Resident phenotypically modulated vascular smooth muscle cells in healthy human arteries

Vascular interstitial cells (VICs) are non‐contractile cells with filopodia previously described in healthy blood vessels of rodents and their function remains unknown. The objective of this study was to identify VICs in human arteries and to ascertain their role. VICs were identified in the wall of human gastro‐omental arteries using transmission electron microscopy. Isolated VICs showed ability to form new and elongate existing filopodia and actively change body shape. Most importantly sprouting VICs were also observed in cell dispersal. RT‐PCR performed on separately collected contractile vascular smooth muscle cells (VSMCs) and VICs showed that both cell types expressed the gene for smooth muscle myosin heavy chain (SM‐MHC). Immunofluorescent labelling showed that both VSMCs and VICs had similar fluorescence for SM‐MHC and αSM‐actin, VICs, however, had significantly lower fluorescence for smoothelin, myosin light chain kinase, h‐calponin and SM22α. It was also found that VICs do not have cytoskeleton as rigid as in contractile VSMCs. VICs express number of VSMC‐specific proteins and display features of phenotypically modulated VSMCs with increased migratory abilities. VICs, therefore represent resident phenotypically modulated VSMCs that are present in human arteries under normal physiological conditions.     

Real time in vitro studies of doxorubicin release from PHEMA nanoparticles

Background  

Many anticancer agents have poor water solubility and therefore the development of novel delivery systems for such molecules has received significant attention. Nanocarriers show great potential in delivering therapeutic agents into the targeted organs or cells and have recently emerged as a promising approach to cancer treatments. The aim of this study was to prepare and use poly-2-hydroxyethyl methacrylate (PHEMA) nanoparticles for the controlled release of the anticancer drug doxorubicin.     

Metarrhizium anisopliae (Metchinkoff) Sorokin., and its host range

Metarrhizium anisopliae (Metchinkoff)Sorokin., can be readily cultured in quantity on a medium containing honey, peptone, and barley.Poecilocerus pictus andHeliothis obsoleta are two additional hosts of the fungus reported for the first time.     

Microhyla laterite sp. nov., A New Species of Microhyla Tschudi, 1838 (Amphibia: Anura: Microhylidae) from a Laterite Rock Formation in South West India

In recent times, several new species of amphibians have been described from India. Many of these discoveries are from biodiversity hotspots or from within protected areas. We undertook amphibian surveys in human dominated landscapes outside of protected areas in south western region of India between years 2013–2015. We encountered a new species of Microhyla which is described here as Microhyla laterite sp. nov. It was delimited using molecular, morphometric and bioacoustics comparisons. Microhyla laterite sp. nov. appears to be restricted to areas of the West coast of India dominated by laterite rock formations. The laterite rock formations date as far back as the Cretaceous-Tertiary boundary and are considered to be wastelands in-spite of their intriguing geological history. We identify knowledge gaps in our understanding of the genus Microhyla from the Indian subcontinent and suggest ways to bridge them.     

5-year review and reappraisal of ultrasound-guided percutaneous transabdominal fine needle aspiration of pancreatic lesions

OBJECTIVE: To reevaluate the efficacy and safety offine needle aspiration cytology (FNAC) of pancreatic lesions performed by transabdominal approach. STUDY DESIGN: Retrospective 5-year (2001-2006) audit of all pancreatic FNA samples. RESULTS: This series includes 267 patients (88 men, 179 women). Seven cases (2.6%) yielded insufficient material for diagnosis; 260 cases were classified as benign (n=118) and malignant (n=142) lesions. Of the 118 benign aspirates, the cytodiagnosis was acute/chronic inflammation in 24, tuberculosis in 1, benign cyst in 10 and a benign aspirate, not otherwise specified, in the remaining 83 cases. Of the 142 malignant aspirates, the cytodiagnosis was adenocarcinoma in 126, neuroendocrine/carcinoid tumor in 7, papillary solid epithelial neoplasm in 2, mucinous cystadenocarcinoma in 2, acinar cell carcinoma in 1 and metastatic small cell carcinoma in lung in 4 cases. Cytohistologic correlation yielded a sensitivity of 81% and specificity of 100%. CONCLUSION: A spectrum of pancreatic lesions can be accurately diagnosed by the technique. The false negative rate can be minimized by proper positioning of the needle under guidance and adequate sampling. No postprocedural complications were encountered, proving that this procedure is safe if carried out by an experienced team in a hospital setting.     

Lactic acid bacteria isolated from yak milk show probiotic potential

Applied Microbiology and Biotechnology - Probiotic industries strive for new, efficient and promising probiotic strains that impart a positive impact on consumer health. Challenges are persisting...     

