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Breast cancer is an ancient disease recognized first by the Egyptians as early as 1600 BC. The first cancer-causing gene in a chicken tumor virus was found in 1970. The United States signed the National Cancer Act in 1971, authorizing federal funding for cancer research. Irrespective of multi-disciplinary approaches, diverting a great deal of public and private resources, breast cancer remains at the forefront of human diseases, affecting as many as one in eight women during their lifetime. Because of overarching challenges and changes in the breast cancer landscape, five-year disease-free survival is no longer considered adequate. The absence of a cure, and the presence of drug resistance, severe side effects, and destruction of the patient's quality of life, as well as the fact that therapy is often expensive, making it unaffordable to many, have created anxiety among patients, families, and friends. One of the reasons for the failure of cancer therapeutics is that the approaches do not consider cancer holistically. Characteristically, all breast cancer cells and their microenvironmental capillary endothelial cells express asparagine-linked (N-linked) glycoproteins with diverse structures. We tested a small biological molecule, Tunicamycin, that blocks a specific step of the protein N-glycosylation pathway in the endoplasmic reticulum (ER), i.e., the catalytic activity of N-acetylglusosaminyl 1-phosphate transferase (GPT). The outcome was overwhelmingly exciting. Tunicamycin quantitatively inhibits angiogenesis in vitro and in vivo, and inhibits the breast tumor progression of multiple subtypes in pre-clinical mouse models with "zero" toxicity. Mechanistic details support ER stress-induced unfolded protein response (upr) signaling as the cause for the apoptotic death of both cancer and the microvascular endothelial cells. Additionally, it interferes with Wnt signaling. We therefore conclude that Tunicamycin can be expected to supersede the current therapeutics to become a glycotherapy for treating breast cancer of all subtypes. [ABSTRACT FROM AUTHOR]

R3004 --> 680.4 --> Neovascularization, a normal physiological process orchestrates through vasculogenesis from mesoderm cell precursors. In healthy adults, the vasculature is quiescent but pathological angiogenesis is evident during wound healing and in tumor growth. Thus, angiogenesis becomes a fundamental step for tumors transitioning from a benign state to malignancy where endothelial cells proliferate 45 times faster. Our laboratory has recently shown that asparagine‐linked (N‐linked) protein glycosylation is essential for enhanced capillary endothelial cell proliferation in situ. Inhibiting N‐glycosylation with Tunicamycin causes G1 arrest and induces cell death by apoptosis due to "ER stress" and is not reversible by VEGF. Tunicamycin also inhibits angiogenesis in vivo in Matrigel™ implants in nude mice as well as in a pre‐clinical mouse model of double negative (ER‐/PR‐/HER2+) breast tumor. Wnt pathway is involved in cellular differentiation to organogenesis, and supports angiogenesis. In "canonical" Wnt/β‐catenin signaling pathway β‐catenin translocates to nucleus and activates Wnt/β‐catenin target genes, including vascular endothelial growth factor A (VEGF‐A). Irrespective of the Wnt ligand and Frizzled being glycoproteins, the cross‐talk between protein N‐glycosylation and the Wntpathway has not been addressed. This led us to hypothesize that decreased expression of Wntprotein(s) in Tunicamycin treated endothelial cell would decrease cellular proliferation. We have used capillary endothelial cells as a model and observed down regulated expression of Wnt proteins in Tunicamycin treated cells. Our results support Tunicamycin‐induced "ER stress" interferes with Wnt signaling and inhibits angiogenesis. We, therefore, conclude molecular understanding of the process would provide a novel target to inhibit angiogenesis and breast tumor progression. [ABSTRACT FROM AUTHOR]

To explore the heat and cold adaptation in Indian goats by the physiological, haematological, blood biochemical parameters and their seasonal variations, this study was conducted on heat- and cold-adapted Indian goats maintained in their natural habitat. Study was carried out in three different phases coincide with the three seasons (winter, spring and summer). The levels of physiological responses, that is rectal temperature, respiration rate and pulse rate, were observed to be significantly (p < 0.01) lower in heat-adapted breeds and higher in cold-adapted breeds, whereas the levels of Hb, PCV and TEC were significantly (p < 0.01) higher in cold-adapted goats. Significantly (p < 0.01) higher levels of plasma thyroid hormones (thyroxine and triiodothyronine) and plasma stress enzyme (AST and ALT) were also observed in cold-adapted goats. Significant (p < 0.01) seasonal variations in physiological responses, haematological and blood biochemical parameters in both heat- and cold-adapted breeds were reported in this study. Physiological responses, plasma enzymes and plasma cortisol levels significantly (p < 0.01) increased during summer in all the goat breeds. The levels of haematological parameters (Hb, PCV, TEC and TLC) and plasma thyroid hormones (thyroxine and triiodothyronine) decreased during summer. The changes in physiological parameters during summer due to heat stress were higher in cold-adapted goats whereas the levels of changes in these parameters during winter due to cold were higher in heat-adapted goats. High neutrophil/lymphocyte ratio during summer in cold-adapted breeds is an indicator of higher level of stress. Decrease in plasma electrolytes (Na and K) during summer also observed in cold-adapted breeds during summer. The variations in physiological, haematological, blood biochemical parameters in heat- and cold-adapted goats may be due to their adaptation to different environmental and geographical conditions essential for their survival. [ABSTRACT FROM AUTHOR]

