Right-Pointing-Pointer Evaluation of different target setups. Black-Right-Pointing-Pointer Different specific activity determination methods in comparison. Black-Right-Pointing-Pointer Detailed description of module operation. Black-Right-Pointing-Pointer Optimized {sup 64}Ni recycling and purification.

  7. A Symmetrical, Planar SOFC Design for NASA's High Specific Power Density Requirements (United States)

    Cable, Thomas L.; Sofie, Stephen W.


    Solid oxide fuel cell (SOFC) systems for aircraft applications require an order of magnitude increase in specific power density (1.0 kW/kg) and long life. While significant research is underway to develop anode supported cells which operate at temperatures in the range of 650-800 C, concerns about Cr-contamination from the metal interconnect may drive the operating temperature down further, to 750 C and lower. Higher temperatures, 900-1000 C, are more favorable for SOFC stacks to achieve specific power densities of 1.0 kW/kg. Since metal interconnects are not practical at these high temperatures and can account for up to 75% of the weight of the stack, NASA is pursuing a design that uses a thin, LaCrO3-based ceramic interconnect that incorporates gas channels into the electrodes. The bi-electrode supported cell (BSC) uses porous YSZ scaffolds, on either side of a 10-20 microns electrolyte. The porous support regions are fabricated with graded porosity using the freeze-tape casting process which can be tailored for fuel and air flow. Removing gas channels from the interconnect simplifies the stack design and allows the ceramic interconnect to be kept thin, on the order of 50 -100 microns. The YSZ electrode scaffolds are infiltrated with active electrode materials following the high temperature sintering step. The NASA-BSC is symmetrical and CTE matched, providing balanced stresses and favorable mechanical properties for vibration and thermal cycling.

  8. Very high specific activity ⁶⁶/⁶⁸Ga from zinc targets for PET. (United States)

    Engle, J W; Lopez-Rodriguez, V; Gaspar-Carcamo, R E; Valdovinos, H F; Valle-Gonzalez, M; Trejo-Ballado, F; Severin, G W; Barnhart, T E; Nickles, R J; Avila-Rodriguez, M A


    This work describes the production of very high specific activity (66/68)Ga from (nat)Zn(p,n) and (66)Zn(p,n) using proton irradiations between 7 and 16 MeV, with emphasis on (66)Ga for use with common bifunctional chelates. Principal radiometallic impurities are (65)Zn from (p,x) and (67)Ga from (p,n). Separation of radiogallium from target material is accomplished with cation exchange chromatography in hydrochloric acid solution. Efficient recycling of Zn target material is possible using electrodeposition of Zn from its chloride form, but these measures are not necessary to achieve high specific activity or near-quantitative radiolabeling yields from natural targets. Inductively coupled plasma mass spectroscopy (ICP-MS) measures less than 2 ppb non-radioactive gallium in the final product, and the reactivity of (66)Ga with common bifunctional chelates, decay corrected to the end of irradiation, is 740 GBq/μmol (20 Ci/μmol) using natural zinc as a target material. Recycling enriched (66)Zn targets increased the reactivity of (66)Ga with common bifunctional chelates. Published by Elsevier Ltd.

  9. Generating Safety-Critical PLC Code From a High-Level Application Software Specification (United States)


    The benefits of automatic-application code generation are widely accepted within the software engineering community. These benefits include raised abstraction level of application programming, shorter product development time, lower maintenance costs, and increased code quality and consistency. Surprisingly, code generation concepts have not yet found wide acceptance and use in the field of programmable logic controller (PLC) software development. Software engineers at Kennedy Space Center recognized the need for PLC code generation while developing the new ground checkout and launch processing system, called the Launch Control System (LCS). Engineers developed a process and a prototype software tool that automatically translates a high-level representation or specification of application software into ladder logic that executes on a PLC. All the computer hardware in the LCS is planned to be commercial off the shelf (COTS), including industrial controllers or PLCs that are connected to the sensors and end items out in the field. Most of the software in LCS is also planned to be COTS, with only small adapter software modules that must be developed in order to interface between the various COTS software products. A domain-specific language (DSL) is a programming language designed to perform tasks and to solve problems in a particular domain, such as ground processing of launch vehicles. The LCS engineers created a DSL for developing test sequences of ground checkout and launch operations of future launch vehicle and spacecraft elements, and they are developing a tabular specification format that uses the DSL keywords and functions familiar to the ground and flight system users. The tabular specification format, or tabular spec, allows most ground and flight system users to document how the application software is intended to function and requires little or no software programming knowledge or experience. A small sample from a prototype tabular spec application is

  10. Tumor-infiltrating B lymphocytes as an efficient source of highly specific immunoglobulins recognizing tumor cells

    Directory of Open Access Journals (Sweden)

    Pelliccia Angela


    Full Text Available Abstract Background There is much evidence that tumor cells elicit a humoral immune response in patients. In most cases, the presence of antibodies in peripheral blood is detected only in small proportion of patients with tumors overexpressing the corresponding antigen. In the present study, we analyzed the significance of local humoral response provided by tumor-infiltrating lymphocytes in breast cancer patients. Methods The ability of a patient's immune system to produce specific antibodies inside tumor tissue, capable of recognizing tumor cells, was explored through analysis of the oligoclonality of antibodies derived from tumor-infiltrating lymphocytes and construction of a series of recombinant antibody libraries in scFv format, derived from breast tumor-infiltrating B lymphocytes. These libraries and one from peripheral blood lymphocytes of a single breast cancer patient were panned against three purified surface tumor antigens, such as CEA, MUC1 and ED-B domain, and against intact MCF7 breast carcinoma cells. Results Application of novel display vector, pKM19, allowed isolation of a large panel of breast cancer-specific antibodies against known tumor antigens, as well as against breast carcinoma cells. Reactivity of novel scFvs was confirmed by ELISA, immunohistochemistry, fluorescence staining and flow cytometry. We demonstrated that seven of ten primary breast tumor specimens, obtained using discarded surgical material, could be exploited as an appropriate source for generation of phage display libraries, giving highly specific antitumor antibodies which recognize heterologous tumor cells. Conclusion Local humoral immune response within tumor tissue in breast cancer patients frequently has an oligoclonal character. Efficient selection of specific antitumor antibodies from recombinant antibody libraries, derived from such oligoclonal tumor-infiltrated B lymphocytes, indicates the presence of natural immune response against tumor antigens

  11. Isotope specific resolution recovery image reconstruction in high resolution PET imaging. (United States)

    Kotasidis, Fotis A; Angelis, Georgios I; Anton-Rodriguez, Jose; Matthews, Julian C; Reader, Andrew J; Zaidi, Habib


    Measuring and incorporating a scanner-specific point spread function (PSF) within image reconstruction has been shown to improve spatial resolution in PET. However, due to the short half-life of clinically used isotopes, other long-lived isotopes not used in clinical practice are used to perform the PSF measurements. As such, non-optimal PSF models that do not correspond to those needed for the data to be reconstructed are used within resolution modeling (RM) image reconstruction, usually underestimating the true PSF owing to the difference in positron range. In high resolution brain and preclinical imaging, this effect is of particular importance since the PSFs become more positron range limited and isotope-specific PSFs can help maximize the performance benefit from using resolution recovery image reconstruction algorithms. In this work, the authors used a printing technique to simultaneously measure multiple point sources on the High Resolution Research Tomograph (HRRT), and the authors demonstrated the feasibility of deriving isotope-dependent system matrices from fluorine-18 and carbon-11 point sources. Furthermore, the authors evaluated the impact of incorporating them within RM image reconstruction, using carbon-11 phantom and clinical datasets on the HRRT. The results obtained using these two isotopes illustrate that even small differences in positron range can result in different PSF maps, leading to further improvements in contrast recovery when used in image reconstruction. The difference is more pronounced in the centre of the field-of-view where the full width at half maximum (FWHM) from the positron range has a larger contribution to the overall FWHM compared to the edge where the parallax error dominates the overall FWHM. Based on the proposed methodology, measured isotope-specific and spatially variant PSFs can be reliably derived and used for improved spatial resolution and variance performance in resolution recovery image reconstruction. The

  12. Comparative analysis of two and three marker immunohistochemical cocktails in the diagnosis of prostata carcinoma

    DEFF Research Database (Denmark)

    Dabir, Parag Deepak; Ottosen, Peter; Hamilton-Dutoit, Stephen Jacques;


    Comparative analysis of two and three marker immunohistochemical cocktails in the diagnosis of prostata carcinoma......Comparative analysis of two and three marker immunohistochemical cocktails in the diagnosis of prostata carcinoma...

  13. Comparative analysis of two and three marker immunohistochemical cocktails in the diagnosis of prostata carcinoma

    DEFF Research Database (Denmark)

    Dabir, Parag Deepak; Ottosen, Peter; Hamilton-Dutoit, Stephen Jacques;


    Comparative analysis of two and three marker immunohistochemical cocktails in the diagnosis of prostata carcinoma......Comparative analysis of two and three marker immunohistochemical cocktails in the diagnosis of prostata carcinoma...

  14. Optimized over-expression of [FeFe] hydrogenases with high specific activity in Clostridium acetobutylicum

    Energy Technology Data Exchange (ETDEWEB)

    von Abendroth, Gregory; Stripp, Sven; Happe, Thomas [Ruhr-Universitaet Bochum, Lehrstuhl fuer Biochemie der Pflanzen, AG Photobiotechnologie, 44780 Bochum (Germany); Silakov, Alexey [Max-Planck-Institut fuer Bioanorganische Chemie, 45470 Muelheim an der Ruhr (Germany); Croux, Christian; Soucaille, Philippe; Girbal, Laurence [UMR5504, UMR792 Ingenierie des Systemes Biologiques et des Procedes, CNRS, INRA, INSA, 31400 Toulouse (France)


    It was previously shown that Clostridium acetobutylicum is capable to over-express various [FeFe] hydrogenases although the protein yield was low. In this study we report on doubling the yield of the clostridial hydrogenase by replacing the native gene hydA1{sub Ca} with a recombinant one via homologous recombination. The purified protein HydA1{sub Ca} shows an unexpected high specific activity (up to 2257 {mu}mol H{sub 2} min{sup -1} mg{sup -1}) for hydrogen evolution. Furthermore, the highly active green algal hydrogenase HydA1{sub Cr} from Chlamydomonas reinhardtii was heterologously expressed in C. acetobutylicum, and purified with increased yield (1 mg protein per liter of cells) and high activity (625 {mu}mol H{sub 2} min{sup -1} mg{sup -1}). EPR studies demonstrate intact H-clusters for homologously and heterologously expressed [FeFe] hydrogenases in the CO-inhibited oxidized redox state, and prove the high efficiency of the C. acetobutylicum expression system. (author)

  15. Photopolymers designed for high resolution laser ablation at a specific irradiation wavelength

    Energy Technology Data Exchange (ETDEWEB)

    Lippert, T.; Bennett, L.S. [Los Alamos National Lab., NM (United States); Kunz, T.; Hahn, C. [Paul Scherrer Institute, Villigen (Switzerland). Dept. of General Energy Research] [and others


    We have developed novel photopolymers based on the triazeno chromophore group. The absorption properties can be tailored for a specific irradiation wavelength (e.g. 308 nm XeCl laser). With the introduction of a photolabile group into the main chain of the polymer we expected a mechanism which is mainly photochemical. This should result in high resolution etching with no thermal damage or chemical / physical modification to the material. The gaseous products of the photochemical decomposition were thought to assist the material removal, and to prevent the re-deposition of solid products which would contaminate the surface. We confirmed (SEM/AFM) that the irradiation of the polymer at 308 mn resulted in high resolution etching. No debris has been found around the etched comers. Maximum ablation rates of about 3 {mu}m / pulse were achieved due to the dynamic absorption behavior (bleaching during the pulse). No physical or chemical modifications of the polymer surface could be detected after irradiation at the tailored absorption wavelength, whereas irradiation at different wavelengths resulted in modified (physical and chemical) surfaces. The etching mechanism can be described as a laser induced microexplosion, revealed by ns-imaging. The etching of the polymer starts and ends with the laser pulse, shown by ns-interferometry, confirming that the acting mechanism is mainly photochemical at high fluences for our polymers. Our results demonstrate that the mechanism of ablation can be controlled by designing special polymers, which can be used as high resolution laser dry etching resists.

  16. Gonyautoxin 1/4 aptamers with high-affinity and high-specificity: From efficient selection to aptasensor application. (United States)

    Gao, Shunxiang; Hu, Bo; Zheng, Xin; Cao, Ying; Liu, Dejing; Sun, Mingjuan; Jiao, Binghua; Wang, Lianghua


    Gonyautoxin 1/4 (GTX1/4) are potent marine neurotoxins with significant public health impact. However, the ethical issues and technical defects associated with the currently applied detection methods for paralytic shellfish toxin GTX1/4 are pressing further studies to develop suitable alternatives in a regulatory monitoring system. This work describes the first successful selection, optimization, and characterization of an aptamer that bind with high affinity and specificity to GTX1/4. Compared to the typical MB-SELEX, GO-SELEX, an advanced screening technology, has significant advantages for small molecular aptamer development. Furthermore, we truncated GTX1/4 aptamer and obtained the aptamer core sequence with a higher Kd of 17.7 nM. The aptamer GO18-T-d was then used to construct a label-free and real-time optical BLI aptasensor for the detection of GTX1/4. The aptasensor showed a broad detection range from 0.2 to 200 ng/mL GTX1/4 (linear range from 0.2 to 90 ng/mL), with a low detection limit of 50 pg/mL. Moreover, the aptasensor exhibited a high degree of specificity for GTX1/4 and no cross reactivity to other marine toxins. The aptasensor was then applied to the detection of GTX1/4 in spiked shellfish samples and showed a good reproducibility and stability. We believe that this novel aptasensor offers a promising alternative to traditional analytical methods for the rapid detection of the marine biotoxin GTX1/4. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Intranasal location and immunohistochemical characterization of the equine olfactory epithelium

    Directory of Open Access Journals (Sweden)

    Alexandra Kupke


    Full Text Available The olfactory epithelium (OE is the only body site where neurons contact directly the environment and are therefore exposed to a broad variation of substances and insults. It can serve as portal of entry for neurotropic viruses which spread via the olfactory pathway to the central nervous system (CNS. For horses, it has been proposed and concluded mainly from rodent studies that different viruses, e.g. Borna disease virus (BoDV, equine herpesvirus 1 (EHV-1, hendra virus, influenza virus, rabies virus, vesicular stomatitis virus (VSV can use this route. However, little is yet known about cytoarchitecture, protein expression and the intranasal location of the equine OE. Revealing differences in cytoarchitecture or protein expression pattern in comparison to rodents, canines or humans might help to explain varying susceptibility to certain intranasal virus infections. On the other hand, disclosing similarities especially between rodents and other species, e.g. horses would help to underscore transferability of rodent models. Analysis of the complete noses of 5 adult horses revealed that in the equine OE two epithelial subtypes with distinct marker expression exist, designated as types a and b which resemble those previously described in dogs. Detailed statistical analysis was carried out to confirm the results obtained on the descriptive level. The equine OE was predominantly located in caudodorsal areas of the nasal turbinates with a significant decline in rostroventral direction, especially for type a. Immunohistochemically, olfactory marker protein (OMP and doublecortin (DCX expression was found in more cells of OE type a, whereas expression of proliferating cell nuclear antigen (PCNA and tropomyosin receptor kinase A (TrkA was present in more cells of type b. Accordingly, type a resembles the mature epithelium, in contrast to the more juvenile type b. Protein expression profile was comparable to canine and rodent OE but equine type a and b were

  18. Intranasal Location and Immunohistochemical Characterization of the Equine Olfactory Epithelium (United States)

    Kupke, Alexandra; Wenisch, Sabine; Failing, Klaus; Herden, Christiane


    The olfactory epithelium (OE) is the only body site where neurons contact directly the environment and are therefore exposed to a broad variation of substances and insults. It can serve as portal of entry for neurotropic viruses which spread via the olfactory pathway to the central nervous system. For horses, it has been proposed and concluded mainly from rodent studies that different viruses, e.g., Borna disease virus, equine herpesvirus 1 (EHV-1), hendra virus, influenza virus, rabies virus, vesicular stomatitis virus can use this route. However, little is yet known about cytoarchitecture, protein expression and the intranasal location of the equine OE. Revealing differences in cytoarchitecture or protein expression pattern in comparison to rodents, canines, or humans might help to explain varying susceptibility to certain intranasal virus infections. On the other hand, disclosing similarities especially between rodents and other species, e.g., horses would help to underscore transferability of rodent models. Analysis of the complete noses of five adult horses revealed that in the equine OE two epithelial subtypes with distinct marker expression exist, designated as types a and b which resemble those previously described in dogs. Detailed statistical analysis was carried out to confirm the results obtained on the descriptive level. The equine OE was predominantly located in caudodorsal areas of the nasal turbinates with a significant decline in rostroventral direction, especially for type a. Immunohistochemically, olfactory marker protein and doublecortin (DCX) expression was found in more cells of OE type a, whereas expression of proliferating cell nuclear antigen and tropomyosin receptor kinase A was present in more cells of type b. Accordingly, type a resembles the mature epithelium, in contrast to the more juvenile type b. Protein expression profile was comparable to canine and rodent OE but equine types a and b were located differently within the nose and

  19. Psychological balance in high level athletes: gender-based differences and sport-specific patterns.

    Directory of Open Access Journals (Sweden)

    Karine Schaal

    Full Text Available OBJECTIVES: Few epidemiological studies have focused on the psychological health of high level athletes. This study aimed to identify the principal psychological problems encountered within French high level athletes, and the variations in their prevalence based on sex and the sport practiced. METHODS: Multivariate analyses were conducted on nationwide data obtained from the athletes' yearly psychological evaluations. RESULTS: A representative sample of 13% of the French athlete population was obtained. 17% of athletes have at least one ongoing or recent disorder, generalized anxiety disorder (GAD being the most prevalent (6%, followed by non-specific eating disorders (4.2%. Overall, 20.2% of women had at least one psychopathology, against 15.1% in men. This female predominance applied to anxiety and eating disorders, depression, sleep problems and self-harming behaviors. The highest rates of GAD appeared in aesthetic sports (16.7% vs. 6.8% in other sports for men and 38.9% vs. 10.3% for women; the lowest prevalence was found in high risk sports athletes (3.0% vs. 3.5%. Eating disorders are most common among women in racing sports (14% vs. 9%, but for men were found mostly in combat sports (7% vs. 4.8%. DISCUSSION: This study highlights important differences in psychopathology between male and female athletes, demonstrating that the many sex-based differences reported in the general population apply to elite athletes. While the prevalence of psychological problems is no higher than in the general population, the variations in psychopathology in different sports suggest that specific constraints could influence the development of some disorders.

  20. Devices and approaches for generating specific high-affinity nucleic acid aptamers (United States)

    Szeto, Kylan; Craighead, Harold G.


    High-affinity and highly specific antibody proteins have played a critical role in biological imaging, medical diagnostics, and therapeutics. Recently, a new class of molecules called aptamers has emerged as an alternative to antibodies. Aptamers are short nucleic acid molecules that can be generated and synthesized in vitro to bind to virtually any target in a wide range of environments. They are, in principal, less expensive and more reproducible than antibodies, and their versatility creates possibilities for new technologies. Aptamers are generated using libraries of nucleic acid molecules with random sequences that are subjected to affinity selections for binding to specific target molecules. This is commonly done through a process called Systematic Evolution of Ligands by EXponential enrichment, in which target-bound nucleic acids are isolated from the pool, amplified to high copy numbers, and then reselected against the desired target. This iterative process is continued until the highest affinity nucleic acid sequences dominate the enriched pool. Traditional selections require a dozen or more laborious cycles to isolate strongly binding aptamers, which can take months to complete and consume large quantities of reagents. However, new devices and insights from engineering and the physical sciences have contributed to a reduction in the time and effort needed to generate aptamers. As the demand for these new molecules increases, more efficient and sensitive selection technologies will be needed. These new technologies will need to use smaller samples, exploit a wider range of chemistries and techniques for manipulating binding, and integrate and automate the selection steps. Here, we review new methods and technologies that are being developed towards this goal, and we discuss their roles in accelerating the availability of novel aptamers.

  1. Development of a highly specific enzyme immunoassay for oxytocin and its use in plasma samples. (United States)

    Haraya, Shiomi; Karasawa, Koji; Sano, Yoshihiro; Ozawa, Kimiko; Kato, Nobumasa; Arakawa, Hidetoshi


    Background The peptide hormone oxytocin acts in the central nervous system and plays an important role in various complex social behaviours. We report the production of a high affinity and specificity antibody for oxytocin and its use in a highly sensitive enzyme immunoassay. Biotin that was chemically bound to oxytocin derivative containing zero to six lysines as bridge was the labelled antigen. Seven labelled antigens were used to develop a highly sensitive enzyme immunoassay. Methods Antioxytocin antiserum was obtained by immunization of oxytocin-bovine thyrogloblin conjugate to rabbit. Oxytocin sample was added to the second antibody-coated microtitre plate and allowed to react overnight at 4℃, then biotinylated oxytocin was added 1 h at 4℃, and horseradish peroxidase-labelled avidin was added and incubated for 1 h at room temperature. The plate was then washed. Horseradish peroxidase activity was measured by a colorimetric method using o-phenylenediamine (490 nm). Results The sensitivity of the enzyme immunoassay improved as the number of lysine residues increased; consequently, biotinylated oxytocin bridged with five lysines was used. A standard curve for oxytocin ranged from 1.0 to 1000 pg/assay. The detection limit of the assay was 2.36 pg, and the reproducibility was 3.6% as CV% ( n = 6). Cross-reactivity with vasopressin and vasotocin was less than 0.01%. Conclusion The sensitivity of the enzyme immunoassay could be improved by increasing the number of lysine residues on the biotin-labelled antigen. The proposed method is sensitive and more specific than conventional immunoassays for oxytocin and can be used to determine plasma oxytocin concentrations.

  2. Analysis of DNA strand-specific differential expression with high density tiling microarrays

    Directory of Open Access Journals (Sweden)

    Antequera Francisco


    Full Text Available Abstract Background DNA microarray technology allows the analysis of genome structure and dynamics at genome-wide scale. Expression microarrays (EMA contain probes for annotated open reading frames (ORF and are widely used for the analysis of differential gene expression. By contrast, tiling microarrays (TMA have a much higher probe density and provide unbiased genome-wide coverage. The purpose of this study was to develop a protocol to exploit the high resolution of TMAs for quantitative measurement of DNA strand-specific differential expression of annotated and non-annotated transcripts. Results We extensively filtered probes present in Affymetrix Genechip Yeast Genome 2.0 expression and GeneChip S. pombe 1.0FR tiling microarrays to generate custom Chip Description Files (CDF in order to compare their efficiency. We experimentally tested the potential of our approach by measuring the differential expression of 4904 genes in the yeast Schizosaccharomyces pombe growing under conditions of oxidative stress. The results showed a Pearson correlation coefficient of 0.943 between both platforms, indicating that TMAs are as reliable as EMAs for quantitative expression analysis. A significant advantage of TMAs over EMAs is the possibility of detecting non-annotated transcripts generated only under specific physiological conditions. To take full advantage of this property, we have used a target-labelling protocol that preserves the original polarity of the transcripts and, therefore, allows the strand-specific differential expression of non-annotated transcripts to be determined. By using a segmentation algorithm prior to generating the corresponding custom CDFs, we identified and quantitatively measured the expression of 510 transcripts longer than 180 nucleotides and not overlapping previously annotated ORFs that were differentially expressed at least 2-fold under oxidative stress. Conclusions We show that the information derived from TMA

  3. Production of Medical Radioisotopes with High Specific Activity in Photonuclear Reactions with $\\gamma$ Beams of High Intensity and Large Brilliance

    CERN Document Server

    Habs, D


    We study the production of radioisotopes for nuclear medicine in $(\\gamma,x{\\rm n}+y{\\rm p})$ photonuclear reactions or ($\\gamma,\\gamma'$) photoexcitation reactions with high flux [($10^{13}-10^{15}$)$\\gamma$/s], small diameter $\\sim (100 \\, \\mu$m$)^2$ and small band width ($\\Delta E/E \\approx 10^{-3}-10^{-4}$) $\\gamma$ beams produced by Compton back-scattering of laser light from relativistic brilliant electron beams. We compare them to (ion,$x$n$ + y$p) reactions with (ion=p,d,$\\alpha$) from particle accelerators like cyclotrons and (n,$\\gamma$) or (n,f) reactions from nuclear reactors. For photonuclear reactions with a narrow $\\gamma$ beam the energy deposition in the target can be managed by using a stack of thin target foils or wires, hence avoiding direct stopping of the Compton and pair electrons (positrons). $(\\gamma,\\gamma')$ isomer production via specially selected $\\gamma$ cascades allows to produce high specific activity in multiple excitations, where no back-pumping of the isomer to the ground st...

  4. Immunohistochemical detection of VHS virus in paraffin-embedded specimens of rainbow trout (Oncorhynchus mykiss); The influence of primary antibody, fixative, and antigen unmasking on method sensitivity

    DEFF Research Database (Denmark)

    Evensen, O.; Olesen, Niels Jørgen


    The influence of the primary antibody, the fixative, and the antigen unmasking technique on the method sensitivity of immunohistochemistry as a method for the identification of viral hemorrhagic septicemia (VHS) virus in paraffin-embedded specimens of naturally infected rainbow trout (Oncorhynchus...... was performed an formalin-, PLP-, and Bouin's fluid-fixed specimens. The presence of virus peptides in situ or N-protein in the artificial antigen substrates was Visualized using an immunohistochemical method based on alkaline phosphatase or peroxidase and one polyclonal and five monoclonal polypeptide......-specific antibodies. VHS virus was identified in situ in specimens with high virus titers (10(7-8) TCID50/ml) regardless of the fixative and without the need of an unmasking procedure. A pronounced masking effect was observed for the cross-linking formalin and PLP fixatives. Regardless of the primary antibodies used...

  5. Communication and Memory Architecture Design of Application-Specific High-End Multiprocessors

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    Yahya Jan


    Full Text Available This paper is devoted to the design of communication and memory architectures of massively parallel hardware multiprocessors necessary for the implementation of highly demanding applications. We demonstrated that for the massively parallel hardware multiprocessors the traditionally used flat communication architectures and multi-port memories do not scale well, and the memory and communication network influence on both the throughput and circuit area dominates the processors influence. To resolve the problems and ensure scalability, we proposed to design highly optimized application-specific hierarchical and/or partitioned communication and memory architectures through exploring and exploiting the regularity and hierarchy of the actual data flows of a given application. Furthermore, we proposed some data distribution and related data mapping schemes in the shared (global partitioned memories with the aim to eliminate the memory access conflicts, as well as, to ensure that our communication design strategies will be applicable. We incorporated these architecture synthesis strategies into our quality-driven model-based multi-processor design method and related automated architecture exploration framework. Using this framework, we performed a large series of experiments that demonstrate many various important features of the synthesized memory and communication architectures. They also demonstrate that our method and related framework are able to efficiently synthesize well scalable memory and communication architectures even for the high-end multiprocessors. The gains as high as 12-times in performance and 25-times in area can be obtained when using the hierarchical communication networks instead of the flat networks. However, for the high parallelism levels only the partitioned approach ensures the scalability in performance.

  6. Specific localization of thallium 201 in human high-grade astrocytoma by microautoradiography. (United States)

    Mountz, J M; Raymond, P A; McKeever, P E; Modell, J G; Hood, T W; Barthel, L K; Stafford-Schuck, K A


    The ability to accurately distinguish remaining or recurrent high-grade astrocytoma from necrosis or edema following treatment is essential to optimal patient management. Thallium 201 planar gamma-camera imaging has been shown to be helpful in detecting recurrent high-grade astrocytoma; however, due to tissue heterogeneity adjacent to and within tumor, the cellular specificity and quantification of 201Tl uptake are largely unknown. In order to determine which tissues are responsible for the radioisotope uptake, microautoradiographic techniques were used to examine multiple tissue sections from five patients with high-grade astrocytoma. Each patient received 5 mCi of 201Tl i.v. 1 h prior to tumor removal. Additionally, all patients received computerized tomographic and 201Tl planar gamma-camera scans prior to surgery. Following surgery, the excised tissue specimens were tentatively classified by gross pathological examination and then immediately processed for dry mount autoradiography; grain density was determined over regions containing tumor, adjacent and uninvolved brain tissue, necrotic tissue, and background. Highly significant differences were found in grain densities (201Tl uptake) between tumor and uninvolved brain tissue, as well as between uninvolved brain tissue and necrotic tissue; there was no significant difference between background grain density and that in necrotic tissue. Mean grain densities (grains/cm2 +/- 1 SD) across patients were: tumor, 102 +/- 23; adjacent, uninvolved brain tissue, 29 +/- 11; necrotic tissue, 6.2 +/- 1.1; and background, 7.0 +/- 4.1. We conclude that the ability of 201Tl to selectively image high-grade astrocytoma is due to its preferential uptake into tumor cells.

  7. The gastric/pancreatic amylase ratio predicts postoperative pancreatic fistula with high sensitivity and specificity. (United States)

    Jin, Shuo; Shi, Xiao-Ju; Sun, Xiao-Dong; Zhang, Ping; Lv, Guo-Yue; Du, Xiao-Hong; Wang, Si-Yuan; Wang, Guang-Yi


    This article aims to identify risk factors for postoperative pancreatic fistula (POPF) and evaluate the gastric/pancreatic amylase ratio (GPAR) on postoperative day (POD) 3 as a POPF predictor in patients who undergo pancreaticoduodenectomy (PD).POPF significantly contributes to mortality and morbidity in patients who undergo PD. Previously identified predictors for POPF often have low predictive accuracy. Therefore, accurate POPF predictors are needed.In this prospective cohort study, we measured the clinical and biochemical factors of 61 patients who underwent PD and diagnosed POPF according to the definition of the International Study Group of Pancreatic Fistula. We analyzed the association between POPF and various factors, identified POPF risk factors, and evaluated the predictive power of the GPAR on POD3 and the levels of serum and ascites amylase.Of the 61 patients, 21 developed POPF. The color of the pancreatic drain fluid, POD1 serum, POD1 median output of pancreatic drain fluid volume, and GPAR were significantly associated with POPF. The color of the pancreatic drain fluid and high GPAR were independent risk factors. Although serum and ascites amylase did not predict POPF accurately, the cutoff value was 1.24, and GPAR predicted POPF with high sensitivity and specificity.This is the first report demonstrating that high GPAR on POD3 is a risk factor for POPF and showing that GPAR is a more accurate predictor of POPF than the previously reported amylase markers.

  8. Barcoded pyrosequencing reveals that consumption of galactooligosaccharides results in a highly specific bifidogenic response in humans.

    Directory of Open Access Journals (Sweden)

    Lauren M G Davis

    Full Text Available Prebiotics are selectively fermented ingredients that allow specific changes in the gastrointestinal microbiota that confer health benefits to the host. However, the effects of prebiotics on the human gut microbiota are incomplete as most studies have relied on methods that fail to cover the breadth of the bacterial community. The goal of this research was to use high throughput multiplex community sequencing of 16S rDNA tags to gain a community wide perspective of the impact of prebiotic galactooligosaccharide (GOS on the fecal microbiota of healthy human subjects. Fecal samples from eighteen healthy adults were previously obtained during a feeding trial in which each subject consumed a GOS-containing product for twelve weeks, with four increasing dosages (0, 2.5, 5, and 10 gram of GOS. Multiplex sequencing of the 16S rDNA tags revealed that GOS induced significant compositional alterations in the fecal microbiota, principally by increasing the abundance of organisms within the Actinobacteria. Specifically, several distinct lineages of Bifidobacterium were enriched. Consumption of GOS led to five- to ten-fold increases in bifidobacteria in half of the subjects. Increases in Firmicutes were also observed, however, these changes were detectable in only a few individuals. The enrichment of bifidobacteria was generally at the expense of one group of bacteria, the Bacteroides. The responses to GOS and the magnitude of the response varied between individuals, were reversible, and were in accordance with dosage. The bifidobacteria were the only bacteria that were consistently and significantly enriched by GOS, although this substrate supported the growth of diverse colonic bacteria in mono-culture experiments. These results suggest that GOS can be used to enrich bifidobacteria in the human gastrointestinal tract with remarkable specificity, and that the bifidogenic properties of GOS that occur in vivo are caused by selective fermentation as well as by

  9. Cartilage canals in newborn dogs: histochemical and immunohistochemical findings

    Directory of Open Access Journals (Sweden)

    A. Di Giancamillo


    Full Text Available Cartilage canals (CCs are microscopic structures involved in secondary ossification centers (SOCs development. The features of CCs were investigated in the humeral and femoral proximal epiphyses of small-sized newborn dogs (from premature to 28 days after birth with histochemical and immunohistochemical approaches. Masson’s Trichrome revealed a ring-shaped area around CCs, which changes in colour from green (immature collagen to red (mature collagen as ossification progresses; perichondrium staining always matched the ring colour. Safranin-O was always negative. Immunohistochemical analysis revealed immunopositivity for both collagen type I and V around the CCs; collagen type II was negative. CCs count showed a tendency to be higher in the humerus than in the femur. This work enlightened for the first time changes in composition of CCs surrounding matrix during SOCs development in dogs, paving the way to further investigations.

  10. Immunohistochemical study of basal cell adenoma in the parotid gland. (United States)

    Hamano, H; Abiko, Y; Hashimoto, S; Inoue, T; Shimono, M; Takagi, T; Noma, H


    Basal cell adenoma of the parotid gland was studied with immunohistochemical methods. We observed cells in the tumor with positive reaction to polyclonal keratin, prekeratin, monoclonal PKK-1, polyclonal S-100 protein, monoclonal S-100 protein (alpha), secretory component, actin and laminin. However, no cells which stained positively with monoclonal KL-1, amylase, carcinoembryonic antigen, or epithelial membrane antigen were recognized. From these immunohistochemical results and our ultrastructural observations reported previously, we conclude that the cells constituting the basal cell adenoma are ductal, myoepithelial, and squamous cells but not secretory ones. It is also suggested that the origins of basal cell ademona as well as those of pleomorphic and clear cell adenoma are undifferentiated cells of intercalated duct.

