Embryos from Prepubertal Hyperglycemic Female Mice Respond Differentially to Oxygen Tension In Vitro.

Reduced oxygen during embryo culture in human ART prevents embryo oxidative stress. Oxidative stress is also the major mechanism by which maternal diabetes impairs embryonic development. This study employed induced hyperglycemia prepubertal mice to mimic childhood diabetes to understand the effects of varying oxygen tension during in vitro embryonic development. The oocytes were fertilized and cultured at low (≈5%) oxygen (LOT) or atmospheric (≈20%) oxygen tension (HOT) for up to 96 h. Embryo development, apoptosis in blastocysts, inner cell mass (ICM) outgrowth proliferation, and Hif1α expression were assessed. Though the oocyte quality and meiotic spindle were not affected, the fertilization rate (94.86 ± 1.18 vs. 85.17 ± 2.81), blastocyst rate (80.92 ± 2.92 vs. 69.32 ± 2.54), and ICM proliferation ability (51.04 ± 9.22 vs. 17.08 ± 3.05) of the hyperglycemic embryos were significantly higher in the LOT compared to the HOT group. On the other hand, blastocysts from the hyperglycemic group, cultured at HOT, had a 1.5-fold increase in apoptotic cells compared to the control and lower Hif1α transcripts in ICM outgrowths compared to the LOT. Increased susceptibility of embryos from hyperglycemic mice to higher oxygen tension warrants the need to individualize the conditions for embryo culture systems in ART clinics, particularly when an endogenous maternal pathology affects the ovarian environment.

EXAMINATION OF THE INCIDENCE OF POOR SLEEP QUALITY AND FACTORS ASSOCIATED FOR POOR SLEEP DURING THE VARIOUS PHASES OF PREGNANCIES.

Objective - to measure poor sleep quality, its components, and the variables that contribute to it in a cohort of pregnant women across time. Four hundred and eighty-six strong singleton pregnancies were collected ahead of the fourteenth gestational week. Data on poor sleep quality were gathered before pregnancy and analyzed five distinct times in each trimester and six months after delivery. "Poor sleep quality (PSQ) was defined as a score of fewer than eight on the Athens Insomnia Scale (AIS), and for each trimester, adjusted odds ratios (aOR) with 95% confidence intervals (CI)"were acquired by use of multivariate logistic regression analysis. Pregnancy prevalence of poor SQ was 6.1 percent, followed by 44.2 percent in the first trimester (TR1), 46.3 percent in the second trimester (TR2), and 63.7 percent in the third trimester (TR3). Poor sleep quality after pregnancy was reported by 33.2 percent of women "(28.2-37.9) (p<0.001 for pre-gestational versus TR1, TR2 vs. TR3, and TR3 vs. post-pregnancy)."Due to a decrease in the quality of their nocturnal sleep, TR3's mean AIS score went from 2.34 before pregnant to 9.87; in contrast, TR1's detrimental impact on daytime functioning was larger. Poor sleep during the previous trimester was linked to poor sleep in TR2 and TR3. Poor SQ during pregnancy was a factor in TR1's poor SQ, and obesity was linked to bad sleep in TR3. The risks of having poor sleep quality in TR3 were instead decreased by moderate physical activity. Poor sleep throughout pregnancy was shown to be much more common than good sleep at any point in the pregnancy. In the latter stages of pregnancy, two out of every three expecting moms suffer poor SQ. Particular attention should be paid to pre-gestational poor SQ prevention and high body mass index.

Correlates of post-traumatic growth among persons bereaved from cancer: A systematic review and meta-analysis.

BACKGROUND: Recent research identified that cancer bereavement can lead to post-traumatic growth (PTG). Although PTG and its correlates are well explored in cancer patients and survivors, persons bereaved from cancer have received scant attention. Therefore, the present review attempts to identify the correlates of PTG among persons bereaved from cancer. METHODS: A systematic search in PubMed, Web of Science, APA PsycNet, Science Direct, Scopus, and Wiley was conducted to identify quantitative studies published in English, resulting in 12 eligible reports being included in the final analysis. JBI critical checklists were employed to appraise the risk of bias. RESULTS: The review identified 17 correlates, which were classified into four categories: demographic factors (age, gender, religious status, level of education), loss-related factors (time since death, quality of death, prolonged grief symptoms), interpersonal factors (relationship to the deceased, social support, attachment style, bereavement behaviours) and intrapersonal factors (resilience, coping, rumination, benevolence, meaningfulness, self-worth). Random effects meta-analyses on six correlates revealed correlation coefficients of age = -0.02 (95% CI: -0.35-0.31), gender = 0.27 (95% CI: 0.08-0.45), time since death = 0.09 (95% CI: -0.02-0.20), quality of death = 0.29 (95% CI: -0.01-0.54), prolonged grief symptoms = 0.22 (95% CI: 0.08-0.35) and relationship to the deceased = 0.13 (95% CI: -0.03-0.29). Fixed effects meta-analysis was performed for social support (r = 0.13, 95% CI: 0.04-0.21). However, PTG was found to be significantly associated with gender, prolonged grief symptoms, and social support. CONCLUSIONS: Very few studies examined PTG among persons bereaved from cancer, highlighting the need for increased attention, understanding, and conceptualisation of PTG in the population.