Abstract: Asparagine-linked (N-linked) protein glycosylation is one of the most important protein modifications. N-glycans with “high mannose”, “hybrid”, or “complex” type sugar chains participate in a multitude of cellular processes. These include cell–cell/cell–matrix/receptor–ligand interaction, cell signaling/growth and differentiation, to name a few. Many diseases such as disorders of blood clotting, congenital disorder of glycosylation, diseases of blood vessels, cancer, neo-vascularization, i.e., angiogenesis essential for breast and other solid tumor progression and metastasis are associated with N-glycan expression. Biosynthesis of N-glycans requires multiple steps and multiple cellular compartments. Following transcription and translation the proteins migrate to the endoplasmic reticulum (ER) lumen to acquire glycan chain(s) with a defined glycoform, i.e., a tetradecasaccharide. These are further modified, i.e., edited in ER lumen and in Golgi prior to moving to their respective destinations. The tetradecasaccharide is pre-assembled on a poly-isoprenoid lipid called dolichol, and becomes an essential component of the supply chain. Therefore, dolichol cycle synthesizing the lipid-linked oligosaccharide (LLO) is a hallmark for all N-linked glycoproteins. It is expected that there is a great deal of cross-talk between the participating glycosyltransferases and any missed step would express defective N-glycans that could have fatal consequences. The positive impact of the structurally altered N-glycans could lead to discovery of an N-glycan signature for a disease and/or help developing glycotherapeutic treating cancer or other human diseases. The purpose of this review is to identify the gaps of N-glycan biology and help developing appropriate technology for biomedical applications. This article is part of a Special Issue entitled Glycoproteomics. [Copyright &y& Elsevier]

To explore the circadian changes in physiological responses and blood ionized sodium, potassium concentrations under thermal exposure in Tharparkar (TP) and Karan Fries (KF) heifers, 12 animals, six each from two breeds were taken and kept first in control thermoneutral (21-26°C and 50-60% RH) and in heat exposed (38°C and 50-60% RH) conditions thereafter. Throughout a 24-hour period physiological responses (rectal temperature, respiration rate and pulse rate) and blood samples were taken at 4-hour intervals, then blood samples were analyzed for sodium (Na+) and potassium (K+) levels. Significant circadian variations in physiological responses and blood Na+ and K+ within 24 hours have been observed. Significant (P < 0.01) variation in physiological responses and blood Na+, K+ concentrations were observed in both Tharparkar and Karan Fries heifers under control and heat exposed conditions. Values of physiological responses were lower in the morning and showed an increasing trend and reached the highest value in the afternoon. Blood Na+ and K+ concentrations decreased from 11.00 hours to 19.00 hours due to heat exposure. The decreases in blood electrolyte levels were greater in Karan Fries in comparison to Tharparkar heifers that expressed less heat tolerance capacity in crossbred than indigenous cattle. This finding suggests that it is important to standardize the time of sample collection in comparative studies, in which more discrete differences may be identified in physiological responses and blood electrolyte concentrations. [ABSTRACT FROM AUTHOR]

To explore the effect of thermal stress on diurnal rhythms of physiological responses, peripheral temperatures, feed and water intake, blood biochemical parameters, stress enzymes and plasma sodium, potassium levels in Tharparkar (TP) and Karan Fries (KF) heifers, 12 animals, six each from two breeds were taken and kept first in control thermoneutral (21-26°C and 50-60% RH) and in heat exposed (38-39°C temperature and 60-70% RH) conditions thereafter. All the data recording and blood sample collections were done twice (morning and evening) a day for a continuous seven days each for control and thermal exposed conditions. Values of physiological responses, peripheral temperatures, blood biochemical parameters, stress enzymes and plasma sodium, potassium levels in both Tharparkar and Karan Fries heifers showed a significant variation (P < 0.05) in the morning and evening. So a diurnal fluctuation of rectal temperature was observed in this experiment. Significant (P < 0.01) variation in all the above parameters was observed in both Tharparkar and Karan Fries heifers in control and heat exposed conditions. Feed intake decreased and water intake increased in the heat exposed condition in both the breeds. Values of physiological responses were lower in the morning and showed an increasing trend, reaching highest values in the afternoon. The changes in physiological parameters were greater in Karan Fries in comparison to Tharparkar heifers that expressed lower heat tolerance capacity in crossbred than indigenous cattle. The correlation coefficient showed a significant relationship among all the parameters studied. This finding suggests that it is important to standardize the time of sample collection in comparative studies, in which more discrete differences may be identified in physiological parameters. Animals are comfortable in thermoneutral conditions and are in stress in thermal exposed conditions, so shade and cooling facilities should be provided during summer to withstand the heat stress and for better health and productivity. [ABSTRACT FROM AUTHOR]

The physiological processes of organisms are regulated by a circadian rhythm. The circadian rhythm is regulated by the wavelength, intensity, timing and duration of the light stimulus. Biological rhythms affect the sleep-wake cycle, migration behaviour in birds, seasonal fattening, hibernation and reproductive cycles in animals. The circadian rhythm is ubiquitous in nature. Circadian rhythms appear to be generated at the cellular level. Daily biological rhythms are endogenously controlled by self-contained circadian clocks. The period length is controlled by a circadian oscillator (clock). The timing of sleep and wakefulness under natural conditions is in synchrony with the circadian control of the sleep cycle and all other circadian-controlled rhythms. Adverse effects may ensue when the sleep-wake cycle is out of phase with the rhythms that are controlled by the circadian clock. Circadian rhythm is related to the light/dark cycle of the solar day but it also persists in constant conditions. The environmental cues entraining the circadian rhythm are called Zeitgebers or circadian synchronizers. Temporal restrictions of feeding (RF) can phase-shift behavioural and physiological circadian rhythms in mammals. Phase advances of circadian rhythms happens for instance, in the liver, kidney, heart, pancreas and some brain structures, uncoupling them from the control of the SCN, whose entrainment to light remains intact. [ABSTRACT FROM AUTHOR]

