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Abstract Sperm transcriptomics provide insights into subtle differences in sperm fertilization competence. For predicting the success of complex traits like male fertility, identification of hub genes involved in various sperm functions are essential. The bulls from the transcriptome profiled samples (n = 21), were grouped into good and poor progressive motility (PM), acrosome integrity (AI), functional membrane integrity (FMI) and fertility rate (FR) groups. The up-regulated genes identified in each group were 87, 470, 1715 and 36, respectively. Gene networks were constructed using up- and down-regulated genes from each group. The top clusters from the upregulated gene networks of the PM, AI, FMI and FR groups were involved in tyrosine kinase (FDR = 1.61E−11), apoptosis (FDR = 1.65E−8), translation (FDR = 2.2E−16) and ribosomal pathway (FDR = 1.98E−21), respectively. From the clusters, the hub genes were identified and validated in a fresh set of semen samples (n = 12) using RT-qPCR. Importantly, the genes (fold change) RPL36AL (14.99) in AI, EIF5A (54.32) in FMI, and RPLP0 (8.55) and RPS28 (13.42) in FR were significantly (p

Abstract The adaptive ability of sperm in the female reproductive tract micromilieu signifies the successful fertilization process. The study aimed to analyze the preparedness of sperm to the prevailing osmotic and pH stressors in the female reproductive tract. Fresh bovine sperm were incubated in 290 (isosmotic-control), 355 (hyperosmotic-uterus and oviduct), and 420 (hyperosmotic-control) mOsm/kg and each with pH of 6.8 (uterus) and 7.4 (oviduct). During incubation, the changes in sperm functional attributes were studied. Sperm kinematics and head area decreased significantly (p

Electrochemical detection of β-nicotinamide adenine dinucleotide (NADH) has fascinated the scientific community due to its significance in pharmaceuticals, food supplements as well as in various biological reactions and pathways. Herein, a facile and low-cost disposable non-enzymatic electrochemical sensor for the selective and sensitive determination of NADH was developed using porous graphene oxide (PGO) modified screen printed carbon electrode (SPE). PGO has received growing attention owing to its high internal porosity, improved conductivity, large surface area, excellent stability, improved active sites, and hence expected to show excellent electrocatalytic activity than graphene and other derivatives of carbon. Firstly, graphene oxide (GO) sheets were synthesized, after which pores were created on these GO sheets through simple base reduction and acid treatment and the pore formation was confirmed using different characterization techniques. The PGO was directly dropcasted on the surface of SPE to form PGO/SPE and the fabricated electrode demonstrated eminent electrocatalytic activity towards NADH oxidation. Further, the fabricated sensor has shown significant response over a broad-range of concentration from 30 to 1071 μM with a good sensitivity of 21.83 μA mM−1 cm−2 and a low detection limit of 8.6 μM. Moreover, the fabricated PGO/SPE has advantages like cost-effectiveness, robustness, disposability, ease of fabrication, high stability, and excellent reproducibility.

Abstract Background The incidence of poor semen quality and sub-fertility/infertility is higher in crossbred as compared to Zebu males. Several attempts have been made to understand the possible reasons for higher incidence of fertility problems in crossbred males, at sperm phenotype, proteome and genome level but with variable results. Since the quality of the ejaculated spermatozoa is determined by the testicular environment, assessing the testicular transcriptome between these breeds would help in identifying the possible mechanisms associated with infertility in crossbred bulls. However, such information is not available. We performed global transcriptomic profiling of testicular tissue from crossbred and Zebu bulls using Agilent Bos taurus GXP 8X60k AMADID: 29411 array. To the best of our knowledge, this is the first study comparing the testicular mRNAs between crossbred and Zebu bulls. Results Out of the 14,419 transcripts detected in bovine testis, 1466 were differentially expressed between crossbred and Zebu bulls, in which 1038 were upregulated and 428 were downregulated in crossbred bulls. PI4KB and DPY19L2 genes, reported to be involved in sperm capacitation and acrosome formation respectively, were among the top 10 downregulated transcripts in crossbred testis. Genes involved in ubiquitination and proteolysis were upregulated, while genes involved in cell proliferation, stem cell differentiation, stem cell population maintenance, steroidogenesis, WNT signalling, protein localization to plasma membrane, endocannabinoid signalling, heparin binding, cAMP metabolism and GABA receptor activity were downregulated in crossbred testis. Among the 10 genes validated using qPCR, expression of CCNYL, SOX2, MSMB, SPATA7, TNP1, TNP2 and CRISP2 followed the same trend as observed in microarray analysis with SPATA7 being significantly downregulated and transition proteins (TNP1, TNP2) being significantly upregulated in crossbred bulls. Conclusions Abundant proteolysis by ubiquitination and downregulation of WNT signaling, cell proliferation, differentiation and steroidogenesis might be associated with higher incidence of poor semen quality and/or sub-fertility/infertility in crossbred bulls as compared to Zebu bulls. Downregulation of SPATA7 (Spermatogenesis Associated 7) and upregulation of transition proteins (TNP1 and TNP2) in crossbred bull testis might be associated with impaired spermatogenesis processes including improper chromatin compaction in crossbred bulls.

Although, it is known that spermatozoa harbor a variety of RNAs that may influence embryonic development, little is understood about sperm transcriptomic differences in relation to fertility, especially in buffaloes. In the present study, we compared the differences in sperm functional attributes and transcriptomic profile between high- and low-fertile buffalo bulls. Sperm membrane and acrosomal integrity were lower (P < 0.05), while protamine deficiency and lipid peroxidation were higher (P < 0.05) in low- compared to high-fertile bulls. Transcriptomic analysis using mRNA microarray technology detected a total of 51,282 transcripts in buffalo spermatozoa, of which 4,050 transcripts were differentially expressed, and 709 transcripts were found to be significantly dysregulated (P < 0.05 and fold change >1) between high- and low-fertile bulls. Majority of the dysregulated transcripts were related to binding activity, transcription, translation, and metabolic processes with primary localization in the cell nucleus, nucleoplasm, and in cytosol. Pathways related to MAPK signaling, ribosome pathway, and oxidative phosphorylation were dysregulated in low-fertile bull spermatozoa. Using bioinformatics analysis, we observed that several genes related to sperm functional attributes were significantly downregulated in low-fertile bull spermatozoa. Validation of the results of microarray analysis was carried out using real-time qPCR expression analysis of selected genes (YBX1, ORAI3, and TFAP2C). The relative expression of these genes followed the same trend in both the techniques. Collectively, this is the first study to report the transcriptomic profile of buffalo spermatozoa and to demonstrate the dysregulation of functionally relevant transcripts in low-fertile bull spermatozoa. The results of the present study open up new avenues for understanding the etiology for poor fertility in buffalo bulls and to identify fertility biomarkers.

