Impact of p16 status on pro- and anti-angiogenesis factors in head and neck cancers.

BACKGROUND: Head and neck cancers (HNC) are aggressive tumours. Overexpression of p16 in HNC correlates with human papilloma virus (HPV)-associated HNC that carry a better prognosis than HPV-negative tumours. Angiogenesis is an important factor in tumour progression. Our aim was to dissect the impact of p16 expression on angiogenesis factors in HNC. METHODS: Eighteen newly diagnosed HNC patients and controls were analysed. Eleven pro- and anti-angiogenesis factors were quantified using multiplex ELISA in HNC patients and controls. Angiogenesis factors were analysed in tumour tissue using immunohistochemistry. RESULTS: Circulating levels of endostatin (anti-angiogenesis factor) were higher in the HNC group compared with healthy donors. Interestingly, the pro-angiogenesis factors angiopoietin-1 and vascular endothelial growth factor (VEGF) were significantly higher in patients with p16-negative compared with p16-positive HNC. Moreover, the major source of VEGF in p16-positive HNC tissue was tumour stromal cells. In contrast, both tumour cells and stromal cells expressed VEGF in p16-negative tissue. CONCLUSIONS: We show that p16-negative tumours associate with increased circulating levels of pro-angiogenic VEGF and angiopoietin-1. Tissue expression of VEGF differs between p16-positive and p16-negative tumours. These findings may explain differences in the biological behaviour of p16-positive and p16-negative HNC. Better understanding of mechanisms by which the p16 status influences tumour angiogenesis may guide the development of targeted therapies

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Not AvailableConserving the mithun (Bos frontalis), a rare bovine of south-east Asia, could benefit from effective ex situ genetic management and breeding programmes including the use of assisted reproduction. Longterm storage of epididymal sperm collected from dead mithun bulls by cryopreservation, with high survival rate, is essential for the establishment of genetic resource banks of this rare species. A study was therefore conducted to (i) characterize the epididymal sperm collected from dead mithun bulls, (ii) investigate the effectiveness of epididymal sperm for cryopreservation and (iii) to study the expression pattern of genes related to motility (TSSK6 and ADAM5P ) and fertility (PRM1, PRM2, PRM3, Tnp1 and Tnp2) in mithun epididymal sperm. For the purpose, sperm collected from caudal epididymis of eight dead mithun bulls were evaluated for concentration, progressive motility, morphological abnormalities, live sperm counts, acrosome integrity, membrane stability (hypo-osmotic swelling test; HOST) and DNA integrity. Epididymal sperm were cryopreserved using tris-egg yolk-glycerol diluent in liquid nitrogen. Post-thaw quality of the cryopreserved sperm in terms of progressive motility, morphological abnormalities, live cell counts, acrosome integrity, membrane stability and DNA integrity were assessed. The RNA extracted from fresh and post-thawed cryopreserved epididymal sperm was reverse transcribed to cDNA and expressions of the genes related to motility and fertility were determined by RT-PCR. The progressive motility, live cell counts, morphological abnormalities and acrosome integrity (normal) of fresh sperm were 89.9±2.7, 88.7±6.7, 7.8±1.07 and 95.3±7.8%, respectively. Fresh sperm that responded to HOST were 88.3±7.2%, and 89.5±6.4% fresh sperm had intact DNA. Following cryopreservation, the percentage of progressive motility (fresh vs frozen-thawed), live cell counts, morphological abnormalities, acrosome integrity, membrane stability and DNA integrity were found to decrease significantly (P<0.01) with a motility recovery rate of 75±8.5%. Transcripts encoding a serine/threonine testis-specific protein kinase (TSSK6) and a metalloproteinase non coding RNA (ADAM5P ) were found to be associated with high-motility status (P<0.01) and their relative expressions were higher in fresh than frozen-thawed sperm. There were no differences in relative expressions for the transcripts related to fertility (PRM1, PRM2, PRM3, Tnp1 and Tnp2) between fresh and frozen-thawed sperm indicating no change of fertility of mithun epididymal sperm after cryopreservation. Taken together, results of our study provide a foundation of collection and cryopreservation of epididymal sperm from dead mithun bulls. Furthermore, the post-thaw quality in terms of percentage of progressive motility, live cell counts, morphological abnormalities, acrosome integrity, membrane stability and DNA integrity were sufficient for genetic improvement of this rare species through AI and conservation of the valuable germplasm for future.Not Availabl

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Not AvailablePresent experiment was conducted to study the effect of dietary protein levels on growth, immunity and anti-oxidative status of Labeo rohita fingerlings during feed deprivation followed by refeeding. Fish (5.44 ± 0.10 g) were deprived of feed for 3 weeks and then re-fed to satiation for 5 weeks with one of the diets containing 25 (25P), 30 (30P), 35 (35P) or 40 (40P) percent crude protein (CP) level. In addition to these groups, a control group (C) was also maintained by feeding to satiation level twice daily with a diet containing 30% CP throughout the experimental period. At the end of 8-weeks' trial, fish were challenged with Aeromonas hydrophila and survival was recorded for the next 7 days. Complete recovery of growth in terms of weight gain percentage was achieved in the fish fed 35 and 40% protein during refeeding. The body indices (condition factor and hepatosomatic index), haematological parameters and serum protein contents at the end of the experimental trial were not significantly different (P > 0.05) among different groups suggesting that the overall health of the fish was not compromised. However, respiratory burst activity and serum lysozyme activity were indicative of a better immune function in the higher protein fed groups (35P and 40P) than the lower protein groups (25P and 30P). Following challenge with Aeromonas hydrophila, survival rate, blood monocyte%, respiratory burst activity, serum lysozyme activity, serum protein and globulin were significantly higher (P < 0.05) in the 35P and 40P groups compared to the other groups. Further, fish fed lower dietary protein were not able to restore the activities of anti-oxidative enzymes (superoxide dismutase and catalase) in the liver. Conclusively, an improved disease resistance capability and immune status was observed in the fish fed a higher dietary protein (35-40%), even out-performing the daily-fed fish.Not Availabl