The fluffy brown sparrows are 15cm in length and distributed all over India up to 4000m in the Himalayas. The disappearance of sparrows has been widely reported in India. The sparrow population in Andhra Pradesh alone had dropped by 80 per cent, and in other states like Kerala, Gujarat and Rajasthan, it had dipped by 20 per cent, while the decline in coastal areas was as sharp as 70 to 80 per cent. But reliable information on sparrow populations is not available. No one is actually counting and keeping a record of the sparrows. The spread of diseases due to decline in sparrow population is an alarming danger. Introduction of unleaded petrol, use of chemically treated seeds, flow of electromagnetic waves from cellphone towers, reducing areas of free growing weeds or reducing numbers of badly maintained buildings, competition for food by other species etc. are possible reasons for this disappearance. The BirdLife International, Royal Society for the Protection of Birds (RSPB), a UK-based organisation and the Bombay Natural History Society (BNHS) have taken plan for the protection of sparrow population. [ABSTRACT FROM AUTHOR]

DPM1 is the structural gene for mannosylphosphodolichol synthase (i.e. Dol-P-Man synthase, DPMS) in Saccharomyces cerevisiae. Earlier studies with cDNA cloning and sequence analysis have established that 31kDa DPMS of S. cerevisiae contains a consensus sequence (YRRVIS141) that can be phosphorylated by cAMP-dependent protein kinase (PKA). We have been studying the up-regulation of DPMS activity by protein kinase A-mediated phosphorylation in higher eukaryotes, and used the recombinant DPMS from S. cerevisiae in this study to advance our knowledge further. DPMS catalytic activity was indeed enhanced severalfold when the recombinant protein was phosphorylated in vitro. The rate as well as the magnitude of catalysis was higher with the phosphorylated enzyme. A similar increase in the catalytic activity was also observed when the in vitro phosphorylated recombinant DPMS was assayed as a function of increasing concentrations of exogenous dolichylmonophosphate (Dol-P). Kinetic studies indicated that there was no change in the Km for GDP-mannose between the in vitro phosphorylated and control recombinant DPMS, but the Vmax was increased by 6-fold with the phosphorylated enzyme. In vitro phosphorylated recombinant DPMS also exhibited higher enzyme turnover (kcat) and enzyme efficiency (kcat/Km). SDS-PAGE followed by autoradiography of the 32P-labeled DPMS detected a 31-kDa phosphoprotein, and immunoblotting with anti-phosphoserine antibody established the presence of a phosphoserine residue in in vitro phosphorylated recombinant DPMS. To confirm the phosphorylation activation of recombinant DPMS, serine 141 in the consensus sequence was replaced with alanine by PCR site-directed mutagenesis. The S141A DPMS mutant exhibited more than half-a-fold reduction in catalytic activity compared with the wild type when both were analyzed after in vitro phosphorylation. Thus, confirming that S. cerevisiae DPMS activity is indeed regulated by the cAMP-dependent protein phosphorylation signal, and the phosphorylation target is serine 141. [ABSTRACT FROM AUTHOR]

Simple Summary: The use of non-therapeutic antibiotics in poultry industry is a common practice to generate desired growth of the birds. Non-therapeutic uses of antibiotics were detected as a culprit for the generation of antibiotic-resistant bacteria in the food chain. Many countries restrict the use of non-therapeutic antibiotics. Consequently, there is an increased demand for alternatives to antibiotic growth promoters (AGPs). The study was conducted to assess the impact of dietary supplementation of a probiotic and a postbiotic (Saccharomyces cerevisiae fermentation product, SCFP) on growth performance, carcass traits, blood haemato-biochemical profile, gut microflora, gut morphology, and immune response in broilers as an alternative to AGP. The study conducted on 324 one-day-old chickens revealed significant improvement of feed conversion ratio in the postbiotic group than the control. Cholesterol levels and concentrations of corticosterone were significantly lowered in the postbiotic group compared to other groups. Pathogenic- and antibiotic-resistant Gram-negative bacterial populations were significantly lower in postbiotic and probiotic groups. Moreover, humoral immunity was significantly improved in postbiotic and probiotic groups than the control birds. It can be concluded that both postbiotics and probiotics could be viable alternatives to antimicrobials in poultry production. Concern for global health security and the environment due to the emergence of antibiotic-resistant bacteria and antibiotic residues in meat and other livestock products has led many countries to restrict the use of antibiotics in animal feed. This experiment was performed to assess the impact of dietary supplementation of a probiotic (Bacillus subtilis) and a postbiotic (Saccharomyces cerevisiae fermentation product) on growth performance, carcass traits, blood haemato-biochemical profile, gut microflora, gut morphology, and immune response in broilers as an alternative to antimicrobials in poultry production system to minimize the effect on global health security. A total of 324 one-day-old Ven Cobb 400 broiler chicks were randomly divided into three dietary groups, each containing 12 replicated pens, and each replicate contained nine chickens. The dietary groups consisted of (1) a basal diet without any growth promoters (T1), (2) the basal diet augmented with Bacillus subtilis at 200 g/MT feed (T2), and (3) the basal diet supplemented with Saccharomyces cerevisiae fermentation product at 1.25 kg/MT feed (T3). To calculate body weight gain, all birds and residual feed were weighed on a replicated basis on days 0, 7, 14, 21, 28, 35, and 42; mortality was recorded daily. At the end of the trial (42 d), two chickens from each replicate were slaughtered for carcass traits, gut microflora, and morphology measurements. Blood samples were collected for the haemato-biochemical profile on 35 d and antibody titer on 28 d and 35 d. Feeding with SCFP (T3 group) significantly improved average daily feed intake (ADFI) and average daily gain (ADG) of chickens compared to the T1 (control) and T2 (probiotic) groups from 1 to 14 days of age. Feed conversion ratio (FCR) was significantly improved in SCFP-fed birds (T3) relative to the control (T1) over the entire experimental period. Carcass traits and blood haemato-biochemical parameters remained unaffected by any diets. However, cholesterol levels and concentrations of corticosterone were significantly lower in T3 compared to T2 and T1 groups. Total E. coli, Enterohaemorrhagic E. coli, ESBL-producing Enterobacteriaceae, and Salmonella counts were significantly lower in T2 and T3 groups compared to T1 group and Salmonella counts were lower in T3 when compared to T2. However, there was no significant difference in Lactobacillus count among treatment groups. A significant increase in villi height and villi-height-to-crypt-depth ratio (VH: CD) was observed in both T3 and T2 groups. On day 28, the T3 and T2 groups exhibited a significant increase in antibody titers against Newcastle disease virus and infectious bursal disease virus. It can be concluded that Saccharomyces cerevisiae fermentation product and Bacillus subtilis probiotic could be viable alternatives to antimicrobials in poultry production considering beneficial impacts in broilers fed an antibiotic-free diet. [ABSTRACT FROM AUTHOR]