The study was carried out in 48 poultry flocks to elucidate the roles of various complicating pathogens involved along with Newcastle disease (ND)/ low pathogenic avian influenza (LPAI) outbreaks. Necropsy was conducted and samples were collected for the isolation of Newcastle disease virus (NDV), Influenza A virus, infectious bronchitis virus (IBV), pathogenic bacteria; molecular detection of infectious laryngotracheitis virus (ILTV), fowl adeno virus (FAV), chicken anaemia virus (CAV), Mycoplasma synoviae (MS) and Mycoplasma gallisepticum (MG). The isolation results confirmed that 18/48 flocks (37%) were positive for the presence of hemagglutinating agents. Out of 18 hemagglutination (HA) positive flocks, 11 flocks (61%) were positive for both avian influenza virus (AIV) and NDV; 4 flocks (22%) were positive for NDV; and 3 flocks (17%) were positive for AIV. Sequence analysis of hemagglutinin and neuraminidase genes of AIV revealed that all were belonging to LPAI-H9N2 subtype. Sequence analysis of F gene of NDV revealed that they belong to virulent type. The PCR results confirmed the presence of three to seven etiological agents (CAV, FAV, ILTV, MG, MS and avian pathogenic E. coli along with LPAI/NDV from all the 18 HA-positive flocks. The detection rate of triple, quadruple, quintuple, sextuple and sevenfold infections was 17% (3 flocks), 28% (5 flocks), 11%, (2 flocks) 28% (5 flocks) and 17% (3 flocks), respectively. In conclusion, the disease complex involved more than one pathogen, primarily resulting from the interplay between LPAI-H9N2 and NDV; subsequently this could be exacerbated by co-infection with other agents which may cause exacerbated outbreaks that may otherwise go undetected in field.

The uniqueness of Zn is that, it is the second most abundant trace element in the animal body but can't be stored in the body, thus regular dietary intake is required. Zinc oxide (ZnO) nanoparticles (NP) particles are being extensively used in paints, skin lotions pigments, food, electronics appliances, biological and pharmaceutical applications and many more. Zinc oxide nanoparticles are the specially prepared mineral salt having particle size of 1 to 100 nm. It promotes growth can act as antibacterial agent, modulates the immunity and reproduction of the animals. Both in lower and higher doses of specifications it has exhibited a variety of effects on animal performances. Apart from being highly bio-available, reports have already pointed out the growth promoting, antibacterial, immuno-modulatory and many more effects of nano zinc (nZn). These can be used at lower doses and can provide better result than the conventional Zn sources and indirectly prevents environmental contamination also. The toxicological studies provide mixed results in animal models. Studies been undertaken in diversified animal species and encouraging effects have been reported with nZn supplementation. However, there is a need to optimize the dose and duration of ZnO NP supplementation for human and livestock, depending on its biological effects. Actual bioavailability of ZnO NP in livestock is still to be worked out. In this review we have attempted to summarize, conclude the beneficial effects of nZnO and its possible usage as mineral supplement to different categories of human and livestock.

A study was conducted to assess the combined effects of heat and nutritional stress on body weight changes and HSP70 gene expressionin Osmanabadi goats. Twenty four adult Osmanabadi bucks were divided into four groups of six animals each viz. C (control), HS (heat stress), NS (nutritional stress) and CS (combined stress). The C and HS bucks had ad libitum access to their feed while NS and CS bucks were subjected to restricted feed (30% intake of C bucks) to induce nutritional stress. The HS and CS bucks were exposed to heat stress in outside environment for six hours a day between 10:00 h to 16:00 h. Both C and HS groups showed significantly higher (P

A mild, efficient, and eco-friendly procedure for the conversion of aliphatic, alicyclic and aromatic carbonyl compounds into the corresponding oximes, was catalyzed by a novel imidazolium based ionic liquid coupled with amino acid (asparagine) (l-AAIL, l-Amino acid functionalized ionic liquid) catalytic system. The quantitative conversion of aryl and alkyl carbonyl compounds into the corresponding oximes was achieved by simply grinding at ambient temperature using 0.05 mmol of catalyst in 50 s. In addition, this L-AAIL catalyst exhibited good reusability for five consecutive trials without significant loss of its catalytic activity.

N-heterocyclic carbenes (NHC) are now commonly encountered in organometallic and inorganic coordination chemistry because of its environmental friendly nature as solvent and catalyst, high activity, selectivity and easily recovered materials that were used to replace the traditional volatile organic solvents which generally suffered from environmental problems. Benzimidazole based ionic liquids (BILs) offering a new possibility for developing environmentally-friendly basic catalyst with transition metal and non-transition metal (Ag, Au, Ru, Rh, Fe, Co, Cu, Cd and Pd). They are flexible, nonvolatile, noncorrosive and immiscible with many organic solvents. In this review, we wish to present an overview of the preparation and applications in various reactions like alkylation, arylation, etherification, benzoin reaction, aldol condensation and metal mediated catalyst in Suzuki, Suzuki–Miyaura, Heck cross coupling and reduction, also in electrochemical application (electrocatalyst), anion sensor, solar cell, proton conduction, malaria parasite, antimicrobial activity, etc.

