Nanofibrous composite from chitosan-casein polyelectrolyte complex for rapid hemostasis in rat models in vivo.

Bleeding causes ∼5.8 million deaths globally; half of the patients die if rapid hemostasis is not achieved. Here, we report a chitosan-casein (CC)-based nanofibrous polyelectrolyte complex (PEC) that could clot blood within 10 s in the rat femoral artery model in vivo. The nanofiber formation by self-assembly was also optimized for process parameters (concentration, mixing ratio, pH, and ultrasonication). Results showed that increasing the concentration of chitosan from 10 % to 90 % in the formulation increased the productivity (r = 0.99) of PECs but led to increased blood clotting time (r = 0.90) due to an increase in zeta potential (r = 0.98), fiber diameter (r = 0.93), and decreased surface porosity (r = -0.99), absorption capacity (r = -0.99). The pH also influenced the zeta potential of PEC, with an optimized pH of 8.0 ± 0.1 yielding clear nanofibers. Sonication improved the segregation of nanofibers by promoting water removal. The optimized PECs containing chitosan and casein in the ratio of 30:70 (CC30) at a pH of 8.0 and dehydration under sonication could clot the blood within 9 ± 2 s in vitro and 9 ± 2 s in rat femoral artery puncture model. The CC30 formulation did not cause any irritation or corrosion on rat skin. Histopathology and immunohistochemistry of various organs showed that CC30 was biocompatible and non-immunogenic under in vivo conditions.

Identification and validation of putative biomarkers by in silico analysis, mRNA expression and oxidative stress indicators for negative energy balance in buffaloes during transition period.

OBJECTIVE: Transition period is considered from 3 weeks prepartum to 3 weeks postpartum, characterized with dramatic events (endocrine, metabolic, and physiological) leading to occurrence of production diseases (negative energy balance/ketosis, milk fever etc). The objectives of our study were to analyze the periodic concentration of serum beta-hydroxy butyric acid (BHBA), glucose and oxidative markers along with identification, and validation of the putative markers of negative energy balance in buffaloes using in-silico and quantitative real time-polymerase chain reaction (qRT-PCR) assay. METHODS: Out of 20 potential markers of ketosis identified by in-silico analysis, two were selected and analyzed by qRT-PCR technique (upregulated; acetyl serotonin o-methyl transferase like and down regulated; guanylate cyclase activator 1B). Additional two sets of genes (carnitine palmotyl transferase A; upregulated and Insulin growth factor; downregulated) that have a role of hepatic fatty acid oxidation to maintain energy demands via gluconeogenesis were also validated. Extracted cDNA (complementary deoxyribonucleic acid) from the blood of the buffaloes were used for validation of selected genes via qRTPCR. Concentrations of BHBA, glucose and oxidative stress markers were identified with their respective optimized protocols. RESULTS: The analysis of qRT-PCR gave similar trends as shown by in-silico analysis throughout the transition period. Significant changes (p<0.05) in the levels of BHBA, glucose and oxidative stress markers throughout this period were observed. This study provides validation from in-silico and qRT-PCR assays for potential markers to be used for earliest diagnosis of negative energy balance in buffaloes. CONCLUSION: Apart from conventional diagnostic methods, this study improves the understanding of putative biomarkers at the molecular level which helps to unfold their role in normal immune function, fat synthesis/metabolism and oxidative stress pathways. Therefore, provides an opportunity to discover more accurate and sensitive diagnostic aids.

Transcriptome Analysis Reveals Spermatogenesis-Related CircRNAs and LncRNAs in Goat Spermatozoa.

Mammalian spermatozoa comprises both coding and non-coding RNAs, which are traditionally believed to be a residual of spermatogenesis. The differential expression level of spermatozoal RNAs is also observed between fertile and infertile human, thereby anticipated as potential molecular marker of male fertility. This study investigated the transcriptome profile of goat (Capra hircus) spermatozoa. The sperm transcriptome was analyzed by three different methods viz. RLM-RACE, long-read RNA sequencing (RNAseq) in Nanopore™ platform, and short-read RNAseq in Illumina™ platform. The Illumina™ sequencing discovered 16,604 transcripts with 357 mRNAs having FPKM (fragments per kilobase per million mapped reads) of more than five. The spermatozoal RNA suite included mRNA (94%), rRNA (3%), miscRNA (1%), circRNA (1%), miRNA (1%), etc. This study also predicted circRNAs (127), lncRNAs (655), and imprinted genes (160) that have potential role in male reproduction. The gene ontology analysis revealed the involvement of spermatozoal RNA in regulating male meiosis (TET3, STAT5B), capacitation (ACRBP, CATSPER4), sperm motility (GAS8, TEKT2), spermatogenesis (ADAMTS2, CREB3L4), etc. The spermatozoal RNA were also associated with different biological pathways viz. Wnt signaling pathway, cAMP signaling pathway, AMPK signaling pathway, and MAPK signaling pathways having potential role in spermatogenesis. Overall, this study enlightened the suite of spRNA transcripts in goat and their relevance in male fertility for diagnostic approach.

