Parthenogenetic activation of buffalo (Bubalus bubalis) oocytes: comparison of different activation reagents and different media on their developmental competence and quantitative expression of developmentally regulated genes.

This study was carried out to compare the efficacy of different methods to activate buffalo A + B and C + D quality oocytes parthenogenetically and to study the in vitro developmental competence of oocytes and expression of some important genes at the different developmental stages of parthenotes. The percentage of A + B oocytes (62.16 ± 5.06%, range 53.8-71.3%) was significantly higher (P < 0.001) compared with that of C + D oocytes (37.8 ± 5.00%, range 28.6-46.1%) retrieved from slaughterhouse buffalo ovaries. Among all combinations, ethanol activation followed by culture in research vitro cleave medium gave the highest cleavage and blastocyst yields for both A + B and C + D grade oocytes. Total cell numbers, inner cell mass/trophectoderm ratio and apoptotic index of A + B group blastocysts were significantly different (P < 0.05) from their C + D counterpart. To determine the status of expression patterns of developmentally regulated genes, the expression of cumulus-oocyte complexes, fertilization, developmental competence and apoptotic-related genes were also studied in parthenogenetically produced buffalo embryos at different stages, and indicated that the differential expression patterns of the above genes had a role in early embryonic development.

Primary hyperparathyroidism presenting as severe hypercalcemia with acute pancreatitis in pregnancy.

Primary hyperparathyroidism presenting first time with severe hypercalcemia is rare in pregnancy. We report a case of primary hyperparathyroidism due to a cystic parathyroid adenoma presenting as severe hypercalcemia with acute pancreatitis in second trimester of pregnancy. Acute pancreatitis was managed by conservative treatment. Hypercalcemia failed to respond to medical management and ultimately responded to ultrasound-guided ethanol ablation of parathyroid adenoma. The delivery was uneventful and patient continues to remain normocalcemic during follow up. As such, ethanol ablation of parathyroid adenoma may be considered during pregnancy in case of failure of response to medical management and when surgical removal of parathyroid adenoma is not safe.

Molecular expression and identification of caprine estrogen receptor gene 1 for fertility status in bucks.

Estrogen and its receptors are essential for sexual development and reproduction. Oestrogen receptor alpha (ERα) is a nuclear receptor activated by the hormone oestrogen. In male, ERα is encoded by the gene ESR1 (oestrogen receptor1) responsible for better fertility. ESR1 is involved in the reabsorption of luminal fluid during the transit of spermatozoa from the testis to the head of the epididymis which is important for their survival and maturation during epididymal storage. The absence of ESR1 leads to reduced epididymal sperm content, reduced sperm motility and fertilizing ability. The present study was undertaken to investigate the expression and presence of ESR1 gene in fertile and low-fertile male goat breeds. We identified ESR1 gene through various molecular tools. Genotyping was carried out by high resonance melting analysis using Roche Light Cycler 480(LC-480) system and found three different genotypes. Genotypic frequency-AA (blue-0.67), BB(Red-0.2), AB(Green-0.08) with allele frequency A(0.71 and B (0.29). The predominance of this gene in head of epididymis in fertile bucks was confirmed by SDS-PAGE, Western blotting and immunohistochemistry. From the results, we corroborated that the present study provides a useful and effective way to predict male fertility in goat breeds, which in turn increases the percentage of fertility in flock leading to more number of offspring in a kidding season.

Genetic analysis of growth parameters and survival potential of Jamunapari goats in semiarid tropics.

Goats are adaptable to varied farming environments and contribute significantly towards sustainable livestock production and food security. Selective breeding of goats for improved performance is an important component of sustainable production. The objectives of the present study were to determine the most appropriate models of analysis for growth traits, to estimate genetic parameters, survival potential of kids from birth to 12 months of age and to estimate genetic trends for growth traits of Jamunapari kids at different ages. Genetic parameter estimates were obtained from 6590 records generated between 1982 and 2012 from 5922 animals with a pedigree covering over 13 generations. The most parsimonious model for early growth traits included permanent environmental effects due to the dam (PE) and litter effects. Similarly, the most appropriate model for early average daily gain (ADG) between birth and 3 or 6 months also included PE and litter effects. The estimates of heritability for survival from birth to 12 months of age ranged from 0.10 to 0.43. The estimates of heritability for ADG varied from 0.04 to 0.41. In general, higher estimates of heritability were observed when a sire model was fitted in the random effect. There was no genetic variation observed for survival between birth and 3 months of age. However, heritability estimates of between 0.18 and 0.39 were observed for survivability during post-weaning period to 12 months of age. The genetic trend at 9 months of age and 12 months of age was 0.144 kg 0.189 kg per year respectively. The genetic trend at all the ages was positive during the study period.