The application of spermatogonial stem cells (SSC) will be more effective and feasible following the successful cryopreservation and transfer of SSCs in livestock. Like other cells, SSCs are also sensitive to cryoinjury; hence composition of the cryomedia and freezing protocols need to be optimized. The present study aims to optimising the best freezing rates by minimising the ice crystallization and dehydration effect in order to maximize the post-thaw SSCs survivability and stemness characteristics. Three different freezing protocols with varied cooling profiles, cooling profile 1 (isopropanol based freezing): 1°C/min from 0°C to -10°C, 0.5°C/min upto -40°C, further reduced to 0.25°C/min upto -50°C and 0.1°C/min to -60°C, cooling profile 2 (using programmable freezer): 1°C/min up to 4°C, 0.3°C/min up to -8°C, and cooled at 0.5°C/min to -50°C, further decrease to -90°C (8°C/min) and cooling profile 3 (uncontrolled rapid freezing): 3.3°C/min from 0°C to -10°C, 5°C/min upto -40°C, 2°C/min to -50°C and 1.2°C/min upto -60°C, were compared for cryopreservation efficiency. The overall viability (91.41 ± 2.00% Vs 74.59 ± 2.34%), stemness activity (1.34 ± 0.095 OD units Vs 0.356 ± 0.026 OD units), and proliferation rate (0.849 ± 0.019 OD units Vs 0.749 ± 0.015 OD units) of post-thaw SSC culture irrespective of the freezing regimes was significantly decreased when compared to pre-freeze SSC culture characteristics. The post-thaw viability was significantly greater in cooling profile 1 (79.64 ± 4.1%) when compared to cooling profile 2 (69.72 ± 2.4%) and cooling profile 3 (75.43 ± 4.8%). Also, cooling profile 1 yielded greater (p<0.05) post-thaw stemness activity (0.456±0.044 OD units) when compared to other methods. The study suggests that the cooling profile 1 using isopropanol based freezing can be recommended for preservation of viability and stemness characteristics of SSCs., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025. Published by Elsevier Inc.)

Colorectal cancer is one of the most prevalent malignancies worldwide and a leading cause of mortality. Chemotherapy medications are often limited in use due to issues like drug resistance, P-glycoprotein efflux, and relapse of chemotherapy. In this study, we formulated a nanosuspension with curcumin and nimbin to address these limitations and assessed its anticancer potential using in silico molecular docking and in vitro MTT assay., Methods: In silico docking and ADMET analyses targeted proteins implicated in colorectal cancer, with doxorubicin as the standard. The docking studies were conducted using AutoDock 4.2, while in vitro anticancer activity was assessed through the MTT assay in HCT 116 cell lines., Results: In silico docking of curcumin and nimbin showed significant interactions with target proteins compared to the standard. ADMET analysis indicated favorable Caco-2 permeability and intestinal absorption of the selected phytoconstituents. The MTT assay demonstrated concentration-dependent cell viability inhibition in HCT 116 cell lines treated with the nanosuspension, with an IC 50 value of 30%., Conclusion: The curcumin-nimbin loaded nanosuspension demonstrated promising anticancer activity against HCT 116 cell lines in both in silico and in vitro studies. Further studies are required to evaluate the anticancer effect of curcumin-nimbin loaded nanosupension through clinical and preclinical studies for the progress of potential formulation in the treatment of colorectal cancer., Competing Interests: The authors declare no conflict of interest., (© 2024 Institute of Bioorganic Chemistry, Polish Academy of Sciences.)

Dolutegravir (DLG) has become a distinctive first-line antiretroviral therapy for the treatment of HIV in most countries due to its affordability, high efficacy, and low drug-drug interactions. However, the evaluation of genotoxic impurities (GTIs) in DLG and their toxicity assessment has not been explored thoroughly. Thus, in this study, a simple, fast, and selective analytical methodology was developed for the identification and determination of 7 GTIs in the comprehensive, explicit route of synthesis for the dolutegravir sodium (DLG-Na) drug. A facile, fast ultrasonication-assisted liquid-liquid extraction procedure was adapted to isolate the GTIs in DLG-Na and then analyzed using the gas chromatography (GC)-electron impact (EI)/mass spectrometer (MS) quantification (using selective ion monitoring mode) technique. This EI-GC/MS method was validated as per the current requirements of ICH Q2 (R1) guidelines. Under optimal method conditions, excellent linearities were achieved with R between 0.9959 and 0.9995, and high sensitivity was obtained in terms of detection limits (LOD) between 0.15 to 0.63 µg/g, and quantification limits (LOQ) between 0.45 to 1.66 µg/g for the seven GTIs in DLG. The obtained recoveries ranged from 98.2 to 104.3 % at LOQ, 15 µg/g, and 18 µg/g concentration levels (maximum daily dose of 100 mg). This developed and validated method is rapid, easy to adopt, specific, sensitive, and accurate in estimating the seven GTIs in a relatively complex sodium matrix of the DLG-Na drug moiety. As a method application, two different manufactured samples of DLG-Na drug substances were analyzed for the fate of the GTIs and drug safety for the intended dosage applications. Moreover, an in-silico QSAR toxicity prediction assessment was carried out to prove scientifically the potential GTI nature of each impurity from the alerting functional groups., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Three new isostructural MOFs (ZnTIA, CoTIA and CdTIA) were synthesized by the solvothermal synthesis of the organic linker 5-triazole isophthalic acid (5-TIA) with the transition metals Zn(II), Co(II) and Cd(II) in the presence of the structure directing agent tetramethyl ammonium chloride (TMA). These three MOFs were characterized thoroughly by ScXRD, PXRD, FT-IR, TGA, BET and SEM. They have excellent thermal and water stabilities. Among all these MOFs mentioned, pristine CoTIA exhibited excellent electrocatalytic activity toward the oxygen evolution reaction (OER). It exhibits a Tafel slope of 68.9 mV dec -1 with an overpotential of 337 mV at 10 mA cm -2 current density. The OER activity of the CoTIA MOF is relatively equivalent to that of the state-of-the-art catalyst (RuO 2 ). Furthermore, the mechanical stability of crystalline ZnTIA, CoTIA and CdTIA MOFs was tested under ball mill pressure. The result showed that all the MOFs exhibit low tolerance to mechanical force because their structure was highly distorted or collapsed under such pressure, which is reflected by their poor electrocatalytic OER activity.