Pre-implantation development of cattle embryos produced from fresh bull semen enriched for X- chromosome-bearing spermatozoa using a monoclonal antibody.

Immunological approaches are gaining attention as a convenient and economical method for sex-sorting mammalian spermatozoa. A monoclonal antibody (WholeMom™) has previously been reported to cause agglutination of Y-chromosome-bearing spermatozoa in frozen-thawed semen for gender preselection. However, its usefulness for gender preselection in fresh semen and subsequent in vitro fertilization (IVF) after freeze-thawing has not been reported. This study investigated the in vitro development of cattle embryos produced from fresh bull semen pre-treated with WholeMom™ monoclonal antibody. Results showed that antibody-treated, non-agglutinated spermatozoa (presumably X-chromosome-bearing spermatozoa) could fertilize cattle oocytes in vitro. However, embryos generated from non-agglutinated (enriched in X-chromosome-bearing spermatozoa) had a lower (p < 0.05) ability to cleave (66.4 ± 2.5% vs. 75.1 ± 3.3%) than those of non-treated control sperm. Nevertheless, the percentage of blastocysts developed from cleaved embryos did not differ (p > 0.05) between the groups (34.8 ± 3.7% vs. 35.8 ± 3.4%). Duplex PCR of blastocysts, using a bovine-specific universal primer pair and a Y-chromosome-specific primer pair, showed a sex ratio of 95.8% females from sex-sorted spermatozoa, which was higher than those of non-treated control spermatozoa (46.4%). In conclusion, the results of the present study suggest that monoclonal antibody-based enrichment of X- chromosome-bearing spermatozoa can be applied to fresh bull semen without compromising their post-fertilization early embryonic development to the blastocyst stage. Future studies should investigate the term development and sex ratio of calves from antibody-treated spermatozoa.

Preparation, characterization, and bioactivity of reinforced monetite with chitosan-gelatin electrospun composite scaffold for bone tissue engineering.

In this study, chitosan-gelatin-monetite (CGM)-based electrospun scaffolds have been developed that closely mimicked the microstructure and chemical composition of the extracellular matrix of natural bone. CGM-based nanofibrous composite scaffolds were prepared with the help of the electrospinning technique, post-cross-linked using ethyl(dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide solution to improve their stability in an aqueous environment. The prepared chitosan/gelatin (CG) scaffold showed an average fiber diameter of 308 ± 17 nm, whereas 5 and 7 wt% monetite containing CGM5and CGM7scaffolds, exhibited an average fiber diameter of 287 ± 13 and 265 ± 9 nm, respectively, revealing the fine distribution of monetite particles on the fibrous surface. The distribution of monetite nanoparticles onto the CG nanofibrous surface was confirmed using x-ray diffraction, Fourier transform infrared, and EDAX. Moreover, the addition of 7 wt% monetite into the CG electrospun matrix increased their ultimate tensile strength from 7.62 ± 0.13 MPa in the CG scaffold to 14.34 ± 0.39 MPa in the CGM7scaffold. Simulated body fluid study and staining with alizarin red S (ARS) confirmed the higher mineralization ability of monetite-containing scaffolds compared to that revealed by the CG scaffold. The monetite incorporation into the CG matrix improved its osteogenic properties, including pre-osteoblast MG-63 cell adhesion, proliferation, and differentiation, when seeded with the cells. A higher degree of cellular adhesion, spreading, and migration was observed on the monetite-incorporated CG scaffold than that on the CG scaffold. From 3-(4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide) MTT assay, alkaline phosphatase activity, ARS staining, and immunocytochemistry study, the cultured cells discovered a more conducive microenvironment to proliferate and subsequently differentiate into osteoblast lineage in contact with CGM7nanofibers rather than that in CGM0and CGM5.In-vitroresults indicated that electrospun CGM-based composite scaffolds could be used as a potential candidate to repair and regenerate new bone tissues.