A continuous fluorescence assay for sulfhydryl oxidase

Flavin-dependent sulfhydryl oxidases represent a newly discovered family of proteins with a range of cellular locations and putative roles. The avian and mammalian proteins can catalyze the direct oxidation of protein cysteine residues to disulfides with the reduction of dioxygen to hydrogen peroxide. Although thiols interfere with the peroxidase-mediated quantitation of hydrogen peroxide, a very sensitive, continuous fluorescence assay of the sulfhydryl oxidases can be devised with careful selection of thiol substrate concentration and fluorogen. Purified avian enzyme (or crude chicken egg white) was used for these experiments. Homovanillic acid was found to be a suitable fluorogen in the presence of 300 microM thiols from either dithiothreitol or reduced ribonuclease A. High concentrations of horseradish peroxidase minimized the effects of contaminating catalase in biological samples. Using fluorescence microcells, the assay could detect 15fmol of avian sulfhydryl oxidase and the rates were linearly dependent on enzyme concentration up to 6nM. Aspects of the interaction among thiols, homovanillic acid, and peroxidase are discussed which limit the sensitivity of the assay and require that care is exercised in the application of this new procedure. Finally, the assay is used to show that there is sulfhydryl oxidase activity in a number of secretory fluids including human tears.     

Synthesis and characterization of hapten-protein conjugates for antibody production against small molecules

For the generation of antibodies against small hapten molecules, the hapten is cross-linked with some carrier protein to make it immunogenic. However, the formation of such conjugates is not always reproducible. This may lead to inconsistent hapten-protein stoichiometries, resulting in large variations in the generation of the desired antibodies. In the study described here the hapten (mercaptopropionic acid derivative of atrazine) was coupled to carrier protein at five different molar ratios. The hapten-protein conjugates prepared were characterized thoroughly by spectrophotometric absorption, fluorescence, matrix-assisted laser desorption ionization (MALDI), and gel electrophoresis methods, before being used for the immunization and assay purposes. Electrophoresis and fluorescence methods were very useful in detecting hapten-protein cross-linking while MALDI-MS and spectrophotometric detection provided qualitatively comparable hapten density. The production of specific antibodies was sought following the generation of appropriate hapten-protein conjugates. A high antibody titer with moderate antibody specificity was obtained with hapten density around 15 molecules per carrier protein. The study proved useful for monitoring the course of hapten-protein conjugation for the production of specific antibodies against small molecules.     

Correlation between VEGF expression and angiogenesis in breast carcinoma

OBJECTIVE: To analyze the role of vascular endothelial growth factor (VEGF) secreted by tumor cells in angiogenesis of breast carcinoma using image morphometry. STUDY DESIGN: Thirty-four cases of node-negative breast carcinoma were used in the study. There were 6 grade 1, 20 grade 2 and 8 grade 3 tumors. For each case, 2 consecutive sections from the same block were cut. Immunostaining for VEGF and CD31 was carried out, and areas of highest staining density were marked. Those marked "hot spots" for CD31 and VEGF for each case were subsequently compared morphometrically. The area and intensity of immunostaining on each slide were also scored. RESULTS: The total scores for VEGF and CD31 were 5.15 and 3.79, respectively. All 34 cases showed cytoplasmic positivity for VEGF within the tumor cells. The average number of hot spots for VEGF and CD31 were 2.41 and 2.47, respectively, and the average number of hot spots that matched between these 2 groups were 0.79. Statistical analysis using Pearson's coefficient of correlation showed no significant match between the hot spots for CD31 and VEGF. Also, there was no significant difference between the total scores of CD31 and VEGF. CONCLUSION: VEGF is expressed in most breast carcinomas. However, the lack of topographic correlation between microvessel density and VEGF expression supports the notion that multiple angiogenic factors may play a role along with VEGF in the angiogenic process.     

Effects of superoxide dismutase and polyclonal tumor necrosis factor-alpha antibodies on chloroacetate-induced cellular death and superoxide anion production by J774.A1 macrophages

Dichloroacetate (DCA) and trichloroacetate (TCA) are by-products that are formed during the process of water chlorination and have been previously shown to induce superoxide anion (SA) production and cellular death when added to J774.A1 macrophage cultures. In this study, the effects of superoxide dismutase (SOD) and polyclonal tumor necrosis factor-alpha (TNF-alpha) antibodies on DCA- and TCA-induced SA production and cellular death have been tested on the J774.A1 macrophage cultures. TCA and DCA were added to different cultures either alone, each at a concentration of 16 mM, or in combination with SOD (2-12 units/ml), or with TNF-alpha antibodies (10 and 25 units/ml). Cells were incubated for 48 h, after which cellular death/viability, lactate dehydrognase (LDH) leakage by the cells, and SA production by the cells were determined. While TCA and DCA caused significant cellular toxicity, indicated by reduction in cellular viability and increases in LDH leakage and SA production, SOD addition resulted in significant reduction of the effects induced by the compounds. On the other hand, addition of TNF-alpha antibodies to the DCA- and TCA-treated cultures resulted in significant reduction of DCA- but not TCA-induced cellular death and SA production by the cells. Although these results suggest a significant role for SA in DCA- and TCA-induced cellular death, they may also suggest two different mechanisms for the chloroacetate-induced SA production by the cells.