The facile fabrication is reported of highly electrochemically active Ti 3 C 2 Tx MXene/MWCNT (3D/1D)-modified screen-printed carbon electrode (SPE) for the efficient simultaneous electrochemical detection of paracetamol, theophylline, and caffeine in human blood samples. 3D/1D Ti 3 C 2 Tx MXene/MWCNT nanocomposite was synthesized using microwave irradiation and ultrasonication processes. Then, the Ti 3 C 2 Tx/MWCNT-modified SPE electrode was fabricated and thoroughly characterized towards its physicochemical and electrochemical properties using XPS, TEM, FESEM, XRD, electrochemical impedance spectroscopy, cyclic voltammetry, and differential pulse voltammetry techniques. As-constructed Ti 3 C 2 Tx-MWCNT/SPE offers excellent electrochemical sensing performance with good detection limits (0.23, 0.57, and 0.43 µM) and wide linear ranges (1.0 ~ 90.1, 2.0 ~ 62.0, and 2.0-90.9 µM) for paracetamol, caffeine, and theophylline, respectively,  in the human samples. Notably, the non-enzymatic electroactive nanocomposite-modified electrode has depicted a semicircle Nyquist plot with low charge transfer resistance (R ct ∼95 Ω), leading to high ionic diffusion and facilitating an excellent electron transfer path. All the above results in efficient stability, reproducibility, repeatability, and sensitivity compared with other reported works, and thus, it claims its practical utilization in realistic clinical applications., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Degradation of antibiotics through electrocatalytic oxidation has recently been comprehended as a promising strategy in wastewater treatment. Herein, nitrogen and sulphur doped graphene oxide (N,S-rGO) nanosheets were synthesized and employed as metal-free anodic material for electrochemical degradation of antibiotics, viz. metronidazole (MNZ) and tetracycline (TC). The synthesized anodic material was characterized using various spectral techniques and further the electrochemical behaviour of N,S-rGO was thoroughly examined. Thereafter, the N,S-rGO material was then employed as the anode material towards the electrocatalytic degradation of antibiotics. Parameters such as initial concentration of the antibiotics and current densities were varied and their effect towards the degradation of MNZ and TC were probed. Notably, the N,S-rGO based anode has shown impressive removal efficiency of 99% and 98.5%, after 120 min of reaction time for MNZ and TC, respectively, under optimized conditions. The obtained results including the kinetic parameters, removal efficiency and electrical efficiency ensure that the prepared anodic material has huge prospective towards real-time application for removal of antibiotics from water., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A novel zeolitic tetrazolate framework (ZTF-8) has been synthesized by solvent-free heat-assisted (70 °C) mechanochemical grinding of zinc acetate and 5-methyl tetrazole in the presence of NaOH powder. The structure of ZTF-8 adopts the zeolitic sodalite (SOD) topology with uncoordinated N-heteroatom sites and resembles the structure of the well-known zeolitic imidazole framework ZIF-8. ZTF-8 is exceptionally stable in 0.1 M aqueous acid and base solutions for 60 days at 25 °C. The unique structure with uncoordinated N-heteroatom active sites and exceptional stability of ZTF-8 facilitated the electrocatalytic oxidation of dopamine to dopamine quinone at neutral pH. Without any postsynthetic modification, ZTF-8 is directly used for the facile electrochemical detection of dopamine over a wide range of concentrations (5-550 μM) with a high sensitivity (2410.8 μA mM -1 cm -2 ). It also demonstrated promising selectivity over other interferents of similar oxidation potential, such as ascorbic acid and uric acid. The DFT study revealed that the ZTF-8 framework has a higher binding energy (-145.07 kJ/mol) and stronger interaction with dopamine than its isostructural ZIF-8 structure (-130.42 kJ/mol).

Sperm antigenicity has been implicated as a regulatory factor for acquiring fertilizing competence in the female reproductive tract. Overt immune response against the sperm proteins leads to idiopathic infertility. Hence, the aim of the study was to evaluate the influence of the auto-antigenic potential of sperm on the antioxidant status, metabolic activities and reactive oxygen species (ROS) in bovine. Semen from Holstein-Friesian bulls ( n  = 15) was collected and classified into higher (HA, n  = 8) and lower (LA, n  = 7) antigenic groups based on micro-titer agglutination assay. The neat semen was subjected to the evaluation of bacterial load, leukocyte count, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lipid peroxidation (LPO) levels. Antioxidant activities in seminal plasma and intracellular ROS levels in the post-thawed sperm were estimated. The number of leukocytes was lower ( p  < .05) in the HA than the LA semen. The percentage of metabolically active sperm was higher ( p  < .05) in HA than the LA group. The activities of total non-enzymatic antioxidant, superoxide dismutase (SOD) and catalase (CAT) were higher ( p  < .05) while glutathione peroxidase activity was lower ( p  < .05) in the seminal plasma of LA group. The LPO levels of neat sperm and the percentage sperm positive for intracellular ROS in the cryopreserved sample were lower ( p  < .05) in the HA group. Auto-antigenic levels were positively correlated with the percentage of metabolically active sperm ( r  = 0.73, p  < .01). However, the seminal auto-antigenicity was negatively ( p  < .05) correlated with the levels of SOD ( r =-0.66), CAT ( r =-0.72), LPO ( r =-0.602) and intracellular ROS ( r =-0.835). The findings were represented in graphical abstract. It is inferred that the higher auto-antigenic levels protect the quality of bovine semen by promoting sperm metabolism and lowering ROS and LPO levels.