Biopolymer-protected graphene-Fe3O4 nanocomposite based wearable microneedle sensor: toward real-time continuous monitoring of dopamine.

Neurological disorders can occur in the human body as a result of nano-level variations in the neurotransmitter levels. Patients affected by neuropsychiatric disorders, that are chronic require continuous monitoring of these neurotransmitter levels for effective disease management. The current work focus on developing a highly sensitive and personalized sensor for continuous monitoring of dopamine. Here we propose a wearable microneedle-based electrochemical sensor, to continuously monitor dopamine in interstitial fluid (ISF). A chitosan-protected hybrid nanomaterial Fe3O4-GO composite has been used as a chemical recognition element protected by Nafion antifouling coating layer. The morphological and physiochemical characterizations of the nanocomposite were carried out with XRD, XPS, FESEM, EDAX and FT-IR. The principle of the developed sensor relies on orthogonal detection of dopamine with square wave voltammetry and chronoamperometric techniques. The microneedle sensor array exhibited an attractive analytical performance toward detecting dopamine in phosphate buffer and artificial ISF. The limit of detection (LOD) of the developed sensor was observed to be low, 90 nM in square wave voltammetry and 0.6 µM in chronoamperometric analysis. The practical applicability of the microneedle sensor array has been demonstrated on a skin-mimicking phantom gel model. The microneedle sensor also exhibited good long-term storage stability, reproducibility, and sensitivity. All of these promising results suggest that the proposed microneedle sensor array could be reliable for the continuous monitoring of dopamine.

Unravelling the Road to Recovery: Mechanisms of Wnt Signalling in Spinal Cord Injury.

Spinal cord injury (SCI) is a complex neurodegenerative pathology that consistently harbours a poor prognostic outcome. At present, there are few therapeutic strategies that can halt neuronal cell death and facilitate functional motor recovery. However, recent studies have highlighted the Wnt pathway as a key promoter of axon regeneration following central nervous system (CNS) injuries. Emerging evidence also suggests that the temporal dysregulation of Wnt may drive cell death post-SCI. A major challenge in SCI treatment resides in developing therapeutics that can effectively target inflammation and facilitate glial scar repair. Before Wnt signalling is exploited for SCI therapy, further research is needed to clarify the implications of Wnt on neuroinflammation during chronic stages of injury. In this review, an attempt is made to dissect the impact of canonical and non-canonical Wnt pathways in relation to individual aspects of glial and fibrotic scar formation. Furthermore, it is also highlighted how modulating Wnt activity at chronic time points may aid in limiting lesion expansion and promoting axonal repair.

CKAP5 stabilizes CENP-E at kinetochores by regulating microtubule-chromosome attachments.

Stabilization of microtubule plus end-directed kinesin CENP-E at the metaphase kinetochores is important for chromosome alignment, but its mechanism remains unclear. Here, we show that CKAP5, a conserved microtubule plus tip protein, regulates CENP-E at kinetochores in human cells. Depletion of CKAP5 impairs CENP-E localization at kinetochores at the metaphase plate and results in increased kinetochore-microtubule stability and attachment errors. Erroneous attachments are also supported by computational modeling. Analysis of CKAP5 knockout cancer cells of multiple tissue origins shows that CKAP5 is preferentially essential in aneuploid, chromosomally unstable cells, and the sensitivity to CKAP5 depletion is correlated to that of CENP-E depletion. CKAP5 depletion leads to reduction in CENP-E-BubR1 interaction and the interaction is rescued by TOG4-TOG5 domain of CKAP5. The same domain can rescue CKAP5 depletion-induced CENP-E removal from the kinetochores. Interestingly, CKAP5 depletion facilitates recruitment of PP1 to the kinetochores and furthermore, a PP1 target site-specific CENP-E phospho-mimicking mutant gets stabilized at kinetochores in the CKAP5-depleted cells. Together, the results support a model in which CKAP5 controls mitotic chromosome attachment errors by stabilizing CENP-E at kinetochores and by regulating stability of the kinetochore-attached microtubules.