  11. The role of immunohistochemical analysis in the diagnosis of parapsoriasis. (United States)

    Bordignon, Matteo; Belloni-Fortina, Anna; Pigozzi, Barbara; Saponeri, Andrea; Alaibac, Mauro


    Parapsoriasis is a chronic dermatosis whose biological distinction from early mycosis fungoides, the most frequent form of cutaneous T-cell lymphoma, is still not clearly defined. Two types of parapsoriasis have been delineated: large-plaque parapsoriasis and small-plaque parapsoriasis. The lack of clinical and histological features, which may allow distinguishing parapsoriasis from early mycosis fungoides has prompted several investigations to assess the role of immunohistochemistry in establishing a conclusive diagnosis of these conditions. However, the additional data obtained by immunohistochemical analysis concerning the CD4/CD8 ratio, the aberrant expression of T-cell antigens and the expression of proliferation markers has not generally helped establish a more definitive diagnosis. This review critically discusses these immunohistochemical markers and their use in diagnosis of parapsoriasis.

  12. Delineation of Chondroid Lipoma: An Immunohistochemical and Molecular Biological Analysis

    Directory of Open Access Journals (Sweden)

    Ronald S. A. de Vreeze


    Full Text Available Aims. Chondroid lipoma (CL is a benign tumor that mimics a variety of soft tissue tumors and is characterized by translocation t(11;16. Here, we analyze CL and its histological mimics. Methods. CL (n=4 was compared to a variety of histological mimics (n=83 for morphological aspects and immunohistochemical features including cyclinD1(CCND1. Using FISH analysis, CCND1 and FUS were investigated as potential translocation partners. Results. All CLs were strongly positive for CCND1. One of 4 myoepitheliomas, CCND1, was positive. In well-differentiated lipomatous tumors and in chondrosarcomas, CCND1 was frequently expressed, but all myxoid liposarcomas were negative. FISH analysis did not give support for direct involvement of CCND1 and FUS as translocation partners. Conclusions. Chondroid lipoma is extremely rare and has several and more prevalent histological mimics. The differential diagnosis of chondroid lipomas can be unraveled using immunohistochemical and molecular support.

  13. Highly specific electronic signal transduction mediated by DNA/metal self-assembly.

    Energy Technology Data Exchange (ETDEWEB)

    Dentinger, Paul M.; Pathak, Srikant


    Highly specific interactions between DNA could potentially be amplified if the DNA interactions were utilized to assemble large scale parts. Fluidic assembly of microsystem parts has the potential for rapid and accurate placement of otherwise difficult to handle pieces. Ideally, each part would have a different chemical interaction that allowed it to interact with the substrate only in specific areas. One easy way to obtain a multiple chemical permutations is to use synthetic DNA oligomers. Si parts were prepared using silicon-on-insulator technology microfabrication techniques. Several surface chemistry protocols were developed to react commercial oligonucleotides to the parts. However, no obvious assembly was achieved. It was thought that small defects on the surface did not allow the microparts to be in close enough proximity for DNA hybridization, and this was. in part, confirmed by interferometry. To assist in the hybridization, plastic, pliable parts were manufactured and a new chemistry was developed. However, assembly was still absent even with the application of force. It is presently thought that one of three mechanisms is preventing the assembly. The surfaces of the two solid substrates can not get in close enough proximity, the surface chemistry lacks sufficient density to keep the parts from separating, or DNA interactions in close proximity on solid substrates are forbidden. These possibilities are discussed in detail.

  14. Aptamer-MIP hybrid receptor for highly sensitive electrochemical detection of prostate specific antigen. (United States)

    Jolly, Pawan; Tamboli, Vibha; Harniman, Robert L; Estrela, Pedro; Allender, Chris J; Bowen, Jenna L


    This study reports the design and evaluation of a new synthetic receptor sensor based on the amalgamation of biomolecular recognition elements and molecular imprinting to overcome some of the challenges faced by conventional protein imprinting. A thiolated DNA aptamer with established affinity for prostate specific antigen (PSA) was complexed with PSA prior to being immobilised on the surface of a gold electrode. Controlled electropolymerisation of dopamine around the complex served to both entrap the complex, holding the aptamer in, or near to, it's binding conformation, and to localise the PSA binding sites at the sensor surface. Following removal of PSA, it was proposed that the molecularly imprinted polymer (MIP) cavity would act synergistically with the embedded aptamer to form a hybrid receptor (apta-MIP), displaying recognition properties superior to that of aptamer alone. Electrochemical impedance spectroscopy (EIS) was used to evaluate subsequent rebinding of PSA to the apta-MIP surface. The apta-MIP sensor showed high sensitivity with a linear response from 100pg/ml to 100ng/ml of PSA and a limit of detection of 1pg/ml, which was three-fold higher than aptamer alone sensor for PSA. Furthermore, the sensor demonstrated low cross-reactivity with a homologous protein (human Kallikrein 2) and low response to human serum albumin (HSA), suggesting possible resilience to the non-specific binding of serum proteins.

  15. Highly specific electronic signal transduction mediated by DNA/metal self-assembly.

    Energy Technology Data Exchange (ETDEWEB)

    Dentinger, Paul M.; Pathak, Srikant


    Highly specific interactions between DNA could potentially be amplified if the DNA interactions were utilized to assemble large scale parts. Fluidic assembly of microsystem parts has the potential for rapid and accurate placement of otherwise difficult to handle pieces. Ideally, each part would have a different chemical interaction that allowed it to interact with the substrate only in specific areas. One easy way to obtain a multiple chemical permutations is to use synthetic DNA oligomers. Si parts were prepared using silicon-on-insulator technology microfabrication techniques. Several surface chemistry protocols were developed to react commercial oligonucleotides to the parts. However, no obvious assembly was achieved. It was thought that small defects on the surface did not allow the microparts to be in close enough proximity for DNA hybridization, and this was. in part, confirmed by interferometry. To assist in the hybridization, plastic, pliable parts were manufactured and a new chemistry was developed. However, assembly was still absent even with the application of force. It is presently thought that one of three mechanisms is preventing the assembly. The surfaces of the two solid substrates can not get in close enough proximity, the surface chemistry lacks sufficient density to keep the parts from separating, or DNA interactions in close proximity on solid substrates are forbidden. These possibilities are discussed in detail.

  16. High and stable substrate specificities of microorganisms in enhanced biological phosphorus removal plants. (United States)

    Kindaichi, Tomonori; Nierychlo, Marta; Kragelund, Caroline; Nielsen, Jeppe Lund; Nielsen, Per Halkjaer


    Microbial communities are typically characterized by conditions of nutrient limitation so the availability of the resources is likely a key factor in the niche differentiation across all species and in the regulation of the community structure. In this study we have investigated whether four species exhibit any in situ short-term changes in substrate uptake pattern when exposed to variations in substrate and growth conditions. Microautoradiography was combined with fluorescence in situ hybridization to investigate in situ cell-specific substrate uptake profiles of four probe-defined coexisting species in a wastewater treatment plant with enhanced biological phosphorus removal. These were the filamentous 'Candidatus Microthrix' and Caldilinea (type 0803), the polyphosphate-accumulating organism 'Candidatus Accumulibacter', and the denitrifying Azoarcus. The experimental conditions mimicked the conditions potentially encountered in the respective environment (starvation, high/low substrate concentration, induction with specific substrates, and single/multiple substrates). The results showed that each probe-defined species exhibited very distinct and constant substrate uptake profile in time and space, which hardly changed under any of the conditions tested. Such niche partitioning implies that a significant change in substrate composition will be reflected in a changed community structure rather than the substrate uptake response from the different species. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  17. Preparation of hydrazine functionalized polymer brushes hybrid magnetic nanoparticles for highly specific enrichment of glycopeptides. (United States)

    Huang, Guang; Sun, Zhen; Qin, Hongqiang; Zhao, Liang; Xiong, Zhichao; Peng, Xiaojun; Ou, Junjie; Zou, Hanfa


    Hydrazide chemistry is a powerful technique in glycopeptides enrichment. However, the low density of the monolayer hydrazine groups on the conventional hydrazine-functionalized magnetic nanoparticles limits the efficiency of glycopeptides enrichment. Herein, a novel magnetic nanoparticle grafted with poly(glycidyl methacrylate) (GMA) brushes was fabricated via reversible addition-fragmentation chain transfer (RAFT) polymerization, and a large amount of hydrazine groups were further introduced to the GMA brushes by ring-opening the epoxy groups with hydrazine hydrate. The resulting magnetic nanoparticles (denoted as Fe3O4@SiO2@GMA-NHNH2) demonstrated the high specificity of capturing glycopeptides from a tryptic digest of the sample comprising a standard non-glycosylated protein bovine serum albumin (BSA) and four standard glycoproteins with a weight ratio of 50 : 1, and the detection limit was as low as 130 fmol. In the analysis of a real complex biological sample, the tryptic digest of hepatocellular carcinoma, 179 glycosites were identified by the Fe3O4@SiO2@GMA-NHNH2 nanoparticles, surpassing that of 68 glycosites by Fe3O4@SiO2-single-NHNH2 (with monolayer hydrazine groups on the surface). It can be expected that the magnetic nanoparticles modified with hydrazine functionalized polymer brushes via RAFT technique will improve the specificity and the binding capacity of glycopeptides from complex samples, and show great potential in the analysis of protein glycosylation in biological samples.

  18. Record-high specific conductance and temperature in San Francisco Bay during water year 2014 (United States)

    Downing-Kunz, Maureen; Work, Paul; Shellenbarger, Gregory


    The U.S. Geological Survey (USGS) has operated a water-quality monitoring network in San Francisco Bay since the late 1980s (Buchanan and others, 2015). This network includes 19 stations in the bay; currently, 8 stations are in operation (fig. 1). All eight stations are equipped with specific conductance (which can be related to salinity) and water-temperature sensors that record measurements at 15-minute intervals. Water quality in the bay constantly changes with the ocean tides and with seasonal and interannual differences in river inflows. Our network was designed to observe and characterize some of these changes in the bay across space and over time. Our data demonstrated a high degree of variability both in specific conductance and temperature at time scales from tidal to annual and also revealed longer term changes that are likely to influence overall environmental health in the bay (San Francisco Estuary Institute, 2014). Figure 1. Locations of fixed water-quality monitoring stations in San Francisco Bay, California, for the 2014 water year (October 1, 2013 to September 30, 2014).

  19. Exploring site-specific chemical interactions at surfaces: a case study on highly ordered pyrolytic graphite (United States)

    Dagdeviren, Omur E.; Götzen, Jan; Altman, Eric I.; Schwarz, Udo D.


    A material’s ability to interact with approaching matter is governed by the structural and chemical nature of its surfaces. Tailoring surfaces to meet specific needs requires developing an understanding of the underlying fundamental principles that determine a surface’s reactivity. A particularly insightful case occurs when the surface site exhibiting the strongest attraction changes with distance. To study this issue, combined noncontact atomic force microscopy and scanning tunneling microscopy experiments have been carried out, where the evolution of the local chemical interaction with distance leads to a contrast reversal in the force channel. Using highly ordered pyrolytic graphite surfaces and metallic probe tips as a model system, we find that at larger tip-sample distances, carbon atoms exhibit stronger attractions than hollow sites while upon further approach, hollow sites become energetically more favorable. For the tunneling current that is recorded at large tip-sample separations during acquisition of a constant-force image, the contrast is dominated by the changes in tip-sample distance required to hold the force constant (‘cross-talk’) at smaller separations the contrast turns into a convolution of this cross-talk and the local density of states. Analysis shows that the basic factors influencing the force channel contrast reversal are locally varying decay lengths and an onset of repulsive forces that occurs for distinct surface sites at different tip-sample distances. These findings highlight the importance of tip-sample distance when comparing the relative strength of site-specific chemical interactions.

  20. Identification of lung cancer with high sensitivity and specificity by blood testing

    Directory of Open Access Journals (Sweden)

    Stephan Bernhard


    Full Text Available Abstract Background Lung cancer is a very frequent and lethal tumor with an identifiable risk population. Cytological analysis and chest X-ray failed to reduce mortality, and CT screenings are still controversially discussed. Recent studies provided first evidence for the potential usefulness of autoantigens as markers for lung cancer. Methods We used extended panels of arrayed antigens and determined autoantibody signatures of sera from patients with different kinds of lung cancer, different common non-tumor lung pathologies, and controls without any lung disease by a newly developed computer aided image analysis procedure. The resulting signatures were classified using linear kernel Support Vector Machines and 10-fold cross-validation. Results The novel approach allowed for discriminating lung cancer patients from controls without any lung disease with a specificity of 97.0%, a sensitivity of 97.9%, and an accuracy of 97.6%. The classification of stage IA/IB tumors and controls yielded a specificity of 97.6%, a sensitivity of 75.9%, and an accuracy of 92.9%. The discrimination of lung cancer patients from patients with non-tumor lung pathologies reached an accuracy of 88.5%. Conclusion We were able to separate lung cancer patients from subjects without any lung disease with high accuracy. Furthermore, lung cancer patients could be seprated from patients with other non-tumor lung diseases. These results provide clear evidence that blood-based tests open new avenues for the early diagnosis of lung cancer.

  1. MOF-Bacteriophage Biosensor for Highly Sensitive and Specific Detection of Staphylococcus aureus. (United States)

    Bhardwaj, Neha; Bhardwaj, Sanjeev K; Mehta, Jyotsana; Kim, Ki-Hyun; Deep, Akash


    To produce a sensitive and specific biosensor for Staphylococcus aureus, bacteriophages have been interfaced with a water-dispersible and environmentally stable metal-organic framework (MOF), NH2-MIL-53(Fe). The conjugation of the MOF with bacteriophages has been achieved through the use of glutaraldehyde as cross-linker. Highly sensitive detection of S. aureus in both synthetic and real samples was realized by the proposed MOF-bacteriophage biosensor based on the photoluminescence quenching phenomena: limit of detection (31 CFU/mL) and range of detection (40 to 4 × 10(8) CFU/mL). This is the first report exploiting the use of an MOF-bacteriophage complex for the biosensing of S. aureus. The results of our study highlight that the proposed biosensor is more sensitive than most of the previous methods while exhibiting some advanced features like specificity, regenerability, extended range of linear detection, and stability for long-term storage (even at room temperature).

  2. High-specificity quantification method for almond-by-products, based on differential proteomic analysis. (United States)

    Zhang, Shiwei; Wang, Shifeng; Huang, Jingmin; Lai, Xintian; Du, Yegang; Liu, Xiaoqing; Li, Bifang; Feng, Ronghu; Yang, Guowu


    A highly specific competitive enzyme-linked immunosorbent assay (ELISA) protocol has been developed to identify and classify almond products based on differential proteomic analysis. We applied two-dimensional electrophoresis to compare the differences between almond and apricot kernels to search for almond-specific proteins. The amino acid of apricot Pru-1 was sequenced and aligned to almond Pru-1. One peptide, RQGRQQGRQQQEEGR, which exists in almond but not in apricot, was used as hapten to prepare monoclonal antibody against almond Pru-1. An optimized ELISA method was established using this antibody. The assay did not exhibit cross-reactivity with the tested apricot kernels and other edible plant seeds. The limit of detection (LOD) was 2.5-100μg/g based on different food samples. The recoveries of fortified samples at levels of twofold and eightfold LOD ranged from 82% to 96%. The coefficients of variation were less than 13.0%. Using 7M urea as extracting solution, the heat-treated protein loss ratios were 2%, 5% and 15% under pasteurization (65°C for 30min), baking (150°C for 30min) and autoclaved sterilization (120°C for 15min), respectively.

  3. High-throughput profiling of N-myristoylation substrate specificity across species including pathogens. (United States)

    Traverso, José A; Giglione, Carmela; Meinnel, Thierry


    One of the most critical modifications affecting the N-terminus of proteins is N-myristoylation. This irreversible modification affects the membrane-binding properties of crucial proteins involved in signal transduction cascades. This cotranslational modification, catalyzed by N-myristoyl transferase, occurs both in lower and higher eukaryotes and is a validated therapeutic target for several pathologies. However, this lipidation proves very difficult to be evidenced in vivo even with state-of-the-art proteomics approaches or bioinformatics tools. A large part of N-myristoylated proteins remains to be discovered and the rules of substrate specificity need to be established in each organism. Because the peptide substrate recognition occurs around the first eight residues, short peptides are used for modeling the reaction in vitro. Here, we provide a novel approach including a dedicated peptide array for high-throughput profiling protein N-myristoylation specificity. We show that myristoylation predictive tools need to be fine-tuned to organisms and that their poor accuracy should be significantly enhanced. This should lead to strongly improved knowledge of the number and function of myristoylated proteins occurring in any proteome.

  4. Immunohistochemical localization of human papilloma virus in conjunctival neoplasias: A retrospective study

    Directory of Open Access Journals (Sweden)

    Sen Seema


    Full Text Available Background: The extent of association of human papilloma virus (HPV in human conjunctival neoplasias has been debated in studies originating from different parts of the world, but no substantial evidence has been generated on Indian subjects. This prompted us to carry out a retrospective study on conjunctival neoplasias diagnosed over the past 12 years. Materials and Methods: Histopathological and immunohistochemical analysis of 65 specimens of ocular neoplasias and 30 normal controls diagnosed between 1991 and 2002 at a tertiary eye care hospital, was undertaken. Formalin-fixed, paraffin-embedded tissues were reviewed for confirming histopathological diagnosis, presence of koilocytosis and changes related to actinic keratosis. Immunohistochemical analysis was done using HPV-specific monoclonal antibodies. Clinicopathological correlation and the association of HPV antigen with the histopathological features were performed. Results: Out of the 65 cases analyzed, 35 were papillomas and 30 were ocular surface squamous neoplasias (OSSN. The mean age was 48 years with a male preponderance. Histologically, koilocytosis was observed in 17.1% of papillomas and 36.6% of OSSN. Actinic keratosis was present in 33% of OSSN. Immunohistochemically 17.1% conjunctival papillomas stained positive for HPV antigen, all cases of OSSN were negative for HPV. There was no correlation between koilocytosis or actinic keratosis and the detection of HPV antigen. Conclusions: The association between HPV and conjunctival neoplasias is variable in different geographical areas and also depends on the methods of detection used. This study warrants the need for applying more advanced techniques at a molecular level to determine the possible etiology of HPV in conjunctival neoplasias among Asian-Indians.

  5. Immunohistochemical localization of human papilloma virus in conjunctival neoplasias: A retrospective study (United States)

    Sharma, Anjana; Panda, Anita


    Background: The extent of association of human papilloma virus (HPV) in human conjunctival neoplasias has been debated in studies originating from different parts of the world, but no substantial evidence has been generated on Indian subjects. This prompted us to carry out a retrospective study on conjunctival neoplasias diagnosed over the past 12 years. Materials and Methods: Histopathological and immunohistochemical analysis of 65 specimens of ocular neoplasias and 30 normal controls diagnosed between 1991 and 2002 at a tertiary eye care hospital, was undertaken. Formalin-fixed, paraffin-embedded tissues were reviewed for confirming histopathological diagnosis, presence of koilocytosis and changes related to actinic keratosis. Immunohistochemical analysis was done using HPV-specific monoclonal antibodies. Clinicopathological correlation and the association of HPV antigen with the histopathological features were performed. Results: Out of the 65 cases analyzed, 35 were papillomas and 30 were ocular surface squamous neoplasias (OSSN). The mean age was 48 years with a male preponderance. Histologically, koilocytosis was observed in 17.1% of papillomas and 36.6% of OSSN. Actinic keratosis was present in 33% of OSSN. Immunohistochemically 17.1% conjunctival papillomas stained positive for HPV antigen, all cases of OSSN were negative for HPV. There was no correlation between koilocytosis or actinic keratosis and the detection of HPV antigen. Conclusions: The association between HPV and conjunctival neoplasias is variable in different geographical areas and also depends on the methods of detection used. This study warrants the need for applying more advanced techniques at a molecular level to determine the possible etiology of HPV in conjunctival neoplasias among Asian-Indians. PMID:17699945

  6. Immunomagnetic Separation of Fat Depot-Specific Sca1high Adipose-Derived Stem Cells (Ascs) (United States)

    Barnes, Richard H; Chun, Tae-Hwa


    The isolation of adipose-derived stem cells (ASCs) is an important method in the field of adipose tissue biology, adipogenesis, and extracellular matrix (ECM) remodeling. In vivo, ECM-rich environment consisting of fibrillar collagens provides a structural support to adipose tissues during the progression and regression of obesity. Physiological ECM remodeling mediated by matrix metalloproteinases (MMPs) plays a major role in regulating adipose tissue size and function1, 2. The loss of physiological collagenolytic ECM remodeling may lead to excessive collagen accumulation (tissue fibrosis), macrophage infiltration, and ultimately, a loss of metabolic homeostasis including insulin resistance3, 4. When a phenotypic change of the adipose tissue is observed in gene-targeted mouse models, isolating primary ASCs from fat depots for in vitro studies is an effective approach to define the role of the specific gene in regulating the function of ASCs. In the following, we define an immunomagnetic separation of Sca1high ASCs. PMID:27583550

  7. Shortening trinucleotide repeats using highly specific endonucleases: a possible approach to gene therapy? (United States)

    Richard, Guy-Franck


    Trinucleotide repeat expansions are involved in more than two dozen neurological and developmental disorders. Conventional therapeutic approaches aimed at regulating the expression level of affected genes, which rely on drugs, oligonucleotides, and/or transgenes, have met with only limited success so far. An alternative approach is to shorten repeats to non-pathological lengths using highly specific nucleases. Here, I review early experiments using meganucleases, zinc-finger nucleases (ZFN), and transcription-activator like effector nucleases (TALENs) to contract trinucleotide repeats, and discuss the possibility of using CRISPR-Cas nucleases to the same end. Although this is a nascent field, I explore the possibility of designing nucleases and effectively delivering them in the context of gene therapy.

  8. Functional specificity for high-level linguistic processing in the human brain. (United States)

    Fedorenko, Evelina; Behr, Michael K; Kanwisher, Nancy


    Neuroscientists have debated for centuries whether some regions of the human brain are selectively engaged in specific high-level mental functions or whether, instead, cognition is implemented in multifunctional brain regions. For the critical case of language, conflicting answers arise from the neuropsychological literature, which features striking dissociations between deficits in linguistic and nonlinguistic abilities, vs. the neuroimaging literature, which has argued for overlap between activations for linguistic and nonlinguistic processes, including arithmetic, domain general abilities like cognitive control, and music. Here, we use functional MRI to define classic language regions functionally in each subject individually and then examine the response of these regions to the nonlinguistic functions most commonly argued to engage these regions: arithmetic, working memory, cognitive control, and music. We find little or no response in language regions to these nonlinguistic functions. These data support a clear distinction between language and other cognitive processes, resolving the prior conflict between the neuropsychological and neuroimaging literatures.


    Energy Technology Data Exchange (ETDEWEB)

    Chen Xu,Charles Reece,Michael Kelley


    The high-field performance of SRF cavities will eventually be limited by the realization of fundamental material limits, whether it is Hc1 or Hsh, or some derivative thereof, at which the superconductivity is lost. Before reaching this fundamental field limit at the macro level, it must be encountered at localized, perhaps microscopic, sites of field enhancement due to local topography. If such sites are small enough, they may produce thermally stabilized normal-conducting regions which contribute non-linear losses when viewed from the macro resonant field perspective, and thus produce degradation in Q0. We have undertaken a calculation of local surface magnetic field enhancement from specific fine topographic structure by conformal mapping method and numerically. A solution of the resulting normal conducting volume has been derived and the corresponding RF Ohmic loss simulated.

  10. Very High Specific Energy, Medium Power Li/CFx Primary Battery for Launchers and Space Probes (United States)

    Brochard, Paul; Godillot, Gerome; Peres, Jean Paul; Corbin, Julien; Espinosa, Amaya


    Benchmark with existing technologies shows the advantages of the lithium-fluorinated carbon (Li/CFx) technology for use aboard future launchers in terms of a low Total Cost of Ownership (TCO), especially for high energy demanding missions such as re-ignitable upper stages for long GTO+ missions and probes for deep space exploration.This paper presents the new results obtained on this chemistry in terms of electrical and climatic performances, abuse tests and life tests. Studies - co-financed between CNES and Saft - looked at a pure CFx version with a specific energy up to 500 Wh/kg along with a medium power of 80 to 100 W/kg.

  11. High specific surface area Mo2C nanoparticles as an efficient electrocatalyst for hydrogen evolution (United States)

    Tang, Chaoyun; Sun, Aokui; Xu, Yushuai; Wu, Zhuangzhi; Wang, Dezhi


    Mo2C nanoparticles with high specific surface area (120 m2 g-1) are successfully synthesized using a typical and low-cost monosaccharide of glucose via a facile calcination and subsequent reduction process. The HER functions of the obtained Mo2C nanoparticles are investigated and the effect of reduction time in hydrogen is also discussed. It is found that η-MoC can be obtained at 800 °C with a reduction time of 10 min, but the formation of β-Mo2C phase requires more than 20 min. Moreover, the β-Mo2C obtained with a reduction time of 20 min exhibits the best HER activity with a small Tafel slope of 55 mV dec-1 and a large current density of 60 mA cm-2 at -200 mV, which is among the best records over Mo2C-based HER catalysts.


    Energy Technology Data Exchange (ETDEWEB)

    Vitro Engineering Company


    This is the specifications for the High Temperature Lattice Test Reactor Building 318 and it is divided into the following 21 divisions or chapters: (1) Excavating, Backfill & Grading; (2) Reinforced Concrete; (3) Masonry; (4) Structural Steel & Miscellaneous Metal Items, Contents - Division V; (5) Plumbing, Process & Service Piping; (6) Welding; (7) Insulated Metal Siding; (8) Roof Decks & Roofing; (9) Plaster Partitions & Ceiling; (10) Standard Doors, Windows & Hardware; (11) Shielding Doors; (12) Sprinkler System & Fire Extinguishers, Contents - Division XIII; (13) Heating, Ventilating & Air Conditioning; (14) Painting, Protective Coating & Floor Covering, Contents - Division XV; (15) Electrical; (16) Communications & Alarm Systems; (17) Special Equipment & Furnishings; (18) Overhead Bridge Crane; (19) Prefabricated Steel Building; (20) Paved Drive; and (21) Landscaping & Irrigation Sprinklers.

  13. High sequence variability among hemocyte-specific Kazal-type proteinase inhibitors in decapod crustaceans. (United States)

    Cerenius, Lage; Liu, Haipeng; Zhang, Yanjiao; Rimphanitchayakit, Vichien; Tassanakajon, Anchalee; Gunnar Andersson, M; Söderhäll, Kenneth; Söderhäll, Irene


    Crustacean hemocytes were found to produce a large number of transcripts coding for Kazal-type proteinase inhibitors (KPIs). A detailed study performed with the crayfish Pacifastacus leniusculus and the shrimp Penaeus monodon revealed the presence of at least 26 and 20 different Kazal domains from the hemocyte KPIs, respectively. Comparisons with KPIs from other taxa indicate that the sequences of these domains evolve rapidly. A few conserved positions, e.g. six invariant cysteines were present in all domain sequences whereas the position of P1 amino acid, a determinant for substrate specificity, varied highly. A study with a single crayfish animal suggested that even at the individual level considerable sequence variability among hemocyte KPIs produced exist. Expression analysis of four crayfish KPI transcripts in hematopoietic tissue cells and different hemocyte types suggest that some of these KPIs are likely to be involved in hematopoiesis or hemocyte release as they were produced in particular hemocyte types or maturation stages only.

  14. Study of Stable Cathodes and Electrolytes for High Specific Density Lithium-Air Battery (United States)

    Hernandez-Lugo, Dionne M.; Wu, James; Bennett, William; Ming, Yu; Zhu, Yu


    Future NASA missions require high specific energy battery technologies, greater than 400 Wh/kg. Current NASA missions are using "state-of-the-art" (SOA) Li-ion batteries (LIB), which consist of a metal oxide cathode, a graphite anode and an organic electrolyte. NASA Glenn Research Center is currently studying the physical and electrochemical properties of the anode-electrolyte interface for ionic liquid based Li-air batteries. The voltage-time profiles for Pyr13FSI and Pyr14TFSI ionic liquids electrolytes studies on symmetric cells show low over-potentials and no dendritic lithium morphology. Cyclic voltammetry measurements indicate that these ionic liquids have a wide electrochemical window. As a continuation of this work, sp2 carbon cathode and these low flammability electrolytes were paired and the physical and electrochemical properties were studied in a Li-air battery system under an oxygen environment.

  15. Epithelial ovarian tumors: Clinicopathological correlation and immunohistochemical study



    Background: Ovarian cancer is the third leading site of cancer among women, trailing behind cervix and breast cancer. Aim: This study was undertaken to analyze the immunohistochemical (IHC) profile of estrogen receptors (ER), progesterone receptors (PR), Ki-67, and p53 in various ovarian epithelial tumors and attempt correlation with clinical and histopathological findings. Materials and Methods: The present study was conducted over a period of 4 years. A technique of manual tissue ar...

  16. Stem cells expanded from the human embryonic hindbrain stably retain regional specification and high neurogenic potency. (United States)

    Tailor, Jignesh; Kittappa, Raja; Leto, Ketty; Gates, Monte; Borel, Melodie; Paulsen, Ole; Spitzer, Sonia; Karadottir, Ragnhildur Thora; Rossi, Ferdinando; Falk, Anna; Smith, Austin


    Stem cell lines that faithfully maintain the regional identity and developmental potency of progenitors in the human brain would create new opportunities in developmental neurobiology and provide a resource for generating specialized human neurons. However, to date, neural progenitor cultures derived from the human brain have either been short-lived or exhibit restricted, predominantly glial, differentiation capacity. Pluripotent stem cells are an alternative source, but to ascertain definitively the identity and fidelity of cell types generated solely in vitro is problematic. Here, we show that hindbrain neuroepithelial stem (hbNES) cells can be derived and massively expanded from early human embryos (week 5-7, Carnegie stage 15-17). These cell lines are propagated in adherent culture in the presence of EGF and FGF2 and retain progenitor characteristics, including SOX1 expression, formation of rosette-like structures, and high neurogenic capacity. They generate GABAergic, glutamatergic and, at lower frequency, serotonergic neurons. Importantly, hbNES cells stably maintain hindbrain specification and generate upper rhombic lip derivatives on exposure to bone morphogenetic protein (BMP). When grafted into neonatal rat brain, they show potential for integration into cerebellar development and produce cerebellar granule-like cells, albeit at low frequency. hbNES cells offer a new system to study human cerebellar specification and development and to model diseases of the hindbrain. They also provide a benchmark for the production of similar long-term neuroepithelial-like stem cells (lt-NES) from pluripotent cell lines. To our knowledge, hbNES cells are the first demonstration of highly expandable neuroepithelial stem cells derived from the human embryo without genetic immortalization.

  17. CLCA2 as a Novel Immunohistochemical Marker for Differential Diagnosis of Squamous Cell Carcinoma from Adenocarcinoma of the Lung

    Directory of Open Access Journals (Sweden)

    Kazuya Shinmura


    Full Text Available Recent progress in targeted therapy for lung cancer has revealed that accurate differential diagnosis between squamous cell carcinoma (SCC and adenocarcinoma (ADC of the lung is essential. To identify a novel immunohistochemical marker useful for differential diagnosis between the two subtypes of lung cancer, we first selected 24 SCC-specific genes and 6 ADC-specific genes using data (case number, 980 from the Cancer Genome Atlas (TCGA database. Among the genes, we chose the CLCA2 gene, which is involved in chloride conductance and whose protein expression in lung cancer is yet to be characterized, and evaluated its protein expression status in 396 cases of primary lung cancer at Hamamatsu University Hospital. Immunohistochemical analysis revealed a significantly higher CLCA2 expression level in the SCCs than in the ADCs (P<0.0001 and also a significantly higher frequency of CLCA2 protein expression in the SCCs (104/161, 64.6% as compared with that in the ADCs (2/235, 0.9% (P<0.0001; sensitivity 64.6%, specificity 99.1%. The CLCA2 protein expression status was associated with the histological tumor grade in the SCCs. These results suggest that CLCA2 might be a novel excellent immunohistochemical marker for differentiating between primary SCC and primary ADC of the lung.

  18. A simplified immunohistochemical classification of skeletal muscle fibres in mouse

    Directory of Open Access Journals (Sweden)

    M. Kammoun


    Full Text Available The classification of muscle fibres is of particular interest for the study of the skeletal muscle properties in a wide range of scientific fields, especially animal phenotyping. It is therefore important to define a reliable method for classifying fibre types. The aim of this study was to establish a simplified method for the immunohistochemical classification of fibres in mouse. To carry it out, we first tested a combination of several anti myosin heavy chain (MyHC antibodies in order to choose a minimum number of antibodies to implement a semi-automatic classification. Then, we compared the classification of fibres to the MyHC electrophoretic pattern on the same samples. Only two anti MyHC antibodies on serial sections with the fluorescent labeling of the Laminin were necessary to classify properly fibre types in Tibialis Anterior and Soleus mouse muscles in normal physiological conditions. This classification was virtually identical to the classification realized by the electrophoretic separation of MyHC. This immunohistochemical classification can be applied to the total area of Tibialis Anterior and Soleus mouse muscles. Thus, we provide here a useful, simple and time-efficient method for immunohistochemical classification of fibres, applicable for research in mouse

  19. Ameloblastoma vs basal cell carcinoma: an immunohistochemical comparison. (United States)

    Jawad, Salam N; Abdullah, Bashar H


    Despite behavioral mimicry of ameloblastoma (AB) and basal cell carcinoma (BCC), they are classified at 2 extremes within pertinent WHO classifications with respect to benign and malignant designation. This study aims to appraise the current allocation of AB in the classification through an immunohistochemical comparison of some aspects of behavior with BCC. Sections from retrospectively retrieved formalin-fixed, paraffin-embedded tissue blocks of AB (n = 37) and BCC (n = 34) were comparatively examined for the immunohistochemical expression for Ki-67, Bcl-2, MMP-2, MMP-9, CD31, and D2-40 monoclonal antibodies. No statistically significant differences between the tumors were found regarding the immunoexpressions of Bcl-2 (P = .252), CD31 microvessel density (P = .895), lymphatic vessel density (P = .642), and MMP-9 stromal expression (P = .083). MMP-2 expression was significantly higher in epithelial and stromal regions of AB (P = .009 and P = .001, respectively), whereas Ki-67 and MMP-9 epithelial expressions were significantly higher in BCC (P < .000 and P = .026, respectively). Within the studied immunohistochemical attributes for tumor behavior, the study accentuated the overall behavioral mimicry of the tumors and indicated that BCCs surmount ABs by the proliferative rate only. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Immunohistochemical investigation of the coma blister and its pathogenesis. (United States)

    Kashiwagi, Masayuki; Ishigami, Akiko; Hara, Kenji; Matsusue, Aya; Waters, Brian; Takayama, Mio; Tokunaga, Itsuo; Nishimura, Akiyoshi; Kubo, Shin-ichi


    The erythematous patches and vesicles that are observed in coma patients, usually from an overdose of medication, are known as coma blisters. However, it is unknown whether the degenerated sweat gland is a necrosis or apoptosis. We immunohistochemically examined such skin lesions to investigate the characteristics and pathogenesis of the coma blister. Skin lesions were obtained from a forensic autopsy case, a woman in her thirties, of caffeine intoxication. Those lesions were observed in the left femoral, the lower left thigh, and the right knee. Histologically, the skin lesions showed that the keratinocytes had necrosed and the epidermis was thin in some areas. Eccrine sweat gland degeneration was observed. Obvious inflammatory cell infiltrations were not detected. Immunohistochemically, we stained each skin lesion against CD3, CD8, CD45RO, cytokeratin, 70 kD heat shock protein, ubiquitin, 150 kD oxygen regulated protein, and caspase-cleaved keratin 18 neo-epitope M30. They were also stained with an in situ apoptosis detection kit. Degenerated sweat glands featured CD45RO and M30 immunoreactivity. Immunohistochemical staining for CD45RO, CK-L, and M30 might be useful to observe sweat gland degeneration in the coma blister. Therefore, the apoptosis might be related to coma blisters and sweat gland degenerations.