The RFamide-related peptides (RFRPs) are the group of neuropeptides synthesized predominantly from the hypothalamus that negatively affects the hypothalamo-hypophyseal-gonadal (hypothalamic-pituitary-gonadal [HPG]) axis. These peptides are first identified in quail brains and emerged as the mammalian orthologs of avian gonadotropin inhibitory hormones. The RFRP-3 neurons in the hypothalamus are present in several mammalian species. The action of RFRP-3 is mediated through a G-protein-coupled receptor called OT7T022. The predominant role of RFRP-3 is the inhibition of HPG axis with several other effects such as the regulation of metabolic activity, stress regulation, controlling of non-sexual motivated behavior, and sexual photoperiodicity in concert with other neuropeptides such as kisspeptin, neuropeptide-Y (NPY), pro-opiomelanocortin, orexin, and melanin. RFamide peptides synthesized in the granulosa cells, interstitial cells, and seminiferous tubule regulate steroidogenesis and gametogenesis in the gonads. The present review is intended to provide the recent findings that explore the role of RFRP-3 in regulating HPG axis and its potential applications in the synchronization of reproduction and its therapeutic interventions to prevent stress-induced amenorrhea. [ABSTRACT FROM AUTHOR]

Ghoongroo pigs have good adaptability in a low input production system with high prolificacy. The present study was conducted on pre-and post-weaned Ghoongroo piglets from 2–3 days to 12 weeks of age to evaluate the effect of a milk replacer and oregano essential oil (EO) on growth, physiological and immunological responses. Thirty six piglets were randomly divided into three groups. The control group (n = 12) was allowed to suck mother's milk. Second group piglets were provided milk replacer (MR) and piglets of the third group were provided milk replacer along with oregano EO at 500 mg/kg diet. After weaning, piglets were provided standard concentrate diets. The results showed that the body weight in MR and MR + EO groups were significantly higher compared with the control. The MR + EO group had better intestinal microbiota, greater nonspecific innate immunity with the phagocytosis efficacy of neutrophils, lower cortisol concentration and more stable thyroid hormones than the other groups. The better haematological status supported the rapid organ development and improved intestinal health status in both the experimental groups. In conclusion, milk replacer, especially with the inclusion of oregano EO, can lower weaning stress, enhance nonspecific immunity and improve growth and health status of piglets. [ABSTRACT FROM AUTHOR]

The adaptive physiological functions of black Bengal goats (Capra hircus) of coastal regions were compared with goats from the plains during winter, spring, and summer seasons. Goats in the coastal zone had significantly (P < 0.01) higher plasma electrolytes levels. Goats in the plains zone had significantly (P < 0.01) higher haemoglobin, PCV, TEC, and erythrocyte indices and lower TLC values. Glucose, total protein, and creatinine were significantly (P < 0.01) lower, whereas cholesterol, BUN, and SGPT were significantly (P < 0.01) higher in goats of coastal zone. Regardless of zones, glucose, total protein, and creatinine levels were higher in winter, cholesterol and SGOT levels were higher in spring, and BUN was higher in summer. Goats from coastal areas had significantly (P < 0.05) higher urinary pH and urea levels. Goats in the coastal zone had significantly (P < 0.05) higher cortisol concentrations. From the above study, we can conclude that salinity of feed, soil and water does not significantly affect the homeostasis of black Bengal goats reared in coastal areas, indicating their adaptability to saline coastal environments and their potential for reproduction in a climate-resilient livestock system. The fodder, water and soil of coastal areas contain higher amounts of sodium, zinc, and iron in the compared to plain areas. Black Bengal goats raised in coastal regions are able to maintain their homeostasis without much altering hemato-biochemical and endocrine profiles. Black Bengal goats can be a good option for climate resilient animal husbandry system. [ABSTRACT FROM AUTHOR]

The present investigation was undertaken on data of 30 Sahiwal cattle and 30 crossbred cattle (HF × Jersey × Sahiwal) from Instructional Dairy Farm (IDF), G.B. Pant University of Agriculture & Technology, Pantnagar, Uttarakhand. Significant (P<0.01) variations were observed in birth weight, first service period, milk yield in first 305 days (first lactation milk yield), second lactation milk yield, average daily milk yield during first lactation and days in milk in first lactation between Sahiwal and crossbred cattle. Non-significant variations were observed in first calving interval between Sahiwal and crossbred cattle. Correlation coefficient showed a significant (P< 0.01) relationship among birth weight, first calving interval, first service period, milk yield in first 305 days (first lactation milk yield), second lactation milk yield, average daily milk yield during first lactation and days in milk in first lactation. Our present investigation showed that the overall performance of crossbred (HF × Jersey × Sahiwal) is better than Sahiwal. [ABSTRACT FROM AUTHOR]