Caring burden and quality of life among the caregivers of people living with dementia - a cross-sectional study in Udupi district of Karnataka.

Dementia is a chronic disorder of the brain that affects cognitive performance. The caregivers of individuals with dementia experience a greater burden that affects their Quality of Life (QoL). This cross-sectional study conducted in India was designed to assess the caring burden and QoL among the caregivers of people with dementia, as well as to ascertain the relationship between QoL scores and burden. Our sample included 80 caregivers of people with dementia. Most of the caregivers (n = 59, 73.8%) had a higher level of caregiver burden. There was a negative correlation between caregiver burden scores and QoL. A higher level of caregiver stress and low QoL were experienced by caregivers of dementia patients. In developing countries like India, counseling, and education on home health care for people with dementia should be provided to reduce the burden and enhance the QoL of caregivers.

Development and Testing of an Electronic Diabetes Diary Integrated With a Hospital Information System for Individuals With Type 2 Diabetes Mellitus: Protocol for a Mixed Methods Study.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is one of the leading noncommunicable diseases that require diabetes self-management (DSM) practices. This study proposes to develop a customized mobile health (mHealth) app integrated with a hospital information system (HIS) to enable real-time, two-way transfer of information between the patient and physician. The captured information in the electronic health record will facilitate physicians to have a chronological account of the patient's diabetes history and enable tweaking of the treatment. OBJECTIVE: The objectives of the study are (1) to develop the HIS-integrated Electronic Diabetes Diary (EDDy) per the end-user expectations at a tertiary care hospital in a south Indian state with a high prevalence of T2DM and (2) to evaluate and test adherence to EDDy in the management of T2DM. METHODS: The study will be carried out in 3 phases. Phase 1 involved in-depth interviews with primary end users to gather information regarding their expectations from the hospital-based EDDy. Phase 2 will use this information to develop a customized mHealth app using an iterative model of software development. Phase 3 will involve a pre- and posttest design; the developed app will be tested among consenting patients, where physicians will receive the patients' data through the HIS-integrated mHealth app. The pre- and posttest values will be analyzed for adherence leading to improvement in patients' self-management of blood glucose, user experience, glycemic control, and clinical utility. RESULTS: Phase 1 was completed on November 28, 2023. Phase 2 commenced in December 2023 and will end in May 2025. Phase 3 will follow afterward. CONCLUSIONS: The proposed app will include a convenient and simple alert system that enables the patient to test glucose values at self-selected intervals, provide grading options to enter diabetic-related complications, enhance patients' knowledge of tracking and managing the complications of diabetes, and help in maintaining the visual representation of glucose values and complications. The simplicity and usability of the modules are its novelty, which may motivate the patients to keep track of their glucose values and help them attain better health outcomes. TRIAL REGISTRATION: Clinical Trial Registry India CTRI/2023/03/051077; http://tinyurl.com/4tau4ndb. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/50732.

Standardizing Minimally Invasive Tissue Sampling of Postmortem Brain Using Bard Monopty Needle in Newborns with Neurological Injury.

INTRODUCTION: Minimally invasive tissue sampling of the brain in newborns using the Bard Monopty needle helps to diagnose various neurological conditions by obtaining relevant brain cores. We designed a modified procedure to provide maximum diagnostic utility in brain tissue biopsies. METHOD: Twenty newborns underwent postmortem minimally invasive tissue sampling of the brain through the anterior fontanelle and posterior approach, using the engraved lines on the needle labeled from mark 0 to 13. The cores were correlated with conventional autopsy findings. RESULTS: Meninges were best obtained at marks 0 and 1 from the anterior fontanelle and mark 1 from posterior fontenelle in 85% of cases. Periventricular brain parenchyma was best obtained from mark 3 and mark 1 from anterior and posterior fontanel, respectively in 90% cases. The sampling success in obtaining brain cores was 100%. DISCUSSION: This modified technique increases the yield of meninges and brain tissue in newborns and aids in diagnosis.

Trends and challenges in electroanalytical biosensing methodologies for infectious viral diseases.