    Institute of Scientific and Technical Information of China (English)

    朱雪琼; 石一复; 陈晓端; 吴裕中


    Objective: To evaluate the diagnostic implications of immunohistochemical markers in uterine smooth muscle tumors. Methods: Formalin-fixed paraffin-embedded tissue blocks were selected from 17 uterine leiomyosarcomas, 40 uterine unusual leiomyomas and 25 uterine usual leiomyomas. Utilizing immunohistochemical techniques with antigen retrieval, serial sections of each tumor for immunoreactivity with myogenic markers, ovarian steroid receptors, CD44v3, proliferating cell nuclear antigen and mast cells were assessed. Results: Although the myogenic markers and CD44v3 showed less frequent positivity in uterine leiomyosarcomas than those in unusual leiomyomas, they were not reliable markers for differentiating leiomyosarcoma from leiomyoma. Uterine leiomyosarcoma tended to have lower ovarian steroid receptors immunoreactivity rates than leiomyoma. Leiomyoma tended to have a higher quantity of intratumoral mast cells than leiomyosarcoma, while the expression of proliferating cell nuclear antigen was lower in them. Conclusion: Because the estimation of mitotic count was subject to significant variation, the immunohistochemical expression of ovarian steroid receptors, mast cells and proliferating cell nuclear antigen seemed to be helpful for the discrimination of unusual leiomyoma from leiomyosarcoma.

  2. Histological and Immunohistochemical Revision of Hepatocellular Adenomas: A Learning Experience

    Directory of Open Access Journals (Sweden)

    S. Fonseca


    Full Text Available Light has been shed on the genotype/phenotype correlation in hepatocellular adenoma (HCA recognizing HNF1α-inactivated HCA (H-HCA, inflammatory HCA (IHCA, and β-catenin-activated HCA (b-HCA. We reviewed retrospectively our surgical HCA series to learn how to recognize the different subtypes histopathologically and how to interpret adequately their immunohistochemical staining. From January 1992 to January 2012, 37 patients underwent surgical resection for HCA in our institution. Nine had H-HCA (25% characterized by steatosis and loss of L-FABP expression; 20 had IHCA (55.5% showing CRP and/or SAA expression, sinusoidal dilatation, and variable inflammation; and 1 patient had both H-HCA and IHCA. In 5 patients (14%, b-HCA with GS and β-catenin nuclear positivity was diagnosed, two already with hepatocellular carcinoma. Two cases (5.5% remained unclassified. One of the b-HCA showed also the H-HCA histological and immunohistochemical characteristics suggesting a subgroup of β-catenin-activated/HNF1α-inactivated HCA, another b-HCA exhibited the IHCA histological and immunohistochemical characteristics suggesting a subgroup of β-catenin-activated/inflammatory HCA. Interestingly, three patients had underlying vascular abnormalities. Using the recently published criteria enabled us to classify histopathologically our retrospective HCA surgical series with accurate recognition of b-HCA for which we confirm the higher risk of malignant transformation. We also underlined the association between HCA and vascular abnormalities.

  3. Reduced immunohistochemical expression of adhesion molecules in vitiligo skin biopsies. (United States)

    Reichert Faria, Adriane; Jung, Juliana Elizabeth; Silva de Castro, Caio César; de Noronha, Lucia


    Because defects in adhesion impairment seem to be involved in the etiopathogenesis of vitiligo, this study aimed to compare the immunohistochemical expression of several adhesion molecules in the epidermis of vitiligo and non lesional vitiligo skin. Sixty-six specimens of lesional and non lesional skin from 33 volunteers with vitiligo were evaluated by immunohistochemistry using anti-beta-catenin, anti-E-cadherin, anti-laminin, anti-beta1 integrin, anti-collagen IV, anti-ICAM-1 and anti-VCAM-1 antibodies. Biopsies of vitiligo skin demonstrated a significant reduction in the expression of laminin and integrin. The average value of the immunohistochemically positive reaction area of the vitiligo specimens was 3053.2μm(2), compared with the observed value of 3431.8μm(2) in non vitiligo skin (p=0.003) for laminin. The immuno-positive area was 7174.6μm(2) (vitiligo) and 8966.7μm(2) (non lesional skin) for integrin (p=0.042). A reduction in ICAM-1 and VCAM-1 expression in the basal layer of the epidermis in vitiligo samples was also observed (p=0.001 and pskin. Our results suggest that an impairment in adhesion exists in vitiligo skin, which is supported by the diminished immunohistochemical expression of laminin, beta1 integrin, ICAM-1 and VCAM-1.

  4. High-mannose N-glycan-specific lectin from the red alga Kappaphycus striatum (Carrageenophyte). (United States)

    Hung, Le Dinh; Sato, Yuichiro; Hori, Kanji


    From a fresh sample (1 kg) of cultivated red alga Kappaphycus striatum, three isolectins, KSA-1 (15.1 mg), KSA-2 (58.0 mg) and KSA-3 (6.9 mg), were isolated by a combination of extraction with aqueous ethanol, ethanol precipitation, and ion exchange chromatography. Isolated KSAs were monomeric proteins of about 28kDa having identical 20N-terminal amino acid sequences to each other. Their hemagglutination activities were not inhibited by monosaccharides, but inhibited by glycoproteins bearing high-mannose N-glycans. In a binding experiment with pyridylaminated oligosaccharides by centrifugal ultrafiltration-HPLC assay, the isolectin KSA-2 was exclusively bound to high-mannose type N-glycans, but not to other glycans. Including complex types and a pentasaccharide core of N-glycans, indicating that it recognized branched oligomannosides. The binding activity of KSA-2 was slightly different among high-mannose N-glycans examined, indicating that the lectin has a higher affinity for those having the exposed (α1-3) Man in the D2 arm. On the other hand, KSA-2 did not bind to a free oligomannose that is a constituent of the branched oligomannosides, implying that the portion of the core GlcNAc residue(s) of the N-glycans is also essential for binding. Thus, KSA-2 appears to recognize the extended carbohydrate structure with a minimal length of a tetrasaccharide, Man(α1-3)Man(α1-6)Man(β1-4)GlcNAc. This study indicates that K. striatum, which has extensively been cultivated as a source of carrageenan, is a good source of a valuable lectin(s) that is strictly specific for high-mannose N-glycans.

  5. Lineage-specific distribution of high levels of genomic 5-hydroxymethylcytosine in mammalian development

    Institute of Scientific and Technical Information of China (English)

    Alexey Ruzov1; Yanina Tsenkina; Andrea Serio; Tatiana Dudnakova; Judy Fletcher; Yu Bai; Tatiana Chebotareva


    Methylation of cytosine is a DNA modification associated with gene repression.Recently,a novel cytosine modification,5-hydroxymethylcytosine (5-hmC) has been discovered.Here we examine 5-hmC distribution during mammalian development and in cellular systems,and show that the developmental dynamics of 5-hmC are different from those of 5-methylcytosine (5-mC); in particular 5-hmC is enriched in embryonic contexts compared to adult tissues.A detectable 5-hmC signal appears in pre-implantation development starting at the zygote stage,where the paternal genome is subjected to a genome-wide hydroxylation of 5-mC,which precisely coincides with the loss of the 5-mC signal in the paternal pronucleus.Levels of 5-hmC are high in cells of the inner cell mass in blastocysts,and the modification colocalises with nestin-expressing cell populations in mouse post-implantation embryos.Compared to other adult mammalian organs,5-hmC is strongly enriched in bone marrow and brain,wherein high 5-hmC content is a feature of both neuronal progenitors and post-mitotic neurons.We show that high levels of 5-hmC are not only present in mouse and human embryonic stem cells (ESCs) and lost during differentiation,as has been reported previously,but also reappear during the generation of induced pluripotent stem cells; thus 5-hmC enrichment correlates with a pluripotent cell state.Our findings suggest that apart from the cells of neuronal lineages,high levels of genomic 5-hmC are an epigenetic feature of embryonic cell populations and cellular pluri- and multi-lineage potency.To our knowledge,5-hmC represents the first epigenetic modification of DNA discovered whose enrichment is so cell-type specific.

  6. Development of the radiosynthesis of high-specific-activity {sup 123}I-NKJ64

    Energy Technology Data Exchange (ETDEWEB)

    Tavares, Adriana Alexandre S., E-mail: [Institute of Neuroscience and Psychology, College of Medical, Veterinary and Life Sciences, University of Glasgow, G12 8QQ Glasgow (United Kingdom); Jobson, Nicola K. [WestCHEM, School of Chemistry, The Joseph Black Building, University of Glasgow, G12 8QQ Glasgow (United Kingdom); Dewar, Deborah [Institute of Neuroscience and Psychology, College of Medical, Veterinary and Life Sciences, University of Glasgow, G12 8QQ Glasgow (United Kingdom); Sutherland, Andrew [WestCHEM, School of Chemistry, The Joseph Black Building, University of Glasgow, G12 8QQ Glasgow (United Kingdom); Pimlott, Sally L. [West of Scotland Radionuclide Dispensary, University of Glasgow and North Glasgow University Hospital NHS Trust, G11 6NT Glasgow (United Kingdom)


    Introduction: {sup 123}I-NKJ64, a reboxetine analogue, is currently under development as a potential novel single photon emission computed tomography radiotracer for imaging the noradrenaline transporter in brain. This study describes the development of the radiosynthesis of {sup 123}I-NKJ64, highlighting the advantages and disadvantages, pitfalls and solutions encountered while developing the final radiolabelling methodology. Methods: The synthesis of {sup 123}I-NKJ64 was evaluated using an electrophilic iododestannylation method, where a Boc-protected trimethylstannyl precursor was radioiodinated using peracetic acid as an oxidant and deprotection was investigated using either trifluoroacetic acid (TFA) or 2 M hydrochloric acid (HCl). Results: Radioiodination of the Boc-protected trimethylstannyl precursor was achieved with an incorporation yield of 92{+-}6%. Deprotection with 2 M HCl produced {sup 123}I-NKJ64 with the highest radiochemical yield of 98.05{+-}1.63% compared with 83.95{+-}13.24% with TFA. However, the specific activity of the obtained {sup 123}I-NKJ64 was lower when measured after using 2 M HCl (0.15{+-}0.23 Ci/{mu}mol) as the deprotecting agent in comparison to TFA (1.76{+-}0.60 Ci/{mu}mol). Further investigation of the 2 M HCl methodology found a by-product, identified as the deprotected proto-destannylated precursor, which co-eluted with {sup 123}I-NKJ64 during the high-performance liquid chromatography (HPLC) purification. Conclusions: The radiosynthesis of {sup 123}I-NKJ64 was achieved with good isolated radiochemical yield of 68% and a high specific activity of 1.8 Ci/{mu}mol. TFA was found to be the most suitable deprotecting agent, since 2 M HCl generated a by-product that could not be fully separated from {sup 123}I-NKJ64 using the HPLC methodology investigated. This study highlights the importance of HPLC purification and accurate measurement of specific activity while developing new radiosynthesis methodologies.

  7. Immunohistochemical localization of glutathione S-transferase-pi in human colorectal polyps

    Institute of Scientific and Technical Information of China (English)


    AIM: To investigate the distribution of the placental form of glutathione-S-transferase (GST) in colon polyps in order to evaluate the role of GST-pi in these tissues. METHODS: Sixteen polyp tissues removed at colon- oscopy were examined. Tissues were investigated his- tologicaUy and ultrastructurally. GST-pi expression was also analysed immunohistochemically, using peroxidase anti-peroxidase (PAP) method and immunogold label- ling method, for light and electron microscope respec- tively. RESULTS: All polyp tissues examined were adenoma of low, mild and high- grade dysplasia as shown in the histopathological reports. Nevertheless, the examina- tion of the above specimens with electron microscope revealed that 3 of 9 adenoma of mild dysplasia had ultrastuctural features similar to high-grade dysplasia adenoma. GST-pi was variably expressed in adenoma, with the lowest relative levels occurring in low-grade adenoma and the highest levels found in high-grade adenoma. GST-pi was located mainly in undifferentiat- ed epithelial cells. GST-pi positive particles were found in the cytoplasm and especially in the nucleus adjacent to the nuclear membrane of these cells. CONCLUSION: The overexpression of GST-pi in mild- grade adenomas with significant subcellular changes and in the majority of high-grade dysplasia adenoma suggests that this might be related to the carcinogenetic proceeding. Immunohistochemical localization of GST-pi in combination with ultrastructural changes indicate that GST-pi might be a sensitive agent for the detection of preneoplastic transformations in adenoma.

  8. Immunohistochemical Study to Evaluate the Prognostic Significance of Four Biomolecular Markers in Radiotherapy of Nasopharyngeal Carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Yeon Joo; Lee, Seung Hee; Wu, Hong Gyun; Go, Heoun Jeong; Jeon, Yoon Kyung [Seoul National University College of Medicine, Seoul (Korea, Republic of)


    We performed an immunohistochemical study with pre-treatment biopsy specimens to evaluate the prognostic significance of four biomolecular markers which can be used as a predictive assay for radiotherapy (RT) treatment of nasopharyngeal carcinoma (NPC). From January 1998 through December 2006, 68 patients were histologically diagnosed as non-metastatic NPC and treated by RT. Only 38 patients had the paraffin block for the immunohistochemical study. Thirty-one patients had undifferentiated carcinoma and 7 patients had squamous cell carcinoma. Thirty two patients (84%) had advanced stage NPC (2002 AJCC Stage III{approx}IV). Immunohistochemical staining was performed for Met, COX-2, nm23-H1, and epidermal growth factor receptor (EGFR) expression using routine methods. The median follow-up time was 30 months (range, 11 to 83 months) for all patients, and 39 months (range, 19 to 83 months) for surviving patients. The 5-year overall survival (OS) rate of the patients with high Met extent ({>=}50%) was significantly lower than that of the patients with low Met extent (48% vs. 84%, p=0.02). In addition, Met extent was also a significant prognostic factor in multivariate analysis (p=0.01). No correlation was observed between Met extent and T stage, N stage, stage group, gender, age, and the response to chemotherapy or RT. Met extent showed moderate correlation with COX-2 expression (Pearson coefficient 0.496, p<0.01), but COX-2 expression did not affect OS. Neither nm23-H1 or EGFR expression was a prognostic factor for OS in this study. High Met extent ({>=}50%) might be an independent prognostic factor that predicts poor OS in NPC treated with RT.

  9. Membrane-based nanocalorimeter for high-resolution measurements of low-temperature specific heat

    CERN Document Server

    Tagliati, S; Rydh, A


    A differential, membrane-based nanocalorimeter for general specific heat studies of very small samples, ranging from 0.5 mg to sub-{\\mu}g in mass, is described. The calorimeter operates over the temperature range from above room temperature down to 0.5 K. It consists of a pair of cells, each of which is a stack of heaters and thermometer in the center of a silicon nitride membrane, in total giving a background heat capacity less than 100 nJ/K at 300 K, decreasing to 10 pJ/K at 1K. The device has several distinctive features: i) The resistive thermometer, made of a Ge_{1-x}Au_{x} alloy, displays a high dimensionless sensitivity |dlnR/dlnT | \\geq 1 over the entire temperature range. ii) The sample is placed in direct contact with the thermometer, which is allowed to self-heat. The thermometer can thus be operated at high dc current to increase the resolution. iii) Data are acquired with a set of eight synchronized lock-in amplifiers measuring dc, 1st and 2nd harmonic signals of heaters and thermometer. This giv...

  10. Investigation of recoil collection method for production of high specific activity nuclear medicine isotopes. (United States)

    Cheng, K Y; Kunze, J F; Ehrhardt, G J


    High specific activity radiopharmaceuticals cannot readily be produced by (n, gamma) reactions in nuclear reactors, because the great abundance of parent atoms remaining have the same chemical characteristics as the produced isotope. We have investigated the effectiveness of using recoil atom collection methods for separating the produced radioisotope. Gold-198, produced from isotopically pure (natural) gold-197, was chosen for these experiments, which were run in the high flux (approximately 10(14) n/cm2 s thermal flux) of the reflector of the University of Missouri Research Reactor. Seven separate experiments were run, with a 2 mm separation between the emitter and the collector. Collection efficiencies were only a few percent of the radioisotope atoms produced in the top atomic layer of the emitter, instead of the 30% range anticipated. Furthermore, the collected radioisotope, instead of being nearly pure, contained a large quantity of the parent. Unless the reason for the "contamination" of product with the parent can be reduced by several orders of magnitude, this "surface hot atom recoil" method appears to offer no practical application for nuclear medicine isotope production.

  11. Penicillin V acylase from Pectobacterium atrosepticum exhibits high specific activity and unique kinetics. (United States)

    Avinash, V S; Ramasamy, Sureshkumar; Suresh, C G; Pundle, Archana


    Penicillin V acylases (PVAs, E.C.3.5.11) belong to the Ntn hydrolase super family of enzymes that catalyze the deacylation of the side chain from phenoxymethyl penicillin (penicillin V). Penicillin acylases find use in the pharmaceutical industry for the production of semi-synthetic antibiotics. PVAs employ the N-terminal cysteine residue as catalytic nucleophile and are structurally and evolutionarily related to bile salt hydrolases (BSHs). Here, we report the cloning and characterization of a PVA enzyme from the Gram-negative plant pathogen, Pectobacterium atrosepticum (PaPVA). The enzyme was cloned and expressed in Escherichia coli attaining a very high yield (250 mg/l) and a comparatively high specific activity (430 IU/mg). The enzyme showed marginally better pH and thermo-stability over PVAs characterized from Gram-positive bacteria. The enzyme also showed enhanced activity in presence of organic solvents and detergents. The enzyme kinetics turned out to be significantly different from that of previously reported PVAs, displaying positive cooperativity and substrate inhibition. The presence of bile salts had a modulating effect on PaPVA activity. Sequence analysis and characterization reveal the distinctive nature of these enzymes and underscore the need to study PVAs from Gram-negative bacteria.

  12. Lightweight Solar Paddle with High Specific Power of 150 W/Kg (United States)

    Shimazaki, Kazunori; Takahashi, Masato; Imaizumi, Mitsuru; Takamoto, Tatsuya; Ito, Takehiko; Nozaki, Yukishige; Kusawake, Hiroaki


    A lightweight solar paddle using space solar sheet (SSS) is currently being developed, which uses glass-type SSS (G-SSS) comprising InGaP/GaAs/InGaAs triple- junction high-efficiency thin-film solar cells. To avoid damage to the G-SSS due to vibration during launch, we adopted a new architecture on a panel. This panel employed a curved frame-type structure, on which the G-SSS is mounted and test models were manufactured to evaluate the vibration tolerance. The dimensions of the 1.0-cm-thick unit panel were about 1.0 × 1.0 m. Acoustic and sine vibration tests were performed on the model and the results demonstrated the high durability of the curved panel in an acoustic and vibration environments. The specific power of the solar paddle using the curved panel is estimated at approximately 150 W/kg at an array power of about 10 kW.

  13. High-resolution mass-selective UV spectroscopy of pseudoephedrine: evidence for conformer-specific fragmentation. (United States)

    Karaminkov, R; Chervenkov, S; Delchev, V; Neusser, H J


    Using resonance-enhanced two-photon ionization spectroscopy with mass resolution of jet-cooled molecules, a low-resolution S(1) ← S(0) vibronic spectrum of pseudoephedrine was recorded at the mass channels of three distinct fragments with m/z = 58, 71, and 85. Two of the fragments, with m/z = 71 and 85, are observed for the first time for this molecule. The vibronic spectra recorded at different mass channels feature different patterns, implying that they originate from different conformers in the cold molecular beam, following conformer-specific fragmentation pathways. Highly resolved spectra of all prominent vibronic features were measured, and from their analysis based on genetic algorithms, the molecular parameters of the conformers giving rise to the respective bands have been determined. Comparing the experimental results with those obtained from high-level ab initio quantum chemistry calculations, the observed prominent vibronic bands have been assigned to originate from four distinct conformers. The conformers are separated into two groups that have different fragmentation pathways determined by the different intramolecular interactions.

  14. Highly sensitive fiber-optic accelerometer by grating inscription in specific core dip fiber. (United States)

    Rong, Qiangzhou; Guo, Tuan; Bao, Weijia; Shao, Zhihua; Peng, Gang-Ding; Qiao, Xueguang


    A highly sensitive fiber-optic accelerometer based on detecting the power output of resonances from the core dip is demonstrated. The sensing probe comprises a compact structure, hereby a short section of specific core (with a significant core dip) fiber stub containing a straight fiber Bragg grating is spliced to another single-mode fiber via a core self-alignment process. The femtosecond laser side-illumination technique was utilized to ensure that the grating inscription region is precisely positioned and compact in size. Two well-defined core resonances were achieved in reflection: one originates from the core dip and the other originates from fiber core. The key point is that only one of these two reflective resonances exhibits a high sensitivity to fiber bend (and vibration), whereas the other is immune to it. For low frequency (core mode reflection. Moreover, the sensor simultaneously provides an inherent power reference to eliminate unwanted power fluctuations from the light source and transmission lines, thus providing a means of evaluating weak seismic wave at low frequency.

  15. Value of Immunohistochemical Methods in Detecting EML4-ALK Fusion Mutations: 
A Meta-analysis

    Directory of Open Access Journals (Sweden)

    Chang LIU


    Full Text Available Background and objective The fusion between echinoderm microtubule-associated protein 4 (EML4 and anaplastic lymphatic tumor kinase (ALK rearrangement is present in approximately 5% of non-small cell lung cancer (NSCLC patients. It has been regarded as another new target gene after epidermal growth factor receptor (EGFR and K-ras. Figures showed that the disease control rate could reach up to 80% in NSCLC patients with EML4-ALK fusion gene after treated with ALK inhibitors. Thus, exploring an accurate and rapid detecting method is the key in screening NSCLC patients with EML4-ALK expressions. The aim of this study is to analyze the specificity and sensitivity of IHC in detecting EML4-ALK fusion mutations. To evaluate the accuracy and clinical value of this method, and then provide basis for individual molecular therapy of NSCLC patients. Methods Using Pubmed database to search all documents required. The deadline of retrieval was February 25, 2015. Then further screening the articles according to the inclusion and exclusion criteria. Using diagnostic test meta-analysis methods to analyze the sensitivity and specificity of the immunohistochemistry (IHC method compared with fluorescence in situ hybridization (FISH method. Results Eleven literatures were added into the meta analysis, there were 3,234 of total cases. The diagnositic odds ratio (DOR was 1,135.00 (95%CI: 337.10-3,821.46; the area under curve (AUC of summary receiver operating characteristic curve (SROC curve was 0.992,3 (SEAUC=0.003,2, the Q* was 0.964,4 (SEQ*=0.008,7. Conclusion Immunohistochemical detection of EML4-ALK fusion gene mutation with specific antibody is feasible. It has high sensitivity and specificity. IHC can be a simple and rapid way in screening EML4-ALK fusion gene mutation and exhibits important clinical values.

  16. Quantitative Mass Spectrometry for Bacterial Protein Toxins — A Sensitive, Specific, High-Throughput Tool for Detection and Diagnosis

    Directory of Open Access Journals (Sweden)

    Suzanne Kalb


    Full Text Available Matrix-assisted laser-desorption time-of-flight (MALDI-TOF mass spectrometry (MS is a valuable high-throughput tool for peptide analysis. Liquid chromatography electrospray ionization (LC-ESI tandem-MS provides sensitive and specific quantification of small molecules and peptides. The high analytic power of MS coupled with high-specificity substrates is ideally suited for detection and quantification of bacterial enzymatic activities. As specific examples of the MS applications in disease diagnosis and select agent detection, we describe recent advances in the analyses of two high profile protein toxin groups, the Bacillus anthracis toxins and the Clostridium botulinum neurotoxins. The two binary toxins produced by B. anthracis consist of protective antigen (PA which combines with lethal factor (LF and edema factor (EF, forming lethal toxin and edema toxin respectively. LF is a zinc-dependent endoprotease which hydrolyzes specific proteins involved in inflammation and immunity. EF is an adenylyl cyclase which converts ATP to cyclic-AMP. Toxin-specific enzyme activity for a strategically designed substrate, amplifies reaction products which are detected by MALDI-TOF-MS and LC-ESI-MS/MS. Pre-concentration/purification with toxin specific monoclonal antibodies provides additional specificity. These combined technologies have achieved high specificity, ultrasensitive detection and quantification of the anthrax toxins. We also describe potential applications to diseases of high public health impact, including Clostridium difficile glucosylating toxins and the Bordetella pertussis adenylyl cyclase.

  17. Thermoanaerobacterium thermosaccharolyticum β-glucosidase: a glucose-tolerant enzyme with high specific activity for cellobiose

    Directory of Open Access Journals (Sweden)

    Pei Jianjun


    Full Text Available Abstract Background β-Glucosidase is an important component of the cellulase enzyme system. It does not only participate in cellulose degradation, it also plays an important role in hydrolyzing cellulose to fermentable glucose by relieving the inhibition of exoglucanase and endoglucanase from cellobiose. Therefore, the glucose-tolerant β-glucosidase with high specific activity for cellobiose might be a potent candidate for industrial applications. Results The β-glucosidase gene bgl that encodes a 443-amino-acid protein was cloned and over-expressed from Thermoanaerobacterium thermosaccharolyticum DSM 571 in Escherichia coli. The phylogenetic trees of β-glucosidases were constructed using Neighbor-Joining (NJ and Maximum-Parsimony (MP methods. The phylogeny and amino acid analysis indicated that the BGL was a novel β-glucosidase. By replacing the rare codons for the N-terminal amino acids of the target protein, the expression level of bgl was increased from 6.6 to 11.2 U/mg in LB medium. Recombinant BGL was purified by heat treatment followed by Ni-NTA affinity. The optimal activity was at pH 6.4 and 70°C. The purified enzyme was stable over pH range of 5.2–7.6 and had a 1 h half life at 68°C. The activity of BGL was significantly enhanced by Fe2+ and Mn2+. The Vmax of 64 U/mg and 120 U/mg were found for p-nitrophenyl-β-D-glucopyranoside (Km value of 0.62 mM and cellobiose (Km value of 7.9 mM, respectively. It displayed high tolerance to glucose and cellobiose. The Kcat for cellobiose was 67.7 s-1 at 60°C and pH 6.4, when the concentration of cellobiose was 290 mM. It was activated by glucose at concentrations lower that 200 mM. With glucose further increasing, the enzyme activity of BGL was gradually inhibited, but remained 50% of the original value in even as high as 600 mM glucose. Conclusions The article provides a useful novel β-glucosidase which displayed favorable properties: high glucose and cellobiose tolerance

  18. Wet autoclave pretreatment for immunohistochemical demonstration of oestrogen receptors in routinely processed breast carcinoma tissue. (United States)

    Bier, B; Bankfalvi, A; Grote, L; Blasius, S; Ofner, D; Böcker, W; Jasani, B; Schmid, K W


    The immunohistochemical demonstration of oestrogen receptor (OR) was performed on 32 randomly selected and routinely processed breast carcinomas after wet autoclave pretreatment of sections. The autoclave method was compared to the OR status found on frozen sections as well as to alternative pretreatment methods such as enzymatic predigestion and microwave irradiation. Using four different monoclonal antibody clones (H222, LH1, CC4-5, 1D5.26), the OR status was evaluated for each of the various pretreatment methods applied. All cases with a high OR content on frozen sections (n = 11) also showed a high OR status on wet autoclave-pretreated paraffin tissues using antibody clones 1D5.26 and CC4-5; in cases with low OR content on frozen sections, no false-negative cases were recorded using only the antibody 1D5.26 neither after wet autoclave nor microwave pretreatment. In addition, with this antibody, OR was detectable after autoclave pretreatment in two cases which were considered to be OR-negative even on frozen sections. When the primary antibody was omitted, no false-positive cases were observed after wet autoclave pretreatment. Thus, in our hands, wet autoclave pretreatment, in combination with the antibody 1D5.26, offers a highly sensitive method for the immunohistochemical demonstration of OR in routinely formalin-fixed, paraffin-embedded sections of breast carcinomas.

  19. Prostate-Specific Antigen Bounce After High-Dose-Rate Monotherapy for Prostate Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Mehta, Niraj H., E-mail: [University of California, Los Angeles, Los Angeles, California (United States); Kamrava, Mitchell; Wang, Pin-Chieh; Steinberg, Michael; Demanes, Jeffrey [University of California, Los Angeles, Los Angeles, California (United States)


    Purpose: To characterize the magnitude and kinetics of prostate-specific antigen (PSA) bounces after high-dose-rate (HDR) monotherapy and determine relationships between certain clinical factors and PSA bounce. Methods and Materials: Longitudinal PSA data and various clinical parameters were examined in 157 consecutive patients treated with HDR monotherapy between 1996 and 2005. We used the following definition for PSA bounce: rise in PSA ≥threshold, after which it returns to the prior level or lower. Prostate-specific antigen failure was defined per the Phoenix definition (nadir +2 ng/mL). Results: A PSA bounce was noted in 67 patients (43%). The number of bounces per patient was 1 in 45 cases (67%), 2 in 19 (28%), 3 in 2 (3%), 4 in 0, and 5 in 1 (1%). The median time to maximum PSA bounce was 1.3 years, its median magnitude was 0.7, and its median duration was 0.75 years. Three patients (2%) were noted to have PSA failure. None of the 3 patients who experienced biochemical failure exhibited PSA bounce. In the fully adjusted model for predicting each bounce, patients aged <55 years had a statistically significant higher likelihood of experiencing a bounce (odds ratio 2.22, 95% confidence interval 1.38-3.57, P=.001). There was also a statistically significant higher probability of experiencing a bounce for every unit decrease in Gleason score (odds ratio 1.52, 95% confidence interval 1.01-2.04, P=.045). Conclusions: A PSA bounce occurs in a significant percentage of patients treated with HDR monotherapy, with magnitudes varying from <1 in 28% of cases to ≥1 in 15%. The median duration of bounce is <1 year. More bounces were identified in patients with lower Gleason score and age <55 years. Further investigation using a model to correlate magnitude and frequency of bounces with clinical variables are under way.

  20. Evolution of the Specific Surface Area of Snow in a High Temperature Gradient Metamorphism (United States)

    Wang, X.; Baker, I.


    The structural evolution of low-density snow under a high temperature gradient over a short period usually takes place in the surface layers during diurnal recrystallization or on a clear, cold night. To relate snow microstructures with their thermal properties, we combined X-ray computed microtomography (micro-CT) observations with numerical simulations. Different types of snow were tested over a large range of TGs (100 K m-1- 500 K m-1). The Specific Surface Area (SSA) was used to characterize the temperature gradient metamorphism (TGM). The magnitude of the temperature gradient and the initial snow type both influence the evolution of SSA. The SSA evolution under TGM was dominated by grain growth and the formation of complex surfaces. Fresh snow experienced a logarithmic decrease of SSA with time, a feature been observed previously by others [Calonne et al., 2014; Schneebeli and Sokratov, 2004; Taillandier et al., 2007]. However, for initial rounded and connected snow structures, the SSA will increase during TGM. Understanding the SSA increase is important in order to predict the enhanced uptake of chemical species by snow or increase in snow albedo. Calonne, N., F. Flin, C. Geindreau, B. Lesaffre, and S. Rolland du Roscoat (2014), Study of a temperature gradient metamorphism of snow from 3-D images: time evolution of microstructures, physical properties and their associated anisotropy, The Cryosphere Discussions, 8, 1407-1451, doi:10.5194/tcd-8-1407-2014. Schneebeli, M., and S. A. Sokratov (2004), Tomography of temperature gradient metamorphism of snow and associated changes in heat conductivity, Hydrological Processes, 18(18), 3655-3665, doi:10.1002/hyp.5800. Taillandier, A. S., F. Domine, W. R. Simpson, M. Sturm, and T. A. Douglas (2007), Rate of decrease of the specific surface area of dry snow: Isothermal and temperature gradient conditions, Journal of Geophysical Research: Earth Surface (2003-2012), 112(F3), doi: 10.1029/2006JF000514.

  1. Rapid and Highly Sensitive Non-Competitive Immunoassay for Specific Detection of Nodularin

    Directory of Open Access Journals (Sweden)

    Sultana Akter


    Full Text Available Nodularin (NOD is a cyclic penta-peptide hepatotoxin mainly produced by Nodularia spumigena, reported from the brackish water bodies of various parts of the world. It can accumulate in the food chain and, for safety reasons, levels of NOD not only in water bodies but also in food matrices are of interest. Here, we report on a non-competitive immunoassay for the specific detection of NOD. A phage display technique was utilized to interrogate a synthetic antibody phage library for binders recognizing NOD bound to an anti-ADDA (3-Amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4(E,6(E-dienoic acid monoclonal antibody (Mab. One of the obtained immunocomplex binders, designated SA32C11, showed very high specificity towards nodularin-R (NOD-R over to the tested 10 different microcystins (microcystin-LR, -dmLR, -RR, -dmRR, -YR, -LY, -LF, -LW, -LA, -WR. It was expressed in Escherichia coli as a single chain antibody fragment (scFv fusion protein and used to establish a time-resolved fluorometry-based assay in combination with the anti-ADDA Mab. The detection limit (blank + 3SD of the immunoassay, with a total assay time of 1 h 10 min, is 0.03 µg/L of NOD-R. This represents the most sensitive immunoassay method for the specific detection of NOD reported so far. The assay was tested for its performance to detect NOD using spiked (0.1 to 3 µg/L of NOD-R water samples including brackish sea and coastal water and the recovery ranged from 79 to 127%. Furthermore, a panel of environmental samples, including water from different sources, fish and other marine tissue specimens, were analyzed for NOD using the assay. The assay has potential as a rapid screening tool for the analysis of a large number of water samples for the presence of NOD. It can also find applications in the analysis of the bioaccumulation of NOD in marine organisms and in the food chain.