Effect of arginine-induced motility and capacitation on RNA population in goat spermatozoa.

INTRODUCTION: In vitro capacitation is essential in assisted reproductive technologies (ART) for embryo production. Recently, arginine has been proven to enhance capacitation in mammalian spermatozoa. However, the detailed mechanism of action of arginine remains elusive. AIM: This study investigated the effect of arginine-induced capacitation and motility enhancement on the spermatozoal RNA (spRNA) population in goats. MATERIAL AND METHODS: Goat spermatozoa were treated with arginine for up to six hours and compared with non-treated or PHE (penicillamine, hypotaurine, and epinephrine)-treated spermatozoa at different intervals (0, 1, 2, 4, and 6 hours). Sperm parameters, including viability, individual motility, capacitation, acrosome reaction, and ROS production, were evaluated. The spRNA population was analyzed by short-read RNA sequencing (RNA-seq). RESULTS: The percentage of capacitated (73.21 ± 4.22%) and acrosome reacted (18.35 ± 0.56%) spermatozoa was highest in arginine treatment, while PHE treatment showed the highest percentage (79.82 ± 4.31%) of motile spermatozoa from 0 to 4 hours of incubation. RNA-seq analysis identified 1,321 differentially expressed genes (DEGs) in arginine-treated spermatozoa compared to the control. The PGK2, RNASE10, ODF1, and ROPN1L genes involved in sperm motility and ACR, DKKL1, KCNJ11, and PRND genes involved in the capacitation process were upregulated in arginine-treated spermatozoa. The DEGs regulate sperm capacitation-related cAMP-PKA, PI3-Akt, calcium, and MAPK signaling pathways. CONCLUSION: The arginine-induced capacitation and enhanced sperm motility were associated with the upregulation of several genes involved in sperm motility and capacitation pathways. The comparative study also suggests that arginine may be used in lieu of PHE for motility enhancement and in vitro capacitation of goat spermatozoa.

Analyzing the effect of heparin on in vitro capacitation and spermatozoal RNA population in goats.

Heparin is a glycosaminoglycan polymer that is commonly used as an anticoagulant. Heparin also induces in vitro capacitation in spermatozoa, although its molecular mechanism is elusive. This study investigated the effect of heparin on in vitro capacitation and spermatozoal RNA (spRNA) population in goats. Goat spermatozoa were treated with 20 µM heparin for 0-6 h and evaluated for motility, capacitation, acrosome reaction, and spRNA population by RNA sequencing (RNA-seq). It was observed that heparin enhanced sperm motility up to 6 h of incubation (p < 0.05). Heparin also induced capacitation and acrosome reaction within 4 h. RNA-seq identified 1254 differentially expressed genes (DEGs) between heparin-treated and control spermatozoa. Most DEGs (1251 nos.) were upregulated and included 1090 protein-coding genes. A few genes (PRND, ITPR1, LLCFC1, and CHRM2) showed >5-fold increased expression in heparin-treated spermatozoa compared to the control. The upregulated genes were found to be involved in cAMP-PKA, PI3-Akt, calcium, MAPK signaling, and oxidative stress pathways. DCFDA staining confirmed the increased oxidative stress in heparin-treated spermatozoa compared to the control (p < 0.05). In conclusion, the results of the present study suggest that heparin enhances sperm motility and induces capacitation by upregulation of the spRNA population and oxidative stress pathway.

Physicochemical, mechanical, dielectric, and biological properties of sintered hydroxyapatite/barium titanate nanocomposites for bone regeneration.