Estimation of genetic parameters and genetic trends for milk yield traits in Jamunapari goats in semiarid tropics.

The Jamunapari goats are reared as dairy animals in semi-arid conditions, therefore it is necessary to evaluate the genetic potential for the genetic improvement of milk production traits. The data comprised of 2217 phenotypic records for milk yield at 90 days (MY90) and 140 days (MY140), total milk yield (TMY) and lactation length (LL) obtained from the progeny of 173 sires and 446 dams during the period 1990-2013. The data were analysed using mixed linear models exploring random effects due to direct additive, maternal and permanent environment variance due to animal. The most appropriate genetic models for milk yield traits were those that included permanent environment effects due to the animal. The direct additive heritability estimates were 0.15 ± 0.05, 0.26 ± 0.07, 0.25 ± 0.08 for MY90, MY140 and TMY, respectively. The additive heritability estimate for LL was low and non-significant at 0.02 ± 0.03. The repeatability estimates were moderate to high ranging from 0.68 to 0.73 for milk yield traits. The repeatability for lactation length was 0.20 ± 0.03. Maternal variances were low ranging from 0.03 for MY90 to 0.13 for TMY. There was an increase in mean milk yield of 0.25, 0.70 and 0.72 kg/year respectively at 90 and 140 days, and for TMY. Genetic trends and phenotypic trends for MY90, MY140 and TMY were positive and indicated significant improvement in milk traits due to selective breeding.

Endometriotic stricture of the sigmoid colon presenting with intestinal obstruction.

Endometriosis, a relatively common condition, rarely involves the bowel; even more rarely does it present as a large-bowel stricture with intestinal obstruction. We report the case of a young woman who presented to an emergency department with intestinal obstruction secondary to an endometriotic stricture of the sigmoid colon, without evidence of disease elsewhere in the peritoneal cavity. Although large-bowel obstruction is usually caused by a malignant tumour, it can sometimes result from rare causes such as endometriosis. Symptoms of a cyclical nature may remind the clinician of this possibility.

Studies on repellent activity of seed oils alone and in combination on mosquito, Aedes aegypti.

The study was undertaken to investigate the relative repellency of Pongamia pinnata and Azadirachta indica seed oils on vector mosquito, Aedes aegypti under laboratory conditions. The repellents were formulated into 3 groups: seed oils, their mixture and combination of seed oils with three carrier oils viz. olive, mustard and coconut oil. Different formulations of each oil were tested at the concentrations of 1% and 5% on human baits. Efficiency was assessed, based on the total protection time; biting rate and percent protection provided by each formulation. Results showed that 5% formulation of the Pongamia pinnata and Azadirachta indica seed oils, mixed in 1:1 ratio exhibited highest percentage repellency of 85%, protection time of 300 min and bite rate of 6%. 5% concentration of A. indica and P. pinnata seed oil in mustard oil base offered 86.36% and 85% protection respectively with total protection time of 230 and 240 min respectively. The study confirms that Azadirachta indica and Pongamia pinnata have mosquito-repellent potential. When mixed in different ratios or with some carrier oil their efficacy increases 2-fold in some cases. These formulations are very promising for topical use (> 5 hrs complete protection) and are comparable to the protection provided by advanced Odomos mosquito repellent cream available commercially and thus are recommended for field trial.

Enterotoxigenic Escherichia coli CS21 pilus contributes to adhesion to intestinal cells and to pathogenesis under in vivo conditions.

Colonization surface antigens (CSs) represent key virulence-associated factors of enterotoxigenic Escherichia coli (ETEC) strains. They are required for gut colonization, the first step of the diarrhoeal disease process induced by these bacteria. One of the most prevalent CSs is CS21, or longus, a type IV pili associated with bacterial self-aggregation, protection against environmental stresses, biofilm formation and adherence to epithelial cell lines. The objectives of this study were to assess the role of CS21 in adherence to primary intestinal epithelial cells and to determine if CS21 contributes to the pathogenesis of ETEC infection in vivo. We evaluated adherence of a CS21-expressing wild-type ETEC strain and an isogenic CS21-mutant strain to pig-derived intestinal cell lines. To determine the role of CS21 in pathogenesis we used the above ETEC strains in a neonatal mice challenge infection model to assess mortality. Quantitative adherence assays confirmed that ETEC adheres to primary intestinal epithelial cells lines in a CS21-dependent manner. In addition, the CS21-mediated ETEC adherence to cells was specific as purified LngA protein, the CS21 major subunit, competed for binding with the CS21-expressing ETEC while specific anti-LngA antibodies blocked adhesion to intestinal cells. Neonatal DBA/2 mice died after intra-stomach administration of CS21-expressing strains while lack of CS21 expression drastically reduced the virulence of the wild-type ETEC strain in this animal model. Collectively these results further support the role of CS21 during ETEC infection and add new evidence on its in vivo relevance in pathogenesis.