Background: The present study focused on the localization and comparative expression analysis of RFRP-3 gene in the hypothalamus and testis of adult male rats. Methods: Brain and testis tissue samples were collected from adult Wister rats and used for immunohistochemistry for identification and localization RFRP-3 producing neurons. The expression level of the RFRP-3 gene was assessed by Real-time PCR and compared with muscle tissue. Result: The results confirmed the presence of RFRP-3 secreting neurons in the hypothalamus and in the testes. Scattered RFRP-3 in cell bodies was identified primarily in the VMH, DMH and surrounding the third ventricle of the brain. In testis, RFRP-3 was detected in Leydig cells, Sertoli cells and germ cells. The expression level of RFRP-3 gene in the hypothalamus was observed to be significantly higher in hypothalamus than in testicular tissue (p<0.01) of adult rats. [ABSTRACT FROM AUTHOR]

Breast cancer is the most frequently diagnosed cancer in women, accounting for one out of every eight women. The etiology of the disease is complex and requires neovascularization for the growth of a malignant tumor. Our laboratory has observed recently that tunicamycin, (a) an antibiotic, (b) a glucosamine-containing pyrimidine nucleoside, and (c) a protein N-glycosylation inhibitor reduces angiogenesis significantly. This was due to an induction of unfolded protein response-mediated cell cycle arrest and apoptosis. The objective of our present study is to test the hypothesis that tunicamyin inhibits MCF-7 cell proliferation. MCF-7 cells ware synchronized by serum deprivation for 48 hours, and treated with tunicamycin (0-10 µg/ml) for 24 hours. The cell morphology was monitored by light microscopy. The cell growth was reduced only 20-30% at 0.1 µg/ml and 1.0 µg/ml of tunicamycin, respectively but the inhibition was >70% at 10 µg/ml of tunicamycin. There was no indication of the cell cycle arrest. Total Bcl-2 expression was increased in the presence of 10 µg/ml and 0.1 pg/ml of tunicamycin as well as at "zero time". Bak expression was not detected but a high level expression of Bax, Bad and p53 was detected at low tunicamycin concentration. There was no detectable expression of either caspase 3 or 9. Therefore, we conclude that tunicamycin-mediated MCF-7 cells death may not be due to an induction apoptosis. Flow cytometric evaluation of the cell cycle also supports such conclusion. [ABSTRACT FROM AUTHOR]

The exact mechanism of switching to a new angiogenic phenotype during tumor growth is currently unknown. But, we have observed a direct relationship between the dolichol cycle and the cell cycle dynamics during in vitro angiogenesis. A non-transformed capillary endothelial cell line when exposed to 2mM 8Br-cAMP the cell proliferation enhanced ∼70% with one-half of the cell population in the S phase. A significant increase in the lipid-linked oligosaccharide (LLO) biosynthesis and turnover, the expression of cell surface N-glycans, and HSP-70 and HSP-90 was also observed. Mannosylphospho dolichol synthase (DPMS) catalyzes a 'key step' in the assembly of LLO. Its activity was upregulated during 8Br-cAMP treatment. The objective of the present study is to address that DPMS undergoes a structural change following cAMP-dependent protein phosphorylation. To test the hypothesis we have isolated a cDNA clone (bDPMS) after RT-PCR screening of a cDNA library from bovine adrenal medullary capillary endothelial cells. DNA sequence indicated that bDPMS has 70% homology with the human DPMS, and alignment of the predicted amino acid sequence supported that bDPMS has a conserved sequence to be phosphorylated by cAMP-dependent protein kinase (PKA). bDPMS has now been expressed as a 33 kDa His-tag protein. We, therefore, conclude that PKA-mediated phosphorylation motif on bDPMS is a potential angiogenic switch. [ABSTRACT FROM AUTHOR]

The preferential expression of heat shock proteins confers transient thermal tolerance during heat or cold stress which is characterized by the heat or cold shock response and adaptations associated with acclimation. Goat occupies wide gamut of thermal niches and this variation relates to the extent of thermotolerance in this species. As the cellular response to thermal challenge is poorly understood in goat, the present study was undertaken in vitro to analyze the heat and cold challenge response in terms of expression of different heat shock protein family genes and HSP70 protein in the peripheral blood mononuclear cells (PBMCs) of goatlings. Total RNA and protein were isolated from the PBMCs after a thermal exposure at three different temperatures of 10°C (cold challenge), 38.5°C (control), and 42°C (heat challenge) for 3h. Quantitative real time polymerase chain reaction was performed to analyze the transcript variants of five HSP70 family genes (HSPA1A, HSPA1L, HSPA2, HSPA6 and HSPA8) and HSP27 in each thermal condition. The results clearly demonstrated that upon heat stress, the expression of all the genes studied except HSPA2 were significantly higher (p <0.0001) as compared to unstressed cells. No significant difference in the expression of HSPA1A, HSPA2, HSPA6 and HSPA8 was observed between cold stressed cells and their normal counterparts. HSP70 protein expression was also increased significantly in the heat challenged PBMCs. The major finding of this study was the increased levels of expression of HSP27 in both heat and cold stress conditions. Hence this study suggests HSP27 as useful indicator for assessing in vitro stress response, and provides the basis for validation of this gene as a thermal stress indicator in animals by further in vivo studies. Additionally, this study forms an essential step in understanding the cellular stress response in terms of HSP genes expression against heat and cold challenge in goat. [ABSTRACT FROM AUTHOR]