The study aimed to assess the influence of cryostress on RNA integrity and functional significance in sperm fertilizing ability. The fresh and post-thawed buffalo sperm (n = 6 each) samples were evaluated for their functional attributes, and sperm total RNA was subjected to transcriptome sequencing followed by validation using real-time PCR and dot blot. Overall, 6911 genes had an expression of FPKM > 1, and among these 431 genes were abundantly expressed (FPKM > 20) in buffalo sperm. These abundantly expressed genes regulate reproductive functions such as sperm motility (TEKT2, SPEM1, and PRM3, FDR = 1.10E-08), fertilization (EQTN, PLCZ1, and SPESP1, FDR = 7.25E-06) and the developmental process involved in reproduction (SPACA1, TNP1, and YBX2, FDR = 7.21E-06). Cryopreservation significantly (p < 0.05) affected the structural and functional membrane integrities of sperm. The expression levels of transcripts that regulate the metabolic activities and fertility-related functions were compromised during cryopreservation. Interestingly, cryostress induces the expression of genes involved (p < 0.05) in chemokine signaling (CX3CL1, CCL20, and CXCR4), G-protein coupled receptor binding (ADRB1, EDN1, and BRS3), translation (RPS28, MRPL28, and RPL18A), oxidative phosphorylation (ND1, ND2, and COX2), response to reactive oxygen species (GLRX2, HYAL2, and EDN1), and immune responses (CX3CL1, CCL26, and TBXA2R). These precociously expressed genes during cryopreservation alter the signaling mechanisms that govern sperm functional competence and can impact fertilization and early embryonic development., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The present study evaluated the effect of dietary supplementation with n-3 polyunsaturated fatty acids (PUFA) on preovulatory follicle (POF) turnover, prolificacy, and endocrine and metabolic milieu in Malpura sheep. Fifty cyclic ewes with 3-3.5 body condition scores on a five-point scale were allocated equally to two groups (n = 25) following estrus synchronization and were supplemented with 0.6 mL/kg body weight of n-3 PUFA-rich fish oil (FO) or palm oil (PO) as control, for 60 d following an acclimatization period of 7 d. All ewes were mated with sexually active rams at the end of the supplementation period. On ultrasonographic ovarian scanning at the last fourth estrus, the mean number of POFs was 77.8% greater (P < 0.01) in FO ewes than in the PO ewes. The proportion of ewes with multiple ovulations two months after the beginning of supplementation was 56% in the FO group as compared to 8% in the PO group. The number of fetuses was 46% higher (P < 0.01) in the FO than in the PO ewes at d 45 of gestation. At lambing, the twinning percent in the FO ewes was three times greater than in the PO ewes (27.3 vs. 9.1%). Plasma cholesterol, estradiol, and insulin concentrations were lower (P < 0.01) in ewes fed with FO than those offered PO group at the end of the feeding period. It was concluded that the dietary supplementation of n-3 PUFA-rich FO in well-fed Malpura ewes improved the number of follicles and ovulation rate which led to an increased prolificacy, accompanied by a reduction of plasma cholesterols, estradiol, and insulin., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Dr Ajit Singh Mahla reports financial support was provided by Science and Engineering Research Board, Department of Science and Technology, Government of India., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Three new metal organic frameworks (ZnTIA-1mc, CuTIA-1mc, and CoTIA-1mc) were synthesized by the mechanochemical grinding (mc) method in the unadulterated form. They compared with their solvothermally synthesized (st) counterparts, where the mixtures of isomeric forms have been isolated. Kinetics study with the function of grinding time during the mechanosynthesis process revealed the formation of new metastable phases. Less crystallinity and short of mechanical defects in the structure of synthesized mc metal organic frameworks showed enhanced electrocatalytic activity toward oxygen evolution reaction (OER). Among all, CoTIA-1mc showed high OER activity with 289 mV overpotential, 10 mA cm -2 current density, and 55.4 mV dec -1 Tafel slope in 1 M KOH which is close to the commercially used RuO 2 .

Halophytic plants can tolerate a high level of salinity through several morphological and physiological adaptations along with the presence of salt tolerant rhizo-microbiome. These microbes release phytohormones which aid in alleviating salinity stress and improve nutrient availability. The isolation and identification of such halophilic PGPRs can be useful in developing bio-inoculants for improving the salt tolerance and productivity of non-halophytic plants under saline conditions. In this study, salt-tolerant bacteria with multiple plant growth promoting characteristics were isolated from the rhizosphere of a predominant halophyte, Sesuvium portulacastrum grown in the coastal and paper mill effluent irrigated soils. Among the isolates, nine halotolerant rhizobacterial strains that were able to grow profusely at a salinity level of 5% NaCl were screened. These isolates were found to have multiple plant growth promoting (PGP) traits, especially 1-aminocyclopropane-1-carboxylic acid deaminase activity (0.32-1.18  μM of α-ketobutyrate released mg -1 of protein h -1 ) and indole acetic acid (9.4-22.8  μg mL -1 ). The halotolerant PGPR inoculation had the potential to improve salt tolerance in Vigna mungo L. which was reflected in significantly ( p  < 0.05) higher germination percentage (89%) compared to un-inoculated seeds (65%) under 2% NaCl. Similarly, shoot length (8.9-14.6 cm) and vigor index (792-1785) were also higher in inoculated seeds. The strains compatible with each other were used for the preparation of two bioformulations and these microbial consortia were tested for their efficacy in salt stress alleviation of Vigna mungo L. under pot study. The inoculation improved the photosynthetic rate (12%), chlorophyll content (22%), shoot length (5.7%) and grain yield (33%) in Vigna mungo L. The enzymatic activity of catalase and superoxide dismutase were found to be lower (7.0 and 1.5%, respectively) in inoculated plants. These results revealed that halotolerant PGPR isolated from S. portulacastrum can be a cost-effective and ecologically sustainable method to improve crop productivity under high saline conditions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 John, Maheswari, Kalaiselvi, Prasanthrajan, Poornachandhra, Rakesh, Gopalakrishnan, Davamani, Kokiladevi and Ranjith.)

Background: Disclosure of tuberculosis (TB) status by patients is a critical step in their treatment cascade of care. There is a lack of systematic assessment of TB disclosure patterns and its positive outcomes which happens dynamically over the disease period of individual patients with their family and wider social network relations., Methods: This prospective observational study was conducted in Chennai Corporation treatment units during 2019-2021. TB patients were recruited and followed-up from treatment initiation to completion. Information on disease disclosures made to different social members at different time points, and outcomes were collected and compared. Bivariate and multi variate analysis were used to identify the patients and contact characteristics predictive of TB disclosure status., Results: A total of 466 TB patients were followed-up, who listed a total of 4039 family, extra familial and social network contacts of them. Maximum disclosures were made with family members (93%) and half of the relatives, occupational contacts and friendship contacts (44-58%) were disclosed within 15 days of treatment initiation. Incremental disclosures made during the 150-180 days of treatment were highest among neighbourhood contacts (12%), and was significantly different between treatment initiation and completion period. Middle aged TB patients (31 years and 46-55 years) were found less likely to disclose (AOR 0.56 and 0.46 respectively; p<0.05) and illiterates were found more likely to disclose their TB status (AOR 3.91; p<0.05). Post the disclosure, family contacts have mostly provided resource support (44.90%) and two third of all disclosed contacts have provided emotional support for TB patients (>71%)., Conclusion: Findings explain that family level disclosures were predominant and disclosures made to extra familial network contacts significantly increased during the latter part of treatment. Emotional support was predominantly received by TB patients from all their contacts post disclosure. Findings could inform in developing interventions to facilitate disclosure of disease status in a beneficial way for TB patients., Competing Interests: The authors declare that no competing interest exists., (Copyright: © 2023 Nagarajan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The effect of dietary calcium (Ca) and magnesium (Mg) supplementation on serum biochemical parameters, steroid hormones, gene expression, and the sex ratio was investigated in female New Zealand white rabbits. A total of 25 rabbits were allocated into five treatment groups: The control group was fed with regular pellet feed, whereas, treatment groups were supplemented with Ca and Mg: T1 (0.40% and 0.01%), T2 (0.60% and 0.02%), T3 (0.80% and 0.03%) and T4 (1.00% and 0.04%), respectively. The rabbits were subjected to three breeding cycles. The T3 group skewed towards females (65.33%) from all three breeding. There was elevated Ca concentration in T3 (15.26 ± 0.77 mg dL -1 ) and T4 (15.61 ± 0.82 mg dL -1 ) groups compared to the control. The concentration of estradiol was significantly high in T3 and T4 groups at 0.5 days post-coitus (dpc) and T2, T3 and T4 groups at 21dpc. Testosterone was significantly high in T4 group at 0.50 dpc and T2 and T4 group at 21dpc. The expression of 13 genes was studied in the oviduct. Genes such as OVGP1, CCT4, ANXA2 and TLR4 were up-regulated and positively correlated with the female sex ratio. The molecular functions and pathways of up-regulated genes were suggestive of their role in fertilization such as sperm selection, sperm storage, immune regulation, implantation and early embryonic development. The variations in the serum electrolytes, steroid hormones and gene expression might have an impact on the skewing process., Competing Interests: There is no conflict of interest., (© 2023 Urmia University. All rights reserved.)