Viral pandemic diseases have disruptive global consequences leading to millions of deaths and a severe impact on the global economy. Inadequate preventative protocols have led to an overwhelming demand for intensive care leading to uncontrollable burdens and even breakdown of healthcare sectors across many countries. The rapid detection of viral disease helps in the understanding of the relevant intricacies, helping to tackle infection with improved guidelines. Portable biosensor devices offer promising solutions by facilitating on-site detection of viral pathogens. This review summarizes the latest innovative strategies reported using electroanalytical methods for the screening of viral antigens. The structural components of viruses and their categories are presented followed by the various recognition elements and transduction techniques involved in biosensors. Core sections focus on biosensors reported for viral genomic detection(DNA and RNA) and antigenic capsid protein. Strategies for addressing the challenges of electroanalytical biosensing of viral components are also presented. The advantages, and disadvantages of biorecognition elements and nanozymes for the detection of viral disease are highlighted. Such technical insights will help researchers working in chemistry, and biochemistry as well as clinicians working in medical diagnostics.

Exploring the Effect of Trataka (A Yogic Cleansing Technique) and Cold Eye Pack on Eye Strain During COVID-19 Pandemic: A Randomized Three-Arm Trial.

Objective: Computer vision syndrome is a major global public health concern affecting >60 million individuals globally. Yoga and naturopathy practices can reduce visual fatigue and strain. The present study attempted to explore the effect of trataka that is, a yogic cleansing technique and cold eye pack on visual strain and fatigue. Subjects: Three hundred volunteers from an IT company were recruited following inclusion and exclusion criteria. Intervention: The subjects were randomly distributed in three groups, that is, trataka, cold eye pack, and waitlist control group with an allocation ratio of 1:1:1. Outcome measure: Visual Fatigue Scale and Visual symptoms checklist (VSC) was administered at baseline and end of 2 weeks. Repeated measures analysis of variance (RM-ANOVA) with Bonferroni corrections was used to test the difference across the groups. Results: All the variables were similar at the baseline among the groups. Significant changes in the within-group analysis occurred in both the trataka and cold eye pack groups. The RM-ANOVA revealed significant differences in the VAS and VSC (p = 0.001) and the post hoc analysis suggested that there were significant differences in both the trataka and cold eye pack group when compared with the control group (p = 0.001); however, there was no differences between the trataka and cold eye pack group in both the scales (p = 1). Conclusions: The results of the present study suggest that a trataka and cold eye pack for 14 days improves self-rated visual strain and fatigue among IT professionals with computer vision syndrome. Clinical Trial registration number: CTRI/2020/11/029003.

Oblique direction reach test: evaluating psychometric properties in stroke population.

Background: Post-stroke individuals are observed to have reduced limits of stability (LOS) in all directions. Functional activities are rarely performed in pure cardinal planes; instead, they are most likely to be performed in an oblique direction. Existing tools are either expensive or sophisticated to assess the LOS in an oblique direction. Therefore, this study's primary objective is to evaluate the intra-rater, inter-rater reliability, and validity of the oblique direction reach test (ODRT) among stroke subjects. Materials & Methods: A total of 96 first-time stroke patients with age, gender, height, and weight-matched healthy controls aged 18-80 years were recruited for the study. Oblique, forward, and lateral reach distances were assessed using the standard procedure of ODRT, Functional Reach Test (FRT), and Lateral Reach Test (LRT), respectively. Validity was tested by correlating the ODRT distance with the Berg Balance Scale (BBS) Score using Spearman's rank correlation coefficient. Intraclass correlation coefficients (ICCs) and Bland Altman analysis were used to establish inter-rater reliability. ICCs were used to find intra-rater reliability. The Mann-Whitney U test was used to establish the mean difference of the FRT, LRT, and ODRT. Spearman's rank correlation coefficient and linear regression were used to correlate the distance of FRT and LRT with ODRT. Results: A high concurrent validity was found between BBS and ODRT with an r-value of 0.905 (p < 0.001). Inter-rater reliability was high with an ICC of 0.997 (95% CI [0.996-0.998]), and intra-rater reliability was highly significant with an ICC of 0.996 (95% CI [0.994-0.998]). The stroke subjects reached a significantly shorter distance than healthy individuals in FRT, ODRT, and LRT. ODRT was highly correlated with FRT (r = 0.985) and LRT (r = 0.978) (p < 0.001) and had an R2 = 0.987. Conclusion: ODRT is a highly valid and reliable tool that can be used to evaluate balance in stroke patients. Individuals who reached less in the forward and lateral directions showed reduced reach distance in the oblique direction.

Navigating Tumour Microenvironment and Wnt Signalling Crosstalk: Implications for Advanced Cancer Therapeutics.