Light trap has often been used in the ecological studies of lepidopteran insect pests in agroecosystems. However, the light trap in Indian agricultural systems is rarely adopted either to monitor the population size or to study the migration of moth pests. In the present study, we have installed light traps in shaded and unshaded tea plantations of North East (NE) India to study (1) the species composition, (2) effect of shade on moth pests, (3) seasonality of major pests and (4) to learn the sex proportion of major pests captured in light traps. The two-year catches in light traps suggested that Hyposidra talaca (Geometridae) is a major pest of tea in NE India. It peaks in number during winter months, with relatively few moths caught in the later parts of the year. Eterutia magnifera (Zygaenidae) is the second major pest of tea that peaks in number during summer months. Hyposidra infixaria, Buzura suppressaria, Ectropis obliqua, Ascotis sp. (all Geometridae) and Arctornis submarginata (Lymantridae) were caught in the light traps, but in low numbers. The catches also suggested that shade status has an important role on moth pest population in tea agroecosystems; shaded plantations received significantly higher number of moths of all major pests. The difference in the catches of H. talaca moths was significant among years. Pair-wise analysis suggests that the difference in the catches of H. talaca in corresponding standard meteorological weeks (SMWs) of two years was not significant for most of the SMWs, which confirms the inter-annual consistency in the life cycle and generations. Male moths constitute more than 77% of the overall catches of H. talaca. Overall, our results highlight that light trap is an effective, bias-free monitoring tool of moth pests of tea, which also captures the variability due to habitat type. [ABSTRACT FROM AUTHOR]

Leptin is a 167-amino acid protein produced by the leptin gene. Mal production of leptin may cause severe hereditary obesity in animals. Regulating through a neuro endocrine pathway it performs an important role in the control of body weight, feed intake, energy expenditure, immune function and reproduction. [ABSTRACT FROM AUTHOR]

Blood samples (5 mL/animal) were collected from male (n = 50) and female (n = 50) black Bengal goat kids to evaluate the alterations in plasma Na, K, Ca and Fe during pre-pubertal growth phase. The plasma sodium concentration significantly (P < 0.01) decreased with the advancement of age in both male and female kids. Plasma K did not differ significantly between age group and sex. There was a significant (P < 0.01) increase in plasma calcium level. There was no significant alteration in iron levels during different days of pre-pubertal period in the kids of both sexes. Baseline data of plasma Na, K, Ca and Fe levels in black Bengal kids of either sexes during different days of pre-pubertal growth stage generated in this study will be required to calculate mineral requirements in goats during growth phase. It can be recommended that during ration formulation for goat kids in early growth stage, mineral levels do not need to be adjusted based on sex, as sex had no influence over the above said minerals. In addition, the net Ca requirements for growth increase and Na requirements decrease as the animals grow. [ABSTRACT FROM AUTHOR]

Aim: The present investigation was intended to generate some basic data on electrocardiography (ECG) parameters in different breeds and their alterations in respect to sex and age. Materials and Methods: The present investigation was carried out on 239 owned dogs of 11 different breeds presented to the Institute Veterinary Clinic during 2018-2019. The animals, irrespective of breed and sex were grouped on the basis of their age groups. Recordings of ECG were performed by a single-channel ECG machine (CardiartR, BPL, India) at 25 mm/s paper speed and 10 mm=1 mV calibration keeping the animals on the right lateral recumbence without any anesthesia. Heart rate, along with the amplitude and duration of different waves and complexes was measured. Results: Heart rate did not vary significantly among breeds, sex, and different age groups. The highest heart rate has been reported in Doberman and the lowest in Beagle. The heart rate was lowest at the age group of 6 months-2.5 years and highest around 10.5-12.5 years irrespective of breed and sex. The incidence of sinus arrhythmia was mostly seen in older dogs. All the waves and complexes did not vary significantly between breeds, age, and sex except P duration which was significantly (p<0.05) higher in Golden retriever and Doberman breeds. Conclusion: The present investigation generated some reference values of ECG in dogs which will help the clinicians to diagnose different cardiac abnormalities through ECG. [ABSTRACT FROM AUTHOR]

The study was carried out to investigate hemato-biochemical profile and blood metabolites during different periods of pre-pubertal growth in summer- and winter-born black Bengal kids of either sexes. The body weight was 8.07 ± 0.21 kg on 180 day with significantly (P < 0.01) higher growth in male kids. The winter born kids exhibited significantly (P < 0.01) higher body weight, packed cell volume (PCV), hemoglobin (Hb), total leukocyte counts (TLC), glucose and cholesterol and significantly (P < 0.01) lower non-esterified fatty acids (NEFA) and α-amino nitrogen (AAN). Both PCV and Hb were lowest on day 15 and then increased significantly (P ≤ 0.01) and reached maximum value at 90 days (PCV) and 180 days (Hb). TEC increased significantly (P ≤ 0.01) from day 15 to 180 days and reached its maximum value at 180 days age. Both glucose and total cholesterol were maximum at day 15 and significantly (P ≤ 0.01) decreased with the advancement of age and found lowest was at day 150. But, total protein was lowest at day 15 and increased significantly (P ≤ 0.01) up to day 150. NEFA and AAN were lowest on day 15 and increased significantly (P ≤ 0.01) with the advancement of age and reached peak value on 180 days. [ABSTRACT FROM AUTHOR]