In the present work, we report the synthesis of wurtzite CuGaS 2 and its composite with MoS 2 and explored their efficacy toward two important applications, viz. electrocatalytic hydrogen evolution reaction (HER) and adsorption of Rhodamine B dye. The CuGaS 2 was synthesized via a low-temperature ethylenediamine-mediated solvothermal method. The obtained products were characterized by various techniques such as X-ray diffraction, field emission scanning electron microscopy, transmission electron microscopy, and X-ray photoelectron spectroscopy to ascertain the phase formation, surface morphology, and elemental oxidation states. The electrocatalytic activity of the wurtzite CuGaS 2 and CuGaS 2 /MoS 2 composites toward HER was investigated, wherein the CuGaS 2 /MoS 2 composite exhibited superior activity when compared to the pristine sample with a small Tafel slope of 56.2 mV dec -1 and an overpotential value of -464 mV at the current density of 10 mA cm -2 . On the other hand, the synthesized CuGaS 2 also showed an impressive adsorption behavior toward Rhodamine B dye with 99% adsorption in 60 min, which is relatively better than that observed with the composite material., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

A simple and facile functional group-specific multilateral derivatization cum extraction method coupled with GC-MS based analytical methodology has been developed for the rapid identification and determination of five potential genotoxic impurities (GTIs), including epichlorohydrin, hydrazine, phenylhydrazine, 3-chloro-1,2-propanediol and 1-(2-chloroethoxy)- 2-methoxybenzene in the carvedilol phosphate (CRV-P) drug active pharmaceutical ingredient (API). A generic synthetic route has been explored to apply the current investigation to the majority of the market available synthetic routes for the carvedilol process. Five significant GTIs were identified, and their toxicity was examined using in-silico model. The pharmacokinetic and pharmacodynamic properties of the impurities were compared with the drug molecule to evince the associated risk of impurities during therapeutic action. Furthermore, a quantitative comparison has been made for each impurity with the drug molecule for their ADMET properties, and the potential nature of the impurities has been thoroughly assessed. The developed method encompasses simple derivatization cum extraction-oriented GC-MS method for the reported GTIs, which was also validated as per current ICH guidelines. The obtained LOD and LOQ for the method were between 0.06 ~ 0.61 µg/g and 0.17 ~ 1.8 µg/g, respectively, and the r 2 values (0.994 ~ 0.997) show that the method is very sensitive and linear over a wide range (LOQ to 120 % of the target concentration). The percentage recoveries and relative standard deviation obtained were between 85.3 and 109.5 and 0.1-4.7, respectively, showing fit for purpose. Moreover, method precision, intermediate precision, and robustness of the method have also been successfully demonstrated. Thus, this method could be directly engaged as the quality forecasting tool for the marketed drug samples aimed at the estimation of the reported GTIs at trace level., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The role of sperm expressed X-linked genes on bull fertility has not been studied in detail. The objective of the present study was to assess the influence of X-linked genes on the sperm functional parameters and field fertility rate in the Holstein Friesian cattle (n = 12) and Murrah buffalo (n = 7) bulls. The enrichment analysis (cattle = 8; buffalo = 8) of the X-linked genes was carried out using retrospective RNA-seq data and mRNA expression levels of functionally relevant genes were validated using the RT-qPCR. The mRNA expression levels of these genes were functionally associated with sperm attributes and field fertility rate. The sperm transcriptome studies revealed that the total number of expressed genes and the transcript content of the X-linked genes in the mature sperm were very low in both species, and only 23.31% of these genes were commonly expressed between them. The transcript pool corresponding to the X-linked genes represents embryonic organ development (p = 0.03) and reproduction (p = 0.02) processes in cattle and buffalo sperm, respectively. The mRNA expression levels of X-linked genes, RPL10 and ZCCHC13 in cattle; AKAP4, TSPAN6, RPL10 and RPS4X in buffalo were significantly (p < 0.05) correlated with sperm kinematics. Importantly, the mRNA expression levels of the genes RPL10 (r = -0.68) and RPS4X (r = 0.81) had a significant correlation with the field fertility rate in cattle and buffalo, respectively. Multivariate regression models and receiver operating curve analysis suggest that the mRNA expression levels of X-linked genes may be useful in predicting bull fertility. The study indicates that sperm-expressed X-linked genes influence semen quality and field fertility rate in both cattle and buffalo., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Maize is an important cereal crop which is severely affected by Spodoptera frugiperda . The study aims to identify endophytic bacteria of maize root and leaf apoplastic fluid with bioprotective traits against S. frugiperda and plant growth promoting properties. Among 15 bacterial endophytic isolates, two strains-namely, RAF5 and LAF5-were selected and identified as Alcaligenes sp. MZ895490 and Bacillus amyloliquefaciens MZ895491, respectively. The bioprotective potential of B. amyloliquefaciens was evaluated through bioassays. In a no-choice bioassay, second instar larvae of S. frugiperda fed on B. amyloliquefaciens treated leaves (B+) recorded comparatively lesser growth (1.10 ± 0.19 mg mg -1 day -1 ) and consumptive (7.16 ± 3.48 mg mg -1 day -1 ) rates. In larval dip and choice bioassay, the same trend was observed. In detached leaf experiment, leaf feeding deterrence of S. frugiperda was found to be greater due to inoculation with B . amyloliquefaciens than Alcaligenes sp. The phenolics content of B. amyloliquefaciens inoculated plant was also found to be greater (3.06 ± 0.09 mg gallic acid g -1 ). However, plant biomass production was more in Alcaligenes sp inoculated treatment. The study thus demonstrates the potential utility of Alcaligenes sp. and B. amyloliquefaciens for improving growth and biotic ( S. frugiperda ) stress tolerance in maize.