Cancer therapeutics face significant challenges due to drug resistance and tumour recurrence. The tumour microenvironment (TME) is a crucial contributor and essential hallmark of cancer. It encompasses various components surrounding the tumour, including intercellular elements, immune system cells, the vascular system, stem cells, and extracellular matrices, all of which play critical roles in tumour progression, epithelial-mesenchymal transition, metastasis, drug resistance, and relapse. These components interact with multiple signalling pathways, positively or negatively influencing cell growth. Abnormal regulation of the Wnt signalling pathway has been observed in tumorigenesis and contributes to tumour growth. A comprehensive understanding and characterisation of how different cells within the TME communicate through signalling pathways is vital. This review aims to explore the intricate and dynamic interactions, expressions, and alterations of TME components and the Wnt signalling pathway, offering valuable insights into the development of therapeutic applications.

Understanding the economic burden of cancer care and the utility of health insurance: An exploratory cross-sectional study in a tertiary care hospital in Karnataka, India.

Background: Families and caregivers of cancer patients experience significant financial challenges associated with out-of-pocket (OOP) expenditures. Objectives: This study aims to assess the OOP expenditure and its impact on the livelihood of patients and families, associated with receiving cancer care from a teaching hospital in Karnataka. Materials and Methods: It focuses on understanding health insurance use and its effects on OOP expenses for cancer care based on data obtained from 271 patients receiving treatment for more than 6 months. A structured questionnaire was developed and used for data collection and focused on obtaining direct costs such as consultation fees, surgery costs, and radiotherapy costs and indirect costs such as travel expenses, food costs, and patient income loss, as well as questions that measure the impact of the financial burden on patients and their associated livelihood. Results: In the present study, the median cost of OOP expense incurred for cancer treatment is estimated to be 3.10 lakh Indian rupees. It was also found that patients enrolled in public health insurance schemes, especially Ayushman Bharath-Arogya Karnataka have lesser OOP expenditure than those with either private health insurance or no health insurance. Conclusions: This indicates the need for effective implementation of various public health insurance schemes and their ability to protect patients from huge OOP expenses and related financial risks.

Nutrition morbidity interactions in pre-school children.

Background & objectives: Research studies in the 1970s reported that in pre-school children, undernutrition increased the risk of infections and infections aggravated undernutrition. Over decades, there has been a reduction in prevalence of undernutrition and improvement in access to healthcare for treatment of infections. A mixed longitudinal study was undertaken to assess whether over time there were any changes from the earlier reported effect of undernutrition prior to infection on the risk of morbidity and effect of morbidity on nutritional status in pre-school children. Methods: Pre-school (0-59 months of age) children from urban low- and middle-income families whose parents were willing to allow their participation in the study were enrolled. Information on sociodemographic profile of the families was collected at enrolment. Weight of all children and length in infants were recorded every month; length/height in children 12-59 months of age was recorded once in three months. Morbidity information was collected through fortnightly visits. Results: 3888 pre-school children were followed up in 74636 home visits. Among these children, underweight and wasting were associated with a small increase in risk of infections. The odds ratio for risk of infection for underweight children was 1.09 (95% CI: 1.02 to 1.16) and for wasting was 1.18 (95% CI: 1.08 to 1.29). The deterioration in Z scores for weight-for-age and body mass index-for-age in children during illness and convalescence was small but significant (P<0.001). Interpretation & conclusions: The increased risk of infections in undernourished children living in overcrowded tenements in areas with poor environmental hygiene was not significant, perhaps because the risk of infection in normally nourished children was also high. The deterioration in nutritional status following infection was small because of the ready access to and utilization of health and nutrition care.

E3-ubiquitin ligase, FBXW7 regulates mitotic progression by targeting BubR1 for ubiquitin-mediated degradation.