GRP78 (an Mr 78 kDa calcium dependent glucose binding protein) is located in ER lumen. It functions as ER chaperone and translocates proteins for glycosylation at the asparagine residue present in the sequon Asn-X-Ser/Thr. Paraffin sections from N-glycosylation inhibitor tunicamycin treated ER-/PR-/HER2+ (double negative) breast tumor in athymic nude mice exhibited reduced Nglycan but increased GRP78 expression. We have evaluated the effect of tunicamycin on cellular localization of GRP78 in metastatic human breast cancer cells MDA-MB-231 (ER-/PR-/HER2-). Tunicamycin inhibited cell proliferation in a time and dose-dependent manner. Nonmetastatic estrogen receptor positive (ER+) MCF-7 breast cancer cells were also equally effective. GRP78 expression (protein and mRNA) was higher in tunicamycin (1.0 μg/mL) treated MCF-7 and MDAMB- 231 cells. GRP78 is an ER stress marker, so we have followed its intracellular localization using immunofluorescence microscopy after subjecting the cancer cells to various stress conditions. Unfixed cells stained with either FITC-conjugated Concanavalin A (Con A) or Texas-red conjugated wheat germ agglutinin (WGA) exhibited surface expression of N-glycans but not GRP78. GRP78 became detectable only after a brief exposure of cells to ice-cold methanol. Western blotting did not detect GRP78 in conditioned media of cancer cells whereas it did for MMP-1. The conclusion, GRP78 is expressed neither on the outer-leaflet of the (ER-/PR-/HER2-) human breast cancer cells nor it is secreted into the culture media during tunicamycin-induced ER stress. Our study therefore suggests strongly that anti-tumorigenic action of tunicamycin can be modeled to develop next generation cancer therapy, i.e., glycotherapy for treating breast and other sold tumors. [ABSTRACT FROM AUTHOR]

The role of lactogenic hormones (prolactin, growth hormone, cortisol and thyroid hormone) on lactation yield in Mithun cows as well as their rhythmicity throughout the lactation cycle were studied in Mizoram (n = 4) and Nagaland (n = 7) strain of mithun (Bos frontalis). Blood samples were collected from all the animals from the day of calving to the complete dry off at an interval of 15 days. All the hormones were estimated in the serum by commercially available ELISA kits. Plasma level of cortisol (μg/dl), growth hormone (GH, in ng/ml), prolactin (PRL, in μIU/ml), triiodothyronine (T3, in nmol/μl) and thyroxin (T4, in ng/ml) were 20.84 ± 0.29, 28.08 ± 0.56, 9.87 ± 0.20, 27.82 ± 0.56 and 51.33 ± 0.48, respectively, in mithun irrespective of strains during the lactation period. Levels of all the hormones varied significantly (p ≤ 0.01) during different days of lactation cycle but, there was no significant difference among strain. Levels of PRL, GH, cortisol and T3were significantly (p < 0.01) higher around calving and declined sharply. The hormones remained in almost steady state during mid-lactation and declined during late lactation. All the hormones stated above were positively correlated with lactational yield thus their role on lactogenesis and galactopoiesis was established. [ABSTRACT FROM PUBLISHER]

In vitro activity of milk leukocytes (viz. neutrophils, lymphocytes and macrophages) was evaluated in forty-eight (48) clinically healthy high-yielding cross-bred cows of mid-lactation stage (100-200 days of lactation), divided into four groups namely 1st parity (n = 12), 2nd parity (n = 12), 3rd parity (n = 12) and 4th and above parity (n = 12). Milk samples were taken (250 ml/cow) were taken. Milk somatic cell counts (SCC) and differential leukocyte counts (DLC) were performed microscopically. In vitro phagocytic index (PI) of milk neutrophils and macrophages was evaluated by colorimetric nitro blue tetrazolium reductive assay. Mitogen-induced milk lymphocyte blastogenic response was measured by colorimetric MTT (tetrazolium) assay after isolation of the milk leukocytes by density gradient centrifugation. Milk SCC differed significantly (p < 0.01) between different parity. Cows of 4 and above parity showed significantly (p < 0.01) higher milk SCC compared to primiparous cows. There was no significant difference in milk DLC during different parities in high-yielding cross-bred cows. There was a significant (p < 0.01) variation in lymphocyte blastogenesis amongst parity. The highest value of lymphocyte blastogenesis was seen at 3rd parity, whereas lowest value was obtained in the cows of both 1st and 4th or above parity. PI of milk neutrophils did not differ significantly between parity. PI of milk macrophages was significantly (p < 0.01) higher in 3rd parity and lower (p < 0.01) in 1st and 4th parities. The study indicated that depressed activity of milk lymphocytes and macropages was lower and SCC was higher in the cows of 4th and above parity indicating more mammary stress and hence susceptible to udder infection and mastitis. Therefore, better care and managemental interventions should be taken around these periods. [ABSTRACT FROM AUTHOR]

Aim: The present study has been conducted to evaluate the variation in electrocardiogram (ECG) parameters among different trained breeds of dogs (viz. Labrador, German Shepherd, and Golden Retriever) used for security reasons. Materials and Methods: The ECG was recorded by single channel ECG at a paper speed of 25 mm/s and calibration of 10 mm=1 mV. The recordings were taken from all the standard bipolar limb leads (Lead-I, II, and III) and unipolar augmented limb leads (Lead-aVR, aVL, and aVF). Results: Heart rate was found to be highest in Labrador and lowest in German Shepherd. P-wave duration was maximum in Golden Retriever breed and lowest in Labrador. Maximum amplitude of P-wave was found in Labrador followed by German Shepherd and Golden Retriever. There was significantly (p<0.05) higher values of PR interval in German Shepherd compared to other breeds. The variation in QRS duration, ST segment duration, T-wave duration, and T-wave amplitude was found to be non-significant among breeds. Inverted T-waves were most common in Golden Retriever and German Shepherd, whereas positive T-waves were found in Labrador. There was significant (p<0.05) variation in mean electrical axis of QRS complex among different breeds and it ranges from +60° to +80°. Conclusion: The present study provides the reference values for different ECG parameters to monitor the cardiac health status among Labrador, German Shepherd, and Golden Retriever breeds. [ABSTRACT FROM AUTHOR]