COVID-19 has become a pandemic, and any new drug for treating the disease could save millions of lives. Several drugs already in use for other diseases and medical conditions are repurposed for treating COVID-19 in an attempt to find treatment for the disease without spending research time on ADME TOX and other studies on side effects. In this exercise, the drugs repurposed are from antiviral, antibiotics, antiviral for HIV and HCV, anti-cancer, natural medicines, etc. Possible repurposing anti-diabetic GPR-120 agonists used as for SAR-CoV-2 is attempted in the study by carrying out docking of 68 GPR-120 agonists. Ten of these compounds were found to have docking scores -8.3 to -8.0, and the best docking score was observed for an arylsulfonamide and a biarylpropanoic acid belonging to GPR120 agonists previously evaluated for the treatment of type II diabetes. These GPR120 agonists could serve as start point for novel inhibitors for the discovery of drugs to treat COVID-19., (© 2022. The Author(s).)

Mineral supplementation has greater impact on male reproduction; however, the mechanism of action has not been studied in detail. The present study was aimed to deal with the lacuna in mechanism of action of mineral supplementation on improvement in sperm characteristics. A group of 40 bucks (aged 5 months) were assigned to 10 groups (4 in each group) based on their body weight and fed with concentrate mixture: basal roughage (minimal diet) in equal proportion to all the bucks. Among the 10 groups, one was considered as control, without any additional mineral supplementation, and the remaining 9 were treatment groups (3 groups each in Zn, Cu, and Zn + Cu). In treatment groups, organic Zn was fed in three different doses as 20, 40, and 60 mg/kg DM; organic Cu was fed in three different doses as 12.5, 25, and 37.5 mg/kg DM; and organic Zn + Cu was combinedly supplied as 20 + 12.5, 40 + 25, and 60 + 37.5 based on their mg/kg DM for 8 months period. The neat semen samples were processed for spermatozoal gene (stress- NOS3, HSP70, HIF1A; fertility- MTF1, MTA1, TIMP2, TNFa, and EGFR) expression studies through qRT-PCR and protein profile changes through single- and two-dimensional gel electrophoresis. Significantly, the stress-responsive genes were downregulated, and fertility-related genes were upregulated in treatment groups. A significant correlation had been noticed among the genes studied: HIF1A with MTA1 (P < 0.05) and MTF1 with EGFR, TIMP2, TNFa, and NOS3 (P < 0.01) respectively. The organic Zn and Cu feeding modulated the expression of stress- and fertility-related genes and protein abundance, thereby improved the sperm characteristics., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

This study aims to systematically review the diagnostic accuracy of a digital blood pressure measurement device compared to the gold standard mercury sphygmomanometer in published studies. Searches were conducted in PubMed, Cochrane, EBSCO, EMBASE and Google Scholar host databases using the specific search strategy and filters from 1st January 2000 to 3rd April 2021. We included studies reporting data on the sensitivity or specificity of blood pressure measured by digital devices and mercury sphygmomanometer used as the reference standard. Studies conducted among children, special populations, and specific disease groups were excluded. We considered published manuscripts in the English language only. The risk of bias and applicability concerns were assessed based on the author's judgment using the QUADAS2 manual measurement evaluation tool. Based on the screening, four studies were included in the final analysis. Sensitivity, specificity, diagnostic odds ratio (DOR), and 95% confidence interval were estimated. The digital blood pressure monitoring has a moderate level of accuracy and the device can correctly distinguish hypertension with a pooled estimate sensitivity of 65.7% and specificity of 95.9%. After removing one study, which had very low sensitivity and very high specificity, the pooled sensitivity estimate was 79%, and the specificity was 91%. The meta-analysis of DOR suggests that the digital blood pressure monitor had moderate accuracy with a mercury sphygmomanometer. This will provide the clinician and patients with accurate information on blood pressure with which diagnostic and treatment decisions could be made., (© 2022. The Author(s).)

Background: Evidence on the extra-household contacts of TB patients who drive disease transmission is scarce., Methods: We conducted a cross-sectional personal social network survey among 300 newly diagnosed index pulmonary TB patients to identify their first-degree extra-household contacts., Results: A significantly higher proportion of neighbourhood (3.5; 95% CI 1.3 to 7.5), occupational (3.2; 95% CI 1.3 to 9.2) and friendship contacts (2.2; 95% CI 0.8 to 4.5) developed TB within 1 y of the index patient's diagnosis than their household contacts (0.7; 95% CI 0.3 to 1.3). Similarly, a higher proportion of extra-household contacts had TB at different time points before the index patient was diagnosed., Conclusion: Extra-household contacts of TB patients could be a potential source of TB or could be at increased risk of TB., (© The Author(s) 2021. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.)