  2. Analysis of the effect of specific vocabulary instruction on high school chemistry students' knowledge and understanding (United States)

    Labrosse, Peggy

    The purpose of this study was to analyze the effects of specific vocabulary instruction on high school chemistry students' knowledge and understanding. Students might be able to formally recite a definition for a term without actually having understood the meaning of the term and its connection to other terms or to related concepts. Researchers (Cassels & Johnstone, 1983; Gabel, 1999; Johnstone, 1991) have been studying the difficulty students have in learning science, particularly chemistry. Gabel (1999) suggests that, "while research into misconceptions (also known as alternative conceptions) and problem-solving has dominated the field for the past 25 years, we are no closer to a solution that would improve the teaching and learning of chemistry" (P. 549). Gabel (1999) relates the difficulty in learning chemistry to use of language. She refers to student difficulty both with words that have more than one meaning in English and with words that are used to mean one idea in chemistry and another idea in every day language. The Frayer Model, a research-based teaching strategy, is a graphic organizer which students use to create meaningful definitions for terms in context (Frayer, Frederick, & Klausmeier, 1969). It was used as the treatment---the specific vocabulary instruction---in this research study. The researcher collected and analyzed data to answer three research questions that focused on the effect of using the Frayer model (a graphic organizer) on high school students' knowledge and understanding of academic language used in chemistry. The research took place in a New England high school. Four intact chemistry classes provided the student participants; two classes were assigned to the treatment group (TG) and two classes were assigned to the control group (CG). The TG received vocabulary instruction on 14 chosen terms using the Frayer Model. The CG received traditional vocabulary instruction with no special attention to the 14 terms selected for this study

  3. Molecular and immunohistochemical detection of Kaposi sarcoma herpesvirus/human herpesvirus-8. (United States)

    Chadburn, Amy; Wilson, Janet; Wang, Y Lynn


    Kaposi sarcoma herpesvirus/human herpesvirus-8 (KSHV/HHV-8) is etiologically related to the development of several human diseases, including Kaposi sarcoma, primary effusion lymphoma (PEL)/extra-cavitary (EC) PEL, multicentric Castleman disease (MCD), and large B-cell lymphoma arising in KSHV/HHV-8-associated multicentric Castleman disease. Although serologic studies can identify persons infected with this virus, molecular genetics, specifically PCR (polymerase chain reaction) and immunohistochemical techniques, are rapid, sensitive, and specific, and are able to more closely link KSHV/HHV-8 to a given disease process. As these KSHV/HHV-8-related diseases cause significant morbidity and mortality in affected individuals, the identification of the virus within lesional tissue will allow for more targeted therapy.

  4. Scrambled eggs: A highly sensitive molecular diagnostic workflow for Fasciola species specific detection from faecal samples (United States)

    Calvani, Nichola Eliza Davies; Windsor, Peter Andrew; Bush, Russell David


    Background Fasciolosis, due to Fasciola hepatica and Fasciola gigantica, is a re-emerging zoonotic parasitic disease of worldwide importance. Human and animal infections are commonly diagnosed by the traditional sedimentation and faecal egg-counting technique. However, this technique is time-consuming and prone to sensitivity errors when a large number of samples must be processed or if the operator lacks sufficient experience. Additionally, diagnosis can only be made once the 12-week pre-patent period has passed. Recently, a commercially available coprological antigen ELISA has enabled detection of F. hepatica prior to the completion of the pre-patent period, providing earlier diagnosis and increased throughput, although species differentiation is not possible in areas of parasite sympatry. Real-time PCR offers the combined benefits of highly sensitive species differentiation for medium to large sample sizes. However, no molecular diagnostic workflow currently exists for the identification of Fasciola spp. in faecal samples. Methodology/Principal findings A new molecular diagnostic workflow for the highly-sensitive detection and quantification of Fasciola spp. in faecal samples was developed. The technique involves sedimenting and pelleting the samples prior to DNA isolation in order to concentrate the eggs, followed by disruption by bead-beating in a benchtop homogeniser to ensure access to DNA. Although both the new molecular workflow and the traditional sedimentation technique were sensitive and specific, the new molecular workflow enabled faster sample throughput in medium to large epidemiological studies, and provided the additional benefit of speciation. Further, good correlation (R2 = 0.74–0.76) was observed between the real-time PCR values and the faecal egg count (FEC) using the new molecular workflow for all herds and sampling periods. Finally, no effect of storage in 70% ethanol was detected on sedimentation and DNA isolation outcomes; enabling


    Directory of Open Access Journals (Sweden)

    M. Pour-Amir


    Full Text Available In this study we produced and purified a high titer of specific and high affin¬ity Fab fragments of anti-digoxin antibody. Immunization of rabbits with a conju¬gate of the cardiac glycoside digoxin, coupled by a periodate oxidation method to the amino group of lysine in bovine serum albumin resulted in the production of this type of high titer digoxin-specific antibodies with exceptionally high affinity (109 L/mol and specificity in immune response. Increase in titer was found in steps of purification ending up with the highest titer for Fab fragment to be at 1.75 ug of purified Fab (for 50% binding of I25I-digoxin. High specificity for antigenic determinants of the steroid nucleus of digoxin was observed such that much less cross-reaction with digoxin (2.3% and no cross-reaction with ouabaine, estradiol, Cortisol, progesterone and testosterone were detected.

  6. Immunohistochemical Expression of Myeloperoxidase in Placental Samples of Systematic Lupus Erythematosus Pregnancies

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    Zahra Heidari


    Full Text Available Objective: Reports of increased level of Myeloperoxidase (MPO in plasma and placental extracts of Systematic Lupus Erythematosus (SLE has been proposed that MPO may have an important role in this pregnancy complication. In present study immunohistochemical MPO expression was investigated in placental samples of SLE women compared with normal controls.Materials and methods: Ten patients with SLE were recruited as case group. Control group was selected from mothers with normal uncomplicated pregnancies. A monoclonal antibody specific for MPO was used for immunohistochemical staining and then the staining was quantified and differences between groups were compared using Mann-Whitney U test.Results: There were significant differences in the expression levels of MPO in the syncytiotrophoblast cells and the extravillous trophoblast cells between the control and SLE groups (p < 0.05. There were no significant differences in the expression level of MPO in the vascular endothelium and the relative number of the MPO-positive leukocytes in placental tissue between SLE and control groups (p > 0.05.Conclusion: The present study showed that MPO expression is increased in syncytiotrophoblast cells and the extravillous trophoblast cells of SLE placentas compared to healthy subjects. It seems that these changes are able to impress structure and function of placenta and survival of the fetus.

  7. Hep par-1: a novel immunohistochemical marker for differentiating hepatocellular carcinoma from metastatic carcinoma. (United States)

    Hanif, Razia; Mansoor, Samina


    To evaluate the diagnostic utility of Hep par-1 in differentiating hepatocellular carcinoma from metastatic carcinoma taking histopathology as a gold standard. Comparative cross-sectional study. Pathology Department, Shaukat Khanum Memorial Cancer Hospital and Research Centre, Lahore, from April 2007 to February 2008. Hep par-1 immunohistochemical stain was performed on 60 cases of liver carcinoma, 30 cases each of metastatic and hepatocellular carcinoma. Information regarding patient age, gender, sign and symptoms, radiographic findings, histological grade of tumour, and expression of Hep par-1 on hepatocellular and metastatic carcinoma were recorded on proforma sheet. Sensitivity, specificity, positive and negative predictive values, and accuracy of Hep par-1 were calculated using the formulas. Hep par-1 expression was noted in 25 out of 30 cases of hepatocellular carcinoma (83%). Out of 30 cases of metastatic carcinoma, only one case expressed staining in Hep par-1 is a reliable immunohistochemical marker for cases of hepatocellular carcinoma (HCC). It can be used along with other markers in morphologically difficult cases when differential diagnosis lies between poorly differentiated HCC and metastatic carcinoma of liver.

  8. Epithelial ovarian tumors: Clinicopathological correlation and immunohistochemical study

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    Pooja S Naik


    Full Text Available Background: Ovarian cancer is the third leading site of cancer among women, trailing behind cervix and breast cancer. Aim: This study was undertaken to analyze the immunohistochemical (IHC profile of estrogen receptors (ER, progesterone receptors (PR, Ki-67, and p53 in various ovarian epithelial tumors and attempt correlation with clinical and histopathological findings. Materials and Methods: The present study was conducted over a period of 4 years. A technique of manual tissue array was employed for cases subjected for IHC. The primary antibodies used were ER, PR, p53, and Ki-67. A correlation was attempted between histopathological and IHC findings. Results were subjected to statistical analysis. Software program "the primer of biostatistics 5.0" was used for calculation of interrelationships between the analyzed ER, PR, p53, and Ki-67 expression and histological factors by Pearson′s Chi-square test. The results were considered to be significant when the P < 0.05. Results: There were 110 cases of surface epithelial ovarian tumors (SEOT encountered over the period of 4 years. The expression of ER was more in malignant tumors (13/16, 81.25% than borderline (9/12, 75% and benign (20/82, 24.39%. As compared to ER, the expression of PR was more in benign (51/82, 62.19% than borderline (8/12, 66.67% and malignant tumors (9/16, 56.25%. The expression of PR was more in benign tumors than borderline and malignant tumors. However, this was not statistically significant (Chi-square = 0.335 with 2 degrees of freedom; P = 0.846. The expression of p53 was less in benign (5/82, 6.1% than borderline (9/12, 75% and malignant tumors (13/16, 81.25%. The expression of Ki-67 was more in malignant (4/82, 4.88% than borderline (10/12, 83.33% and benign tumors (15/16, 93.75%. In all the above cases, the difference was statistically significant (P < 0.05. There was statistically significant difference in the expression of ER, PR, p53, and Ki-67 in the patients with

  9. Evaluation of a Direct Rapid Immunohistochemical Test (dRIT) for Rapid Diagnosis of Rabies in Animals and Humans

    Institute of Scientific and Technical Information of China (English)

    Shampur Narayan Madhusudana; Sundaramurthy Subha; Ullas Thankappan; Yajaman Belludi Ashwin


    Presently the gold standard diagnostic technique for rabies is the direct immunofluorescence assay (dFA) which is very expensive and requires a high level of expertise.There is a need for more economical and user friendly tests,particularly for use in developing countries.We have established one such test called the direct rapid immunohistochemical test (dRIT) for diagnosis of rabies using brain tissue.The test is based on capture of rabies nucleoprotein (N) antigen in brain smears using a cocktail of biotinylated monoclonal antibodies specific for the N protein and color development by streptavidin peroxidase-amino ethyl carbazole and counter staining with haematoxollin.The test was done in parallel with standard FAT dFA using 400 brain samples from different animals and humans.The rabies virus N protein appears under light microscope as reddish brown particles against a light blue background.There was 100 % correlation between the results obtained by the two tests.Also,interpretation of results by dRIT was easier and only required a light microscope.To conclude,this newly developed dRIT technique promises to be a simple,cost effective diagnostic tool for rabies and will have applicability in field conditions prevalent in developing countries.

  10. Mid-Season High-Resolution Satellite Imagery for Forecasting Site-Specific Corn Yield

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    Nahuel R. Peralta


    Full Text Available A timely and accurate crop yield forecast is crucial to make better decisions on crop management, marketing, and storage by assessing ahead and implementing based on expected crop performance. The objective of this study was to investigate the potential of high-resolution satellite imagery data collected at mid-growing season for identification of within-field variability and to forecast corn yield at different sites within a field. A test was conducted on yield monitor data and RapidEye satellite imagery obtained for 22 cornfields located in five different counties (Clay, Dickinson, Rice, Saline, and Washington of Kansas (total of 457 ha. Three basic tests were conducted on the data: (1 spatial dependence on each of the yield and vegetation indices (VIs using Moran’s I test; (2 model selection for the relationship between imagery data and actual yield using ordinary least square regression (OLS and spatial econometric (SPL models; and (3 model validation for yield forecasting purposes. Spatial autocorrelation analysis (Moran’s I test for both yield and VIs (red edge NDVI = NDVIre, normalized difference vegetation index = NDVIr, SRre = red-edge simple ratio, near infrared = NIR and green-NDVI = NDVIG was tested positive and statistically significant for most of the fields (p < 0.05, except for one. Inclusion of spatial adjustment to model improved the model fit on most fields as compared to OLS models, with the spatial adjustment coefficient significant for half of the fields studied. When selected models were used for prediction to validate dataset, a striking similarity (RMSE = 0.02 was obtained between predicted and observed yield within a field. Yield maps could assist implementing more effective site-specific management tools and could be utilized as a proxy of yield monitor data. In summary, high-resolution satellite imagery data can be reasonably used to forecast yield via utilization of models that include spatial adjustment to

  11. A systematic comparison of quantitative high-resolution DNA methylation analysis and methylation-specific PCR (United States)

    Claus, Rainer; Wilop, Stefan; Hielscher, Thomas; Sonnet, Miriam; Dahl, Edgar; Galm, Oliver; Jost, Edgar; Plass, Christoph


    Assessment of DNA methylation has become a critical factor for the identification, development and application of methylation based biomarkers. Here we describe a systematic comparison of a quantitative high-resolution mass spectrometry-based approach (MassARRAY), pyrosequencing and the broadly used methylation-specific PCR (MSP) technique analyzing clinically relevant epigenetically silenced genes in acute myeloid leukemia (AML). By MassARRAY and pyrosequencing, we identified significant DNA methylation differences at the ID4 gene promoter and in the 5′ region of members of the SFRP gene family in 62 AML patients compared with healthy controls. We found a good correlation between data obtained by MassARRAY and pyrosequencing (correlation coefficient R2 = 0.88). MSP-based assessment of the identical samples showed less pronounced differences between AML patients and controls. By direct comparison of MSP-derived and MassARRAY-based methylation data as well as pyrosequencing, we could determine overestimation of DNA methylation data by MSP. We found sequence-context dependent highly variable cut-off values of quantitative DNA methylation values serving as discriminator for the two MSP methylation categories. Moreover, good agreements between quantitative methods and MSP could not be achieved for all investigated loci. Significant correlation of the quantitative assessment but not of MSP-derived methylation data with clinically important characteristics in our patient cohort demonstrated clinical relevance of quantitative DNA methylation assessment. Taken together, while MSP is still the most commonly applied technique for DNA methylation assessment, our data highlight advantages of quantitative approaches for precise characterization and reliable biomarker use of aberrant DNA methylation in primary patient samples, particularly. PMID:22647397

  12. Aspergillus fumigatus SidA is a highly specific ornithine hydroxylase with bound flavin cofactor. (United States)

    Chocklett, Samuel W; Sobrado, Pablo


    Ferrichrome is a hydroxamate-containing siderophore produced by the pathogenic fungus Aspergillus fumigatus under iron-limiting conditions. This siderophore contains N(5)-hydroxylated l-ornithines essential for iron binding. A. fumigatus siderophore A (Af SidA) catalyzes the flavin- and NADPH-dependent hydroxylation of l-ornithine in ferrichrome biosynthesis. Af SidA was recombinantly expressed and purified as a soluble tetramer and is the first member of this class of flavin monooxygenases to be isolated with a bound flavin cofactor. The enzyme showed typical saturation kinetics with respect to l-ornithine while substrate inhibition was observed at high concentrations of NADPH and NADH. Increasing amounts of hydrogen peroxide were measured as a function of reduced nicotinamide coenzyme concentration, indicating that inhibition was caused by increased uncoupling. Af SidA is highly specific for its amino acid substrate, only hydroxylating l-ornithine. An 8-fold preference in the catalytic efficiency was determined for NADPH compared to NADH. In the absence of substrate, Af SidA can be reduced by NADPH, and a C4a-(hydro)peroxyflavin intermediate is observed. The decay of this intermediate is accelerated by l-ornithine binding. This intermediate was only stabilized by NADPH and not by NADH, suggesting a role for NADP(+) in the stabilization of intermediates in the reaction of Af SidA. NADP(+) is a competitive inhibitor with respect to NADPH, demonstrating that Af SidA forms a ternary complex with NADP(+) and l-ornithine during catalysis. The data suggest that Af SidA likely proceeds by a sequential kinetic mechanism.

  13. Identification of High-Impact cis-Regulatory Mutations Using Transcription Factor Specific Random Forest Models.

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    Dmitry Svetlichnyy


    Full Text Available Cancer genomes contain vast amounts of somatic mutations, many of which are passenger mutations not involved in oncogenesis. Whereas driver mutations in protein-coding genes can be distinguished from passenger mutations based on their recurrence, non-coding mutations are usually not recurrent at the same position. Therefore, it is still unclear how to identify cis-regulatory driver mutations, particularly when chromatin data from the same patient is not available, thus relying only on sequence and expression information. Here we use machine-learning methods to predict functional regulatory regions using sequence information alone, and compare the predicted activity of the mutated region with the reference sequence. This way we define the Predicted Regulatory Impact of a Mutation in an Enhancer (PRIME. We find that the recently identified driver mutation in the TAL1 enhancer has a high PRIME score, representing a "gain-of-target" for MYB, whereas the highly recurrent TERT promoter mutation has a surprisingly low PRIME score. We trained Random Forest models for 45 cancer-related transcription factors, and used these to score variations in the HeLa genome and somatic mutations across more than five hundred cancer genomes. Each model predicts only a small fraction of non-coding mutations with a potential impact on the function of the encompassing regulatory region. Nevertheless, as these few candidate driver mutations are often linked to gains in chromatin activity and gene expression, they may contribute to the oncogenic program by altering the expression levels of specific oncogenes and tumor suppressor genes.

  14. High performance wash-free magnetic bioassays through microfluidically enhanced particle specificity. (United States)

    Bechstein, Daniel J B; Lee, Jung-Rok; Ooi, Chin Chun; Gani, Adi W; Kim, Kyunglok; Wilson, Robert J; Wang, Shan X


    Magnetic biosensors have emerged as a sensitive and versatile platform for high performance medical diagnostics. These magnetic biosensors require well-tailored magnetic particles as detection probes, which need to give rise to a large and specific biological signal while showing very low nonspecific binding. This is especially important in wash-free bioassay protocols, which do not require removal of particles before measurement, often a necessity in point of care diagnostics. Here we show that magnetic interactions between magnetic particles and magnetized sensors dramatically impact particle transport and magnetic adhesion to the sensor surfaces. We investigate the dynamics of magnetic particles' biomolecular binding and magnetic adhesion to the sensor surface using microfluidic experiments. We elucidate how flow forces can inhibit magnetic adhesion, greatly diminishing or even eliminating nonspecific signals in wash-free magnetic bioassays, and enhancing signal to noise ratios by several orders of magnitude. Our method is useful for selecting and optimizing magnetic particles for a wide range of magnetic sensor platforms.

  15. Highly abundant and stage-specific mRNAs in the obligate pathogen Bremia lactucae. (United States)

    Judelson, H S; Michelmore, R W


    Germinating spores of the obligate pathogen Bremia lactucae (lettuce downy mildew) contain several unusually abundant species of mRNA. Thirty-nine cDNA clones corresponding to prevalent transcripts were isolated from a library synthesized using poly(A)+ RNA from germinating spores; these clones represented only five distinct classes. Each corresponding mRNA accounted for from 0.4 to 9 percent by mass of poly(A)+ RNA from germinating spores and together represented greater than 20 percent of the mRNA. The expression of the corresponding genes, and a gene encoding Hsp70, was analyzed in spores during germination and during growth in planta. The Hsp70 mRNA and mRNA from one abundant cDNA clone (ham34) were expressed constitutively. Two clones (ham9 and ham12) hybridized only to mRNA from spores and germinating spores. Two clones (ham37 and ham27) showed hybridization specific to germinating spores. Quantification of the number of genes homologous to each cDNA clone indicated that four clones corresponded to one or two copies per haploid genome, and one hybridized to an approximately 11-member family of genes. A sequence of the gene corresponding to ham34 was obtained to investigate its function and to identify sequences conferring high levels of gene expression for use in constructing vectors for the transformation of B. lactucae.

  16. High-Specificity Targeted Functional Profiling in Microbial Communities with ShortBRED. (United States)

    Kaminski, James; Gibson, Molly K; Franzosa, Eric A; Segata, Nicola; Dantas, Gautam; Huttenhower, Curtis


    Profiling microbial community function from metagenomic sequencing data remains a computationally challenging problem. Mapping millions of DNA reads from such samples to reference protein databases requires long run-times, and short read lengths can result in spurious hits to unrelated proteins (loss of specificity). We developed ShortBRED (Short, Better Representative Extract Dataset) to address these challenges, facilitating fast, accurate functional profiling of metagenomic samples. ShortBRED consists of two components: (i) a method that reduces reference proteins of interest to short, highly representative amino acid sequences ("markers") and (ii) a search step that maps reads to these markers to quantify the relative abundance of their associated proteins. After evaluating ShortBRED on synthetic data, we applied it to profile antibiotic resistance protein families in the gut microbiomes of individuals from the United States, China, Malawi, and Venezuela. Our results support antibiotic resistance as a core function in the human gut microbiome, with tetracycline-resistant ribosomal protection proteins and Class A beta-lactamases being the most widely distributed resistance mechanisms worldwide. ShortBRED markers are applicable to other homology-based search tasks, which we demonstrate here by identifying phylogenetic signatures of antibiotic resistance across more than 3,000 microbial isolate genomes. ShortBRED can be applied to profile a wide variety of protein families of interest; the software, source code, and documentation are available for download at

  17. High-Specificity Targeted Functional Profiling in Microbial Communities with ShortBRED.

    Directory of Open Access Journals (Sweden)

    James Kaminski


    Full Text Available Profiling microbial community function from metagenomic sequencing data remains a computationally challenging problem. Mapping millions of DNA reads from such samples to reference protein databases requires long run-times, and short read lengths can result in spurious hits to unrelated proteins (loss of specificity. We developed ShortBRED (Short, Better Representative Extract Dataset to address these challenges, facilitating fast, accurate functional profiling of metagenomic samples. ShortBRED consists of two components: (i a method that reduces reference proteins of interest to short, highly representative amino acid sequences ("markers" and (ii a search step that maps reads to these markers to quantify the relative abundance of their associated proteins. After evaluating ShortBRED on synthetic data, we applied it to profile antibiotic resistance protein families in the gut microbiomes of individuals from the United States, China, Malawi, and Venezuela. Our results support antibiotic resistance as a core function in the human gut microbiome, with tetracycline-resistant ribosomal protection proteins and Class A beta-lactamases being the most widely distributed resistance mechanisms worldwide. ShortBRED markers are applicable to other homology-based search tasks, which we demonstrate here by identifying phylogenetic signatures of antibiotic resistance across more than 3,000 microbial isolate genomes. ShortBRED can be applied to profile a wide variety of protein families of interest; the software, source code, and documentation are available for download at

  18. High specific surface gold electrode on polystyrene substrate: Characterization and application as DNA biosensor. (United States)

    Yang, Zhiliu; Liu, Yichen; Lu, Wei; Yuan, Qingpan; Wang, Wei; Pu, Qiaosheng; Yao, Bo


    In the past decades, many efforts have been made to improve the sensitivity and specificity of electrochemical DNA biosensors. However, it is still strongly required to develop disposable and reliable DNA biosensors for wide and practical application. In this article, we reported superior electrochemical properties of an integrated plastic-gold electrode (PGE) fabricated in-house by chemical plating on polystyrene substrate. PGEs were found having extremely high capacity of DNA immobilization compared with gold electrodes fabricated by standard sputtering based photolithography. Unique nano-structured surface was observed on PGEs through morphology techniques, which would to some extend give an explanation to higher capacity of DNA immobilization on PGEs. A probable mechanism of carboxylic acid produced on polystyrene substrate after exposure to UV irradiation was proposed and discussed for the first time. This biosensor was applied to detection and manipulate of DNA hybridization. Detection limit of 7.2×10(-11) M and 1-500 nM of linearity range was obtained.

  19. Operating Cooperatively (OC sensor for highly specific recognition of nucleic acids.

    Directory of Open Access Journals (Sweden)

    Evan M Cornett

    Full Text Available Molecular Beacon (MB probes have been extensively used for nucleic acid analysis because of their ability to produce fluorescent signal in solution instantly after hybridization. The indirect binding of MB probe to a target analyte offers several advantages, including: improved genotyping accuracy and the possibility to analyse folded nucleic acids. Here we report on a new design for MB-based sensor, called 'Operating Cooperatively' (OC, which takes advantage of indirect binding of MB probe to a target analyte. The sensor consists of two unmodified DNA strands, which hybridize to a universal MB probe and a nucleic acid analyte to form a fluorescent complex. OC sensors were designed to analyze two human SNPs and E. coli 16S rRNA. High specificity of the approach was demonstrated by the detection of true analyte in over 100 times excess amount of single base substituted analytes. Taking into account the flexibility in the design and the simplicity in optimization, we conclude that OC sensors may become versatile and efficient tools for instant DNA and RNA analysis in homogeneous solution.

  20. Specific high-affinity binding of fatty acids to epidermal cytosolic proteins

    Energy Technology Data Exchange (ETDEWEB)

    Raza, H.; Chung, W.L.; Mukhtar, H. (Department of Dermatology, University Hospitals of Cleveland, Case Western Reserve University, OH (USA))


    Cytosol from rat, mouse, and human skin or rat epidermis was incubated with (3H)arachidonic acid, (14C)retinoic acid, (14C)oleic acid, (3H)leukotriene A4, (3H)prostaglandin E2 (PGE2) or (3H) 15-hydroxyeicosatetraenoic acid (15-HETE), and protein-bound ligands were separated using Lipidex-1000 at 4C to assess the binding specificity. The binding of oleic acid and arachidonic acid with rat epidermal cytosol was rapid, saturable, and reversible. Binding of oleic acid was competed out with the simultaneous addition of other ligands and found to be in the following order: arachidonic acid greater than oleic acid greater than linoleic acid greater than lauric acid greater than leukotriene A4 greater than 15-HETE = PGE1 greater than PGE2 = PGF2. Scatchard analysis of the binding with arachidonic acid, oleic acid, and retinoic acid revealed high-affinity binding sites with the dissociation constant in the nM range. SDS-PAGE analysis of the oleic acid-bound epidermal cytosolic protein(s) revealed maximum binding at the 14.5 kDa region. The presence of the fatty acid-binding protein in epidermal cytosol and its binding to fatty acids and retinoic acid may be of significance both in the trafficking and the metabolism of fatty acids and retinoids across the skin.

  1. High fat diet deviates PtC-specific B1 B cell phagocytosis in obese mice. (United States)

    Vo, Hung; Chiu, Joanna; Allaimo, Danielle; Mao, Changchuin; Wang, Yaqi; Gong, Yuefei; Ow, Hooisweng; Porter, Tyrone; Zhong, Xuemei


    Phagocytosis had been attributed predominantly to "professional" phagocytes such as macrophages, which play critical roles in adipose tissue inflammation. However, recently, macrophage-like phagocytic activity has been reported in B1 B lymphocytes. Intrigued by the long-established correlation between high fat diet (HFD)-induced obesity and immune dysfunction, we investigated how HFD affects B1 B cell phagocytosis. A significant number of B1 B cells recognize phosphatidylcholine (PtC), a common phospholipid component of cell membrane. We report here that unlike macrophages, B1 B cells have a unique PtC-specific phagocytic function. In the presence of both PtC-coated and non-PtC control fluorescent nano-particles, B1 B cells from healthy lean mice selectively engulfed PtC-coated beads, whereas B1 B cells from HFD-fed obese mice non-discriminately phagocytosed both PtC-coated and control beads. Morphologically, B1 B cells from obese mice resembled macrophages, displaying enlarged cytosol and engulfed more beads. Our study suggests for the first time that HFD can affect B1 B cell phagocytosis, substantiating the link of HFD-induced obesity and immune deviation.

  2. High-k Dielectric Passivation: Novel Considerations Enabling Cell Specific Lysis Induced by Electric Fields. (United States)

    Wassermann, Klemens J; Barth, Sven; Keplinger, Franz; Noehammer, Christa; Peham, Johannes R


    A better understanding of the electrodynamic behavior of cells interacting with electric fields would allow for novel scientific insights and would lead to the next generation of cell manipulation, diagnostics, and treatment. Here, we introduce a promising electrode design by using metal oxide high-k dielectric passivation. The thermally generated dielectric passivation layer enables efficient electric field coupling to the fluid sample comprising cells while simultaneously decoupling the electrode ohmically from the electrolyte, allowing for better control and adjustability of electric field effects due to reduced electrochemical reactions at the electrode surface. This approach demonstrates cell-size specific lysis with electric fields in a microfluidic flow-through design resulting in 99.8% blood cell lysis at 6 s exposure without affecting the viability of Gram-positive and Gram-negative bacterial spike-ins. The advantages of this new approach can support next-generation investigations of electrodynamics in biological systems and their exploitation for cell manipulation in multiple fields of medicine, life science, and industry.

  3. A dominant EV71-specific CD4+ T cell epitope is highly conserved among human enteroviruses.

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    Ruicheng Wei

    Full Text Available CD4+ T cell-mediated immunity plays a central role in determining the immunopathogenesis of viral infections. However, the role of CD4+ T cells in EV71 infection, which causes hand, foot and mouth disease (HFMD, has yet to be elucidated. We applied a sophisticated method to identify promiscuous CD4+ T cell epitopes contained within the sequence of the EV71 polyprotein. Fifteen epitopes were identified, and three of them are dominant ones. The most dominant epitope is highly conserved among enterovirus species, including HFMD-related coxsackieviruses, HFMD-unrelated echoviruses and polioviruses. Furthermore, the CD4+ T cells specific to the epitope indeed cross-reacted with the homolog of poliovirus 3 Sabin. Our findings imply that CD4+ T cell responses to poliovirus following vaccination, or to other enteroviruses to which individuals may be exposed in early childhood, may have a modulating effect on subsequent CD4+ T cell response to EV71 infection or vaccine.

  4. A new highly specific and robust yeast androgen bioassay for the detection of agonists and antagonists. (United States)

    Bovee, Toine F H; Helsdingen, Richard J R; Hamers, Astrid R M; van Duursen, Majorie B M; Nielen, Michel W F; Hoogenboom, Ron L A P


    Public concern about the presence of natural and anthropogenic compounds which affect human health by modulating normal endocrine functions is continuously growing. Fast and simple high-throughput screening methods for the detection of hormone activities are thus indispensable. During the last two decades, a panel of different in vitro assays has been developed, mainly for compounds with an estrogenic mode of action. Here we describe the development of an androgen transcription activation assay that is easy to use in routine screening. Recombinant yeast cells were constructed that express the human androgen receptor and yeast enhanced green fluorescent protein (yEGFP), the latter in response to androgens. Compared with other reporters, the yEGFP reporter protein is very convenient because it is directly measurable in intact living cells, i.e., cell wall disruption and the addition of a substrate are not needed. When yeast was exposed to 17beta-testosterone, the concentration where half-maximal activation is reached (EC(50)) was 50 nM. The relative androgenic potencies, defined as the ratio between the EC(50) of 17beta-testosterone and the EC(50) of the compound, of 5alpha-dihydrotestosterone, methyltrienolone, and 17beta-boldenone are 2.3, 1.4, and 0.15 respectively. The results presented in this paper demonstrate that this new yeast androgen bioassay is fast, sensitive, and very specific and also suited to detect compounds that have an antiandrogenic mode of action.

  5. Immunohistochemical localization of chondroitin sulfate, chondroitin sulfate proteoglycan, heparan sulfate proteoglycan, entactin, and laminin in basement membranes of postnatal developing and adult rat lungs

    DEFF Research Database (Denmark)

    Sannes, P L; Burch, K K; Khosla, J


    Histologic preparations of lungs from 1-, 5-, 10-, 18-, and 25-day-old postnatal and adult rats were examined immunohistochemically with antibodies specific against chondroitin sulfate (CS), basement membrane chondroitin sulfate proteoglycan (BM-CSPG), heparan sulfate proteoglycan (HSPG), entactin...

  6. Highly sensitive real-time PCR for specific detection and quantification of Coxiella burnetii

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    Linke Sonja


    Full Text Available Abstract Background Coxiella burnetii, the bacterium causing Q fever, is an obligate intracellular biosafety level 3 agent. Detection and quantification of these bacteria with conventional methods is time consuming and dangerous. During the last years, several PCR based diagnostic assays were developed to detect C. burnetii DNA in cell cultures and clinical samples. We developed and evaluated TaqMan-based real-time PCR assays that targeted the singular icd (isocitrate dehydrogenase gene and the transposase of the IS1111a element present in multiple copies in the C. burnetii genome. Results To evaluate the precision of the icd and IS1111 real-time PCR assays, we performed different PCR runs with independent DNA dilutions of the C. burnetii Nine Mile RSA493 strain. The results showed very low variability, indicating efficient reproducibility of both assays. Using probit analysis, we determined that the minimal number of genome equivalents per reaction that could be detected with a 95% probability was 10 for the icd marker and 6.5 for the IS marker. Plasmid standards with cloned icd and IS1111 fragments were used to establish standard curves which were linear over a range from 10 to 107 starting plasmid copy numbers. We were able to quantify cell numbers of a diluted, heat-inactivated Coxiella isolate with a detection limit of 17 C. burnetii particles per reaction. Real-time PCR targeting both markers was performed with DNA of 75 different C. burnetii isolates originating from all over the world. Using this approach, the number of IS1111 elements in the genome of the Nine Mile strain was determined to be 23, close to 20, the number revealed by genome sequencing. In other isolates, the number of IS1111 elements varied widely (between seven and 110 and seemed to be very high in some isolates. Conclusion We validated TaqMan-based real-time PCR assays targeting the icd and IS1111 markers of C. burnetii. The assays were shown to be specific, highly

  7. Development of an unmanned aerial vehicle-based spray system for highly accurate site-specific application (United States)

    Application of crop production and protection materials is a crucial component in the high productivity of American agriculture. Agricultural chemical application is frequently needed at a specific time and location for accurate site-specific management of crop pests. Piloted aircrafts that carry ...

