Comprehensive characterisation and cryopreservation optimisation of buffalo (Bubalus bubalis) adipose tissue-derived mesenchymal stem cells.

Over half of the world's buffalo (Bubalus bubalis) inhabit India, and buffaloes frequently encounter health challenges that resist conventional treatments, prompting the exploration of alternative therapeutic strategies. One promising approach is stem cell therapy, particularly multipotent mesenchymal/stromal stem cells (MSCs). These cells have shown significant efficacy in addressing various diseases in livestock that exhibit resistance to conventional therapies. Adipose tissue-derived MSCs (ADSCs) have garnered attention due to their accessibility and robust expansion potential. The current study comprehensively characterises buffalo ADSCs (bADSCs), confirming their identity as MSCs capable of differentiating into diverse cell lineages-the identified characteristics position bADSCs as promising candidates for applications in regenerative medicine, applicable in veterinary contexts. Notably, the study established that a cryoprotective solution comprising 10 % dimethyl sulfoxide and 90 % fetal bovine serum is optimal for preserving bADSCs. This cryoprotective solution maintains vital parameters, including viability, apoptosis, senescence, cell adherence, adherent cell viability, metabolic and clonogenic efficiency, and the activity of reactive oxygen species and trilineage differentiation potential following thawing. These findings lay the foundation for developing a cryo-banking system for bADSCs. Subsequent research efforts are focused on exploring the therapeutic potential of bADSCs in specific disease models and clinical settings. The outcomes of such investigations may pave the way for innovative and effective treatments, further enhancing our understanding of the regenerative capabilities of bADSCs.

Comprehensive assessment of goat adipose tissue-derived mesenchymal stem cells cultured in different media.

Mesenchymal stem cells (MSCs) have demonstrated potential in treating livestock diseases that are unresponsive to conventional therapies. MSCs derived from goats, a valuable model for studying orthopaedic disorders in humans, offer insights into bone formation and regeneration. Adipose tissue-derived MSCs (ADSCs) are easily accessible and have a high capacity for expansion. Although the choice of culture media significantly influences the biological properties of MSCs, the optimal media for goat ADSCs (gADSCs) remains unclear. This study aimed to assess the effects of four commonly used culture media on gADSCs' culture characteristics, stem cell-specific immunophenotype, and differentiation. Results showed that MEM, DMEM/F12, and DMEM-LG were superior in maintaining cell morphology and culture parameters of gADSCs, such as cell adherence, metabolic activity, colony-forming potential, and population doubling. Conversely, DMEM-HG exhibited poor performance across all evaluated parameters. The gADSCs cultured in DMEM/F12 showed enhanced early proliferation and lower apoptosis. The cell surface marker distribution exhibited superior characteristics in gADSCs cultured in MEM and DMEM/F12. In contrast, the distribution was inferior in gADSCs cultured in DMEM-LG. DMEM/F12 and DMEM-LG culture media demonstrated a significantly higher potential for chondrogenic differentiation and DMEM-LG for osteogenic differentiation. In conclusion, DMEM/F12 is a suitable culture medium for propagating gADSCs as it effectively maintains cell morphology, growth parameters, proliferation and lower apoptosis while exhibiting desirable expression patterns of MSC-specific markers. These findings contribute to optimising culture conditions for gADSCs, enhancing their potential applications in disease treatment and regenerative medicine.

Polycaprolactone/graphene oxide/acellular matrix nanofibrous scaffolds with antioxidant and promyelinating features for the treatment of peripheral demyelinating diseases.

Peripheral demyelinating diseases entail damage to axons and Schwann cells in the peripheral nervous system. Because of poor prognosis and lack of a cure, this group of diseases has a global impact. The primary underlying cause of these diseases involves the inability of Schwann cells to remyelinate the damaged insulating myelin around axons, resulting in neuronal death over time. In the past decade, extensive research has been directed in the direction of Schwann cells focusing on their physiological and neuroprotective effects on the neurons in the peripheral nervous system. One cause of dysregulation in the remyelinating function of Schwann cells has been associated with oxidative stress. Tissue-engineered biodegradable scaffolds that can stimulate remyelination response in Schwann cells have been proposed as a potential treatment strategy for peripheral demyelinating diseases. However, strategies developed to date primarily focussed on either remyelination or oxidative stress in isolation. Here, we have developed a multifunctional nanofibrous scaffold with material and biochemical cues to tackle both remyelination and oxidative stress in one matrix. We developed a nanofibrous scaffold using polycaprolactone (PCL) as a foundation loaded with antioxidant graphene oxide (GO) and coated this bioscaffold with Schwann cell acellular matrix. In vitro studies revealed both antioxidant and remyelination properties of the developed bioscaffold. Based on the results, the developed multifunctional bioscaffold approach can be a promising biomaterial approach for treating demyelinating diseases.

Multicenter study of uterine tears and other reproductive complications in periparturient goats presented to veterinary teaching hospitals.