Bone implants fabricated using nanocomposites containing hydroxyapatite (HA) and barium titanate (BT) show osteoconductive, osteoinductive, osteointegration, and piezoelectricity properties for bone regeneration applications. In our present study, HA and BT nanopowders were synthesized using high-energy ball-milling-assisted solid-state reaction with precursors of calcium carbonate and ammonium dihydrogen phosphate, and barium carbonate and titanium oxide powder mixtures, respectively. Hexagonal HA and tetragonal BT phases were formed after calcination at 700 and 1000 °C, respectively. Subsequently, hydroxyapatite/barium titanate (HA/BT) nanocomposites with different weight percentages of HA and BT were prepared by ball-milling, then compacted and sintered at two different temperatures to endow these bioceramics with better mechanical, dielectric, and biological properties for bone regeneration. Microstructure, crystal phases, and molecular structure characterizations of these sintered HA/BT nanocomposite compacts (SHBNCs) were performed using field-emission scanning electron microscopy, x-ray diffraction, and Fourier-transform infrared spectroscopy, respectively. Bulk density was evaluated using the Archimedes method. HA/BT nanocomposites with increased BT content showed enhanced dielectric properties, and the dielectric constant (ϵr) value for 5HA/95BT was ∼182 at 100 Hz. Mechanical properties such as Vicker's hardness, fracture toughness, yield strength, and diametral tensile strength were also investigated. The hemolysis assay of SHBNCs exhibited hemocompatibility. The effect of these SHBNCs as implants on thein vitrocytocompatibility and cell viability of MG-63 osteoblast-like cells was assessed by MTT assay and live/dead staining, respectively. 15HA/85BT showed increased metabolic activity with a higher number of live cells than BT after the culture period. Overall, the SHBNCs can be used as orthopedic implants for bone regeneration applications.

Siamese-Based Architecture for Cross-Lingual Plagiarism Detection in English-Hindi Language Pairs.

The cross-lingual plagiarism detection (CLPD) is a challenging problem in natural language processing. Cross-lingual plagiarism is when a text is translated from any other language and used as it is without proper acknowledgment. Most of the existing methods provide good results for monolingual plagiarism detection, whereas the performances of existing methods for the CLPD are very limited. The reason for this is that it is difficult to represent the text from two different languages in a common semantic space. In this article, a novel Siamese architecture-based model is proposed to detect the cross-lingual plagiarism in English-Hindi language pairs. The proposed model combines the convolutional neural network (CNN) and bidirectional long short-term memory (Bi-LSTM) network to learn the semantic similarity among the cross-lingual sentences for the English-Hindi language pairs. In the proposed model, the CNN model learns the local context of words, whereas the Bi-LSTM model learns the global context of sentences in forward and backward directions. The performances of the proposed models are evaluated on the benchmark data set, that is, Microsoft paraphrase corpus, which is converted in the English-Hindi language pairs. The proposed model outperforms other models giving 67%, 72%, and 67% weighted average precision, recall, and F1-measure scores. The experimental results show the effectiveness of the proposed models over the baseline models because the proposed model is very efficient in representing the cross-lingual text very efficiently.

Gelatin/monetite electrospun scaffolds to regenerate bone tissue: Fabrication, characterization, and in-vitro evaluation.

This work is dedicated to combining nanotechnology with bone tissue engineering to prepare and characterize electrospun gelatin/monetite nanofibrous scaffold with improved physicochemical, mechanical, and biological properties. Nanofibrous scaffolds possessing fiber diameter in the range of 242-290 nm were prepared after incorporating varying content of monetite nanoparticles up to 7 wt % into the gelatin matrix using the electrospinning technique. Cross-linking of gelatin chains in the scaffold was performed using 0.25 wt% glutaraldehyde as indicated by imine (-CN-) bond formation in the FTIR analysis. With an increase in monetite addition up to 7 wt%, a decrease in swelling ratio and bio-degradability of cross-linked gelatin scaffolds was observed. Gelatin scaffold with 7 wt% monetite content registered the highest values of tensile strength and tensile modulus of 18.8 MPa and 170 MPa, as compared to 0% and 5 wt% monetite containing scaffolds respectively. Cell viability and differentiation were studied after culturing MG-63 cells onto the scaffolds from confocal microscopy of live and dead cells images, MTT assay, and alkaline phosphatase assay for a cell culture period of up to 21 days. It was observed that 7 wt % monetite containing gelatin scaffold exhibited better MG-63 cell adhesion, proliferation, higher biomineralization, and ALP activity compared to 0% and 5 wt% monetite containing electrospun scaffolds studied here.

Prediction of potential molecular markers of bovine mastitis by meta-analysis of differentially expressed genes using combined p value and robust rank aggregation.