  8. Differential expression of HSPA1 and HSPA2 proteins in human tissues; tissue microarray-based immunohistochemical study. (United States)

    Scieglinska, Dorota; Piglowski, Wojciech; Chekan, Mykola; Mazurek, Agnieszka; Krawczyk, Zdzisław


    In the present study we determined the expression pattern of HSPA1 and HSPA2 proteins in various normal human tissues by tissue-microarray based immunohistochemical analysis. Both proteins belong to the HSPA (HSP70) family of heat shock proteins. The HSPA2 is encoded by the gene originally defined as testis-specific, while HSPA1 is encoded by the stress-inducible genes (HSPA1A and HSPA1B). Our study revealed that both proteins are expressed only in some tissues from the 24 ones examined. HSPA2 was detected in adrenal gland, bronchus, cerebellum, cerebrum, colon, esophagus, kidney, skin, small intestine, stomach and testis, but not in adipose tissue, bladder, breast, cardiac muscle, diaphragm, liver, lung, lymph node, pancreas, prostate, skeletal muscle, spleen, thyroid. Expression of HSPA1 was detected in adrenal gland, bladder, breast, bronchus, cardiac muscle, esophagus, kidney, prostate, skin, but not in other tissues examined. Moreover, HSPA2 and HSPA1 proteins were found to be expressed in a cell-type-specific manner. The most pronounced cell-type expression pattern was found for HSPA2 protein. In the case of stratified squamous epithelia of the skin and esophagus, as well as in ciliated pseudostratified columnar epithelium lining respiratory tract, the HSPA2 positive cells were located in the basal layer. In the colon, small intestine and bronchus epithelia HSPA2 was detected in goblet cells. In adrenal gland cortex HSPA2 expression was limited to cells of zona reticularis. The presented results clearly show that certain human tissues constitutively express varying levels of HSPA1 and HSPA2 proteins in a highly differentiated way. Thus, our study can help designing experimental models suitable for cell- and tissue-type-specific functional differences between HSPA2 and HSPA1 proteins in human tissues.

  9. Histopathologic and Immunohistochemical Correlates of Confocal Descriptors in Pigmented Facial Macules on Photodamaged Skin. (United States)

    Gómez-Martín, Ignacio; Moreno, Sara; Andrades-López, Evelyn; Hernández-Muñoz, Inma; Gallardo, Fernando; Barranco, Carlos; Pujol, Ramon M; Segura, Sonia


    Pigmented facial macules on photodamaged skin are a clinical, dermoscopic, and histopathologic challenge. To clinically and dermoscopically characterize, by means of reflectance confocal microscopy (RCM), ambiguous pigmented facial macules and establish a correlation between RCM, histopathologic, and immunohistochemical findings. A prospective study of ambiguous pigmented facial macules on photodamaged skin was conducted in a tertiary referral center for dermatology between January 1, 2009, and December 31, 2015. Sixty-one patients with 63 ambiguous pigmented facial macules and 12 control photodamaged facial areas were included in the study. Melanocyte density in 1-mm basal layers was determined in skin biopsy specimens from all lesions stained with hematoxylin-eosin and immunohistochemical markers (melan-A, microphthalmia-associated transcription factor, and SRY-related HMG-box gene 10). Dermoscopic, RCM images, and histopathologic preparations were systematically evaluated for the presence of lentigo maligna (LM) criteria. Confocal evaluation was blinded to clinical and dermoscopic diagnosis. Sensitivity and specificity of RCM for LM diagnosis and κ value to establish correlations between dermoscopy, RCM, and histopathology were performed. Sensitivity and specificity of RCM for LM diagnosis. Of the 61 patients included in the study, 31 (51%) were women; mean (SD) age was 71.8 (13.1) years. Twenty-four of the 63 (38%) lesions were diagnosed as LM or LM melanoma (LMM) and 39 (62%) as benign pigmented lesions. Reflectance confocal microscopy enhanced the diagnosis of pigmented facial macules with 91.7% sensitivity and 86.8% specificity. Multivariate analysis showed 2 dermoscopic and 2 confocal features associated with LM or LMM: (1) asymmetric follicular pigmentation and targetlike structures, and (2) round, large pagetoid cells and follicular localization of atypical cells, respectively. Continuous proliferation of atypical melanocytes was found in 21 (88%) LM or

  10. Evaluation of two sets of immunohistochemical and Western blot confirmatory methods in the detection of typical and atypical BSE cases

    Directory of Open Access Journals (Sweden)

    Greenlee Justin J


    Full Text Available Abstract Background Three distinct forms of bovine spongiform encephalopathy (BSE, defined as classical (C-, low (L- or high (H- type, have been detected through ongoing active and passive surveillance systems for the disease. The aim of the present study was to compare the ability of two sets of immunohistochemical (IHC and Western blot (WB BSE confirmatory protocols to detect C- and atypical (L- and H-type BSE forms. Obex samples from cases of United States and Italian C-type BSE, a U.S. H-type and an Italian L-type BSE case were tested in parallel using the two IHC sets and WB methods. Results The two IHC techniques proved equivalent in identifying and differentiating between C-type, L-type and H-type BSE. The IHC protocols appeared consistent in the identification of PrPSc distribution and deposition patterns in relation to the BSE type examined. Both IHC methods evidenced three distinct PrPSc phenotypes for each type of BSE: prevailing granular and linear tracts pattern in the C-type; intraglial and intraneuronal deposits in the H-type; plaques in the L-type. Also, the two techniques gave comparable results for PrPSc staining intensity on the C- and L-type BSE samples, whereas a higher amount of intraglial and intraneuronal PrPSc deposition on the H-type BSE case was revealed by the method based on a stronger demasking step. Both WB methods were consistent in identifying classical and atypical BSE forms and in differentiating the specific PrPSc molecular weight and glycoform ratios of each form. Conclusions The study showed that the IHC and WB BSE confirmatory methods were equally able to recognize C-, L- and H-type BSE forms and to discriminate between their different immunohistochemical and molecular phenotypes. Of note is that for the first time one of the two sets of BSE confirmatory protocols proved effective in identifying the L-type BSE form. This finding helps to validate the suitability of the BSE confirmatory tests for BSE

  11. Calcifying Odontogenic Cyst with Complex Odontoma: Histological and Immunohistochemical Features

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    Nooshin Mohtasham


    Full Text Available The calcifying odontogenic cyst (COC is a rare odontogenic cyst. Only 2% of all odontogenic cysts and tumors are COC. COC associated with odontoma (COCaO reported in 24% of COCs. COCaO presents a greater incidence in female, with a ratio of 2 to 1. The highest incidence of COCaO occurs during the second decade with a mean age of 16 years, most frequently occurring in the maxilla (61.5%. Here, we describe a classic case of COCaO of the maxillary incisor-canine region in 17-year-old girl, and discuss the clinicopathological features and immunohistochemical finding of this tumor.

  12. Calcifying Odontogenic Cyst with Complex Odontoma: Histological and Immunohistochemical Features

    Directory of Open Access Journals (Sweden)

    Mohsen Merati


    Full Text Available The calcifying odontogenic cyst (COC is a rare odontogenic cyst. Only 2% of all odontogenic cysts and tumors are COC. COC associated with odontoma (COCaO reported in 24% of COCs. COCaO presents a greater incidence in female, with a ratio of 2 to 1. The highest incidence of COCaO occurs during the second decade with a mean age of 16 years, most frequently occurring in the maxilla (61.5%. Here, we describe a classic case of COCaO of the maxillary incisor-canine region in 17-year-old girl, and discuss the clinicopathological features and immunohistochemical finding of this tumor.

  13. Immunohistochemical studies of the periodontal membrane in primary teeth

    DEFF Research Database (Denmark)

    Bille, Marie-Louise Bastholm; Nolting, Dorrit; Kjær, Inger


    Objectives. To describe the periodontal membrane of human primary teeth immunohistochemically, while focusing on the epithelial layer of Malassez, fibers, and peripheral nerves, and to compare the findings with those of a previous study of human permanent teeth. Material and methods. Nineteen human...... could be identical to those in regions with no resorption. Conclusion. In regions without resorption, spatial organization of the periodontal membrane of primary teeth was similar to that of permanent teeth, although the number and distribution of epithelial cells and fibers differed. In regions...

  14. Histopathological and immunohistochemical diagnosis of infectious bursal disease in poultry birds

    Directory of Open Access Journals (Sweden)

    J. Singh


    Full Text Available Aim: The aim of the present study was to diagnose infectious bursal disease (IBD using gross, histopathological, and immunopathological approaches and to compare efficacy of immunohistochemical techniques with conventional diagnostic techniques. Materials and Methods: A total of 33 samples were collected from the six different poultry farms from Ludhiana and the nearby districts. Upon gross analysis of the necropsied birds, the relevant tissue samples such as bursa, kidney, junction of proventriculus and gizzard, heart, and muscles were then processed for histopathological and immunohistochemical studies. Results: Varied macroscopic changes were noted in bursa, characterized as swollen, hemorrhages to atrophy in size. Nonetheless, hemorrhages over thigh muscles were rarely seen. Histologically, the bursa showed prominent fibrotic and atrophic changes. Rarefaction of bursal follicles with intermittent infiltration of lympho-mononuclear cells with chronic cystic changes was additional changes, considered to be paramount for IBD. Expression and localization of IBD specific viral antigens were noticed mainly intracellular to the rarefied areas of bursal follicle section(s, in conjunction to inner lining of the cystic cavities of affected follicles. In addition, the junction of proventriculus and gizzard, the heart muscle, respiratory ciliated epithelium, and proventriculus also revealed positive expression to IBD virus (IBDV antigen. Advanced immunopathological techniques, i.e., immunofluorescence further testified the evidence of antigen as positive green signal within affected follicles. Further consideration to the reliability of various techniques employed, positive correlation (r=0.64623 was emerged out with conventional pathological scoring. Conclusion: It is concluded that the bursa acts as an organ of choice for demonstrating IBDV antigen for specific diagnosis of disease using immunohistochemistry (IHC, and IHC staining is a precise

  15. Malignant Peripheral Nerve Sheath Tumors: Differentiation Patterns and Immunohistochemical Features - A Mini-Review and Our New Findings


    Guo, Aitao; Liu,Aijun; Wei, Lixin; Song, Xin


    Malignant peripheral nerve sheath tumors (MPNST) represent a group of highly heterogeneous human malignancies often with multiple histological origins, divergent differentiation patterns, and diverse immunohistochemical presentations. The differential diagnosis of MPNST from other spindle cell neoplasms poses great challenges for pathologists. This report provides a mini-review of these unique features associated with MPNST and also presents the first cases of MPNST with six differentiation p...

  16. Malignant Peripheral Nerve Sheath Tumors: Differentiation Patterns and Immunohistochemical Features - A Mini-Review and Our New Findings

    Directory of Open Access Journals (Sweden)

    Aitao Guo, Aijun Liu, Lixin Wei, Xin Song


    Full Text Available Malignant peripheral nerve sheath tumors (MPNST represent a group of highly heterogeneous human malignancies often with multiple histological origins, divergent differentiation patterns, and diverse immunohistochemical presentations. The differential diagnosis of MPNST from other spindle cell neoplasms poses great challenges for pathologists. This report provides a mini-review of these unique features associated with MPNST and also presents the first cases of MPNST with six differentiation patterns.

  17. The prion-related protein (testis-specific) gene (PRNT) is highly polymorphic in Portuguese sheep. (United States)

    Mesquita, P; Garcia, V; Marques, M R; Santos Silva, F; Oliveira Sousa, M C; Carolino, I; Pimenta, J; Fontes, C M G A; Horta, A E M; Prates, J A M; Pereira, R M


    The objective of this study was to search for polymorphisms in the ovine prion-related protein (testis-specific) gene (PRNT). Sampling included 567 sheep from eight Portuguese breeds. The PRNT gene-coding region was analyzed by single-strand conformation polymorphism and sequencing, allowing the identification of the first ovine PRNT polymorphisms, in codons 6, 38, 43 and 48: c.17C>T (p.Ser6Phe, which disrupts a consensus arginine-X-X-serine/threonine motif); c.112G>C (p.Gly38>Arg); c.129T>C and c.144A>G (synonymous) respectively. Polymorphisms in codons 6, 38 and 48 occur simultaneously in 50.6% of the animals, 38.8% presenting as heterozygous. To study the distribution of the polymorphism in codon 43, a restriction fragment length polymorphism analysis was performed. Polymorphic variant c.129C, identified in 89.8% of the animals with 32.8% presented as heterozygous, was considered the wild genotype in Portuguese sheep. Eight different haplotypes which have comparable distribution in all breeds were identified for the PRNT gene. In conclusion, the PRNT coding region is highly polymorphic in sheep, unlike the prion protein 2 dublet gene (PRND), in which we previously found only one synonymous substitution (c.78G>A), in codon 26. The absence or reduced number of PRND heterozygotes (c.78G>A) was significantly associated with three PRNT haplotypes (17C-112G-129T-144A,17CT-112GC-129CT-144AG and 17T-112C-129C-144G), and the only three animals found homozygous at c.78A had the 17C-112G-129C-144A PRNT haplotype. These results constitute evidence of an association between polymorphic variation in PRND and PRNT genes, as has already been observed for PRND and prion protein gene (PRNP). © 2015 Stichting International Foundation for Animal Genetics.

  18. [{sup 11}C]GR103545: novel one-pot radiosynthesis with high specific activity

    Energy Technology Data Exchange (ETDEWEB)

    Nabulsi, Nabeel B., E-mail: nabeel.nabulsi@yale.ed [Department of Diagnostic Radiology, PET Center, Yale School of Medicine, PO Box 208048, New Haven, CT 06520-8048 (United States); Zheng Mingqiang; Ropchan, Jim; Labaree, David; Ding Yushin [Department of Diagnostic Radiology, PET Center, Yale School of Medicine, PO Box 208048, New Haven, CT 06520-8048 (United States); Blumberg, Laura [Pfizer Global R and D, Groton, CT 06340 (United States); Huang Yiyun [Department of Diagnostic Radiology, PET Center, Yale School of Medicine, PO Box 208048, New Haven, CT 06520-8048 (United States)


    Introduction: GR103545 is a potent and selective kappa-opioid receptor agonist. Previous studies in non-human primates demonstrated favorable properties of [{sup 11}C]GR103545 as a positron emission tomography tracer for in vivo imaging of cerebral kappa-opioid receptor. Nonetheless, advancement of [{sup 11}C]GR103545 to imaging studies in humans was hampered by difficulties of its multiple-step radiosynthesis, which produces a final product with low specific activity (SA), which in turn could induce undesirable physiological side effects resulting from the mass associated with an injected amount of radioactivity. We report herein an alternative radiosynthesis of [{sup 11}C]GR103545 with higher SA and radiochemical yields. Methods: The TRACERLab FXC automated synthesis module was used to carry out the two-step, one-pot procedure. In the first step, the desmethoxycarbonyl precursor was converted to the carbamic acid intermediate desmethyl-GR103545 via transcarboxylation with the zwitterionic carbamic complex, 1,8-diazabicyclo[5.4.0]undec-7-ene-carbon dioxide, in the presence and/or absence of cesium carbonate and tetrabutylammonium triflate. In the second step, the intermediate was radiolabeled at the carboxyl oxygen with [{sup 11}C]methyl trifluoromethanesulfonate to give [{sup 11}C]GR103545. Results: This novel synthesis produced [{sup 11}C]GR103545 with {>=}90% chemical and radiochemical purities and an SA of 290.45{+-}99.9 MBq/nmol at the end of synthesis (n=26). Injectable radioactivity was 1961{+-}814 GBq/{mu}mol with 43 min of average synthesis time from the end of beam. Conclusion: We have developed a practical one-pot method for the routine production of [{sup 11}C]GR103545 with reliably high SA and radiochemical yield, thus allowing the advancement of this radiotracer to imaging applications in humans.

  19. High-Sensitivity Monoclonal Antibodies Specific for Homoserine Lactones Protect Mice from Lethal Pseudomonas aeruginosa Infections (United States)

    Downham, Christina; Broadbent, Ian; Charlton, Keith; Porter, Andrew J.


    A number of bacteria, including pathogens like Pseudomonas aeruginosa, utilize homoserine lactones (HSLs) as quorum sensing (QS) signaling compounds and engage in cell-to-cell communication to coordinate their behavior. Blocking this bacterial communication may be an attractive strategy for infection control as QS takes a central role in P. aeruginosa biology. In this study, immunomodulation of HSL molecules by monoclonal antibodies (MAbs) was used as a novel approach to prevent P. aeruginosa infections and as tools to detect HSLs in bodily fluids as a possible first clue to an undiagnosed Gram-negative infection. Using sheep immunization and recombinant antibody technology, a panel of sheep-mouse chimeric MAbs were generated which recognized HSL compounds with high sensitivity (nanomolar range) and cross-reactivity. These MAbs retained their nanomolar sensitivity in complex matrices and were able to recognize HSLs in P. aeruginosa cultures grown in the presence of urine. In a nematode slow-killing assay, HSL MAbs significantly increased the survival of worms fed on the antibiotic-resistant strain PA058. The therapeutic benefit of these MAbs was further studied using a mouse model of Pseudomonas infection in which groups of mice treated with HSL-2 and HSL-4 MAbs survived, 7 days after pathogen challenge, in significantly greater numbers (83 and 67%, respectively) compared with the control groups. This body of work has provided early proof-of-concept data to demonstrate the potential of HSL-specific, monoclonal antibodies as theranostic clinical leads suitable for the diagnosis, prevention, and treatment of life-threatening bacterial infections. PMID:24185854

  20. Layer-specific high-frequency spiking in the prefrontal cortex of awake rats

    Directory of Open Access Journals (Sweden)

    Zimbo Saroeni Raymond Maria Boudewijns


    Full Text Available Cortical pyramidal neurons show irregular in vivo action potential (AP spiking with high frequency bursts occurring on sparse background activity. Somatic APs can backpropagate from soma into basal and apical dendrites and locally generate dendritic calcium spikes. The critical AP frequency for generation of such dendritic calcium spikes can be very different depending on cell-type or brain area involved. Previously, it was shown in vitro that calcium electrogenesis can also be induced in L(ayer 5 pyramidal neurons of prefrontal cortex (PFC. It remains an open question whether somatic burst spiking and resulting dendritic calcium electrogenesis also occur in morphologically more compact L2/3 pyramidal neurons. Furthermore, it is not known whether critical frequencies that trigger dendritic calcium electrogenesis occur in PFC under awake conditions in vivo. Here, we addressed these issues and found that pyramidal neurons in both PFC L2/3 and L5 in awake rats spike APs in short bursts, but with different probabilities. The critical frequency for calcium electrogenesis in vitro was layer-specific and lower in L5 neurons compared to L2/3. Taking the in vitro critical frequency as predictive measure for dendritic electrogenesis during in vivo spontaneous activity, supracritical bursts in vivo were observed in a larger fraction of L5 neurons compared to L2/3 neurons but with similar incidence within these subpopulations. Together, these results show that in PFC of awake rats, AP spiking occurs at frequencies that are relevant for dendritic calcium electrogenesis and suggest that in awake rat PFC, dendritic calcium electrogenesis may be involved in neuronal computation.

  1. Determination of plasma lactic acid concentration and specific activity using high-performance liquid chromatography. (United States)

    Bleiberg, B; Steinberg, J J; Katz, S D; Wexler, J; LeJemtel, T


    Assessment of lactate metabolism is of particular interest during exercise and in disease states such as diabetes, shock, and absorptive abnormalities of short-chain fatty acids by the colon. We describe an analytical method that introduces radio-active tracers and high-performance liquid chromatography (HPLC) to simultaneously analyze concentrations and specific activities (SAs) of plasma lactate. The HPLC conditions included separation on a reversed-phase column (octadecylsilane) and an isocratic buffer (30% acetonitrile in water). [3H]Acetate served as an internal standard. Lactate and acetate were extracted from plasma samples with diethyl ether following a pH adjustment to less than 1.0 and back-extracted into a hydrophilic phase with sodium carbonate (2 mM, pH greater than 10.0). Lactate is detected in the ultraviolet range (242 and 320 nm) by derivatization with alpha-bromoacetophenone. Control plasma samples were studied after an overnight fast for precision and analytical recovery. Calibration curves were linear in the range 0.18-6.0 mM (r = 0.92). The precision was 3% and the analytical recovery was 87%. The detection limit of the method was 36 pmol. Determination of lactate metabolism was performed in a patient with chronic congestive heart failure who was administered primed-continuous L-[U-14C]lactate (10 microCi bolus and 0.3 microCi/min continuously) during a 60-min rest period. Mean arterial lactate concentration and SA were 1.69 +/- 0.2 mM and 253.8 +/- 22 dpm/mumol, respectively. Systemic lactate turnover was 25.65 mumol/kg per min. Lactic acid systemic turnover, organ uptake and release rates can be accurately determined by isocratic HPLC.

  2. Communication: High pressure specific heat spectroscopy reveals simple relaxation behavior of glass forming molecular liquid

    DEFF Research Database (Denmark)

    Roed, Lisa Anita; Niss, Kristine; Jakobsen, Bo


    The frequency dependent specific heat has been measured under pressure for the molecular glass forming liquid 5-polyphenyl-4-ether in the viscous regime close to the glass transition. The temperature and pressure dependences of the characteristic time scale associated with the specific heat...

  3. Iron titanium phosphates as high-specific-capacity electrode materials for lithium ion batteries

    Energy Technology Data Exchange (ETDEWEB)

    Essehli, R., E-mail: [Laboratory of Mineral Solid and Analytical Chemistry (LMSAC), Department of Chemistry, Faculty of Sciences, University Mohamed I, PO. Box 717, 60000 Oujda (Morocco); ESECO SYSTEMS 270 rue Thomas Edison, Atelier Relais No 6, 34400 Lunel (France); El Bali, B. [Laboratory of Mineral Solid and Analytical Chemistry (LMSAC), Department of Chemistry, Faculty of Sciences, University Mohamed I, PO. Box 717, 60000 Oujda (Morocco); Faik, A. [CIC energigune, Parque Tecnológico de Álava, Albert Einstein 48, 01510 Miñano, Álava (Spain); Naji, M. [CNRS, UPR3079 CEMHTI, 1D avenue de la Recherche Scientifique, 45071 Orléans cedex 2 (France); Benmokhtar, S. [LCPGM, Laboratoire de Chimie-Physique Générale des Matériaux, Département de Chimie, Université Hassan II-Mohammedia, Faculté des Sciences Ben M’Sik, Casablanca (Morocco); Zhong, Y.R.; Su, L.W.; Zhou, Z. [Institute of New Energy Material Chemistry, Synergetic Innovation Center of Chemical Science and Engineering (Tianjin), Nankai University, Tianjin 300071 (China); Kim, J.; Kang, K. [Department of Materials Science and Engineering, Seoul National University, Gwanak-ro, Gwanak-gu, Seoul 151-742 (Korea, Republic of); Dusek, M. [Institute of Physics of the ASCR, v.v.i., Na Slovance 2, 182 21 Praha 8 (Czech Republic)


    Highlights: • Iron Titanium Phosphates as High-Specific-Capacity. • Electrode Materials for Lithium ion Batteries. • During the following cycles, good reversible capacity retention and better cyclabilit. • Ex-situ XRD analysis during the first discharge shows an amorphization of this anode material. -- Abstract: Two iron titanium phosphates, Fe{sub 0.5}TiOPO{sub 4} and Fe{sub 0.5}Ti{sub 2}(PO{sub 4}){sub 3}, were prepared, and their crystal structures and electrochemical performances were compared. The electrochemical measurements of Fe{sub 0.5}TiOPO{sub 4} as an anode of a lithium ion cell showed that upon the first discharge down to 0.5 V, the cell delivered a capacity of 560 mA h/g, corresponding to the insertion of 5 Li’s per formula unit Fe{sub 0.5}TiOPO{sub 4}. Ex-situ XRD reveals a gradual evolution of the structure during cycling of the material, with lower crystallinity after the first discharge cycle. By correlating the electrochemical performances with the structural studies, new insights are achieved into the electrochemical behaviour of the Fe{sub 0.5}TiOPO{sub 4} anode material, suggesting a combination of intercalation and conversion reactions. The Nasicon-type Fe{sub 0.5}Ti{sub 2}(PO{sub 4}){sub 3} consists of a three-dimensional network made of corners and edges sharing [TiO{sub 6}] and [FeO{sub 6}] octahedra and [PO{sub 4}] tetrahedra leading to the formation of trimmers [FeTi{sub 2}O{sub 12}]. The first discharge of lithium ion cells based on Fe{sub 0.5}Ti{sub 2}(PO{sub 4}){sub 3} materials showed electrochemical activity of Ti{sup 4+}/Ti{sup 3+} and Fe{sup 2+}/Fe{sup 0} couples in the 2.5–1 V region. Below this voltage, the discharge profiles are typical of phosphate systems where Li{sub 3}PO{sub 4} is a product of the electrochemical reaction with lithium; moreover, the electrolyte solvent is reduced. An initial capacities as high as 1100 mA h g{sup −1} can be obtained at deep discharge. However, there is an irreversible capacity

  4. High efficiency recovery and epitope specific sorting of an scFv yeast display library

    Energy Technology Data Exchange (ETDEWEB)

    Siegel, Robert W.; Coleman, James R.; Miller, Keith D.; Feldhaus, Michael


    In order to more productively utilize the rich source of antigen specific reagents present in the previously described non-immune scFv yeast display library (Feldhaus et al., 2003) one must be able to efficiently isolate and characterize clones within the library. To this end, we have developed and validated a magnetic bead sorting technique utilizing the Miltenyi MacsTm system to recover greater than 90% of the antigen specific clones present in the library. In combination with flow cytometry, we rapidly reduced diversity and enriched for antigen specific clones in three rounds of selection. Furthermore, we demonstrate the use of pre-existing monoclonal antibodies (mAbs) for antigen labeling and subsequent flow cytometric sorting and characterization of epitope specific scFv. Combining these two improvements in library screening allowed isolation and characterization of 3 epitope specific scFv (including a previously uncharacterized epitope) to a 6 kd protein, epidermal growth factor EGF.

  5. Pancreatic Ductal Adenocarcinoma With High Radiotracer Uptake in 68Ga-Prostate-Specific Membrane Antigen PET/CT. (United States)

    Sahbai, Samine; Rieping, Petra; Pfannenberg, Christina; la Fougére, Christian; Reimold, Matthias


    Prostate-specific membrane antigen imaging with PET/CT is increasingly used in prostate cancer and was shown to have a high diagnostic performance. We report a clinical case of a 67-year-old man with previous history of operated prostate cancer and increasing prostate-specific antigen blood level. Ga-HBED-CC prostate-specific membrane antigen PET/CT imaging was indicated for the assessment of local recurrence and lymph node metastases of prostate cancer. In addition, a soft tissue mass in the body of the pancreas with high radiotracer uptake was detected. Histopathology confirmed a pancreatic ductal adenocarcinoma.

  6. A control strategy to investigate the relationship between specific productivity and high-mannose glycoforms in CHO cells. (United States)

    Zalai, Dénes; Hevér, Helga; Lovász, Krisztina; Molnár, Dóra; Wechselberger, Patrick; Hofer, Alexandra; Párta, László; Putics, Ákos; Herwig, Christoph


    The integration of physiological knowledge into process control strategies is a cornerstone for the improvement of biopharmaceutical cell culture technologies. The present contribution investigates the applicability of specific productivity as a physiological control parameter in a cell culture process producing a monoclonal antibody (mAb) in CHO cells. In order to characterize cell physiology, the on-line oxygen uptake rate (OUR) was monitored and the time-resolved specific productivity was calculated as physiological parameters. This characterization enabled to identify the tight link between the deprivation of tyrosine and the decrease in cell respiration and in specific productivity. Subsequently, this link was used to control specific productivity by applying different feeding profiles. The maintenance of specific productivity at various levels enabled to identify a correlation between the rate of product formation and the relative abundance of high-mannose glycoforms. An increase in high mannose content was assumed to be the result of high specific productivity. Furthermore, the high mannose content as a function of cultivation pH and specific productivity was investigated in a design of experiment approach. This study demonstrated how physiological parameters could be used to understand interactions between process parameters, physiological parameters, and product quality attributes.

  7. Primary oral melanoma: A histopathological and immunohistochemical study of 22 cases of Latin America (United States)

    Toral-Rizo, Víctor H.; León, Jorge E.; Contreras, Elisa; Carlos, Román; Delgado-Azañero, Wilson; Mosqueda-Taylor, Adalberto; de-Almeida, Oslei P.


    Objective: The aim of this study was to analyze the histopathological and immunohistochemical characteristics of 22 cases of primary oral melanomas (OM). Study Design: Twenty two cases of primary oral melanoma were analyzed by description of their histopathological features and immunohistochemical study using the antibodies S-100, HMB-45, Melan-A and Ki-67. Results: The mean age was 58 years and 14 cases were female. The main affected sites were the hard palate, followed by the upper gingiva. Microscopically, 15 cases presented level III of invasion, 2 cases were amelanotic and 13 showed a mixed epithelioid and plasmacytoid or spindle cells composition. Some cases showed necrosis, perivascular and perineural invasion. S-100 and HMB-45 were positive in all cases, but 3 cases were negative for Melan-A. The proliferative index with Ki-67 was high, with labeling index ranging from 15.51% to 63% of positive cells. Conclusion: S-100 and HMB-45 are more frequently expressed than Melan-A in primary oral melanomas and these markers are helpful to confirm the diagnosis. Key words:Oral melanoma, histopathology, immunohistochemistry. PMID:22143732

  8. Immunohistochemical expression of cathepsin L in atopic dermatitis and lichen planus

    Directory of Open Access Journals (Sweden)

    Zeinab A El Ashmawy


    Full Text Available Background: Cathepsin L is a member of papain superfamily. It seems to promote T-cell survival, selection maturation in the thymus and enhance the antigen presentation. Cathepsin L plays an important role in tumor necrosis factors (TNF-α induced cell death. Also it degrades the tight junction between cornedesomses in the epidermis. Elevated expression of cathepsin L has been found in many inflammatory and neoplastic diseases. Objective: The aim of this study was to determine immunohistochemical expression of cathepsin L in atopic dermatitis (AD and lichen planus (LP patients in order to evaluate its role in the pathogenesis of both diseases. Materials and Methods: This study included 15 patients with AD (Group I, 15 patients with LP (Group II, in addition to 10 healthy skin specimens served as controls (Group III. Punch biopsies were taken from lesional skin of the patients and controls for immunohistochemical detection of cathepsin L expression. Results: Highly significant increase was found in cathepsin L expression in AD and LP patients compared to controls [P = 0.001]. Conclusion: Cathepsin L could be implicated as an important protease in the pathogenesis of AD and LP. It could be a useful marker for assessing AD severity.

  9. High Specific Power Multiple-Cylinder Free-Piston Alpha Stirling Project (United States)

    National Aeronautics and Space Administration — NASA seeks highly efficient, long life solar dynamic power conversion systems. The requirements for these missions emphasize low mass and high conversion...

  10. System-wide immunohistochemical analysis of protein co-localization.

    Directory of Open Access Journals (Sweden)

    Minjung Kim

    Full Text Available The analysis of co-localized protein expression in a tissue section is often conducted with immunofluorescence histochemical staining which is typically visualized in localized regions. On the other hand, chromogenic immunohistochemical staining, in general, is not suitable for the detection of protein co-localization. Here, we developed a new protocol, based on chromogenic immunohistochemical stain, for system-wide detection of protein co-localization and differential expression.In combination with a removable chromogenic stain, an efficient antibody stripping method was developed to enable sequential immunostaining with different primary antibodies regardless of antibody's host species. Sections were scanned after each staining, and the images were superimposed together for the detection of protein co-localization and differential expression. As a proof of principle, differential expression and co-localization of glutamic acid decarboxylase67 (GAD67 and parvalbumin proteins was examined in mouse cortex.All parvalbumin-containing neurons express GAD67 protein, and GAD67-positive neurons that do not express parvalbumin were readily visualized from thousands of other neurons across mouse cortex. The method provided a global view of protein co-localization as well as differential expression across an entire tissue section. Repeated use of the same section could combine assessments of co-localization and differential expression of multiple proteins.

  11. Histochemical and Immunohistochemical Study of Peripolar Cells in Sheep

    Directory of Open Access Journals (Sweden)

    Passos Joana


    Full Text Available Peripolar cells are granulated cells located in the vascular pole of the renal corpuscle. Even though these cells have already been described, there are still many unknown histological and physiological characteristics. We carried out histochemical and immunohistochemical analyses of peripolar cells in sheep and compared their number in both normal and injured kidneys, discriminating according to the age of the animal. We tested HE, Toluidine Blue, PAS, and Masson's Trichrome stains to select the best stain for identification and quantification. Masson Trichrome yielded the best results and was selected for this purpose. We identified the cells by the presence of cytoplasmatic granules and by their position in the vascular pole. We found no statistically significant association between the number of peripolar cells and the age of the animal or the occurrence of lesions. In the immunohistochemical analysis, we found that the cells were positive to α-smooth muscle actin and less consistently positive to NSE and S100 protein. Chromogranin A, cyclooxygenase-2, AE1/AE3, and Wide Spectrum Cytokeratin and desmin yielded negative results. We conclude that although there was evidence of a contractile function, there was no evidence to support that peripolar cells have either a neuroendocrine or an epithelial nature.

  12. Occurrence of trichoepithelioma in a cat: Histopathologic and immunohistochemical study. (United States)

    Tavasoli, Abbas; Golshahi, Hannaneh; Rad, Maryam Nobakht; Taymouri, Afra


    Trichoepitheliomas are benign follicular appendage tumors with differentiation to all three segments of the hair follicle. A 2 years old female domestic short hair cat presented with a mass on the tail. The mass was surgically excised and for histopathologic and immunohistochemical studies, was sent to Department of Pathology. Histologically, the tumor was encapsulated and consisted of many islands of follicular epithelium and also cysts structures which varied in size and shape. The cells of epithelium islands were round to oval and had variable amounts of slightly, eosinophilic cytoplasm and euchromatic nuclei. The cystic structures were lined by a complex layer of squamous epithelium. Often, cells under went an abrupt transition between basal layers and keratinization without the development of a granular cell layer. No tendency of malignancy was seen in this case. According to mentioned characteristics, trichoepithelioma was diagnosed. By immunohistochemical study it was confirmed that this tumor had epithelial origin because squamous tumor cells reacted with the pan-cytokeratin antibody. The expression of β-catenin was predominately cytoplasmic and also together with numerous positive nuclei but membranous expression was inconsistenet. Distribution of neoplastic cells with β-catenin expression was more than 75% and labeling intensity was strong in both cytoplasm and nuclei. According to author's knowledge, this is the first report of trichoepithelioma in cat in Iran and also investigation of β-catenin expression in feline trichoepithelioma in veterinary literature.