BACKGROUND: Goats are increasingly popular as both production animals and pets. The frequency of and factors associated with periparturient reproductive complications in goats are largely unreported. OBJECTIVES: (1) To report the frequency of periparturient reproductive complications in does presented to university veterinary hospitals and (2) to identify factors associated with uterine tears in the study population. ANIMALS: A total of 198 periparturient does presented to 9 university veterinary hospitals from October 2021 to June 2022. METHODS: Multicenter, cross-sectional study, with data collected from questionnaires completed by attending veterinarians. Logistic regression was used to identify factors associated with diagnosis of uterine tears. RESULTS: Ninety-three (47%) does had at least 1 periparturient reproductive complication. Periparturient complications included retained fetal membranes (n = 38, 26%), vaginal or perineal trauma (n = 33, 19%), uterine tears (n = 32, 18%), metritis (n = 22, 13%), uterine or vaginal hemorrhage (n = 8, 5%), Cesarean section complications (n = 8, 8%), and uterine prolapse (n = 1, 0.5%). A positive interaction effect was found between small breeds (Nigerian Dwarf and Pygmy) and manipulation on the farm by a layperson upon diagnosis of uterine tears (odd ratios [OR], 5.48; 95% confidence interval [CI], 1.41, 21.25; P = .01). CONCLUSIONS AND CLINICAL IMPORTANCE: Periparturient reproductive complications were common. Small breed combined with manipulation on the farm by layperson was associated with diagnosis of uterine tears. Clients should be educated that in the event of dystocia, small breed does are at greater risk of uterine tears and prompt veterinary intervention is critical.

Causes of equine perinatal mortality.

The peripartum period is critical in equine medicine for maintaining healthy mares, and ensuring the delivery of healthy neonatal foals. The field of perinatal mortality in horses is continuously evolving, with several advances being recently made in causes of perinatal fetal and foal loss. This review details the main causes of perinatal loss in horses, through late pregnancy, parturition and the neonatal period. Recent advances in identification of infectious organisms and indicators of survival in neonatal foals will be discussed. Continued advances in reproductive and neonatal medicine will aid improved survival of foals through fewer pregnancy losses, and improved management of high-risk pregnancies and critically ill neonatal foals.

Utility of cell-free DNA concentrations and illness severity scores to predict survival in critically ill neonatal foals.

Plasma cell-free DNA (cfDNA) levels have been associated with disease and survival status in septic humans and dogs. To date, studies investigating cfDNA levels in association with critical illness in foals are lacking. We hypothesized that cfDNA would be detectable in the plasma of foals, that septic and sick-nonseptic foals would have significantly higher cfDNA levels compared to healthy foals, and that increased cfDNA levels would be associated with non-survival. Animals used include 80 foals of 10 days of age or less admitted to a tertiary referral center between January and July, 2020 were stratified into three categories: healthy (n = 34), sick non-septic (n = 11) and septic (n = 35) based on specific criteria. This was a prospective clinical study. Blood was collected from critically ill foals at admission or born in hospital for cfDNA quantification and blood culture. Previously published sepsis score (SS) and neonatal SIRS score (NSIRS) were also calculated. SS, NSIRS, blood culture status and cfDNA concentrations were evaluated to predict survival. Continuous variables between groups were compared using Kruskal-Wallis ANOVA with Dunn's post hoc test. Comparisons between two groups were assessed using the Mann-Whitney U-test or Spearman rank for correlations. The performance of cfDNA, sepsis score and NSIRS score to predict survival was assessed by receiver operator characteristic (ROC) curve analysis including area under the curve, sensitivity and specificity using cutoffs. Plasma cfDNA was detectable in all foals. No significant differences in cfDNA concentration were detected between healthy foals and septic foals (P = 0.65) or healthy foals and sick non-septic foals (P = 0.88). There was no significant association between cfDNA and culture status, SS, NSIRS or foal survival. SS (AUC 0.85) and NSIRS (AUC 0.83) were superior to cfDNA (AUC 0.64) in predicting survival. Although cfDNA was detectable in foal plasma, it offers negligible utility to diagnose sepsis or predict survival in critical illness in neonates.

Nucleobase analogs for degenerate hybridization devised through conformational pairing analysis.

A conformational pairing analysis was used to devise nucleobase analogs capable of forming nonselective and energetically favorable base pairs opposite either the purine or the pyrimidine constituents of nucleic acids. 5-methylisocytosine and isoguanine were conceived as a degenerate pyrimidine and a degenerate purine, respectively. Data from previous DNA duplex melting experiments verified that the analogs can act as degenerate nucleobases as hypothesized. Isoguanine also formed unusually stable base pairs with guanine. A quantitative PCR assay yielding equivalent results across hepatitis C virus (HCV) subtypes was created with this system, despite the use of a single probe targeted to a polymorphic region. Amplification curves using probes with 5-methylisocytosine or isoguanine opposite appropriate ambiguous target positions exhibited more signal than curves from similar probes containing common degenerate nucleobase hypoxanthine.

Mother-child attachment in later middle childhood: assessment approaches and associations with mood and emotion regulation.

This study examined the overlap and validity of several measures of mother - child attachment developed for preadolescents. Validity was assessed in part by examining how attachment is related to children's mood and emotion regulation. Mother - child attachment was assessed in a sample of 9 to 11 year-old children using a story stem interview technique and questionnaires. Positive and negative mood were scored from daily logs completed by children. Emotion regulation was assessed with mothers' reports of constructive coping and teacher reports of children's ability to tolerate frustration. Interview and questionnaire measures of attachment were not consistently related to one another, although both were related to mood and emotion regulation. As expected, secure attachment and maternal secure base support were related to higher levels of positive mood, more constructive coping, and better regulation of emotion in the classroom, with effects stronger for emotion regulation than for mood. Children classified Disorganized or Ambivalent displayed the most negative mood. All effects remained significant after controlling for child temperament.