An unusual complication of tooth exfoliation and osteonecrosis following herpes zoster infection of trigeminal nerve: a case report and literature review.

Osteonecrosis following herpes zoster infection is a rare but severe complication, and clinicians' awareness is important for early detection and management of this condition. A case of herpes zoster of the left maxillary division of the trigeminal nerve is reported in a young female having no concurrent predisposing factors, with accompanying rare complications of alveolar bone necrosis and rapid tooth exfoliation. Acyclovir was used to manage the case effectively. The previously reported similar cases in the literature have been reviewed and the pathophysiology of tooth exfoliation and osteonecrosis by varicella zoster viruses is discussed.

Optimized, simple and selective analytical tool for determination of carcinogenic aniline in wastewater.

Here, we explored a low-cost, rapid, sensitive, and selective practical approach to manifest carcinogenic anilines by using oxone as an oxidant to mitigate its presence in wastewater. The calibration curves for OAP & PAP indicate reasonable sensitivity, percent recovery & correlation in the range 2.18-9.28 µg/mL. The Sandell's sensitivity for OAP & PAP was found to be 0.010-0.018 & 0.009-0.018 µg/mL, respectively. The range of correlation coefficient 0.979 to 0.998 shows the involvement of high precision and perfect correlation of the data. Values of coefficient of determination (r2) denote the change in absorbance or (dA/dt)i or kobs by 95.8-99.6%. The percent recovery was found to be 98.05-99.91% for six parallel determinations. Furthermore, we also analyzed some wastewater samples to further validate the proposed method. The reproducible results, good percent recovery, and reasonable sensitivity proposed a feasible approach for the estimation of aminophenol in wastewater.

RGO supported ZnO/SnO2 Z-scheme heterojunctions with enriched ROS production towards enhanced photocatalytic mineralization of phenolic compounds and antibiotics at low temperature.

In the present study, the excellent photocatalytic activity of n-ZnO/n-SnO2 heterojunction integrated with reduced graphene oxide nanosheets was explored towards the elimination of different organic pollutants viz. p-bromophenol, bisphenol A, and ofloxacin from water. n-ZnO/n-SnO2 heterojunction was decorated with a different weight percentage of reduced graphene oxide via a facile refluxing method. The structural, morphological and optical properties of the as-prepared n-ZnO/n-SnO2 heterojunction-reduced graphene oxide nanocomposites were investigated systematically. XRD, Raman and FT-IR confirmed the hexagonal wurtzite and tetragonal rutile structures of ZnO and SnO2 crystals in different nanocomposites. Cube and spherical-shaped surface structures were demonstrated by TEM and FE-SEM analysis for ZnO and SnO2, respectively. The maximum photocatalytic productivity of nanocomposite with 5 wt% reduced graphene oxide was observed at about 98.64 % and 98.50 % towards the elimination of p-bromophenol and bisphenol A, respectively after 180 min exposure of UV light. Similarly, this productivity was also observed at about 99.13 % towards the elimination of ofloxacin after 120 min irradiation of UV light. The outstanding photocatalytic activity of nanocomposite with 5 wt% reduced graphene oxide has been proven by the presence of homotypic n-ZnO/n-SnO2 and reduced graphene oxide nanosheets owing to the synergistic effect amongst them resulting in remarkable separation of charge carriers, which is responsible for the larger rate of reactive oxygen species generation and enhanced photodegradation of p-bromophenol, bisphenol A and ofloxacin. In this study, the results illustrated that the photocatalytic degradation of p-bromophenol, bisphenol A and ofloxacin using n-ZnO/n-SnO2 heterojunction-reduced graphene oxide nanocomposites is predominantly based on the hydroxyl radicals and superoxide radical anion as main reactive oxygen species as compared to 1O2. A reasonable photodegradation mechanism using prepared nanocomposites under investigation has also been proposed.

In vitro culture and morphological characterization of prepubertal buffalo (Bubalus bubalis) putative spermatogonial stem cell.