  13. Collagenase-3 expression in periapical lesions: an immunohistochemical study. (United States)

    Bhalla, G; Astekar, M S; Ramesh, G; Kaur, P; Sowmya, G V


    Collagenase-3 (matrix metalloproteinase-13) is a metalloproteinase (MMP) that is associated with bone lesions and exhibits variable expression patterns in odontogenic cysts; it may play a role in regulating focal proliferation and maturation of jaw cyst epithelium. We studied the localization, staining intensity and distribution of collagenase-3 in 13 periapical granulomas with epithelium, 16 periapical granulomas without epithelium and 10 radicular cysts using archived formalin fixed, paraffin embedded tissues. A monoclonal antibody against human collagenase-3 was used to evaluate its expression. Immunohistochemical staining intensities of collagenase-3 in all periapical lesions were (-), 4 (10%); (+), 1 (3%); (++), 22 (56%) and (+++), 12 (31%); differences were not statistically significant. Immunohistochemical distribution of collagenase-3 in epithelial cells was (-), 17 (44%); (+), 17 (44%); (++), 5 (13%); in fibroblasts it was (-), 8 (20%); (+), 23 (59%); (++), 8 (21%); in plasma cells it was (-), 7 (18%); (+), 22 (56%); (++), 10 (26%); in macrophages it was (-), 7 (18%); (+), and 15 (38%); and (++), 17 (44%). Statistically significant differences were found in epithelial cells (p = 0.00) and fibroblasts (p = 0.02), whereas differences were not statistically significant for plasma cells and macrophages. Collagenase-3 may play a role in the conversion of a periapical granuloma with epithelium to radicular cyst. MMP's influence not only epithelial rest cell migration, but also invasion of various stromal cells into granulomatous tissue.

  14. Comparison of immunohistochemical, histochemical and immunochemical methods for the detection of wheat protein allergens in meat samples and cooked, dry, raw and fermented sausage samples. (United States)

    Lukášková, Z Řezáčová; Tremlová, B; Pospiech, M; Renčová, E; Randulová, Z; Steinhauser, L; Reichová, A; Bednář, J


    Nowadays is it a common practice to add vegetable protein in the production of meat products. Because of the possible substitution of high-quality raw meat with vegetable protein without the labelling the product package and because of the allergenic potential of many vegetable proteins, it is important to develop accurate methods for its detection. The objective of the study was to compare histochemical, immunochemical (ELISA, ALERT gliadin screening test) and immunohistochemical methods for the detection of wheat protein in meat samples and sausages. Histochemical methods were useful for the detection of flour in meat samples, but the immunohistochemical method was better for the detection of wheat protein. ALERT gliadin screening test detected gliadin from 10 mg kg(-1), while an immunohistochemical method detected wheat protein concentrations from 1 g kg(-1) and an ELISA method detected wheat protein concentrations from 4 g kg(-1). ALERT gliadin screening test showed results within 1 day, whilst an ELISA detection method took 2 days, and an immunohistochemical procedure took 5 days at the soonest, all including sample preparation. This study also focused on optimisation of an immunohistochemical method for samples of cooked sausage. In addition, three samples were sufficient for wheat protein detection at a concentration of 1 g kg(-1) (and greater) with a confidence level greater than 95%.

  15. 78 FR 21584 - Atlantic Highly Migratory Species; 2013 Atlantic Bluefin Tuna Quota Specifications (United States)


    ... not necessary to prepare an EA for quota specifications alone (in accordance with the approach... category. This constitutes the best available information regarding the universe of permits and permit...

  16. Discriminant analysis to classify glioma grading using dynamic contrast-enhanced MRI and immunohistochemical markers

    Energy Technology Data Exchange (ETDEWEB)

    Awasthi, Rishi [Sanjay Gandhi Post Graduate Institute of Medical Sciences, Department of Radiodiagnosis, Lucknow (India); Rathore, Ram K.S.; Sahoo, Prativa [Indian Institute of Technology, Department of Mathematics and Statistics, Kanpur (India); Soni, Priyanka; Husain, Nuzhat [Chhatrapati Sahuji Maharj Medical University, Department of Pathology, Lucknow (India); Awasthi, Ashish; Pandey, Chandra M. [Sanjay Gandhi Post Graduate Institute of Medical Sciences, Department of Biostatistics, Lucknow (India); Behari, Sanjay; Singh, Rohit K. [Sanjay Gandhi Post Graduate Institute of Medical Sciences, Department of Neurosurgery, Lucknow (India); Gupta, Rakesh K. [Sanjay Gandhi Post Graduate Institute of Medical Sciences, MR Section, Department of Radiodiagnosis, Lucknow, UP (India)


    The purpose of the present study was to look for the possible predictors which might discriminate between high- and low-grade gliomas by pooling dynamic contrast-enhanced (DCE)-perfusion derived indices and immunohistochemical markers. DCE-MRI was performed in 76 patients with different grades of gliomas. Perfusion indices, i.e., relative cerebral blood volume (rCBV), relative cerebral blood flow (rCBF), permeability (k{sup trans} and k{sub ep}), and leakage (v{sub e}) were quantified. MMP-9-, PRL-3-, HIF-1{alpha}-, and VEGF-expressing cells were quantified from the excised tumor tissues. Discriminant function analysis using these markers was used to identify discriminatory variables using a stepwise procedure. To look for correlations between immunohistochemical parameters and DCE metrics, Pearson's correlation coefficient was also used. A discriminant function for differentiating between high- and low-grade tumors was constructed using DCE-MRI-derived rCBV, k{sub ep}, and v{sub e}. The form of the functions estimated are ''D{sub 1} = 0.642 x rCBV + 0.591 x k{sub ep} - 1.501 x v{sub e} - 1.550'' and ''D{sub 2} = 1.608 x rCBV + 3.033 x k{sub ep} + 5.508 x v{sub e} - 8.784'' for low- and high-grade tumors, respectively. This function classified overall 92.1% of the cases correctly (89.1% high-grade tumors and 100% low-grade tumors). In addition, VEGF expression correlated with rCBV and rCBF, whereas MMP-9 expression correlated with k{sub ep}. A significant positive correlation of HIF-1{alpha} with rCBV and VEGF expression was also found. DCE-MRI may be used to differentiate between high-grade and low-grade brain tumors non-invasively, which may be helpful in appropriate treatment planning and management of these patients. The correlation of its indices with immunohistochemical markers suggests that this imaging technique is useful in tissue characterization of gliomas. (orig.)

  17. p40、天门冬氨酸蛋白酶、甲状腺转录因子-1在肺鳞癌与腺癌表达敏感性和特异性%p40、Napsin-A and TTF-1 ---the specific immunohistochemical mark to identify squamous cell carcinoma and adenocarcinoma of non-small cell Lung Cancer

    Institute of Scientific and Technical Information of China (English)



    Objective:To study the significant immunohistochemical mark to identifiy squamous cell carcinoma and adenocarcinoma of non-small cell lung cancer. Methods:The expression of p63, p40, CK5/6, TTF-1, CK7 and Napsin-A was detected in 51 cases with non-small cell lung cancer by using immunohistochemistry of SP. Results:In 51 cases with non-small cell lung cancer, there were 20 cases with squamous cell carcinoma and 31 cases with adenocarcinoma. The positive expression rates of p63,p40 and CK5/6 in lung squamous cell carcinoma was 100%(20/20), 95%(19/20) and 90%(18/20)respectively ; the sensitivity of p63, p40 and CK5/6 was similar (P=0.355),but the specificity of p40 was higher than that of p63 and CK5/6(P=0.040). The positive expression rate of TTF-1, CK7 and Napsin-A in lung adenocarcino-ma was 87.1%(27/31), 80.6%(25/31) and 90.3%(28/31)respectively, the sensitivity of TTF-1,CK7 and Napsin-A was similar, too(P=0.538). The specificity of TTF-1、Napsin-A was higher than that of CK7(P=0.002). Conclusion:The sensi-tivity of p40, Napsin-A and TTF-1 is higher in non-small cell lung cancer, and they are the specific marks to identify squamous cell carcinoma and adenocarcinoma in the lungs. Detection of p40, Napsin-A and TTF-1 together can be more accurate in differential diagnosis of non-small cell lung cancer and provides a theoretical basis for clinical treatment.%目的:探讨肺非小细胞癌组织中鉴别鳞癌和腺癌有意义的免疫组化检测标记。方法:采用免疫组织化学SP法检测非小细胞肺癌51例组织中p63、p40、CK5/6、甲状腺转录因子-1(TTF-1)、CK7、天门冬氨酸蛋白酶(Napsin-A)的表达。结果:(1)p40、p63、CK5/6在非小细胞肺癌中表达:非小细胞肺癌51例中,p40、p63、CK5/6在鳞状细胞癌20例中阳性表达为20例(100.0%),19例(95.0%),18例(90.0%),在腺癌31例组织中阳性表达为0例(0.0%)、6例(19.4%)、3例(9.7%)。p40在31例腺癌中无1

  18. Pregnancy-specific stress, preterm birth, and gestational age among high-risk young women. (United States)

    Cole-Lewis, Heather J; Kershaw, Trace S; Earnshaw, Valerie A; Yonkers, Kimberly Ann; Lin, Haiqun; Ickovics, Jeannette R


    There is evidence that pregnancy-specific stress is associated with preterm birth. The purpose of this study is to examine the association between change in pregnancy-specific stress over the course of pregnancy and birth outcomes (i.e., preterm birth and gestational age) in an understudied but vulnerable group using a theoretically derived model. Multivariate linear and logistic regression techniques were used to examine the association between pregnancy-specific stress (measured in second and third trimester) and length of gestation (i.e., preterm birth and gestational age) among a sample of 920 Black and/or Latina adolescent and young women. Second trimester pregnancy-specific stress was not associated with preterm birth or gestational age. Third trimester pregnancy-specific stress was associated with preterm birth but not with gestational age. Change in pregnancy-specific stress between second and third trimester was significantly associated with increased likelihood of preterm delivery and shortened gestational age, even after controlling for important biological, behavioral, psychological, interpersonal, and sociocultural risk factors. Findings emphasize the importance of measuring pregnancy-specific stress across pregnancy, as the longitudinal change from second to third trimester was significantly associated with length of gestation measured both as a dichotomous variable (preterm birth) and a continuous variable (gestational age). Furthermore, this is the first study to observe the association of pregnancy-specific stress with length of gestation in this understudied population-unique in age, race, and ethnicity. (PsycINFO Database Record (c) 2014 APA, all rights reserved).

  19. Productive High Performance Parallel Programming with Auto-tuned Domain-Specific Embedded Languages (United States)


    ECL Embedded Common Lisp EM Expectation-Maximization FFI Foreign Function Interface FFT Fast Fourier Transform FFTW Fastest Fourier Transform in the...results to the user. 3.3 Domain-Specific Embedded Languages Domain-specific embedded languages are used in many programming languages such as Lisp ...designed to be extensible using metaprogramming, including Haskell and variants of Lisp . These DSELs generally transformed host language code into

  20. Immunohistochemical changes in the expression of HSP27 in exercised human vastus lateralis muscle

    DEFF Research Database (Denmark)

    Folkesson, M; Mackey, Abigail; Holm, L;


    . METHODS: Two different exercise protocols were used: (1) one-leg ergometer cycling (EC, n = 6) consisting of two 30-min bouts at 40% and 75% of peak oxygen uptake, respectively, and (2) leg extension resistance exercise (RE, n = 9) including 10 sets of eight repetitions at a load corresponding to 70......AIM: The role of HSP27 in the adaptive process of skeletal muscle to exercise, especially in humans, is not well understood. The objective of this study was to investigate immunohistochemical changes in HSP27 expression in human vastus lateralis muscle following resistance and endurance exercises......% of one maximal repetition (1RM). Immunohistochemistry using specific monoclonal antibodies was used to determine the location of HSP27 protein in muscle biopsies from human vastus lateralis. RESULTS: Our results show that RE, but not EC, induced a significant appearance of scattered accumulations of HSP...

  1. So-called embryonal hyperplasia of Bowman's capsular epithelium: an immunohistochemical and ultrastructural study. (United States)

    Ogata, K; Hajikano, H; Sakaguchi, H


    The so-called embryonal hyperplasia of Bowman's capsular epithelium (EHBCE) is a rather specific lesion occurring in kidneys of patients maintained on chronic dialysis. It consists of poorly differentiated cells proliferating around sclerosed or obsolescent glomeruli. In this study, immunohistochemical and ultrastructural characterization of EHBCE was performed. The poorly differentiated cells in the lesion exhibited a positive reaction for vimentin and a negative one for cytokeratin (PKK 1) and epithelial membrane antigen. On ultrastructural examination, specialized junctions between adjoining cells, microvilli-like structures on their surfaces, and immature basal folds were observed. These observations suggest that the cells of EHBCE may be associated with the anlage of glomerular epithelium. The background in which neoplasms like renal cell carcinoma or atypical epithelium of cyst wall develop in end-stage kidneys of adult patients on long-term dialysis may cause such a proliferation of poorly differentiated cells in young or paediatric age group patients.

  2. So-called "papillary and cystic neoplasm of the pancreas." An immunohistochemical and ultrastructural study. (United States)

    Kamisawa, T; Fukayama, M; Koike, M; Tabata, I; Okamoto, A


    A case of so-called "papillary and cystic neoplasm of the pancreas" (PCNP) was reported and investigated immunohistochemically and ultrastructurally. A tumor of the pancreatic head in a 21-year-old female was curatively resected. The tumor was cystic and histologically consisted of uniform cells in papillary and solid structure. Although there was no immunoreactivity for pancreaticogut hormones or secretory products of the pancreas in the tumor cells, most of the tumor cells were diffusely immunoreactive for neuron-specific enolase (NSE). Some neurosecretory granules were detected in the tumor cells ultrastructurally. Both facts suggested endocrine cell character of the tumor. Certain cases of PCNP might show a differentiation to endocrine cells.

  3. Intestinal candidiasis in a loggerhead sea turtle (Caretta caretta): an immunohistochemical study. (United States)

    Orós, J; Arencibia, A; Fernández, L; Jensen, H E


    Post mortem examination of a juvenile loggerhead sea turtle (Caretta caretta) stranded in the Canary Islands revealed a fishing-line in the small intestine. Histologically, severe necrotic enteritis, multiple haemorrhages, and marked oedema of the intestinal submucosa were observed. Yeast cells and fungal hyphae were seen in the lamina propria of the intestinal mucosa and in the connective tissue of the submucosa. Because fungal cultures were not taken at the time of necropsy, an immunohistochemical study was performed in order to identify the fungus involved. Specific monoclonal and heterologously absorbed polyclonal antibodies served as the primary reagents for identification of aspergillosis, candidiasis, fusariosis, geotricosis, scedosporiosis, and zygomycosis, using an indirect immunofluorescence staining technique. The fungal elements were strongly stained only by a polyclonal antibody against Candida albicans and a monoclonal antibody against C. albicans. There are no known previous reports of Candida sp. causing skin disease or systemic mycotic infection in sea turtles.

  4. Primary retroperitoneal mucinous cystadenoma with a sarcoma-like mural nodule: an immunohistochemical study with histogenetic considerations and literature review. (United States)

    Demirel, Dilaver; Gun, Ismet; Kucukodaci, Zafer; Balta, Ahmet Ziya; Ramzy, Ibrahim


    Primary retroperitoneal mucinous cystadenomas (PRMCs) are extremely rare tumors and their association with sarcoma-like mural nodules (SLMNs) has not been described thoroughly. The aim of this study is to characterize the gross and microscopic features and the immunohistochemical profile of the first case of PRMC with SLMN and to discuss the differential diagnosis of SLMNs. The literature related to primary retroperitoneal mucinous tumors is reviewed in an attempt to clarify the histogenesis of the epithelial and sarcomatoid components of the associated mural nodules. A 34-yr-old woman presented with a 14-cm retroperitoneal cystic lesion with a 6-cm mural nodule. An immunohistochemical study with a panel of 19 antibodies and a histochemical study for mucin stains were performed. The epithelial component of the PRMC showed positive staining for cytokeratin (CK) 7, CK AE1/3, epithelial membrane antigen, carcinoembryonic antigen, and calretinin. The neoplasm was not immunoreactive for CK 20, CK 5/6, and the other antibodies used in this study. In addition, it stained positively for mucin by mucicarmine, periodic acid-Schiff, and Alcian blue. The stromal cells of the cyst showed estrogen receptor positivity. SLMN cells were negative for all CKs and other epithelial markers used in the study, but they showed diffuse positive staining for vimentin and CD68, and positive staining for Ki-67 was demonstrated in 25% of these cells. The immunohistochemical and histochemical profiles of PRMC were similar to those of ovarian mucinous neoplasms and the mesothelium. The formation of SLMNs seems to be related to subepithelial hemorrhage and some reactive epithelial changes near the mural nodules. The specific immunohistochemical and morphologic features of SLMNs are helpful in differentiating them from malignant mural nodules, including true sarcomas, osteoclast-rich undifferentiated carcinomas, and carcinosarcomas. Such a differentiation is critical in view of its significant

  5. Patient-specific dose calculation methods for high-dose-rate iridium-192 brachytherapy (United States)

    Poon, Emily S.

    In high-dose-rate 192Ir brachytherapy, the radiation dose received by the patient is calculated according to the AAPM Task Group 43 (TG-43) formalism. This table-based dose superposition method uses dosimetry parameters derived with the radioactive 192Ir source centered in a water phantom. It neglects the dose perturbations caused by inhomogeneities, such as the patient anatomy, applicators, shielding, and radiographic contrast solution. In this work, we evaluated the dosimetric characteristics of a shielded rectal applicator with an endocavitary balloon injected with contrast solution. The dose distributions around this applicator were calculated by the GEANT4 Monte Carlo (MC) code and measured by ionization chamber and GAFCHROMIC EBT film. A patient-specific dose calculation study was then carried out for 40 rectal treatment plans. The PTRAN_CT MC code was used to calculate the dose based on computed tomography (CT) images. This study involved the development of BrachyGUI, an integrated treatment planning tool that can process DICOM-RT data and create PTRAN_CT input initialization files. BrachyGUI also comes with dose calculation and evaluation capabilities. We proposed a novel scatter correction method to account for the reduction in backscatter radiation near tissue-air interfaces. The first step requires calculating the doses contributed by primary and scattered photons separately, assuming a full scatter environment. The scatter dose in the patient is subsequently adjusted using a factor derived by MC calculations, which depends on the distances between the point of interest, the 192Ir source, and the body contour. The method was validated for multicatheter breast brachytherapy, in which the target and skin doses for 18 patient plans agreed with PTRAN_CT calculations better than 1%. Finally, we developed a CT-based analytical dose calculation method. It corrects for the photon attenuation and scatter based upon the radiological paths determined by ray tracing


    Directory of Open Access Journals (Sweden)

    Rema Priyadarsini


    Full Text Available BACKGROUND Adenocarcinoma of prostate is the most common form of cancer in men accounting for 29% of cancers in developed nations and the incidence of prostatic cancer is 6.4% in males of Trivandrum District. MATERIALS AND METHODS All prostatic biopsies taken per rectally and stained by haematoxylin and eosin. In suspected cases of malignancy immunohistochemical markers, the AMACR P504S and high molecular weight cytokeratin 34E12 were done. RESULTS The total number of cases studied were 142. The final diagnosis with histomorphological features show that maximum cases were prostatic carcinoma constituting 45.5% of the samples received. CONCLUSION All prostatic carcinomas were graded by revised Gleason’s grade (ISUP 2005 and the use of immunohistochemical markers in arriving at a definite diagnosis in carcinoma prostate was confirmed.

  7. Diagnostic utility of p53 and CK20 immunohistochemical expression grading urothelial malignancies (United States)


    Introduction Current grading system in application by WHO/ISUP divides urothelial malignancies in low and high grade by morphologic criteria while strict segregation may become cumbersome in limited tissue specimens. As grading these carcinomas are of utmost prognostic significance after depth of invasion, therefore we evaluated the role of immunohistochemical expression of p53 and cytokeratin 20 as an adjuctive tool in grading urothelial carcinoma. Methods The study was conducted in Aga khan university hospital, Histopathology section from December 2010 till June 2011 for duration of six months. It involved 95 cases of urothelial carcinomas diagnosed on trans-uretheral resection specimens of bladder growth. Immunohistochemical expression of p53 and cytokeratin 20 was performed according to standard protocols and correlated with grade and depth of invasion. Results There were 48 cases (50.5%) of low grade and 47 cases (49.5%) of high grade urothelial carcinoma included in the study. Male to female ratio was 4.3:1. Majority of patients (80%) were seen in 45 to 90 years age group. Diffuse positive expression of cytokerain 20 was noted in 33 cases (68.8%) of high grade and 19 (40.4%) low grade tumors. Strong positive expression of p53 was seen in 35 cases (72.9%) of high grade while only 17 cases (36.2%) of low grade tumors showed strong p53 expression. Conclusion Significant difference in expression of Cytokeratin 20 and p53 was found between low and high grade urothelial carcinoma. Therefore we suggest combined use of these markers may be helpful in assigning grade to urothelial carcinoma especially when histologic features are borderline. PMID:25089155

  8. New Nanostructured Li 2 S/Silicon Rechargeable Battery with High Specific Energy

    KAUST Repository

    Yang, Yuan


    Rechargeable lithium ion batteries are important energy storage devices; however, the specific energy of existing lithium ion batteries is still insufficient for many applications due to the limited specific charge capacity of the electrode materials. The recent development of sulfur/mesoporous carbon nanocomposite cathodes represents a particularly exciting advance, but in full battery cells, sulfur-based cathodes have to be paired with metallic lithium anodes as the lithium source, which can result in serious safety issues. Here we report a novel lithium metal-free battery consisting of a Li 2S/mesoporous carbon composite cathode and a silicon nanowire anode. This new battery yields a theoretical specific energy of 1550 Wh kg ?1, which is four times that of the theoretical specific energy of existing lithium-ion batteries based on LiCoO2 cathodes and graphite anodes (∼410 Wh kg?1). The nanostructured design of both electrodes assists in overcoming the issues associated with using sulfur compounds and silicon in lithium-ion batteries, including poor electrical conductivity, significant structural changes, and volume expansion. We have experimentally realized an initial discharge specific energy of 630 Wh kg ?1 based on the mass of the active electrode materials. © 2010 American Chemical Society.

  9. Antigen-Specific Inhibition of High-Avidity T Cell Target Lysis by Low-Avidity T Cells via Trogocytosis

    Directory of Open Access Journals (Sweden)

    Brile Chung


    Full Text Available Current vaccine conditions predominantly elicit low-avidity cytotoxic T lymphocytes (CTLs, which are non-tumor-cytolytic but indistinguishable by tetramer staining or enzyme-linked immunospot from high-avidity CTLs. Using CTL clones of high or low avidity for melanoma antigens, we show that low-avidity CTLs can inhibit tumor lysis by high-avidity CTLs in an antigen-specific manner. This phenomenon operates in vivo: high-avidity CTLs control tumor growth in animals but not in combination with low-avidity CTLs specific for the same antigen. The mechanism involves stripping of specific peptide-major histocompatibility complexes (pMHCs via trogocytosis by low-avidity melanoma-specific CTLs without degranulation, leading to insufficient levels of specific pMHC on target cell surface to trigger lysis by high-avidity CTLs. As such, peptide repertoire on the cell surface is dynamic and continually shaped by interactions with T cells. These results describe immune regulation by low-avidity T cells and have implications for vaccine design.

  10. Pathologic, immunohistochemical, and electron microscopic findings in naturally occurring virulent systemic feline calicivirus infection in cats. (United States)

    Pesavento, P A; MacLachlan, N J; Dillard-Telm, L; Grant, C K; Hurley, K F


    Infection with feline calicivirus (FCV) is a common cause of upper respiratory and oral disease in cats. FCV infection is rarely fatal, however, virulent, systemic strains of FCV (VS-FCV) that cause alopecia, cutaneous ulcers, subcutaneous edema, and high mortality in affected cats have recently been described. Seven cats with natural VS-FCV infection all had subcutaneous edema and ulceration of the oral cavity, with variable ulceration of the pinnae, pawpads, nares, and skin. Other lesions that were present in some affected cats included bronchointerstitial pneumonia, and pancreatic, hepatic, and splenic necrosis. Viral antigen was present within endothelial and epithelial cells in affected tissues as determined by immunohistochemical staining with a monoclonal antibody to FCV. Mature intranuclear and intracytoplasmic virions in necrotic epithelial cells were identified by transmission electron microscopy. VS-FCV infection causes epithelial cell cytolysis and systemic vascular compromise in susceptible cats, leading to cutaneous ulceration, severe edema, and high mortality.

  11. Approach in High Precision Topic-Specific Resource Discovery on the Web

    Institute of Scientific and Technical Information of China (English)

    Ye Wei-guo; Lu Zheng-ding


    The Internet presents numerous sources of useful information nowadays. However, these resources are drowning under the dynamic Web, so accurate finding user-specific information is very difficult. In this paper we discuss a Semantic Graph Web Search (SGWS) algorithm in topic-specific resource discovery on the Web. This method combines the use of hyperlinks, characteristics of Web graph and semantic term weights. We implement the algorithm to find Chinese medical information from the Internet. Our study showed that it has better precision than traditional IR (Information Retrieval) methods and traditional search engines.

  12. Method for Predicting the Energy Characteristics of Li-Ion Cells Designed for High Specific Energy (United States)

    Bennett, William, R.


    Novel electrode materials with increased specific capacity and voltage performance are critical to the NASA goals for developing Li-ion batteries with increased specific energy and energy density. Although performance metrics of the individual electrodes are critically important, a fundamental understanding of the interactions of electrodes in a full cell is essential to achieving the desired performance, and for establishing meaningful goals for electrode performance in the first place. This paper presents design considerations for matching positive and negative electrodes in a viable design. Methods for predicting cell-level performance, based on laboratory data for individual electrodes, are presented and discussed.

  13. The account of sagging of wires at definition of specific potential factors of air High-Voltage Power Transmission Lines



    The opportunity approached is shown, but more exact as it is usually accepted, the account of sagging of wires at definition of specific potential factors air High-Voltage Power Transmission Lines. The technique of reception of analytical expressions is resulted. For an opportunity of comparison traditional expressions for specific potential factors are resulted also. Communication of the offered and traditional analytical expressions is shown. Offered analytical expressions are not difficult...

  14. Relationship Between Dual Time Point FDG PET and Immunohistochemical Parameters in Preoperative Colorectal Cancer: Preliminary Study

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Jai Hyuen; Lee, Won Ae; Park, Seok Gun; Park, Dong Kook; Namgung, Hwan [Dankook Univ. College of Medicine, Cheonan (Korea, Republic of)


    The clinical availability of 2 deoxy 2 [18F] fluoro D glucose (FDG) dual time point positron emission tomography/computerized tomography (DTPP) has been investigated in diverse oncologic fields. The aim of this preliminary study was to evaluate the relationship between various immunohistopathologic markers reflecting disease progression of colorectal cancer and parameters extracted from FDG DTPP in colorectal cancer patients. Forty seven patients with histologically confirmed colorectal cancer were analyzed in this preliminary study. FDG DTPP consisted of an early scan 1 h after FDG injection and a delayed scan 1.5 h after the early scan. Based on an analysis of FDG DTPP, we estimated the maximum standardized uptake value (SUV) of tumors on the early and delayed scans (SUV{sup earlya}nd SUV{sup delayed,} respectively). The retention index (RI) was calculated as follows: (SUV{sup delayed-} SUV{sup early)} x 100/ SUV{sup early.} The clinicopathological findings (size and T and N stages) and immunohistochemical factors [glucose transporter 1 (GLUT 1), hexokinase 2 (HK 2), p53, P504S, and {beta} catenin] were analyzed by visual analysis. The RIs calculated from the SUVs ranged from -1.8 to 73.4 (31.8{+-}15.5). The RIs were significantly higher in patients with high T stages (T3 and T4) than with low T stages (T1 and T2; P<0.05). Among the immunohistochemical analytic markers, GLUT 1 had the highest positive staining rate (93.6%) compared to other markers. Based on unvariable analysis, it was shown that the RI of high level GLUT 1 expression was significantly higher than low level GLUT 1 expression (p=0.01), and the RI of high level p53 expression (p=0.08). Multivariate analysis to investigate a link between RI and clinico pathologic parameters of colorectal carcinoma showed that GLUT 1, p53, and T staging were independently connected with increased RIs (p<0.05, total) using backward selection methods. There was no significant statistical relationship between SUV

  15. The account of sagging of wires at definition of specific potential factors of air High-Voltage Power Transmission Lines

    Directory of Open Access Journals (Sweden)

    Suslov V.M.


    Full Text Available The opportunity approached is shown, but more exact as it is usually accepted, the account of sagging of wires at definition of specific potential factors air High-Voltage Power Transmission Lines. The technique of reception of analytical expressions is resulted. For an opportunity of comparison traditional expressions for specific potential factors are resulted also. Communication of the offered and traditional analytical expressions is shown. Offered analytical expressions are not difficult for programming on a personal computer of any class and besides they allow to make an estimation of an error of traditional expressions by means of parallel definition of specific potential factors by both ways.

  16. Going against the Grain: Gender-Specific Media Education in Catholic High Schools (United States)

    Lapayese, Yvette V.


    The Catholic Church has addressed the power of media, as well as the critical importance of understanding and educating Catholic youth on the media's role and place in modern culture. In this article, the narratives of female Catholic teachers are prioritized to illustrate how gender-specific media education influences the schooling experiences of…

  17. High-throughput development of genome-wide locus-specific informative SSR markers in wheat (United States)

    Although simple sequence repeat (SSR) markers are not new, they are still useful and often used markers in molecular mapping and marker-assisted breeding, particularly in developing countries. However, locus-specific SSR markers could be more useful and informative in wheat breeding and genetic stud...

  18. Highly integrated application specific MMICs for active phased array radar applications

    NARCIS (Netherlands)

    Bogaart, F.L.M. van den


    Application specific MMIC solutions for active array radar, developed at TNO-FEL, are presented. The use and application of these MMICs in their respective radar systems will be shown. These MMICs address the needs for current and future phased-array topologies as for example the concept of "smart

  19. Highly Integrated Application Specific MMICS for Active Phased Array Radar Applications

    NARCIS (Netherlands)

    Bogaart, F.L.M. van den


    Application specific MMIC solutions for active array radar, developed at TNO-FEL. are presented. The use and application of these MMICs in their respective radar systems will be shown. These MMICs address the needs for current and future phased-array topologies as for example the concept of "smart

  20. Rapid ABO genotyping by high-speed droplet allele-specific PCR using crude samples. (United States)

    Taira, Chiaki; Matsuda, Kazuyuki; Takeichi, Naoya; Furukawa, Satomi; Sugano, Mitsutoshi; Uehara, Takeshi; Okumura, Nobuo; Honda, Takayuki


    ABO genotyping has common tools for personal identification of forensic and transplantation field. We developed a new method based on a droplet allele-specific PCR (droplet-AS-PCR) that enabled rapid PCR amplification. We attempted rapid ABO genotyping using crude DNA isolated from dried blood and buccal cells. We designed allele-specific primers for three SNPs (at nucleotides 261, 526, and 803) in exons 6 and 7 of the ABO gene. We pretreated dried blood and buccal cells with proteinase K, and obtained crude DNAs without DNA purification. Droplet-AS-PCR allowed specific amplification of the SNPs at the three loci using crude DNA, with results similar to those for DNA extracted from fresh peripheral blood. The sensitivity of the methods was 5%-10%. The genotyping of extracted DNA and crude DNA were completed within 8 and 9 minutes, respectively. The genotypes determined by the droplet-AS-PCR method were always consistent with those obtained by direct sequencing. The droplet-AS-PCR method enabled rapid and specific amplification of three SNPs of the ABO gene from crude DNA treated with proteinase K. ABO genotyping by the droplet-AS-PCR has the potential to be applied to various fields including a forensic medicine and transplantation medical care. © 2017 Wiley Periodicals, Inc.

  1. Synthesis of high-specific activity [15,16-{sup 3}H{sub 2}]buprenorphine

    Energy Technology Data Exchange (ETDEWEB)

    Oetvoes, F.; Toth, G. [Hungarian Academy of Sciences, Szeged (Hungary). Isotope Lab.; Hosztafi, S.; Simon, C. [Alkaloida Chemical Factory Ltd., Tiszavasvari (Hungary)


    Tritium labelling of buprenorphine, a mixed agonist-antagonist opioid ligand, was performed with a specific activity of 2.35 TBq/mmol (63.6 Ci/mmol) starting with 15,16-didehydrobuprenorphine. Labels at positions 15 and 16 of the morphine skeleton proved to be sufficiently stable under strong acidic or basic conditions. (Author).

  2. Isotope specific resolution recovery image reconstruction in high resolution PET imaging

    NARCIS (Netherlands)

    Kotasidis, Fotis A.; Angelis, Georgios I.; Anton-Rodriguez, Jose; Matthews, Julian C.; Reader, Andrew J.; Zaidi, Habib

    Purpose: Measuring and incorporating a scanner-specific point spread function (PSF) within image reconstruction has been shown to improve spatial resolution in PET. However, due to the short half-life of clinically used isotopes, other long-lived isotopes not used in clinical practice are used to

  3. Highly Integrated Application Specific MMICS for Active Phased Array Radar Applications

    NARCIS (Netherlands)

    Bogaart, F.L.M. van den


    Application specific MMIC solutions for active array radar, developed at TNO-FEL. are presented. The use and application of these MMICs in their respective radar systems will be shown. These MMICs address the needs for current and future phased-array topologies as for example the concept of "smart s

  4. Highly integrated application specific MMICs for active phased array radar applications

    NARCIS (Netherlands)

    Bogaart, F.L.M. van den


    Application specific MMIC solutions for active array radar, developed at TNO-FEL, are presented. The use and application of these MMICs in their respective radar systems will be shown. These MMICs address the needs for current and future phased-array topologies as for example the concept of "smart s

  5. Work-related fatigue: the specific case of highly educated women in the Netherlands. (United States)

    Verdonk, Petra; Hooftman, Wendela E; van Veldhoven, Marc J P M; Boelens, Louise R M; Koppes, Lando L J


    This study aims to establish the prevalence of high work-related fatigue (need for recovery, NFR) among employees and to explain group differences categorized by gender, age, and education. The study particularly aims to clarify prevalence and explanatory factors in highly educated women. In 2005 and 2006, large representative samples of 80,000 Dutch employees (net response rate 33.0%; N = 47,263) received the Netherlands working conditions survey questionnaire. First, we calculated the prevalence of high NFR for men and women with different age and education levels. The average prevalence of high NFR was 28.8% and was highest among highly educated women (35.2%) in particular those aged 50-64 years (40.3%). Second, logistic regression analyses were used to compare subgroups' NFR in relation to situational factors, working conditions, and health. Three comparisons were made: (1) highly educated women versus men; (2) highly educated versus lower educated women and; (3) older highly educated versus younger highly educated women. The situational, working conditions and health factors in our model did not explain the gender differences among highly educated employees (OR = 1.37; CI = 1.3-1.5, adjusted for all factors OR = 1.32; CI = 1.2-1.5). Despite that lower autonomy and workplace violence explained highly educated women's NFR, working fewer hours counterbalanced this. Time pressure in work largely explained the differences in NFR among women at different education levels (crude OR 1.44; CI = 1.4-1.5, adjusted OR 1.14; CI = 1.0-1.3). In the age comparison, lower health ratings, more adverse working conditions, and working as a teacher explained older highly educated women's high prevalence of high NFR (crude OR 1.32; CI = 1.2-1.5, adjusted OR 0.94; CI = 0.8-1.2). NFR has high prevalence in highly educated women (35.2%) in particular those aged 50-64 years (40.3%). Our model did not explain gender differences in NFR, because working fewer hours counterbalanced the

  6. Preliminary Observations on Sensitivity and Specificity of Magnetization Transfer Asymmetry for Imaging Myelin of Rat Brain at High Field. (United States)

    Kim, Jae-Woong; Choi, Jiye; Cho, Janggeun; Lee, Chulhyun; Jeon, Daejong; Park, Sung-Hong


    Magnetization transfer ratio (MTR) has been often used for imaging myelination. Despite its high sensitivity, the specificity of MTR to myelination is not high because tissues with no myelin such as muscle can also show high MTR. In this study, we propose a new magnetization transfer (MT) indicator, MT asymmetry (MTA), as a new method of myelin imaging. The experiments were performed on rat brain at 9.4 T. MTA revealed high signals in white matter and significantly low signals in gray matter and muscle, indicating that MTA has higher specificity than MTR. Demyelination and remyelination studies demonstrated that the sensitivity of MTA to myelination was as high as that of MTR. These experimental results indicate that MTA can be a good biomarker for imaging myelination. In addition, MTA images can be efficiently acquired with an interslice MTA method, which may accelerate clinical application of myelin imaging.