The removal of crossbred bulls from semen collection programs due to the production of poor quality semen causes substantial monetary losses to the dairy industry. Seminal quality, a quantitative trait, is greatly influenced by genome level variations. Deletion and/or duplication of Y chromosomal genes and subsequent changes in gene copy number have a major role in determining spermatogenic efficiency and, therefore, seminal quality. In this study, copy numbers of three Y chromosomal genesTSPY, DDX3Y, andUSP9Yin genomic DNA were estimated and compared in two groups of crossbred (Bos taurus × Bos indicus) bulls of ten each, superior and inferior quality semen producing bulls, which were classified based on their seminal quality parameters. ForTSPYgene, the inferior quality semen donor group has significantly lower copy number than superior quality semen donor group (p < 0.05). No significant difference was found inDDX3YandUSP9Ygene copy numbers between two groups (p > 0.05). In conclusion, this study demonstrates that the copy number ofTSPY, a Y chromosomal spermatogenesis related gene, may be an important determinant to predict the quality of bull semen, facilitating better selection of bulls in a herd for semen collection program. [ABSTRACT FROM AUTHOR]

Asparagine-linked protein glycosylation is a hallmark for glycoprotein structure and function. Its impairment by tunicamycin [a competitive inhibitor of N-acetylglucosaminyl 1-phosphate transferase (GPT)] has been known to inhibit neo-vascularization (i.e., angiogenesis) in humanized breast tumor due to an induction of endoplasmic reticulum (ER) stress-mediated unfolded protein response (UPR). The studies presented here demonstrate that (i) tunicamycin inhibits capillary endothelial cell proliferation in a dose-dependent manner; (ii) treated cells are incapable of forming colonies upon its withdrawal; and (iii) tunicamycin treatment causes nuclear fragmentation. Tunicamycin-induced ER stress-mediated UPR event in these cells was studied with the aid of Raman spectroscopy, in particular, the interpretation of bands at 1672, 1684, and 1694 cm-1, which are characteristics of proteins and originate from C=O stretching vibrations of mono-substituted amides. In tunicamycintreated cells, these bands decreased in area as follows: at 1672 cm-1 by 41.85 % at 3 h and 55.39 % at 12 h; at 1684 cm-1 by 20.63 % at 3 h and 40.08 % at 12 h; and also at 1994 cm-1 by 33.33 % at 3 h and 32.92 % at 12 h, respectively. Thus, in the presence of tunicamycin, newly synthesized protein chains fail to arrange properly into their final secondary and/or tertiary structures, and the random coils they form had undergone further degradation. [ABSTRACT FROM AUTHOR]

Up-regulation of the dolichol pathway, a "hallmark" of asparagine-linked protein glycosylation, enhances angiogenesis in vitro. The dynamic relationship between these two processes is now evaluated with tunicamycin. Capillary endothelial cells treated with tunicamycin were growth inhibited and could not be reversed with exogenous VEGF165. Inhibition of angiogenesis is supported by down-regulation of (i) phosphorylated VEGFR1 and VEGFR2 receptors; (ii) VEGF165-specific phosphotyrosine kinase activity; and (iii) MatrigelTM invasion and chemotaxis. In vivo, tunicamycin prevented the vessel development in MatrigelTM implants in athymic Balb/c (nu/nu) mice. Immunohistochemical analysis of CD34 (p < 0.001) and CD144 (p < 0.001) exhibited reduced vascularization. A 3.8-fold increased expression of TSP-1, an endogenous angiogenesis inhibitor in MatrigelTM implants correlated with that in tunicamycin (32 h)-treated capillary endothelial cells. Intravenous injection of tunicamycin (0.5 mg/kg to 1.0 mg/kg) per week slowed down a double negative (MDA-MB-435) grade III breast adenocarcinoma growth by ~50-60% in 3 weeks. Histopathological analysis of the paraffin sections indicated significant reduction in vessel size, the microvascular density and tumor mitotic index. Ki-67 and VEGF expression in tumor tissue were also reduced. A significant reduction of N-glycan expression in tumor microvessel was also observed. High expression of GRP-78 in CD144-positive cells supported unfolded protein response-mediated ER stress in tumor microvasculature. ~65% reduction of a triple negative (MDA-MB-231) breast tumor xenograft in 1 week with tunicamycin (0.25 mg/kg) given orally and the absence of systemic and/or organ failure strongly supported tunicamycin's potential for a powerful glycotherapeutic treatment of breast cancer in the clinic. [ABSTRACT FROM AUTHOR]

Mannosylphosphodolichol synthase (DPMS) plays a critical role in Glc3Man9GlcNAc2-PP-Dol (lipid-linked oligosaccha-ride, LLO) biosynthesis, an essential intermediate in asparagine-linked ( N-linked) protein glycosylation. We observed earlier that phosphorylation of DPMS increases the catalytic activity of the enzyme by increasing the Vmax as well as the enzyme turnover ( kcat) without significantly changing the Km for GDP-mannose. As a result, LLO biosynthesis, turnover and protein N-glycosylation are increased. This is manifested in increased proliferation of capillary endothelial cells, i.e. angiogenesis. We have then asked, if the phosphorylation event or the upregulation of DPMS due to overproduction of the enzyme is the key factor in upregulating angiogenesis? This question has been answered by isolating a stable capillary endothelial cell clone overexpressing the gene encoding DPMS. Our results indicate that the DPMS-overexpressing clone has a high level of DPMS mRNA as judged by QRT-PCR. The clone also expresses nearly four times more DPMS protein than the clone transfected with pEGFP-N1 vector only (i.e. control) as analyzed by Western blotting. Most importantly, the overexpressing DPMS clone has ∼108% higher DPMS activity than the vector control. Immunofluorescence microscopy with Texas Red-conjugated wheat germ agglutinin indicates a high level of expression of (GlcNAc-β-(1,4)-GlcNAc) 1-4-β-GlcNAc-NeuAc glycans on the external surface of the capillary endothelial cells overexpressing DPMS. Increased cellular proliferation and accelerated healing of the wound induced by mechanical stress of the DPMS-overexpressing clone unequivocally supports a role of DPMS in angiogenesis. [ABSTRACT FROM AUTHOR]