Bovine mastitis causes significant economic loss to the dairy industry by affecting milk quality and quantity. Escherichia coli and Staphylococcus aureus are the two common mastitis-causing bacteria among the consortia of mastitis pathogens, wherein E. coli is an opportunistic environmental pathogen, and S. aureus is a contagious pathogen. This study was designed to predict molecular markers of bovine mastitis by meta-analysis of differentially expressed genes (DEG) in E. coli- or S. aureus-infected mammary epithelial cells (MECs) using p value combination and robust rank aggregation (RRA) methods. High-throughput transcriptome of bovine MECs, infected with E. coli or S. aureus, were analyzed, and correlation of z-scores were computed for the expression datasets to identify the lineage profile and functional ontology of DEGs. Key pathways enriched in infected MECs were deciphered by Gene Set Enrichment Analysis (GSEA), following which combined p value and RRA were used to perform DEG meta-analysis to limit type I error in the analysis. The miRNA-gene networks were then built to uncover potential molecular markers of mastitis. Lineage profiling of MECs showed that the gene expression levels were associated with mammary tissue lineage. The up-regulated genes were enriched in immune-related pathways, whereas down-regulated genes influenced the cellular processes. GSEA analysis of DEGs deciphered the involvement of Toll-like receptor (TLR), and NF-kappa B signaling pathway during infection. Comparison after meta-analysis yielded with genes ZC3H12A, RND1, and MAP3K8 having significant expression levels in both E. coli and S. aureus dataset, and on evaluating miRNA-gene network, 7 pairs were common to both sets identifying them as potential molecular markers.

Nanoceria laden decellularized extracellular matrix-based curcumin releasing nanoemulgel system for full-thickness wound healing.

Decellularized extracellular matrix (ECM) has been widely used for wound healing. But, ECM failed to integrate tissue and restore the tissue function properly, when elevated levels of free radicals and biofilm formation occur at the wound site. Here, nanoemulgel systems were fabricated, considering the combinatorial approach of nanotechnology (nanoceria and curcumin nanoemulsion) and ECM gel of goat small intestine submucosa. The curcumin was encapsulated in the nanoemulgel system to enhance bioavailability in terms of antibacterial, antioxidant, sustained release and permeation at the wound site. Nanoceria was also incorporated to enhance the antibacterial, antioxidant and wound healing properties of the fabricated nanoemulgel formulation. All the formulations were porous, hydrophilic, biodegradable, antioxidant, antibacterial, hemocompatible, biocompatible, and showed enhanced wound healing rate. The formulation (DG-SIS/Ce/NC) showed the highest free radicals scavenging capacity and antibacterial property with prolonged curcumin release (62.9% in 96 h), skin permeability (79.7% in 96 h); showed better cell growth under normal and oxidative-stressed conditions: it also showed full-thickness wound contraction (97.33% in 14 days) with highest collagen synthesis at the wound site (1.61 µg/mg in 14 days). The outcomes of this study suggested that the formulation (DG-SIS/Ce/NC) can be a potential nanoemulgel system for full-thickness wound healing application.

Applications of Omics Technology for Livestock Selection and Improvement.

Conventional animal selection and breeding methods were based on the phenotypic performance of the animals. These methods have limitations, particularly for sex-limited traits and traits expressed later in the life cycle (e.g., carcass traits). Consequently, the genetic gain has been slow with high generation intervals. With the advent of high-throughput omics techniques and the availability of multi-omics technologies and sophisticated analytic packages, several promising tools and methods have been developed to estimate the actual genetic potential of the animals. It has now become possible to collect and access large and complex datasets comprising different genomics, transcriptomics, proteomics, metabolomics, and phonemics data as well as animal-level data (such as longevity, behavior, adaptation, etc.,), which provides new opportunities to better understand the mechanisms regulating animals' actual performance. The cost of omics technology and expertise of several fields like biology, bioinformatics, statistics, and computational biology make these technology impediments to its use in some cases. The population size and accurate phenotypic data recordings are other significant constraints for appropriate selection and breeding strategies. Nevertheless, omics technologies can estimate more accurate breeding values (BVs) and increase the genetic gain by assisting the section of genetically superior, disease-free animals at an early stage of life for enhancing animal productivity and profitability. This manuscript provides an overview of various omics technologies and their limitations for animal genetic selection and breeding decisions.

Evidence mapping and review of long-COVID and its underlying pathophysiological mechanism.