  7. Immunohistochemical detection of transgene expression in the brain using small epitope tags

    Directory of Open Access Journals (Sweden)

    Debyser Zeger


    Full Text Available Abstract Background In vivo overexpression of proteins is a powerful approach to study their biological function, generate disease models or evaluate gene therapy approaches. In order to investigate an exogenously expressed protein, specific and sensitive detection is essential. Unfortunately, antibodies that allow histological detection of the protein of interest are not always readily available. The use of an epitope tag fused to the protein can circumvent this problem as well as provide the possibility to discriminate endogenous from overexpressed proteins. In order to minimize impact on the bioactivity and biodistribution of the overexpressed protein, preference is given to small tags. Results In the present study, we evaluated several small epitope tags together with corresponding anti-tag antibodies for the detection of overexpressed proteins in rat brain, using eGFP as a reference. We generated several lentiviral vectors encoding eGFP with different N-terminally fused small epitope tags (AU1, flag, 3flag, HA, myc and V5. After confirmation of their functionality in cell culture, we injected these lentiviral vectors stereotactically into the striatum of rats and prepared paraformaldehyde fixed floating sections for immunohistochemical analysis. Using multiple antibodies and antibody dilutions per epitope tag, we extensively assessed the efficiency of several anti-tag antibodies for chromogenic immunohistochemical detection of the epitope tagged eGFPs by determining the proportion of immunoreactivity detected by anti-tag antibodies compared to anti-GFP antibody. Using fluorescence immunohistochemistry and confocal microscopy, we also quantified the proportion of eGFP-positive cells detected by anti-tag antibodies. Our results show that all the examined small epitope tags could be detected by anti-tag antibodies both in cell extracts as well as in vivo, although to varying degrees depending on the tag and antibody used. Using the presented

  8. Synovial sarcoma. An immunohistochemical study of the epithelial component

    DEFF Research Database (Denmark)

    Jørgensen, L J; Lyon, H; Myhre-Jensen, O


    Twenty-five synovial sarcomas were studied with a battery of antibodies directed against keratin and epithelial membrane antigen (EMA). The keratin antibody MNF 116 showed reactivity in 24 tumors. In addition, 22 tumors showed reactivity with the antibody Keratin Wide Spectrum, 20 with the antibody...... Keratin 56, 64, and 19 with CAM 5.2. Seventeen tumors showed reactivity with EMA. The keratin and EMA reactivity was present in cells lining obvious cleft-like structures in biphasic tumors. In the spindle cell areas of both biphasic and monophasic fibrous tumors, we found clusters of a few reacting cells...... apparently located around small clefts. In the synovioblastic tumors, clusters of plump tumor cells reactive for both the keratins and EMA were present. In conclusion, we found that proper identification of epithelial differentiation in synovial sarcomas is facilitated by an immunohistochemical application...

  9. Parvovirus infection: an immunohistochemical study using fetal and placental tissue. (United States)

    Li, Jing Jing; Henwood, Tony; Van Hal, Sebastian; Charlton, Amanda


    Parvovirus B19 infection causes 5% to 15% of cases of nonimmune hydrops fetalis. The aim of our study was to evaluate the use of immunohistochemistry in diagnosing parvovirus infection in fetal and placental tissue during routine fetal and perinatal autopsies. Histology slides of 20 cases of confirmed parvovirus infection were reviewed, and immunohistochemistry was applied to selected blocks of fetal and placental tissue. Immunohistochemistry was positive in all 20 cases, and histologic viral inclusions were seen in 19 cases. Immunohistochemical staining was closely correlated with histology and was more sensitive than histology in detecting virally infected cells, especially in autolyzed tissue. All cases also had confirmatory evidence of parvovirus infection by polymerase chain reaction of fetal liver and positive maternal serology, where it was available. We conclude that parvovirus immunohistochemistry is a reliable method for diagnosing parvovirus infection, especially in autolyzed tissue where histologic assessment may be suboptimal.

  10. Immunohistochemical Characteristics of Bone Forming Cells in Pleomorphic Adenoma

    Directory of Open Access Journals (Sweden)

    Keisuke Nakano, Takehiro Watanabe, Takako Shimizu, Toshiyuki Kawakami


    Full Text Available Histopathological and immunohistochemical examinations were carried out in a case of pleomorphic adenoma with bone formation, occurring in the chin of a 34-year-old Japanese man. Examination results showed the modified neoplastic myoepithelial cells reacted positively to S-100 protein. The S-100-positive modified neoplastic myoepithelial cells were proliferated in the closely related area of the bone tissue. Furthermore, positive reaction was detected in the bone forming cells: osteoblasts and osteocytes. These cells also reacted positively to Runx2 as a marker of bone forming cells. These results suggest that the origin of the bone forming cells in this case of pleomorphic adenoma was modified neoplastic myoepithelial cells.

  11. A histopathological and immunohistochemical analysis of ameloblastic fibrodentinoma. (United States)

    Bologna-Molina, Ronell; Salazar-Rodríguez, Sirced; Bedoya-Borella, Ana María; Carreón-Burciaga, Ramón Gil; Tapia-Repetto, Gabriel; Molina-Frechero, Nelly


    Ameloblastic fibrodentinoma (AFD) is considered a mixed odontogenic tumor that is characterized by conserved epithelial and ectomesenchymal neoplastic components. AFD is composed of long narrow cords and islands of odontogenic epithelium; the epithelial strands lie in a myxoid cell-rich ectomesenchymal tissue with stellate-shaped fibroblasts that exhibit long slender cytoplasmic extensions that resemble dental papilla. The lesions show the presence of dysplastic dentin. Although AFD is a rare entity and its very existence is not completely accepted, based on the extent of histodifferentiation, it is considered to represent a stage between ameloblastic fibroma and ameloblastic fibroodontoma. This study aimed to provide a histopathological and immunohistochemical characterization of this infrequent tumor. A large panel of antibodies including amelogenin, Ck 19, calretinin, syndecan-1, E-cadherin, MSH2, histone H3, and Ki-67 was used to illustrate the nature of the tumor.

  12. A Histopathological and Immunohistochemical Analysis of Ameloblastic Fibrodentinoma

    Directory of Open Access Journals (Sweden)

    Ronell Bologna-Molina


    Full Text Available Ameloblastic fibrodentinoma (AFD is considered a mixed odontogenic tumor that is characterized by conserved epithelial and ectomesenchymal neoplastic components. AFD is composed of long narrow cords and islands of odontogenic epithelium; the epithelial strands lie in a myxoid cell-rich ectomesenchymal tissue with stellate-shaped fibroblasts that exhibit long slender cytoplasmic extensions that resemble dental papilla. The lesions show the presence of dysplastic dentin. Although AFD is a rare entity and its very existence is not completely accepted, based on the extent of histodifferentiation, it is considered to represent a stage between ameloblastic fibroma and ameloblastic fibroodontoma. This study aimed to provide a histopathological and immunohistochemical characterization of this infrequent tumor. A large panel of antibodies including amelogenin, Ck 19, calretinin, syndecan-1, E-cadherin, MSH2, histone H3, and Ki-67 was used to illustrate the nature of the tumor.

  13. Ebola Virus Disease in Pregnancy: Clinical, Histopathologic, and Immunohistochemical Findings. (United States)

    Muehlenbachs, Atis; de la Rosa Vázquez, Olimpia; Bausch, Daniel G; Schafer, Ilana J; Paddock, Christopher D; Nyakio, Jean Paul; Lame, Papys; Bergeron, Eric; McCollum, Andrea M; Goldsmith, Cynthia S; Bollweg, Brigid C; Prieto, Miriam Alía; Lushima, Robert Shongo; Ilunga, Benoit Kebela; Nichol, Stuart T; Shieh, Wun-Ju; Ströher, Ute; Rollin, Pierre E; Zaki, Sherif R


    Here we describe clinicopathologic features of Ebola virus disease in pregnancy. One woman infected with Sudan virus in Gulu, Uganda, in 2000 had a stillbirth and survived, and another woman infected with Bundibugyo virus had a live birth with maternal and infant death in Isiro, the Democratic Republic of the Congo in 2012. Ebolavirus antigen was seen in the syncytiotrophoblast and placental maternal mononuclear cells by immunohistochemical analysis, and no antigen was seen in fetal placental stromal cells or fetal organs. In the Gulu case, ebolavirus antigen localized to malarial parasite pigment-laden macrophages. These data suggest that trophoblast infection may be a mechanism of transplacental ebolavirus transmission. Published by Oxford University Press for the Infectious Diseases Society of America 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  14. The effect of high temperature induced variation of specific heat on the hypersonic turbulent boundary layer and its computation

    Institute of Scientific and Technical Information of China (English)


    For hypersonic vehicles,as the temperature in its boundary layer usually exceeds 600 K,for which the molecular vibrational degree of freedom is excited,the perfect gas model is no longer valid.In this paper,the effect of high temperature induced variation of specific heat on the hypersonic turbulent boundary layer of flat plates is investigated by direct numerical simulations with the perfect gas model,i.e.with constant specific heat,as well as with a variable specific heat gas model.The comparison of the results from the two gas models has found that the effect of the variation of specific heat on the velocity of the turbulent boundary layers is relatively small,while its effect on temperature,such as the mean temperature,the temperature fluctuations,is appreciable.It is also found that the mean specific heat is quite close to the specific heat calculated by using the mean temperature,indicating that it is possible to do turbulence modeling.The modeling is done under the variable specific heat gas model with the mean temperature as the variable.The feasibility of such consideration is verified by applying the SST model for variable specific heat turbulence computation.

  15. The spectrum of pulmonary sarcoidosis: Variations of high-resolution CT findings and clues for specific diagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Nishino, Mizuki; Lee, Karen S. [Department of Radiology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, MA 02215 (United States); Itoh, Harumi [Department of Radiology, University of Fukui, Faculty of Medical Sciences, Matsuoka-cho, Yoshida-gun, Fukui (Japan); Hatabu, Hiroto [Department of Radiology, Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, MA 02215 (United States)], E-mail:


    Sarcoidosis is a systemic disease of unknown cause, characterized by widespread non-caseating granulomas. There is a wide spectrum of radiologic manifestations in pulmonary sarcoidosis, providing challenges to radiologists. However, recognition of the key features of sarcoidosis with knowledge of its pathologic background can often allow for specific diagnosis. In this review, we describe the variety of high-resolution CT findings in pulmonary sarcoidosis along with its pathologic features as the basis for radiographic manifestations, and discuss the key features on high-resolution CT for the specific diagnosis of pulmonary sarcoidosis.

  16. Clinical, pathological, immunohistochemical and molecular characterization of feline chronic gingivostomatitis. (United States)

    Rolim, Veronica Machado; Pavarini, Saulo Petinatti; Campos, Fabrício Souza; Pignone, Viviam; Faraco, Cláudia; Muccillo, Marcelo de Souza; Roehe, Paulo Michel; da Costa, Fernanda Viera Amorim; Driemeier, David


    Objectives This study presents the clinical, pathological, immunohistochemical and molecular characterization of 26 cats with feline chronic gingivostomatitis (FCG). Methods Oral mucosal biopsies, blood and swabs were collected from cats presenting with oral lesions. The tissue sections were submitted for histopathology and immunohistochemical analysis for feline calicivirus (FCV), feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV). The swabs were subjected to PCR analysis for FCV, and blood for FeLV and FIV. Results The main clinical findings were dysphagia (88.2%), halitosis (76.5%), sialorrhea (47.1%), weight loss (41.2%), intense oral discomfort (35.3%), oral hemorrhage (17.6%), and lackluster and fragile coat (11.8%). Gross inspection revealed bilateral lesions across the palatoglossal fold to the lateral tongue base. The lesions were diffuse, proliferative, intensely red and friable, and bled easily upon examination in 80.8% of cases. In 23.1% of cases, the lesions were multifocal to coalescent, at times forming multiple vesicles on a reddened, edematous palatoglossal fold. Microscopic examination showed that 15.4% of lesions had moderate (grade 2) and 84.6% had severe (grade 3) inflammation. Immunohistochemistry revealed the presence of FeLV antigens in the epithelium and the inflammatory infiltrate of 30.8% of the cats with FCG. FCV antigens were not detected in the FCG lesions. Conclusions and relevance The FCG cases analyzed could not be correlated with FCV. It is possible that FeLV plays a role as a causal agent of lesions in cases where the presence of the virus has been confirmed by immunohistochemistry in epithelial samples.

  17. Estrogen receptors in human thyroid gland. An immunohistochemical study. (United States)

    Arain, Shoukat A; Shah, Munawar H; Meo, Sultan A; Jamal, Qamar


    Thyroid diseases affect women approximately 3 times more frequently than men. It has been suggested that the female sex steroids stimulate thyroid growth such as in the breast. Seventeen beta-estradiol, the major estrogen in the body acts via estrogen receptors (ER) present in the nucleus of the cell. The aim of the study is to determine the ER status in the thyroid gland tissues. Our study was based on immunohistochemical staining for ER. Fifty previously diagnosed cases of various thyroid lesions were selected from the Surgical Pathology Records of Pathology Department, Basic Medical Sciences Institute, Jinnah Postgraduate Medical Center, Karachi, Pakistan between March and August 2000. The staining was performed on formalin-fixed paraffin embedded tissues using monoclonal anti-ER antibody (clone 1D5). Out of 50 cases, 8 were nodular goiter, 9 cases of adenoma, 19 papillary carcinoma, 10 follicular and 4 cases were of medullary carcinoma. Surrounding normal tissue was available in 25 (50%) cases, 4 non-neoplastic and 21 neoplastic lesions. Out of 50 cases, 10 (20%) were males and 40 (80%) were females, the youngest patient was a 14-year-old female and the eldest patient was a 56-year-old male. Despite the availability of normal thyroid tissue and a wide range of lesions, none of our cases showed positive staining. In contrary to many earlier reports by immunohistochemical method using monoclonal antibody (clone 1D5) on formalin-fixed paraffin-embedded thyroid tissues, the ER are not detectable. The effect of estrogen on thyroid gland may be indirect one.

  18. Immunohistochemical Analysis Using Antipodocalyxin Monoclonal Antibody PcMab-47 Demonstrates Podocalyxin Expression in Oral Squamous Cell Carcinomas. (United States)

    Itai, Shunsuke; Yamada, Shinji; Kaneko, Mika K; Harada, Hiroyuki; Kato, Yukinari


    Podocalyxin is a CD34-related type I transmembrane protein that is highly glycosylated with N-glycan, O-glycan, and keratan sulfate. Podocalyxin was originally found in the podocytes of rat kidney and is reportedly expressed in many types of tumors, including brain tumors, colorectal cancers, and breast cancers. Overexpression of podocalyxin is an independent predictor of progression, metastasis, and poor outcome. We recently immunized mice with recombinant human podocalyxin, which was produced using LN229 glioblastoma cells, and produced a novel antipodocalyxin monoclonal antibody (mAb), PcMab-47, which reacts with endogenous podocalyxin-expressing cancer cell lines and normal cell lines independent of glycosylation in Western blot, flow cytometry, and immunohistochemical analyses. In this study, we performed immunohistochemical analysis against oral cancers using PcMab-47. PcMab-47-stained oral squamous cell carcinoma cells in a cytoplasmic pattern and detected 26/38 (68.4%) of oral squamous cell carcinoma cells on tissue microarrays. These results indicate that PcMab-47 is useful in detecting podocalyxin of oral cancers for immunohistochemical analysis.

  19. A novel detection platform for parallel monitoring of DNA hybridization with high sensitivity and specificity

    DEFF Research Database (Denmark)

    Yi, Sun; Perch-Nielsen, Ivan R.; Wang, Zhenyu;

    We developed a high-sensitive platform to monior multiple hybridization events in real time. By creating a microoptical array in a polymeric chip, the system combine the excellent discriminative power of supercritical angle fluorescence (SAF) microscopy with high-throughput capabilities...

  20. Trichoderma harzianum transformant has high extracellular alkaline proteinase expression during specific mycoparasitic interactions

    Directory of Open Access Journals (Sweden)

    Goldman Maria Helena S.


    Full Text Available The mycoparasite Trichoderma harzianum produces an alkaline proteinase that may be specifically involved in mycoparasitism. We have constructed transformant strains of this fungus that overexpress this alkaline proteinase. Some of the transformants were assessed for alkaline proteinase activity, and those with higher activity than the wild type were selected for further studies. One of these transformant strains produced an elevated and constitutive pbr1 mRNA level during mycoparasitic interactions with Rhizoctonia solani.

  1. A new ethylene glycol-silane monolayer for highly-specific DNA detection on Silicon Chips



    Monolayer thin films with ethylene-glycol function onto gold surfaces by using thiols have been extensively investigated. They have been proposed as precursors for applications to bio-detection, where their hydrophilic character improves both specificity and sensitivity. The aim of this letter is to characterize ethylene-glycol monolayer precursors formed onto silicon chips by using silanes. The importance of the ethylene-glycol function is demonstrated by comparing with the well known 3-Amin...

  2. Continuous-wave high specific output power Ar-He-Xe laser with transverse RF excitation

    NARCIS (Netherlands)

    Udalov, Yu.B.; Peters, P.J.M.; Heeman-Ilieva, M.B.; Witteman, W.J.; Ochkin, V.N.


    A transverse RF excited gas discharge has been successfully used to produce a CW Ar-He-Xe laser. A maximum output power of 330 mW has been obtained from an experimental device with 37 cm active length and a 2.25 (DOT) 2.25 cm2 cross-section. This corresponds to a specific output power of about 175 m

  3. A fluorescent probe which allows highly specific thiol labeling at low pH

    DEFF Research Database (Denmark)

    Nielsen, Jonas W.; Jensen, Kristine Steen; Hansen, Rosa E.


    and properties of a thiol-specific reagent, fluorescent cyclic activated disulfide (FCAD), which includes the fluorescein moiety as fluorophore and utilizes a variation of thiol-disulfide exchange chemistry. The leaving-group character of FCAD makes it reactive at pH 3, allowing modification at low pH, limiting...... thiol-disulfide exchange. Different applications are demonstrated including picomolar thiol detection, determination of redox potentials, and in-gel detection of labeled proteins....

  4. Accurate determination of specific heat at high temperatures using the flash diffusivity method (United States)

    Vandersande, J. W.; Zoltan, A.; Wood, C.


    The flash diffusivity method of Parker et al. (1961) was used to measure accurately the specific heat of test samples simultaneously with thermal diffusivity, thus obtaining the thermal conductivity of these materials directly. The accuracy of data obtained on two types of materials (n-type silicon-germanium alloys and niobium), was + or - 3 percent. It is shown that the method is applicable up to at least 1300 K.

  5. High-speed droplet-allele-specific polymerase chain reaction for genotyping of single nucleotide polymorphisms. (United States)

    Matsuda, Kazuyuki; Honda, Takayuki


    Single nucleotide alternations such as single nucleotide polymorphisms (SNPs) or single nucleotide mutations are useful genetic markers for molecular diagnosis, prognosis, drug response, and predisposition to diseases. Rapid identification of SNPs or mutations is clinically important, especially for determining drug responses and selection of molecular-targeted therapy. Here, we describe a rapid genotyping assay based on the allele-specific polymerase chain reaction (AS-PCR) by using our droplet-PCR machine (droplet-AS-PCR).

  6. A Methodology for Writing High Quality Requirements Specification and Evaluating Existing Ones (United States)

    Rosenberg, Linda; Hammer, Theodore


    Requirements development and management have always been critical in the implementation of software systems; engineers are unable to build what analysts can't define. It is generally accepted that the earlier in the life cycle potential risks are identified the easier it is to eliminate or manage the conditions that introduce that risk. Problems that are not found until testing are approximately 14 times more costly to fix than if the problem was found in the requirement phase. The requirements specification, as the first tangible representation of the capability to be produced, establishes the basis for all of the project's engineering management and assurance functions. If the quality of the requirements specification is poor it can give rise to risks in all areas of the project. Recently, automated tools have become available to support requirements management. The use of these tools not only provides support in the definition and tracing of requirements, but it also opens the door to effective use of metrics in characterizing and assessing the quality of the requirement specifications.

  7. Analysis of sex-specific injury patterns and risk factors in young high-level athletes. (United States)

    Frisch, A; Seil, R; Urhausen, A; Croisier, J L; Lair, M L; Theisen, D


    This study analyzed sex-specific injury patterns and risk factors in young athletes (n=256) from 12 sport disciplines practicing at the national or the international level in the Grand-Duchy of Luxembourg. Injury occurrence as a result of sport practice was analyzed retrospectively over the year 2006 using a standardized self-administered questionnaire. Overall incidence was not different between girls and boys (1.20 and 1.21 injuries/1000 athlete-hours, respectively), but in the context of team sport competition girls tended to be at a greater risk (rate ratio 2.05, P=0.053). Girls had a higher proportion of injuries in the ankle/foot region compared with boys (34.8% vs 16.8%). No sex-related differences were found regarding injury severity. Multivariate logistic regression (controlling for age and practice volume) revealed that girls' team sports were associated with a greater injury risk compared with individual sports [odds ratio (OR) of 4.76], while in boys this was observed for racket sports (OR=3.31). Furthermore, physical or emotional stress tended to be a specific risk factor in girls. There was a tendency for injury outside sports to be coupled to a higher injury risk in girls and boys. Consideration of sex-specific injury patterns and risk factors could be of importance for effective injury prevention.

  8. Activity and High-Order Effective Connectivity Alterations in Sanfilippo C Patient-Specific Neuronal Networks (United States)

    Canals, Isaac; Soriano, Jordi; Orlandi, Javier G.; Torrent, Roger; Richaud-Patin, Yvonne; Jiménez-Delgado, Senda; Merlin, Simone; Follenzi, Antonia; Consiglio, Antonella; Vilageliu, Lluïsa; Grinberg, Daniel; Raya, Angel


    Summary Induced pluripotent stem cell (iPSC) technology has been successfully used to recapitulate phenotypic traits of several human diseases in vitro. Patient-specific iPSC-based disease models are also expected to reveal early functional phenotypes, although this remains to be proved. Here, we generated iPSC lines from two patients with Sanfilippo type C syndrome, a lysosomal storage disorder with inheritable progressive neurodegeneration. Mature neurons obtained from patient-specific iPSC lines recapitulated the main known phenotypes of the disease, not present in genetically corrected patient-specific iPSC-derived cultures. Moreover, neuronal networks organized in vitro from mature patient-derived neurons showed early defects in neuronal activity, network-wide degradation, and altered effective connectivity. Our findings establish the importance of iPSC-based technology to identify early functional phenotypes, which can in turn shed light on the pathological mechanisms occurring in Sanfilippo syndrome. This technology also has the potential to provide valuable readouts to screen compounds, which can prevent the onset of neurodegeneration. PMID:26411903

  9. Nontargeted modification-specific metabolomics study based on liquid chromatography-high-resolution mass spectrometry. (United States)

    Dai, Weidong; Yin, Peiyuan; Zeng, Zhongda; Kong, Hongwei; Tong, Hongwei; Xu, Zhiliang; Lu, Xin; Lehmann, Rainer; Xu, Guowang


    Modifications of genes and proteins have been extensively studied in systems biology using comprehensive analytical strategies. Although metabolites are frequently modified, these modifications have not been studied using -omics approaches. Here a general strategy for the nontargeted profiling of modified metabolites, which we call "nontargeted modification-specific metabolomics", is reported. A key aspect of this strategy was the combination of in-source collision-induced dissociation liquid chromatography-mass spectrometry (LC-MS) and global nontargeted LC-MS-based metabolomics. Characteristic neutral loss fragments that are specific for acetylation, sulfation, glucuronidation, glucosidation, or ribose conjugation were reproducibly detected using human urine as a model specimen for method development. The practical application of this method was demonstrated by profiling urine samples from liver cirrhosis patients. Approximately 900 features were identified as modified endogenous metabolites and xenobiotics. Moreover, this strategy supports the identification of compounds not included in traditional metabolomics databases (HMDB, Metlin, and KEGG), which are currently referred to as "unknowns" in metabolomics projects. Nontargeted modification-specific metabolomics opens a new perspective in systems biology.

  10. Atrogin-1, a muscle-specific F-box protein highly expressed during muscle atrophy (United States)

    Gomes, M. D.; Lecker, S. H.; Jagoe, R. T.; Navon, A.; Goldberg, A. L.


    Muscle wasting is a debilitating consequence of fasting, inactivity, cancer, and other systemic diseases that results primarily from accelerated protein degradation by the ubiquitin-proteasome pathway. To identify key factors in this process, we have used cDNA microarrays to compare normal and atrophying muscles and found a unique gene fragment that is induced more than ninefold in muscles of fasted mice. We cloned this gene, which is expressed specifically in striated muscles. Because this mRNA also markedly increases in muscles atrophying because of diabetes, cancer, and renal failure, we named it atrogin-1. It contains a functional F-box domain that binds to Skp1 and thereby to Roc1 and Cul1, the other components of SCF-type Ub-protein ligases (E3s), as well as a nuclear localization sequence and PDZ-binding domain. On fasting, atrogin-1 mRNA levels increase specifically in skeletal muscle and before atrophy occurs. Atrogin-1 is one of the few examples of an F-box protein or Ub-protein ligase (E3) expressed in a tissue-specific manner and appears to be a critical component in the enhanced proteolysis leading to muscle atrophy in diverse diseases.

  11. Activity and High-Order Effective Connectivity Alterations in Sanfilippo C Patient-Specific Neuronal Networks

    Directory of Open Access Journals (Sweden)

    Isaac Canals


    Full Text Available Induced pluripotent stem cell (iPSC technology has been successfully used to recapitulate phenotypic traits of several human diseases in vitro. Patient-specific iPSC-based disease models are also expected to reveal early functional phenotypes, although this remains to be proved. Here, we generated iPSC lines from two patients with Sanfilippo type C syndrome, a lysosomal storage disorder with inheritable progressive neurodegeneration. Mature neurons obtained from patient-specific iPSC lines recapitulated the main known phenotypes of the disease, not present in genetically corrected patient-specific iPSC-derived cultures. Moreover, neuronal networks organized in vitro from mature patient-derived neurons showed early defects in neuronal activity, network-wide degradation, and altered effective connectivity. Our findings establish the importance of iPSC-based technology to identify early functional phenotypes, which can in turn shed light on the pathological mechanisms occurring in Sanfilippo syndrome. This technology also has the potential to provide valuable readouts to screen compounds, which can prevent the onset of neurodegeneration.

  12. Selective radiolabeling of cell surface proteins to a high specific activity

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, J.A.; Lau, A.L.; Cunningham, D.D.


    A procedure was developed for selective radiolabeling of membrane proteins on cells to higher specific activities than possible with available techniques. Cell surface amino groups were derivatized with /sup 125/I-(hydroxyphenyl)propionyl groups via /sup 125/I-sulfosuccinimidyl (hydroxyphenyl)propionate (/sup 125/II-sulfo-SHPP). This reagent preferentially labeled membrane proteins exposed at the cell surface of erythrocytes as assessed by the degree of radiolabel incorporation into erythrocyte ghost proteins and hemoglobin. Comparison with the lactoperoxidase-(/sup 125/I)iodide labeling technique revealed that /sup 125/I-sulfo-SHPP labeled cell surface proteins to a much higher specific activity and hemoglobin to a much lower specific activity. Additionally, this reagent was used for selective radiolabeling of membrane proteins on the cytoplasmic face of the plasma membrane by blocking exofacial amino groups with uniodinated sulfo-SHPP, lysing the cells, and then incubating them with /sup 125/I-sulfo-SHPP. Exclusive labeling of either side of the plasma membrane was demonstrated by the labeling of some marker proteins with well-defined spacial orientations on erythroctyes. Transmembrane proteins such as the epidermal growth factor receptor on cultured cells could also be labeled differentially from either side of the plasma membrane.

  13. High efficiency ex vivo cloning of antigen-specific human effector T cells.

    Directory of Open Access Journals (Sweden)

    Michelle A Neller

    Full Text Available While cloned T cells are valuable tools for the exploration of immune responses against viruses and tumours, current cloning methods do not allow inferences to be made about the function and phenotype of a clone's in vivo precursor, nor can precise cloning efficiencies be calculated. Additionally, there is currently no general method for cloning antigen-specific effector T cells directly from peripheral blood mononuclear cells, without the need for prior expansion in vitro. Here we describe an efficient method for cloning effector T cells ex vivo. Functional T cells are detected using optimised interferon gamma capture following stimulation with viral or tumour cell-derived antigen. In combination with multiple phenotypic markers, single effector T cells are sorted using a flow cytometer directly into multi-well plates, and cloned using standard, non antigen-specific expansion methods. We provide examples of this novel technology to generate antigen-reactive clones from healthy donors using Epstein-Barr virus and cytomegalovirus as representative viral antigen sources, and from two melanoma patients using autologous melanoma cells. Cloning efficiency, clonality, and retention/loss of function are described. Ex vivo effector cell cloning provides a rapid and effective method of deriving antigen-specific T cells clones with traceable in vivo precursor function and phenotype.

  14. Activity and High-Order Effective Connectivity Alterations in Sanfilippo C Patient-Specific Neuronal Networks. (United States)

    Canals, Isaac; Soriano, Jordi; Orlandi, Javier G; Torrent, Roger; Richaud-Patin, Yvonne; Jiménez-Delgado, Senda; Merlin, Simone; Follenzi, Antonia; Consiglio, Antonella; Vilageliu, Lluïsa; Grinberg, Daniel; Raya, Angel


    Induced pluripotent stem cell (iPSC) technology has been successfully used to recapitulate phenotypic traits of several human diseases in vitro. Patient-specific iPSC-based disease models are also expected to reveal early functional phenotypes, although this remains to be proved. Here, we generated iPSC lines from two patients with Sanfilippo type C syndrome, a lysosomal storage disorder with inheritable progressive neurodegeneration. Mature neurons obtained from patient-specific iPSC lines recapitulated the main known phenotypes of the disease, not present in genetically corrected patient-specific iPSC-derived cultures. Moreover, neuronal networks organized in vitro from mature patient-derived neurons showed early defects in neuronal activity, network-wide degradation, and altered effective connectivity. Our findings establish the importance of iPSC-based technology to identify early functional phenotypes, which can in turn shed light on the pathological mechanisms occurring in Sanfilippo syndrome. This technology also has the potential to provide valuable readouts to screen compounds, which can prevent the onset of neurodegeneration.

  15. Simultaneous measurements of auto-immune and infectious disease specific antibodies using a high throughput multiplexing tool.

    Directory of Open Access Journals (Sweden)

    Atul Asati

    Full Text Available Considering importance of ganglioside antibodies as biomarkers in various immune-mediated neuropathies and neurological disorders, we developed a high throughput multiplexing tool for the assessment of gangliosides-specific antibodies based on Biolpex/Luminex platform. In this report, we demonstrate that the ganglioside high throughput multiplexing tool is robust, highly specific and demonstrating ∼100-fold higher concentration sensitivity for IgG detection than ELISA. In addition to the ganglioside-coated array, the high throughput multiplexing tool contains beads coated with influenza hemagglutinins derived from H1N1 A/Brisbane/59/07 and H1N1 A/California/07/09 strains. Influenza beads provided an added advantage of simultaneous detection of ganglioside- and influenza-specific antibodies, a capacity important for the assay of both infectious antigen-specific and autoimmune antibodies following vaccination or disease. Taken together, these results support the potential adoption of the ganglioside high throughput multiplexing tool for measuring ganglioside antibodies in various neuropathic and neurological disorders.

  16. A centre-triggered magnesium fuelled cathodic arc thruster uses sublimation to deliver a record high specific impulse (United States)

    Neumann, Patrick R. C.; Bilek, Marcela; McKenzie, David R.


    The cathodic arc is a high current, low voltage discharge that operates in vacuum and provides a stream of highly ionised plasma from a solid conducting cathode. The high ion velocities, together with the high ionisation fraction and the quasineutrality of the exhaust stream, make the cathodic arc an attractive plasma source for spacecraft propulsion applications. The specific impulse of the cathodic arc thruster is substantially increased when the emission of neutral species is reduced. Here, we demonstrate a reduction of neutral emission by exploiting sublimation in cathode spots and enhanced ionisation of the plasma in short, high-current pulses. This, combined with the enhanced directionality due to the efficient erosion profiles created by centre-triggering, substantially increases the specific impulse. We present experimentally measured specific impulses and jet power efficiencies for titanium and magnesium fuels. Our Mg fuelled source provides the highest reported specific impulse for a gridless ion thruster and is competitive with all flight rated ion thrusters. We present a model based on cathode sublimation and melting at the cathodic arc spot explaining the outstanding performance of the Mg fuelled source. A further significant advantage of an Mg-fuelled thruster is the abundance of Mg in asteroidal material and in space junk, providing an opportunity for utilising these resources in space.