Faithful chromosome segregation requires correct attachment of kinetochores with the spindle microtubules. Erroneously-attached kinetochores recruit proteins to activate Spindle assembly checkpoint (SAC), which senses the errors and signals cells to delay anaphase progression for error correction. Temporal control of the levels of SAC activating-proteins is critical for checkpoint activation and silencing, but its mechanism is not fully understood. Here, we show that E3 ubiquitin ligase, SCF-FBXW7 targets BubR1 for ubiquitin-mediated degradation and thereby controls SAC in human cells. Depletion of FBXW7 results in prolonged metaphase arrest with increased stabilization of BubR1 at kinetochores. Similar kinetochore stabilization is also observed for BubR1-interacting protein, CENP-E. FBXW7 induced ubiquitination of both BubR1 and the BubR1-interacting kinetochore-targeting domain of CENP-E, but CENP-E domain degradation is dependent on BubR1. Interestingly, Cdk1 inhibition disrupts FBXW7-mediated BubR1 targeting and further, phospho-resistant mutation of Cdk1-targeted phosphorylation site, Thr 620 impairs BubR1-FBXW7 interaction and FBXW7-mediated BubR1 ubiquitination, supporting its role as a phosphodegron for FBXW7. The results demonstrate SCF-FBXW7 as a key regulator of spindle assembly checkpoint that controls stability of BubR1 and its associated CENP-E at kinetochores. They also support that upstream Cdk1 specific BubR1 phosphorylation signals the ligase to activate the process.

The role of Wnt signaling in mesenchymal stromal cell-driven angiogenesis.

Development, growth, and remodeling of blood vessels occur through an intricate process involving cell differentiation, proliferation, and rearrangement by cell migration under the direction of various signaling pathways. Recent reports highlight that resident and exogenous mesenchymal stromal cells (MSCs) have the potential to regulate the neovascularization process through paracrine secretion of proangiogenic factors. Recent research has established that the vasculogenic potential of MSCs is regulated by several signaling pathways, including the Wnt signaling pathway, and their interplay. These findings emphasize the complex nature of the vasculogenic process and underscore the importance of understanding the underlying molecular mechanisms for the development of effective cell-based therapies in regenerative medicine. This review provides an updated briefing on the canonical and non-canonical Wnt signaling pathways and summarizes the recent reports of both in vitro and in vivo studies with the involvement of MSCs of various sources in the vasculogenic process mediated by Wnt signaling pathways. Here we outline the current understanding of the plausible role of the Wnt signaling pathway, specifically in MSC-regulated angiogenesis.

Use of sensitivity-enhanced nuclear magnetic resonance spectroscopy equipped with a 1.7-mm cryogenically cooled micro-coil probe in identifying human sperm intracellular metabolites.

CONTEXT: The clinical value of human sperm metabolites has not been established due to the technical complexity in detecting these metabolites when sperm numbers are low. AIMS: To detect endogenous intracellular metabolites in fresh and post-thaw human spermatozoa using 800MHz nuclear magnetic resonance (NMR) spectroscopy equipped with a 1.7-mm cryo-probe. METHODS: Processed spermatozoa from 25 normozoospermic ejaculates were subjected to extraction of intracellular metabolites and then profiled by sensitivity-enhanced NMR spectroscopy equipped with a 1.7-mm cryogenically cooled micro-coil probe. In parallel, some of the processed sperm fractions were subjected to freeze-thawing and were then analysed for intracellular metabolites. KEY RESULTS: Twenty-three metabolites were profiled from only 1.25million sperm cells. Comparison of the metabolomic signature of pre-freeze and post-thaw sperm cells did not show significant changes in the levels of metabolites. CONCLUSIONS: Sensitivity-enhanced NMR spectroscopy equipped with a 1.7-mm cryogenically cooled micro-coil probe is a potential tool for identifying intracellular metabolites when sperm number is low. IMPLICATIONS: Use of sensitivity-enhanced NMR spectroscopy opens up the opportunity to test for endogenous metabolites in samples with a limited number of spermatozoa, to understand the patho-physiology of infertility.

Impact of prepubertal bovine ovarian tissue pre-freeze holding duration on follicle quality.

Ovarian tissue cryopreservation prior to gonadotoxic treatment is the only recommended option for fertility preservation in prepubertal girls. Due to the technical complexity of this technique, limited number of centres across the world are equipped to offer the facility. Hence, the retrieved ovarian tissue needs to be maintained at hypothermic temperature (4 °C) for long time during shipment. The time taken between tissue retrieval and cryopreservation could influence the functionality of cells during fertility restoration. This study explored the tissue integrity and follicle quality of ovarian cortical slices subjected to pre-freeze holding for various time durations in vitro. Prepubertal bovine ovarian tissue from < 12 months old animals were handled at hypothermic holding (4 °C) for 0, 24, 48 and 72 h. The tissues were assessed for follicle viability through confocal analysis of live-dead labelled samples, and follicle quality and tissue integrity through histology. Results have shown that follicle viability, and overall follicle quality were not significantly affected at the end of 72 h hypothermic holding. Though, the observation reassures extended hypothermic holding prior to freezing, findings need to be validated in human tissue prior to use in clinical fertility preservation programs.