PURPOSE: Spermatogonial stem cells (SSCs) have the unique ability both to self-renew and to produce progeny that undergo differentiation to spermatozoa. The present study has been carried out to develop a method to purify and enrich the pure populations of spermatogonial stem cell like cells in buffalo. METHODS: The spermatogonial cells were isolated from testes of 3-7 month old buffalo calves and disaggregated by double enzymatic digestion. Mixed population of isolated cells were then plated on Datura stramonium agglutinin (DSA) lectin coated dishes for attachment of Sertoli cells. The desired cells were obtained from suspension medium after 18 h of incubation and then loaded on discontinuous density gradient using percoll (20-65 %) and different types of spermatogonia cells were obtained at interface of each layer. These cells were cultured in vitro. RESULTS: Spermatogonial cells isolated have spherical outline and two or three eccentrically placed nucleoli, created a colony after proliferation during first week or immediately after passage. After 7-10 days of culture, the resulted developed colonies of spermatogonial cells expressed the spermatogonial specific genes like Plzf and VASA; and other pluripotency related markers viz. alkaline phosphtase, DBA, CD9, CD90, SSEA-1, OCT-4, NANOG and REX-1. CONCLUSION: Our results show that the isolated putative spermatogonial stem cells exhibit the expression of pluripotency related and spermatogonial specific genes. This study may help to establish a long term culture system for buffalo spermatogonia.

ZnO-CdO nanocomposites incorporated with graphene oxide nanosheets for efficient photocatalytic degradation of bisphenol A, thymol blue and ciprofloxacin.

The widespread existence of different organic contaminants mostly phenolic compounds, organic dyes and antibiotics in water bodies initiated by the various industrial wastes that raised great scientific concern and public awareness as well recently owing to their prospective capability to spread these contaminants resistant gene and pose hazard to human. In the present study, a series of nanostructured ZnO-CdO incorporated with reduced graphene oxide (ZCG nanocomposites) were successfully synthesized by a simple refluxing method and characterized by using the X-ray diffraction (XRD), Raman spectroscopy, FT-IR spectroscopy, photoluminescence spectroscopy, field emission-scanning microscope (FE-SEM) and UV-visible diffused reflectance spectroscopy (DRS) for the photocatalytic degradation of bisphenol A (BPA), thymol blue (ThB) and ciprofloxacin (CFn) with illumination of UV light. The maximum degradation and mineralization of BPA, ThB and CFn was achieved around 98.5%, 98.38% and 99.28% over the ZCG-5 nanocomposite photocatalyst after UV light irradiation for 180 min, 120 min and 75 min, respectively. The superior photocatalytic activity of ZCG-5 ascribed to enhance adsorption capacity, effective separation of charge carriers consequential for the production of more ROS after incorporation of RGO nanosheets with ZnO-CdO in photocatalyst. The conceivable photocatalytic degradation mechanism of BPA, ThB and CFn was elucidated through ROS identification and the assessment of photocatalyst stability by reusability, EEO (kwh/m3order) and UV light dose (mJ/cm2) were evaluated. The plausible photocatalytic degradation pathways were proposed for the degradation of BPA, ThB and CFn via GC-MS analysis. The present work investigates the efficient removal of BPA, ThB and CFn using ZCG nanocomposites as photocatalyst.

Combined Theoretical, Bioinformatic, and Biochemical Analyses of RNA Editing by Adenine Base Editors.

Adenine base editors (ABEs) have been subjected to multiple rounds of mutagenesis with the goal of optimizing their function as efficient and precise genome editing agents. Despite an ever-expanding data set of ABE mutants and their corresponding DNA or RNA-editing activity, the molecular mechanisms defining these changes remain to be elucidated. In this study, we provide a systematic interpretation of the nature of these mutations using an entropy-based classification model that relies on evolutionary data from extant protein sequences. Using this model in conjunction with experimental analyses, we identify two previously reported mutations that form an epistatic pair in the RNA-editing functional landscape of ABEs. Molecular dynamics simulations reveal the atomistic details of how these two mutations affect substrate-binding and catalytic activity, via both individual and cooperative effects, hence providing insights into the mechanisms through which these two mutations are epistatically coupled.

Evaluation of post-COVID health status using the EuroQol-5D-5L scale.