  17. Diagnostic value of immunohistochemical staining of GP73, GPC3, DCP, CD34, CD31, and reticulin staining in hepatocellular carcinoma. (United States)

    Yao, Shuzhe; Zhang, Jianping; Chen, Haiyan; Sheng, Yan; Zhang, Xiaoying; Liu, Zhiyan; Zhang, Cuijuan


    It has been reported that Golgi protein-73 (GP73), glypican-3 (GPC3), and des-γ-carboxy prothrombin (DCP) could serve as serum markers for the early detection of hepatocellular carcinoma (HCC). This study aimed to evaluate a panel of immunostaining markers (including GP73, GPC3, DCP, CD34, and CD31) as well as reticulin staining to distinguish HCC from the mimickers. Our results revealed that CD34 immunostaining and reticulin staining were highly sensitive for the diagnosis of HCC. A special immunoreaction pattern of GP73--a diffuse coarse-block pattern in a perinuclear region or a concentrated cluster-like or cord-like pattern in a certain part of the cytoplasm--was observed in HCC cells, in contrast to the cytoplasmic fine-granular pattern in surrounding non-tumor cells and non-malignant nodules. This coarse-block pattern correlated significantly with less differentiated HCC. In comparison, GPC3 displayed a good advantage in diagnosing well-differentiated HCC. In our study, DCP and CD31 showed little diagnostic value for HCC as an immunostaining marker. When GP73, GPC3, and CD34 were combined, the specificity improved to 96.6%. Our findings demonstrate for the first time that the immunohistochemical panel of GP73, GPC3, and CD34 as well as reticulin staining is highly specific for the pathological diagnosis of HCC.

  18. A survey of immunohistochemical biomarkers for basal-like breast cancer against a gene expression profile gold standard. (United States)

    Won, Jennifer R; Gao, Dongxia; Chow, Christine; Cheng, Jinjin; Lau, Sherman Y H; Ellis, Matthew J; Perou, Charles M; Bernard, Philip S; Nielsen, Torsten O


    Gene expression profiling of breast cancer delineates a particularly aggressive subtype referred to as 'basal-like', which comprises ∼15% of all breast cancers, afflicts younger women and is refractory to endocrine and anti-HER2 therapies. Immunohistochemical surrogate definitions for basal-like breast cancer, such as the clinical ER/PR/HER2 triple-negative phenotype and models incorporating positive expression for CK5 (CK5/6) and/or EGFR are heavily cited. However, many additional biomarkers for basal-like breast cancer have been described in the literature. A parallel comparison of 46 proposed immunohistochemical biomarkers of basal-like breast cancer was performed against a gene expression profile gold standard on a tissue microarray containing 42 basal-like and 80 non-basal-like breast cancer cases. Ki67 and PPH3 were the most sensitive biomarkers (both 92%) positively expressed in the basal-like subtype, whereas CK14, IMP3 and NGFR were the most specific (100%). Among biomarkers surveyed, loss of INPP4B (a negative regulator of phosphatidylinositol signaling) was 61% sensitive and 99% specific with the highest odds ratio (OR) at 108, indicating the strongest association with basal-like breast cancer. Expression of nestin, a common marker of neural progenitor cells that is also associated with the triple-negative/basal-like phenotype and poor breast cancer prognosis, possessed the second highest OR at 29 among the 46 biomarkers surveyed, as well as 54% sensitivity and 96% specificity. As a positively expressed biomarker, nestin possesses technical advantages over INPP4B that make it a more ideal biomarker for identification of basal-like breast cancer. The comprehensive immunohistochemical biomarker survey presented in this study is a necessary step for determining an optimized surrogate immunopanel that best defines basal-like breast cancer in a practical and clinically accessible way.

  19. High salt primes a specific activation state of macrophages, M(Na) (United States)

    Zhang, Wu-Chang; Zheng, Xiao-Jun; Du, Lin-Juan; Sun, Jian-Yong; Shen, Zhu-Xia; Shi, Chaoji; Sun, Shuyang; Zhang, Zhiyuan; Chen, Xiao-qing; Qin, Mu; Liu, Xu; Tao, Jun; Jia, Lijun; Fan, Heng-yu; Zhou, Bin; Yu, Ying; Ying, Hao; Hui, Lijian; Liu, Xiaolong; Yi, Xianghua; Liu, Xiaojing; Zhang, Lanjing; Duan, Sheng-Zhong


    High salt is positively associated with the risk of many diseases. However, little is known about the mechanisms. Here we showed that high salt increased proinflammatory molecules, while decreased anti-inflammatory and proendocytic molecules in both human and mouse macrophages. High salt also potentiated lipopolysaccharide-induced macrophage activation and suppressed interleukin 4-induced macrophage activation. High salt induced the proinflammatory aspects by activating p38/cFos and/or Erk1/2/cFos pathways, while inhibited the anti-inflammatory and proendocytic aspects by Erk1/2/signal transducer and activator of transcription 6 pathway. Consistent with the in vitro results, high-salt diet increased proinflammatory gene expression of mouse alveolar macrophages. In mouse models of acute lung injury, high-salt diet aggravated lipopolysaccharide-induced pulmonary macrophage activation and inflammation in lungs. These results identify a novel macrophage activation state, M(Na), and high salt as a potential environmental risk factor for lung inflammation through the induction of M(Na). PMID:26206316

  20. Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry

    DEFF Research Database (Denmark)

    Gallina, Serafina; Saletti, Rosaria; Cunsolo, Vincenzo;


    A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography......, electrospray mass spectrometry, and high collision dissociation fragmentation. The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281...

  1. Diagnostic value of immunohistochemical IMP3 expression in core needle biopsies of pancreatic ductal adenocarcinoma. (United States)

    Wachter, David Lukas; Schlabrakowski, Anne; Hoegel, Josef; Kristiansen, Glen; Hartmann, Arndt; Riener, Marc-Oliver


    The oncofetal protein, insulin-like growth factor II messenger ribonucleic acid-binding protein 3 (IMP3), has been analyzed in many different tumors. Various studies have found that IMP3 is a marker for malignancy and is correlated with increased tumor aggressiveness and reduced overall survival. The diagnosis of pancreatic ductal adenocarcinoma (PDAC) in core needle biopsies can be challenging, and immunohistochemical markers are needed. We studied IMP3 expression in 177 core needle biopsies of the pancreas, including 112 PDACs, 55 cases with chronic sclerosing pancreatitis, and 10 biopsies with tumor-free pancreatic tissue without inflammation. An additional 18 biopsies of PDAC metastases (16 liver biopsies and 2 lymph node biopsies) were analyzed. To study IMP3 expression in large tissue sections, 45 pancreatic resection specimens (26 with PDAC and 19 with chronic sclerosing pancreatitis) were investigated. In contrast to normal or inflamed pancreatic tissue, which was negative in 47 of 65 (72.3%) cases and weakly positive in 15 of 65 (23.1%) cases, strong IMP3 expression was found in 99 of 112 (88.4%) PDACs. Therefore, sensitivity and specificity of IMP3 expression in the differential diagnosis of PDAC and chronic sclerosing pancreatitis using core needle biopsies were found to be 88.4% and 94.6%, respectively. These results were confirmed in the pancreas resection specimens. Furthermore, strong IMP3 expression was found in 17 of 18 (94.4%) of the PDAC metastases that were analyzed. Our study shows that IMP3 is an easy to use and potentially new immunohistochemical marker for the diagnosis of PDAC in core needle biopsies.

  2. Immunohistochemical study on effects of [sup 60]Co [gamma] irradiation on salivary gland ducts of rat

    Energy Technology Data Exchange (ETDEWEB)

    Saitoh, Kazuhiko (Meikai Univ., Sakato, Saitama (Japan). School of Dentistry)


    Cytokeratin distribution in salivary glands was detected by use of polyclonal antikeratin antiserum (TK) and monoclonal antibodies (KL 1, RGE 53, and RPN 1164). The salivary glands of male rats received either 17.82 Gy or 27.97 Gy [sup 60]Co [gamma] irradiation in a single exposure and were then compared immunohistochemically with those of normal rats. Polyclonal anti-keratin antiserum (TK), which reacts with 41-65 KD keratins, stained almost all ducts in normal glands. RPN 1164 (no. 8 keratin) staining was negative in intercalated ducts of normal parotid and submandibular glands, but strongly positive in both striated and excretory ducts of these glands. Monoclonal antibody KL 1 (55-57 KD keratins) and RGE 53 (no. 18 keratin) did not bind to any ductal or acinar epithelia. Only in the sublingual gland were acini positive for TK staining, possibly indicating myoepithelial cells. No effects of [sup 60]Co [gamma] irradiation were apparent regarding keratin distribution in the intercalated in apical cytoplasm by [sup 60]Co [gamma] irradiation. Also, in the excretory duct, the basal side of the cells exhibited weakened staining following [sup 60]Co [gamma] irradiation were the most significant in the parotid and the least in the sublingual gland. Also this reaction depended upon the doses of [sup 60]Co used. The present findings suggested that negative or weakened staining at the basal and perinuclear portions of striated duct cells specifically reflects the primary or secondary cell damage produced by [sup 60]Co [gamma] irradiation. Since the distribution of cytoskeletal proteins in the cytoplasm reflects certain pathological conditions, immunohistochemical detection of these proteins seem to have a diagnostic value with respect to cellular injury. (J.P.N.) 77 refs.

  3. EGFR and gastrointestinal stromal tumor: an immunohistochemical and FISH study of 82 cases. (United States)

    Lopes, Lisandro F; Bacchi, Carlos E


    Gastrointestinal stromal tumor is the most common mesenchymal neoplasm of the gastrointestinal tract. Mutually exclusive KIT or platelet-derived growth factor receptor-alpha mutations are key events in gastrointestinal stromal tumor pathogenesis, and specific treatment targeting KIT/platelet-derived growth factor receptor-alpha activation is available. Epidermal growth factor receptor plays an important role in cancer biology and also constitutes a promising molecular target of therapy. Very few reports have been published in the literature about the relationship between gastrointestinal stromal tumor and epidermal growth factor receptor. The aim of this study was to investigate epidermal growth factor receptor immunohistochemical expression and epidermal growth factor receptor gene amplification in 82 consecutive gastrointestinal stromal tumor cases using tissue microarray technique. Hematoxylin- and eosin-stained sections and clinical information were reviewed, and expression of CD117 (KIT), CD34 and epidermal growth factor receptor was investigated by immunohistochemistry. Epidermal growth factor receptor gene copy number was determined using fluorescence in situ hybridization. Immunohistochemistry revealed that CD117 and CD34 were expressed in 96 and 57% of tumors, respectively. Variable epidermal growth factor receptor protein immunohistochemical overexpression was detected in 96% of gastrointestinal stromal tumor cases, but none of the 75 cases with represented tumor tissue cores and countable fluorescence signals exhibited epidermal growth factor receptor gene amplification by fluorescence in situ hybridization. These results show that there is no correlation between epidermal growth factor receptor protein overexpression by immunohistochemistry and epidermal growth factor receptor gene amplification by fluorescence in situ hybridization. Considering that the mechanisms of epidermal growth factor receptor protein overexpression are not well understood and

  4. High Specific-Strength C-Zr(O)C / C-Ablator TPS for CEV Project (United States)

    National Aeronautics and Space Administration — During the Phase I NASA SBIR, MATECH GSM (MG) has developed and evaluated the world's first ultra-high temperature (UHT) Zr-(O)-C ceramic fiber pre-form / organic...

  5. High specific-strength C-Zr(O)C / C-ablator TPS for CEV Project (United States)

    National Aeronautics and Space Administration — This Phase I NASA SBIR Proposal seeks to demonstrate a new zero-erosion ablative thermal protection system (TPS) technology that is based upon an ultra-high...

  6. Polymer nanomicelles for efficient mucus delivery and antigen-specific high mucosal immunity. (United States)

    Noh, Young-Woock; Hong, Ji Hyun; Shim, Sang-Mu; Park, Hye Sun; Bae, Hee Ho; Ryu, Eun Kyoung; Hwang, Jung Hwan; Lee, Chul-Ho; Cho, Seong Hun; Sung, Moon-Hee; Poo, Haryoung; Lim, Yong Taik


    Micelles for mucosal immunity: A mucosal vaccine system based on γ-PGA nanomicelles and viral antigens was synthesized. The intranasal administration of the vaccine system induces a high immune response both in the humoral and cellular immunity (see picture).

  7. Advanced Lithium-ion Batteries with High Specific Energy and Improved Safety for Nasa's Missions (United States)

    West, William; Smart, Marshall; Soler, Jess; Krause, Charlie; Hwang, Constanza; Bugga, Ratnakumar


    High Energy Materials ( Cathodes, anodes and high voltage and safe electrolyte are required to meet the needs of the future space missions. A. Cathodes: The layered layered composites of of Li2MnO3 and LiMO2 are promising Power capability of the materials, however requires further improvement. Suitable morphology is critical for good performance and high tap (packing) density. Surface coatings help in the interfacial kinetics and stability. B. Electrolytes: Small additions of Flame Retardant Additives improves flammability without affecting performance (Rate and cycle life). 1.0 M in EC+EMC+TPP was shown to have good performance against the high voltage cathode; Performance demonstrated in large capacity prototype MCMB- LiNiCoO2 Cells. Formulations with higher proportions are looking promising. Still requires further validation through abuse tests (e.g., on 18650 cells).

  8. Advanced Lithium-ion Batteries with High Specific Energy and Improved Safety for Nasa's Missions (United States)

    West, William; Smart, Marshall; Soler, Jess; Krause, Charlie; Hwang, Constanza; Bugga, Ratnakumar


    High Energy Materials ( Cathodes, anodes and high voltage and safe electrolyte are required to meet the needs of the future space missions. A. Cathodes: The layered layered composites of of Li2MnO3 and LiMO2 are promising Power capability of the materials, however requires further improvement. Suitable morphology is critical for good performance and high tap (packing) density. Surface coatings help in the interfacial kinetics and stability. B. Electrolytes: Small additions of Flame Retardant Additives improves flammability without affecting performance (Rate and cycle life). 1.0 M in EC+EMC+TPP was shown to have good performance against the high voltage cathode; Performance demonstrated in large capacity prototype MCMB- LiNiCoO2 Cells. Formulations with higher proportions are looking promising. Still requires further validation through abuse tests (e.g., on 18650 cells).

  9. Advanced Nanostructured Cathode for Ultra High Specific Energy Lithium Ion Batteries Project (United States)

    National Aeronautics and Space Administration — Integrate advanced nanotechnology with energy storage technology to develop advanced cathode materials for use in Li-ion batteries while maintaining a high level of...

  10. Specific mold filling characteristics of highly filled phenolic injection molding compounds


    Scheffler, Thomas; Englich, Sascha; Gehde, Michael


    Thermosets show excellent mechanical properties and chemical resistance (for most automotive fluids) even at high temperatures up to 300 °C. Furthermore they can be highly efficient processed by injection molding. So they should be particularly suited for e.g. under the bonnet applications. However, the reality shows that thermosets are, except fiber reinforced composites, heavily underrepresented in technical applications. E.g. thermosetting components only account 0,2 % to a vehicle’s weigh...

  11. Immunohistochemical analyses of a case of extralobar pulmonary sequestration with chest pain in an adult

    Directory of Open Access Journals (Sweden)

    Yuji Ohtsuki


    Full Text Available Computed tomography of a Japanese man in his mid-forties with a complaint of right-side chest pain showed a dome-shaped smooth-surfaced mediastinal mass, which was extirpated. The cut surface was highly hemorrhagic and necrotic and not related to the original pulmonary tissues. Although routine sectioning detected bronchial cartilage, immunohistochemical analyses clearly showed the presence of alveolar type II cells; only the alveolar type II cells located at the periphery of this mass showed positive staining for cytokeratins, thyroid transcription factor 1, surfactant protein A, epithelial membrane antigen and Krebs von den Lungen-6. Thus, these analyses are useful for the detection of pulmonary components, even in severely hemorrhagic and necrotic tissues with marked sequestration. The clinical diagnosis was a rare, adult type of extralobar pulmonary sequestration accompanied by chest pain.

  12. Immunohistochemical investigation of pulmonary surfactant-associated protein A in fire victims. (United States)

    Zhu, B L; Ishida, K; Oritani, S; Quan, L; Taniguchi, M; Li, D R; Fujita, M Q; Maeda, H


    To evaluate the forensic pathological significance of the immunohistochemical distribution of pulmonary surfactant-associated protein A (SP-A) in determining the cause of death in fires, 57 fire victims were examined by scoring the staining intensity. The highest SP-A score with dense granular deposits (aggregates) in the intra-alveolar space was frequently observed in cases with a lower blood carboxyhemoglobin (COHb) level (fires. High SP-A scores showed a relation to the serum level and in part related to the bloody exudate in the lower airway. These observations suggested that the increase in SP-A in fire victims may be mainly related to pulmonary alveolar injury due to the inhalation of hot air and/or irritant gases rather than hypoxia.

  13. High specificity of human secretory class II phospholipase A2 for phosphatidic acid. (United States)

    Snitko, Y; Yoon, E T; Cho, W


    Lysophosphatidic acid (LPA) is a potent lipid second messenger which stimulates platelet aggregation, cell proliferation and smooth-muscle contraction. The phospholipase A2 (PLA2)-catalysed hydrolysis of phosphatidic acid (PA) is thought to be a primary synthetic route for LPA. Of the multiple forms of PLA2 present in human tissues, human secretory class-II PLA2 (hs-PLA2) has been implicated in the production of LPA from platelets and whole blood cells challenged with inflammatory stimuli. To explore further the possibility that hs-PLA2 is involved in the production of LPA, we rigorously measured the phospholipid head group specificity of hs-PLA2 by a novel PLA2 kinetic system using polymerized mixed liposomes. Kinetic analysis of recombinant hs-PLA2 demonstrates that hs-PLA2 strongly prefers PA as substrate over other phospholipids found in the mammalian plasma membrane including phosphatidylserine (PS), phosphatidylcholine (PC) and phosphatidylethanolamine (PE). The order of preference is PA > PE approximately PS > PC. To identify amino acid residues of hs-PLA2 that are involved in its unique substrate specificity, we mutated two residues, Glu-56 and Lys-69, which were shown to interact with the phospholipid head group in the X-ray-crystallographic structure of the hs-PLA2-transition-state-analogue complex. The K69Y mutant showed selective inactivation toward PA whereas the E56K mutant displayed a most pronounced inactivation to PE. Thus it appears that Lys-69 is at least partially involved in the PA specificity of hs-PLA2 and Glu-56 in the distinction between PE and PC. In conjunction with a recent cell study [Fourcade, Simon, Viode, Rugani, Leballe, Ragab, Fournie, Sarda and Chap (1995) Cell 80, 919-927], these studies suggest that hs-PLA2 can rapidly hydrolyse PA molecules exposed to the outer layer of cell-derived microvesicles and thereby produce LPA.

  14. A low-cost lead-acid battery with high specific-energy

    Indian Academy of Sciences (India)

    S K Martha; B Hariprakash; S A Gaffoor; D C Trivedi; A K Shukla


    Lightweight grids for lead-acid battery grids have been prepared from acrylonitrile butadiene styrene (ABS) copolymer followed by coating with lead. Subsequently, the grids have been electrochemically coated with a conductive and corrosion-resistant layer of polyaniline. These grids are about 75% lighter than those employed in conventional lead-acid batteries. Commercial-grade 6V/3.5Ah (C20-rate) lead-acid batteries have been assembled and characterized employing positive and negative plates constituting these grids. The specific energy of such a lead-acid battery is about 50 Wh/kg. The batteries can withstand fast charge-discharge duty cycles.

  15. High-throughput identification of antigen-specific TCRs by TCR gene capture

    DEFF Research Database (Denmark)

    Linnemann, Carsten; Heemskerk, Bianca; Kvistborg, Pia;


    The transfer of T cell receptor (TCR) genes into patient T cells is a promising approach for the treatment of both viral infections and cancer. Although efficient methods exist to identify antibodies for the treatment of these diseases, comparable strategies to identify TCRs have been lacking. We...... the quantitative nature of TCR gene capture, we show the feasibility of identifying antigen-specific TCRs in oligoclonal T cell populations from either human material or TCR-humanized mice. Finally, we demonstrate the ability to identify tumor-reactive TCRs within intratumoral T cell subsets without knowledge...

  16. Identification of antigen-specific human monoclonal antibodies using high-throughput sequencing of the antibody repertoire. (United States)

    Liu, Ju; Li, Ruihua; Liu, Kun; Li, Liangliang; Zai, Xiaodong; Chi, Xiangyang; Fu, Ling; Xu, Junjie; Chen, Wei


    High-throughput sequencing of the antibody repertoire provides a large number of antibody variable region sequences that can be used to generate human monoclonal antibodies. However, current screening methods for identifying antigen-specific antibodies are inefficient. In the present study, we developed an antibody clone screening strategy based on clone dynamics and relative frequency, and used it to identify antigen-specific human monoclonal antibodies. Enzyme-linked immunosorbent assay showed that at least 52% of putative positive immunoglobulin heavy chains composed antigen-specific antibodies. Combining information on dynamics and relative frequency improved identification of positive clones and elimination of negative clones. and increase the credibility of putative positive clones. Therefore the screening strategy could simplify the subsequent experimental screening and may facilitate the generation of antigen-specific antibodies.

  17. Experimental Study on Specific Heat of Concrete at High Temperatures and Its Influence on Thermal Energy Storage

    Directory of Open Access Journals (Sweden)

    Jianwen Pan


    Full Text Available Using concrete as a thermal energy storage (TES material is a promising option for large-scale solar-thermal resource development and utilization. Specific heat is one of the most important characteristics for TES performance. In this paper, the half-open dynamic method based on the mixing principle is proposed and applied to measure concrete-specific heat at temperatures up to 600 °C. Measurement of the specific heat of corundum ceramic (99% Al2O3 is first performed, and the test results illustrate the accuracy and efficiency of the proposed test method. Furthermore, concrete-specific heat tests are carried out at high temperatures. It found that the specific heat increases as the temperature rises, especially, linearly in the range of 300–600 °C, in which the concrete TES module is expected to be in operation. Finally, the effect of concrete-specific heat changes with temperature on its TES capacity is investigated, demonstrating that specific heat is of great significance for concrete TES design for concentrating solar power.

  18. Bottom-up Design of Three-Dimensional Carbon-Honeycomb with Superb Specific Strength and High Thermal Conductivity. (United States)

    Pang, Zhenqian; Gu, Xiaokun; Wei, Yujie; Yang, Ronggui; Dresselhaus, Mildred S


    Low-dimensional carbon allotropes, from fullerenes, carbon nanotubes, to graphene, have been broadly explored due to their outstanding and special properties. However, there exist significant challenges in retaining such properties of basic building blocks when scaling them up to three-dimensional materials and structures for many technological applications. Here we show theoretically the atomistic structure of a stable three-dimensional carbon honeycomb (C-honeycomb) structure with superb mechanical and thermal properties. A combination of sp(2) bonding in the wall and sp(3) bonding in the triple junction of C-honeycomb is the key to retain the stability of C-honeycomb. The specific strength could be the best in structural carbon materials, and this strength remains at a high level but tunable with different cell sizes. C-honeycomb is also found to have a very high thermal conductivity, for example, >100 W/mK along the axis of the hexagonal cell with a density only ∼0.4 g/cm(3). Because of the low density and high thermal conductivity, the specific thermal conductivity of C-honeycombs is larger than most engineering materials, including metals and high thermal conductivity semiconductors, as well as lightweight CNT arrays and graphene-based nanocomposites. Such high specific strength, high thermal conductivity, and anomalous Poisson's effect in C-honeycomb render it appealing for the use in various engineering practices.

  19. The population genetics of Pseudomonas aeruginosa isolates from different patient populations exhibits high-level host specificity

    NARCIS (Netherlands)

    van Mansfeld, R.; Jongerden, I.P.; Bootsma, M.C.; Buiting, A.; Bonten, M.J.M.; Willems, R.J.L.


    Objective To determine whether highly prevalent P. aeruginosa sequence types (ST) in Dutch cystic fibrosis (CF) patients are specifically linked to CF patients we investigated the population structure of P. aeruginosa from different clinical backgrounds. We first selected the optimal genotyping meth

  20. Sequence-specific high mobility group box factors recognize 10-12-base pair minor groove motifs

    DEFF Research Database (Denmark)

    van Beest, M; Dooijes, D; van De Wetering, M


    Sequence-specific high mobility group (HMG) box factors bind and bend DNA via interactions in the minor groove. Three-dimensional NMR analyses have provided the structural basis for this interaction. The cognate HMG domain DNA motif is generally believed to span 6-8 bases. However, alignment...

  1. The population genetics of Pseudomonas aeruginosa isolates from different patient populations exhibits high-level host specificity

    NARCIS (Netherlands)

    van Mansfeld, R.; Jongerden, I.P.; Bootsma, M.C.|info:eu-repo/dai/nl/304830305; Buiting, A.; Bonten, M.J.M.; Willems, R.J.L.


    Objective To determine whether highly prevalent P. aeruginosa sequence types (ST) in Dutch cystic fibrosis (CF) patients are specifically linked to CF patients we investigated the population structure of P. aeruginosa from different clinical backgrounds. We first selected the optimal genotyping meth

  2. Immunohistochemical classification and prognosis of diffuse large B-cell lymphoma in China

    Institute of Scientific and Technical Information of China (English)



    Objective To study the immunohistochemical classification and prognosis of diffuse large B-cell lymphoma(DLBCL).Methods A total of 148 cases of DLBCL were classified into germinal center B-cell-like(GCB)and non-GCB/activated B-cell-like(ABC)subtypes by Hans,Choi and Tally immunohistochemical stain algorithms.The clinical features and survival data of GCB

  3. Hot water and dilute acid pretreatment of high and low specific gravity Populus deltoides clones. (United States)

    Martin, Elizabeth M; Bunnell, Kris A; Lau, Ching-Shuan; Pelkki, Matthew H; Patterson, David W; Clausen, Edgar C; Smith, James A; Carrier, Danielle Julie


    Populus sp. are hardwood feedstocks that grow in forest management areas that are logged for softwoods; however, they are also being considered as an energy-destined feedstock. The objective of this work was to determine the effect of xylose yield from dilute acid and hot water pretreatments performed in unstirred batch stainless steel reactors at temperatures ranging from 140 to 200°C. Populus deltoides clones S13C20 and S7C15 used in this study originated from Eastern Texas and were cultivated for 14 years in Pine Tree, AR. P. deltoides clones S13C20 and S7C15 had specific gravities of 0.48 and 0.40, respectively. Bark and wood were examined separately. As expected, hot water pretreatments, in the tested temperature range, resulted in very little direct xylose recovery. However, the 140°C dilute acid pretreatment of the lower specific gravity clone, S7C15, wood yielded the highest average xylose recovery of 56%. This condition also yielded the highest concentration of furfural, 9 mg/g sample, which can be inhibitory to the fermentation step. The highest xylose recovery from bark samples, 31%, was obtained with clone S7C15, using the 160°C dilute acid pretreatment for 60 min.

  4. Comparative VEGF receptor tyrosine kinase modeling for the development of highly specific inhibitors of tumor angiogenesis. (United States)

    Schmidt, Ulrike; Ahmed, Jessica; Michalsky, Elke; Hoepfner, Michael; Preissner, Robert


    The Vascular Endothelial Growth Factor receptors (VEGF-Rs) play a significant role in tumor development and tumor angiogenesis and are therefore interesting targets in cancer therapy. Targeting the VEGF-R is of special importance as the feed of the tumor has to be reduced. In general, this can be carried out by inhibiting the tyrosine kinase function of the VEGF-R. Nevertheless, there arise some problems with the specificity of known kinase inhibitors: they bind to the ATP-binding site and inhibit a number of kinases, moreover the so far most specific inhibitors act at least on these three major types of VEGF-Rs: Flt-1, Flk-1/KDR, Flt-4. The goal is a selective VEGF-R-2 (Flk-1/KDR) inhibitor, because this receptor triggers rather unspecific signals from VEGF-A, -C, -D and -E. Here, we describe a protocol starting from an established inhibitor (Vatalanib) with 2D-/3D-searching and property filtering of the in silico screening hits and the "negative docking approach". With this approach we were able to identify a compound, which shows a fourfold higher reduction of the proliferation rate of endothelial cells compared to the reduction effect of the lead structure.

  5. Joint-Angle Specific Strength Adaptations Influence Improvements in Power in Highly Trained Athletes

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    Rhea Matthew R.


    Full Text Available Purpose. The purpose of this study was to examine the influence of training at different ranges of motion during the squat exercise on joint-angle specific strength adaptations. Methods. Twenty eight men were randomly assigned to one of three training groups, differing only in the depth of squats (quarter squat, half squat, and full squat performed in 16-week training intervention. Strength measures were conducted in the back squat pre-, mid-, and post-training at all three depths. Vertical jump and 40-yard sprint time were also measured. Results. Individuals in the quarter and full squat training groups improved significantly more at the specific depth at which they trained when compared to the other two groups (p < 0.05. Jump height and sprint speed improved in all groups (p < 0.05; however, the quarter squat had the greatest transfer to both outcomes. Conclusions. Consistently including quarter squats in workouts aimed at maximizing speed and jumping power can result in greater improvements.

  6. Site-specific chromosomal integration in mammalian cells: highly efficient CRE recombinase-mediated cassette exchange. (United States)

    Feng, Y Q; Seibler, J; Alami, R; Eisen, A; Westerman, K A; Leboulch, P; Fiering, S; Bouhassira, E E


    Expression of experimental constructs in mammalian cells or transgenic animals is difficult to control because it is markedly influenced by position effects. This has limited both the analysis of cis -DNA regulatory elements for transcription and replication, and the physiological analysis of proteins expressed from transgenes. We report here two new methods based on the concept of recombinase-mediated cassette exchange (RMCE) to perform site-specific chromosomal integration. The first method permits the exchange of a negative selectable marker pre-localized on the chromosome with a transgene via a CRE-mediated double recombination between inverted Lox sites. Integration efficiency is close to 100 % of negatively selected mouse erythroleukemia cells and ranges from 10 to 50 % in embryonic stem cells. The second method allows RMCE with no selection at all except for cells that have taken up plasmid transiently. While less efficient, this technique permits novel experimental approaches. We find that integration of a transgene at a given genomic site leads to reproducible expression. RMCE should be useful to develop artificial genetic loci that impart specific and reproducible regulation of transgenes in higher eukaryotes. This should facilitate the analysis of cis -regulatory DNA elements governing expression and position effects, improve our control over the physiological effects of transgenes, and accelerate the development of animal models for complex human diseases.

  7. Evaluation of cell proliferative activity after irradiation using immunohistochemical approach and flow cytometry

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    Tamada, Takashi (Okayama Univ. (Japan). School of Medicine)


    To evaluate a proliferative activity of post-irradiated malignant cells, we studied the kinetics of HeLa cells using immunohistochemical approach and flow cytometry. HeLa cells were stained with two proliferation-associated monoclonal antibodies, Ki-67 and anti-DNA polymerase {alpha} antibody. Nucleoli of non-irradiated cells were granularly stained with Ki-67. After irradiation, only the center of nuclei was diffusely stained with Ki-67. One hundred forty-four hours after low-dose irradiation, the staining patterns became the same as the control. On the other hand, after high-dose irradiation, the center of nuclei was weakly stained. DNA polymerase {alpha} was diffusely labelled with nuclei of the control. It was located around the border of nuclei of low-dose irradiated cells like a ring. But after high-dose irradiation, it was granularly distributed in the periphery of nuclei. FITC conjugated Ki-67/PI two parameter analysis was done by a single laser flow cytometer. Twenty-four hours after irradiation, DNA-histograms showed the accumulation to G{sub 2}/M phase and the increase of DNA content of G{sub 2}/M cells, as exposure dose was increased. Two parameter analysis showed the increase of FITC uptake of G{sub 2}/M phase as dose increased. These changes of flow cytometry were remarkably observed after 24 hours' incubation. It was shown that the difference of Ki-67 antigen and DNA polymerase {alpha} appearance depended on the irradiation dose. These findings suggest that immunohistochemical staining with Ki-67 or anti-DNA polymerase {alpha} antibody and flow cytometry using Ki-67 are available to evaluate cell damages after irradiation. (author).

  8. Decreasing trend in prostate cancer with high serum prostate-specific antigen levels detected at first prostate-specific antigen-based population screening in Japan

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    Yasuhide Kitagawa


    Full Text Available To clarify the recent trends in prostate-specific antigen (PSA distribution in men in Japan, we analyzed the PSA distributions of men undergoing PSA-based population screening. We summarized the annual individual data of PSA-based population screening in Kanazawa, Japan, from 2000 to 2011, and analyzed baseline serum PSA values of the participants at the first population screening. Serum PSA distributions were estimated in all participants and those excluding prostate cancer patients according to age. From 2000 to 2011, 19 620 men participated aged 54-69 years old in this screening program. Mean baseline serum PSA level of all participants at the first screening was 2.64 ng ml−1 in 2000, and gradually decreased to approximately 1.30 ng ml−1 in 2006. That of participants excluding prostate cancer patients was 1.46 ng ml−1 in 2000, and there was no remarkable change during the study period. The 95 th percentiles in the participants excluding prostate cancer patients detected at the first population screening of men aged 54-59, 60-64, and 65-69 years old were 2.90, 3.60, and 4.50 ng ml−1 , respectively. After the commencement of population screening, the proportion of prostate cancer patients with high serum PSA levels decreased. However, there were no changes in serum PSA levels in men without prostate cancer. Age-specific PSA reference level of men without prostate cancer in Japan was similar to that in China and Korea.

  9. High specificity ZnO quantum dots for diagnosis and treatment in bacterial infection (United States)

    Zhang, Min; Qian, Zhiyu; Gu, Yueqing


    Early diagnosis and effective treatment of bacterial infection has become increasingly important. Herein, we developed a fluorescent nano-probe MPA@ZnO-PEP by conjugating SiO2-stabilized ZnO quantum dot (ZnO@SiO2) with bacteria-targeting peptide PEP, which was encapsulated with MPA, a near infrared (NIR) dye. The nanoprobe MPA@ZnO-PEP showed excellent fluorescence property and could specifically distinguish bacterial infection from sterile inflammation both in vitro and in vivo. The favorable biocompatability of MPA@ZnO-PEP was verified by MTT assay. This probe was further modified with antibiotic methicillin to form the theranostic nanoparticle MPA/Met@ZnO-PEP with amplified antibacterial activity. These results promised the great potential of MPA@ZnO-PEP for efficient non-invasive early diagnosis of bacterial infections and effective bacterial-targeting therapy.

  10. Site-specific Quantification of Bone Quality using Highly Nonlinear Solitary Waves

    CERN Document Server

    Yang, Jinkyu; Borkowski, Sean; Silvestro, Claudio; De Nardo, Luigi; Daraio, Chiara; Ebramzadeh, Edward


    Osteoporosis is a well recognized problem affecting millions of individuals worldwide. Consequently, the need to effectively, efficiently, and affordably diagnose and identify those at risk is essential; moreover, site-spe