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Not AvailableA study was conducted to assess the effect of liquid semen preservation on semen quality parameters (SQPs) such as forward progressive motility, livability, acrosomal integrity and total morphological abnormality at refrigerated temperature in mithun. Good quality ejaculates (50) were collected and preserved at refrigerated temperature (5°C) for 72 h using tris egg yolk citrate based extender (TEYCE). Seminal attributes, viz. colour, consistency, volume, mass activity and sperm concentration of fresh semen samples were creamy white, medium, 1.8±0.08 ml, 3.4±0.09 (5 point scale) and 478±15 (×106/ml), respectively. SQPs were assessed after initial dilution, after final dilution and up to 72 h at 6 h intervals. The progressive motility, live sperm with intact acrosome decreased significantly over the period of storage and were below 20% after 48 h of storage. Total morphological abnormalities increased significantly at the time of 30% progressive motility. The study concluded that the mithun sperm can be preserved in tris based egg yolk extender in liquid storage at refrigeration temperature (5°C) for 36 h.Not Availabl

Water soaking and exogenous enzyme treatment of plant-based diets: effect on growth performance, whole-body composition, and digestive enzyme activities of rohu, Labeo rohita (Hamilton), fingerlings

A 2 x 2 x 2 factorial experiment was conducted to delineate the main effect of water soaking of plant ingredients, phytase, cellulase, and their interactions on the growth and digestive enzyme activities of Labeo rohita fingerlings. Two basal diets were prepared using water-soaked (S) or unsoaked (US) plant-based ingredients. Feed of US ingredients was supplemented with phytase (U kg(-1)) and cellulase (%) at the level of 0, 0 (C-us); 500, 0 (T-1); 0, 0.2 (T-2); 500, 0.2 (T-3), and feed of S ingredients at 0, 0 (C-s); 500, 0 (T-4); 0, 0.2 (T-5), and 500, 0.2 (T-6), respectively. Three hundred and sixty fingerlings were randomly distributed into eight treatments, each with three replicates. Soaking of the ingredients for 24 h significantly reduced the tannin content. However, feeding of S diets did not improve the fish growth. Highest performance was recorded in the T3 group. A significant interaction between dietary phytase and cellulase was observed for apparent net protein utilization. Tissue crude protein, ether extract, and ash content of the fingerlings were observed highest in the T-3 group. Activities of amylase, protease, and lipase were recorded highest in the T-3 group. Results suggested that soaking of plant-based ingredients reduces tannin content; however, growth and digestive enzyme activities of group fed soaked diet were not improved, possibly due to leaching of soluble nutrients. Probably, a shorter duration soaking may be effective in reducing tannin content and avoiding nutrients leaching

sperm cryopreservation glycerol mithun

Not AvailableThe effect of concentration and addition method of glycerol on the quality of cryopreserved mithun (Bos frontalis) spermatozoa was investigated. Semen samples were collected from five healthy mithun bulls through rectal massage method and cryopreserved in liquid nitrogen. The samples were diluted in Tris–egg yolk–glycerol extender, equilibrated for 4 h at 4 °C and loaded into 0.50‐ml straws. The straws were then frozen in liquid nitrogen vapour for 10 min and finally plunged into liquid nitrogen for storage. The required amount of glycerol was added into the diluted samples either in a single dose (3%, 4%, 5%, 6% or 7%; added at 37 °C immediately before equilibration) or in split doses (5%, 6% or 7%; the total amount was divided into four equal parts, and a part was added at 37 °C immediately before equilibration, and the remaining parts were added subsequently at 1, 2 and 3 h of equilibration at 4 °C). In the single‐dose addition method, following freeze‐thawing, greater (p < 0.05) motility (%) and proportion of live spermatozoa with intact acrosome (LSIA, %) in 5% glycerol (40.6 ± 1.7 and 43.4 ± 1.8 respectively) and lesser (p < 0.05) total morphological abnormalities (%) in 5% (14.1 ± 0.8) and 6% (13.7 ± 1.0) glycerol were observed compared to the other glycerol concentrations. In the split‐dose addition method, following freeze‐thawing, greater (p < 0.05) motility (%) and LSIA proportion (%) were found in 5% (50.2 ± 1.9 and 53.3 ± 1.8 respectively) compared to 6% or 7% glycerol, but the total morphological abnormalities were not different among the glycerol concentrations. In addition, in all the glycerol concentrations, better (p < 0.05) post‐freeze‐thaw motility and LSIA proportions were observed when glycerol was added in split doses compared to a single dose. In conclusion, Tris–egg yolk extender with 5% glycerol added in split doses was found most suitable for cryopreserving mithun sperm.Not Availabl