Chicken poultry industry produces a vast amount of feather waste and is often disposed into landfills, creating environmental pollution. Therefore, we explored the valorization of chicken feather waste into lipids and keratinous sludge biomass. This study demonstrates the successful utilization of keratinous sludge biomass as a unique precursor for the facile preparation of novel keratinous sludge biomass-derived carbon-based molybdenum oxide (KSC@MoO 3 ) nanocomposite material using two-step (hydrothermal and co-pyrolysis) processes. The surface morphology and electrochemical properties of as-prepared nanocomposite material were analyzed using HR-SEM, XRD, XPS, and cyclic voltammetric techniques. KSC@MoO 3 nanocomposite exhibited prominent electrocatalytic behavior to simultaneously determine hydroquinone (HQ) and catechol (CC) in environmental waters. The as-prepared electrochemical sensor showed excellent performance towards the detection of HQ and CC with broad concentration ranges between 0.5-176.5 μM (HQ and CC), and the detection limits achieved were 0.063 μM (HQ) and 0.059 μM (CC). Furthermore, the developed modified electrode has exhibited excellent stability and reproducibility and was also applied to analyze HQ and CC in environmental water samples. Results revealed that chicken feather waste valorization could result in sustainable biomass conversion into a high-value nanomaterial to develop a cost-effective electrochemical environmental monitoring sensor and lipids for biofuel., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Terminally differentiated mammalian sperm are exposed to gradients of viscosity, pH, and osmolality both in the male and female reproductive tract during their perilous journey to quest the ovum. The complex physicochemical factors play an integral role in preparing sperm for the fertilization process., Objectives: To elucidate the influence of the reproductive tract microenvironment especially viscosity, pH, and osmolality in regulating sperm functional and fertilization competence., Materials and Methods: The data used in this review were collected from the research papers and online databases focusing on the influence of viscosity, pH, and osmolality on sperm function., Discussion: The gradients of viscosity, pH, and osmolality exist across various segments of the male and female reproductive tract. The changes in the viscosity create a physical barrier, pH aid in capacitation and hyperactivation, and the osmotic stress selects a progressive sperm subpopulation for accomplishing fertilization. The sperm function tests are developed based on the concept that the male genotype is the major contributor to the reproductive outcome. However, recent studies demonstrate the significance of sperm genotype-environment interactions that are essentially contributing to reproductive success. Hence, it is imperative to assess the impact of physicochemical stresses and the adaptive ability of the terminally differentiated sperm, which in turn would improve the outcome of the assisted reproductive technologies and male fertility assessment., Conclusion: Elucidating the influence of the reproductive tract microenvironment on sperm function provides newer insights into the procedures that need to be adopted for selecting fertile males for breeding, and ejaculates for the assisted reproductive technologies., (© 2021 American Society of Andrology and European Academy of Andrology.)

Background: The National Tuberculosis Elimination Programme (NTEP) of India is aiming to eliminate TB by 2025. The programme has increased its services and resources to strengthen the accurate and early detection of TB. It is important to estimate the cost of TB diagnosis in India considering the advancement and implementation of new diagnostic tools under the NTEP. The objective of this study was to estimate the unit costs of providing TB diagnostic services at different levels of public health facilities with different algorithms implemented under the NTEP in Chennai, Tamil Nadu, South India., Methods: This costing study was conducted from the perspective of the health system. This study used only secondary data and information that were available in the public domain. Data were collected with the approval of health authorities. The patient's diagnostic path from the point of registration until the final diagnosis was considered in the costing exercise. The unit costs of different diagnostic tools used in the NTEP implemented by Chennai Corporation were calculated., Results: We estimated the unit cost of the eight laboratory tests (Ziehl-Neelsen [ZN], fluorescence microscopy [FM], x-ray, digital x-ray, gene Xpert MTB/RIF (cartridge-based nucleic acid amplification test [NAAT] that identifies rifampicin resistant Mycobacterium Tuberculosis) Mycobacterium Tuberculosis/Rifampicin [MTB/RIF], mycobacteria growth indicator tube [MGIT], line probe assay [LPA] and Lowenstein Jensen [LJ] culture) for diagnosis of drug-sensitive and drug-resistant TB. The unit costs included fixed and variable costs for smear examination by ZN microscopy (₹ [Indian Rupee] 326 [US${\$}$4.72], FM (₹104 [US${\$}$1.5]), x-ray (₹218 [US${\$}$3.15]), digital X-ray (₹281 [US${\$}$4.07]), gene Xpert MTB/RIF (₹1137 [US${\$}$16.47]), MGIT (₹7038 [US${\$}$102]), LPA (₹6448 [US${\$}$93.44]) and LJ culture (₹4850 [US${\$}$70.28]). Out of 10 diagnostic algorithms used for TB diagnosis, algorithms using only smear microscopy had the lowest cost, followed by smear microscopy with x-ray for drug-sensitive TB (₹104 [US${\$}$1.5] to ₹544 [US${\$}$7.88]). Diagnostic algorithms for drug-resistant TB involving LPA and gene Xpert MTB/RIF were the most expensive., Conclusions: Understanding the various costs contributing to TB diagnosis in India provides crucial evidence for policymakers, programme managers and researchers to optimise programme spending and efficiently use resources., (© The Author(s) 2021. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.)

To understand the effects of urea on sperm functional attributes, fresh bull semen (n = 12) was subjected to four different concentrations (mg/mL) of urea to mimic the physiological (0.04 and 0.13), supraphysiological (0.43) concentrations and control (0 mg/mL). Sperm membrane integrity, kinematics, chromatin integrity, and mitochondrial membrane potential were assessed at different time points (before incubation, 0, 1, 2, and 4 h) of incubation. The concentration of urea in serum and seminal plasma was estimated and correlated with the ejaculate rejection rate and sperm functional attributes. The relative expression of urea transporter gene transcripts (UT-A and UT-B) was assessed in sperm and testis (control) using real-time PCR. The supraphysiological concentration of urea affected sperm kinematics, viability, functional membrane integrity, and acrosome integrity within 1 h of incubation (p < 0.05). Sperm head area decreased (p < 0.05) at 0 h and subsequently increased at 1 h of incubation in all media except supraphysiological (0.43 mg/dL) concentration of urea. Seminal plasma urea concentration showed a significant negative correlation with sperm motility, membrane integrity, and mitochondrial membrane potential (p < 0.05), but had a positive correlation with the ejaculate rejection rate (r = 0.69). Relative expression of the urea transporter genes revealed that UT-A was expressed only in the testis. In contrast, UT-B was expressed in both the testis and sperm, suggesting UT-B's role in regulating urea transport in sperm. At a supraphysiological level, urea adversely affected sperm functional attributes, osmoadaptation and may affect fertility., Competing Interests: Declaration of competing interest Authors declare no conflict of interest., (Copyright © 2021 Elsevier Inc. All rights reserved.)