PURPOSE: Apart from the global disease burden of acute COVID-19 disease, the health complications arising after recovery have been recognized as a long-COVID or post-COVID-19 syndrome. Evidences of long-COVID symptoms involving various organ systems are rapidly growing in literature. The objective was to perform a rapid review and evidence mapping of systemic complications and symptoms of long-COVID and underlying pathophysiological mechanisms. METHODS: Publications reporting clinical trials, observational cohort studies, case-control studies, case-series, meta-analysis, and systematic reviews, focusing on the squeal of the disease, consequences of COVID-19 treatment/hospitalization, long-COVID, chronic COVID syndrome, and post acute COVID-19 were reviewed in detail for the narrative synthesis of frequency, duration, risk factors, and pathophysiology. RESULTS: The review highlights that pulmonary, neuro-psychological, and cardiovascular complications are major findings in most epidemiological studies. However, dysfunctional gastrointestinal, endocrine, and metabolic health are recent findings for which underlying pathophysiological mechanisms are poorly understood. Analysis of the clinical trial landscape suggests that more than 50% of the industry-sponsored trials are focused on pulmonary symptoms. In contrast to the epidemiological trends and academic trials, cardiovascular complications are not a focus of industry-sponsored trials, suggestive of the gaps in the research efforts. CONCLUSION: The gap in epidemiological trends and academic trials, particularly concerning cardiovascular complications not being a focus of industry-sponsored trials is suggestive of the gaps in research efforts and longer follow-up durations would help identify other long-COVID-related health issues such as reproductive health and fertility.

Intelligent Diagnostic Prediction and Classification Models for Detection of Kidney Disease.

Kidney disease is a major public health concern that has only recently emerged. Toxins are removed from the body by the kidneys through urine. In the early stages of the condition, the patient has no problems, but recovery is difficult in the later stages. Doctors must be able to recognize this condition early in order to save the lives of their patients. To detect this illness early on, researchers have used a variety of methods. Prediction analysis based on machine learning has been shown to be more accurate than other methodologies. This research can help us to better understand global disparities in kidney disease, as well as what we can do to address them and coordinate our efforts to achieve global kidney health equity. This study provides an excellent feature-based prediction model for detecting kidney disease. Various machine learning algorithms, including k-nearest neighbors algorithm (KNN), artificial neural networks (ANN), support vector machines (SVM), naive bayes (NB), and others, as well as Re-cursive Feature Elimination (RFE) and Chi-Square test feature-selection techniques, were used to build and analyze various prediction models on a publicly available dataset of healthy and kidney disease patients. The studies found that a logistic regression-based prediction model with optimal features chosen using the Chi-Square technique had the highest accuracy of 98.75 percent. White Blood Cell Count (Wbcc), Blood Glucose Random (bgr), Blood Urea (Bu), Serum Creatinine (Sc), Packed Cell Volume (Pcv), Albumin (Al), Hemoglobin (Hemo), Age, Sugar (Su), Hypertension (Htn), Diabetes Mellitus (Dm), and Blood Pressure (Bp) are examples of these traits.

Curcumin in decellularized goat small intestine submucosa for wound healing and skin tissue engineering.

Biomaterials derived from extracellular matrices (ECMs) were extensively used for skin tissue engineering and wound healing. ECM is a complex network of biomolecules (e.g., proteins), which provide organizational support to cells for growth. Thus, ECM could be an ideal biomaterial for fabricating the scaffold. However, oxidative stress and biofilm formation at the wound site remains a major challenge that could be neutralized using herbal ingredients (e.g., curcumin). In this study, ECM was extracted from the biowaste of the goat abattoir by using decellularization. The goat small intestine submucosa (G-SIS) is decellularized to obtain the decellularized G-SIS (DG-SIS) and curcumin (in different concentrations) was incorporated in the DG-SIS to fabricate curcumin-embedded DG-SIS scaffolds. Changes brought by increasing the concentrations of the curcumin in DG-SIS were observed in various properties, including free radical scavenging and antibacterial properties. Results depicted that the scaffolds are porous, biodegradable, biocompatible, antibacterial, and hydrophilic and showed sustained release of curcumin. Besides, it showed free radicals scavenging property. The porosity and hydrophilicity of the scaffolds were decreased with an increase in the curcumin content. However, biodegradability, free radical scavenging, biocompatibility, and antibacterial properties of the scaffolds increased with an increase in the curcumin content. The DG-SIS scaffold containing 1 wt % of curcumin may be a potential biomaterial for wound-healing and skin tissue engineering.