SARS-CoV-2 has had a lasting effect on the overall health of recovered patients, called 'long COVID'. Currently, there is a lack of a validated standard questionnaire to assess post-COVID health status. A retrospective observational study involving the recovered COVID patients admitted to a secondary care hospital in India between June to December 2020 (n = 123), was conducted using the EuroQol-5D-5L scale at discharge, 4 weeks and 8 weeks post-discharge. A significant difference in anxiety/depression scores was found (χ2 = 65.6, p < 0.000) among the 3 categories of time (discharge, 4 weeks and 8 weeks). The anxiety/depression dimension scores showed a significant change (p < 0.0001) between discharge and 8 weeks, using paired t-test. Age had a significant relationship with the anxiety/depression dimension at 4 weeks (OR = 5.617, 95% CI = 1.0320-30.5746, p < 0.05). A significant difference was found using Kruskal-Wallis rank-sum test on mean index scores (χ2 = 60.0, p < 0.000) among the three categories of time (discharge, 4 weeks and 8 weeks). There was a statistically significant difference of time on EQ Index scores as determined by one-way repeated measures ANOVA (F(2,375) = 18.941, p = <0.00001). Our study found time to have a statistically significant impact on the mean index scores, level sum scores and dimension scores. Smoking was found to be significantly associated with usual activity scores at 4 weeks. The most remarkable changes occurred in the anxiety/depression dimension. Overall, there was a general trend of health improvement.

Restriction analysis and partial sequencing of the 16S rRNA gene as index for rapid identification of Bacillus species.

Restriction fragment length analysis of PCR amplified 16S rDNA with AluI revealed the presence of a 265 bp fragment in all species of Bacillus with the exception of B. cereus and B. thuringiensis, which contains two restriction sites within this fragment which results in three smaller fragments totalling to 265 bp. Some distant species of Bacillus with no evidence of this fragment could be delineated into other genera based on phenotypic and genotypic parameters. BLAST search for homologous sequences of individual species revealed that it is a highly conserved region. Multiple alignment of the fragment suggests that a region between 160 and 265 bp of the 265 bp fragment was a hypervariable region and were highly species-specific. A set of primers was designed for amplification of this hypervariable region. Partial sequencing of the hypervariable region within the 265 bp fragment seems an index for identification of Bacillus species.

Giant parathyroid adenoma.

Primary Hyperparathyroidism is most commonly caused by parathyroid adenoma. Parathyroid adenoma is generally suspected by certain symptoms and biochemical abnormalities. They rarely attain large size to be evident clinically. We report a rare case of giant parathyroid adenoma measuring 5 x 4 x 3 cm and weighing 35 gm.

rGO-ZnO nanocomposites as efficient photocatalyst for degradation of 4-BP and DEP using high temperature refluxing method in in-situ condition.

Persistent organic pollutants (mainly aromatic compounds) such as bromophenol and diethyl phthalate are dangerous and act as primary contaminants in aqueous system. In this study, efficient reduced graphene oxide zinc oxide (rGO-ZnO) nanocomposites were synthesized by using a simple and facile method for photocatalytic degradation of 4-Bromophenol (4-BP) and diethyl phthalate (DEP). The rGO-ZnO (rGZ) nanocomposites (NCs) with different weight ratio of rGO and ZnO (coded as rGZ-1, rGZ-2, rGZ-5 and rGZ-10) were synthesized via high temperature refluxing method. The crystalline structure and phase, surface morphological study, optical properties, crystal defects and existence of functional groups in rGZ NCs were studied by X-ray diffraction (XRD), field-emission-scanning electron microscope (FE-SEM), UV-Visible diffuse reflectance spectroscopy (DRS), Raman spectroscopy and FT-IR analysis, respectively. The elimination of 4-BP and DEP from water by UV-light exposure was considered to estimate the photocatalytic efficiency of prepared rGZ NCs. The maximum elimination of 4-BP and DEP via photodegradation (advanced oxidation process) was found about 99.04% and 98.63% over rGZ NCs after 180 min UV irradiation, respectively. The photodegradation study was examined by using high performance liquid chromatography (HPLC) technique. This study confirms the efficient photocatalytic activity of rGZ-5 towards degradation of 4-BP and DEP. Finally, degradation mechanism has been proposed for the degradation of 4-BP and DEP.

Optimization and computational studies evaluating molecular dynamics of EDA cored polymeric dendrimer.

In this work we report the results acquired from molecular dynamics simulations as well as the optimization of different generations of polyamidoamine dendrimer. The analysis data revealed synthesized dendrimer as a suitable nanostructured candidate suitable for neutral as well as charged molecule delivery due to the presence of both electrostatic potential and van der Waals forces. The methyl ester terminating groups of half-generation dendrimers with characteristic IR peaks for carbonyl at 1670.41 cm-1 tends to shift to 1514.17 cm-1 on conversion to amide group of full-generation dendrimer. The study includes the usage of detailed analysis, demonstrating how molecular dynamics affect the dendrimer complexation. The present investigations provide an unprecedented insight into the computational and experimental system that may be of general significance for the clinical application of dendrimers.