Solid surface vitrification of goat testicular cell suspension enriched for spermatogonial stem cells.

This study reports solid surface vitrification (SSV) of goat testicular cell suspensions (TCS) enriched for spermatogonial stem cells (SSCs). The TCS was isolated from pre-pubertal goat testis by enzymatic digestion, enriched for SSCs by filtration and differential plating, and were vitrified-warmed by SSV. The study showed that SSV could successfully vitrify goat TCS although the percentage of live cells in the vitrified-warmed group was lower (74.8 ± 4.1%) than in non-vitrified control (80.6 ± 6.27%). The vitrified-warmed TCS formed putative SSC colonies upon their in vitro culture, but the colony size of vitrified-warmed cells (24.3 ± 1.8 µm) was smaller than those of non-vitrified warmed cells (58.4 ± 2.5 µm). Mitochondrial activity (0.40 vs. 0.38 A U.), population doubling time (33.45 ± 1.25 h vs. 31.86 ± 1.90 h), and the cell proliferation rate (0.72 ± 0.10 vs. 0.75 ± 0.11 per day) of total cells (including putative SSCs and other somatic cells) did not differ (p > 0.05) between control and SSV vitrified-warmed groups. However, during in vitro culture for 96 h, vitrified-warmed cells showed significantly lower (0.75 vs. 1.33 A U.; p < 0.05) mitochondrial activity than non-vitrified controls. The DCFDA assay showed that ROS activity was significantly (p < 0.05) higher in vitrified-warmed cells (52.8 ± 4.1 A U) than non-vitrified control cells (32.8 ± 2.1 AU). In conclusion, our results suggest that SSC-enriched goat TCS could be successfully cryopreserved by SSV. However, ROS-induced damages to cell cytoplasmic components reduce their cellular proliferation and require further improvement in the protocol. To the best of our knowledge, this study is the first report on the SSV of SSC-enriched goat TCS.

Development of decellularization protocol for caprine small intestine submucosa as a biomaterial.

Decellularized animal tissues have been proven to be promising biomaterials for various tissue engineering (TE) applications. Among various animal tissues, small intestine submucosa (SIS) has gained attention of many researchers due to its easy availability from the abattoir waste, excellent physicochemical and biological characteristics of a good biomaterial. In this study, Caprine SIS was decellularized to get decellularized caprine SIS (DG-SIS). For decellularization, several physical, chemical and enzymatic protocols have been described in the literature. To optimize the decellularization of caprine SIS, several decellularization protocol (DP), including an in-house developed by us, had been attempted, and effect of the different DPs on the obtained DG-SIS were assessed in terms of decellularization, physiochemical and biological properties. All the DPs differ in terms of decellularization, but three DPs where ionic detergent like sodium dodecyl sulphate (SDS) has been used, largely affect the native composition (e.g. glycosaminoglycans (GAGs)), biological properties and other physiochemical properties of the G-SIS as compared to the DP that uses hypertonic solution of potassium iodide (KI) and non-ionic detergent (TritonX-100). The obtained DG-SISs were fibrous, hemocompatible, biocompatible, hydrophilic, biodegradable and exhibited significant antibacterial activity. Therefore, the DG-SIS will be a prospective biomaterial for TE applications.

Significance and Relevance of Spermatozoal RNAs to Male Fertility in Livestock.

Livestock production contributes to a significant part of the economy in developing countries. Although artificial insemination techniques brought substantial improvements in reproductive efficiency, male infertility remains a leading challenge in livestock. Current strategies for the diagnosis of male infertility largely depend on the evaluation of semen parameters and fail to diagnose idiopathic infertility in most cases. Recent evidences show that spermatozoa contains a suit of RNA population whose profile differs between fertile and infertile males. Studies have also demonstrated the crucial roles of spermatozoal RNA (spRNA) in spermatogenesis, fertilization, and early embryonic development. Thus, the spRNA profile may serve as unique molecular signatures of fertile sperm and may play pivotal roles in the diagnosis and treatment of male fertility. This manuscript provides an update on various spRNA populations, including protein-coding and non-coding RNAs, in livestock species and their potential role in semen quality, particularly sperm motility, freezability, and fertility. The contribution of seminal plasma to the spRNA population is also discussed. Furthermore, we discussed the significance of rare non-coding RNAs (ncRNAs) such as long ncRNAs (lncRNAs) and circular RNAs (circRNAs) in spermatogenic events.