Genistein effect in cultured ovine ovarian granulosa cells.

Genistein, an isoflavone has the potential to mimic, augment, or dysregulate the steroid hormone production pathways. We hypothesized that genistein affects the granulosa cell (GCs) functions through a series of biochemical, molecular, and genomic cascades. The present study was conducted to evaluate the impact of genistein exposure on GCs viability, apoptosis, and steroidogenesis. The present study involved 3/5 days of exposure to genistein on GCs collected from abattoir-derived ovine ovaries at doses of 0, 1, 10, 25, 50, and 100 µM. The harvested GCs were used for growth, cytotoxicity, and gene expression studies related to apoptosis, growth, and steroidogenesis. We observed that genistein had both stimulatory at 10 and 25 µM levels as well as inhibitory effects at 50 and 100 µM levels on the growth and proliferation of GCs. Genistein significantly decreased the levels of 17ß-estradiol at higher exposure (50 and 100 µM), whereas the progesterone level increased significantly as the genistein exposure increased. Additionally, genistein could also alter the mRNA expression of the steroidogenic receptor, enzymes, proteins, and growth-related genes suggesting that genistein could potentially alter the steroidogenic pathways. We conclude that genistein can interfere with cell survival and steroidogenesis by exhibiting a dose-dependent biphasic response on the viability, growth-related parameters, and the synthesis of 17ß-estradiol in the cultured GCs.

From Gut to Hormones: Unraveling the Role of Gut Microbiota in (Phyto)Estrogen Modulation in Health and Disease.

The human gut microbiota regulates estrogen metabolism through the "estrobolome," the collection of bacterial genes that encode enzymes like ß-glucuronidases and ß-glucosidases. These enzymes deconjugate and reactivate estrogen, influencing circulating levels. The estrobolome mediates the enterohepatic circulation and bioavailability of estrogen. Alterations in gut microbiota composition and estrobolome function have been associated with estrogen-related diseases like breast cancer, enometrial cancer, and polycystic ovarian syndrome (PCOS). This is likely due to dysregulated estrogen signaling partly contributed by the microbial impacts on estrogen metabolism. Dietary phytoestrogens also undergo bacterial metabolism into active metabolites like equol, which binds estrogen receptors and exhibits higher estrogenic potency than its precursor daidzein. However, the ability to produce equol varies across populations, depending on the presence of specific gut microbes. Characterizing the estrobolome and equol-producing genes across populations can provide microbiome-based biomarkers. Further research is needed to investigate specific components of the estrobolome, phytoestrogen-microbiota interactions, and mechanisms linking dysbiosis to estrogen-related pathology. However, current evidence suggests that the gut microbiota is an integral regulator of estrogen status with clinical relevance to women's health and hormonal disorders.

Efficacy and safety of short- vs. standard-course antibiotics for culture-negative neonatal sepsis: a systematic review and meta-analysis.

OBJECTIVES: To conduct a systematic review and meta-analysis of evidence from randomized controlled trials (RCTs) comparing a short course of antibiotics (2-4 days), to a standard course (5-7 days), for the treatment of culture-negative neonatal sepsis. METHODS: Relevant databases were searched for RCTs comparing short- vs. standard-course of antibiotics for culture-negative sepsis. The primary outcomes were mortality and treatment failure, defined as the reappearance of clinical signs suggestive of sepsis within 7 days of stoppage of antibiotics. Secondary outcomes included neurological impairment, duration of hospital stay, need for oxygen, respiratory support and double-volume exchange transfusion (DVET). RESULTS: Seven RCTs were included in the review with 729 neonates >30 weeks gestational age at birth. No mortality occurred in either of the groups (2 studies; 276 neonates). Treatment failure rates were similar in the short- and standard-course antibiotic groups [7 studies; 729 neonates; risk ratio (RR) = 1.01; 95% confidence interval (CI), 0.55 to 1.86; very low certainty]. The short course of antibiotics resulted in a shorter hospital stay [3 studies; 293 neonates; mean difference (MD), -2.46 days; 95% CI, -3.16 to -1.75]. There was no difference in the need for oxygen supplementation (2 studies; 258 neonates; RR, 1.40; 95% CI, 0.40 to 4.91), any respiratory support (2 studies; 258 neonates; RR, 1.04; 95% CI, 0.92 to 1.17) or DVET (2 studies; 258 neonates; RR, 1.29; 95% CI, 0.56 to 2.95). CONCLUSION: Very-low certainty evidence suggests that a short antibiotic course, compared to a standard course, does not affect treatment failure rates in culture-negative neonatal sepsis. There is a need for well-designed RCTs powered enough to assess critical outcomes such as mortality and neurological sequelae to generate stronger evidence and inform guidelines. PROSPERO REGISTRATION NUMBER: CRD42023437199.

Prolonged antibiotic usage has been associated with increased mortality and morbidity in neonates. The standard practice in culture-negative neonatal sepsis has been to administer antibiotics for 5­7 days, based on expert consensus. In this systematic review, a short course of antibiotics (2­4 days), in comparison to a standard course (5­7 days), did not affect the treatment failure rates in culture-negative neonatal sepsis. However, the certainty of evidence was too low to make robust conclusions. There is a need for well-designed large trials to generate stronger evidence and inform guidelines.

Effect of Splint Application on the Functional Duration of Peripheral Intravenous Cannulation in Neonates: A Systematic Review and Meta-analysis.

BACKGROUND: The application of splints is one of the most used methods to prolong the life span of peripheral intravenous cannulation (PIVC). OBJECTIVE: To assess the effect of splint application on the functional duration of PIVC in neonates. METHODS: This systematic review and meta-analysis identified, appraised, and synthesized available evidence from randomized and quasi-randomized controlled trials (RCT) related to the effects of splint application compared to no splinting on the functional duration of PIVC and its associated complications in term and preterm neonates. Data were pooled using RevMan 5.4. The quality of evidence for predefined outcomes was analyzed by GRADE. RESULTS: Available evidence (5 RCTs, 826 neonates) showed a significantly lesser functional duration of PIVC in the splint group compared to no-splint [Mean Difference (MD) 95% Confidence Interval (CI) -3.07 (-5.63, -0.51); Low Certainty of Evidence (CoE)]. On gestation-based subgroup analysis, PIVC duration remained significantly lesser in the splint group in preterm neonates [MD (95% CI), -5.09 (-9.53, -0.65), 2 studies, n = 220; Low CoE], whereas it was comparable in the term neonates [MD (95% CI), 3.92 (-4.27, 12.10), 2 studies, n = 89; Very low CoE]. The overall complications were comparable between the groups [Risk Ratio (95% CI), 1.02 (1.00, 1.05), 5 studies, n = 826; Very low CoE]. CONCLUSION: Based on the very low to low CoE found in this systematic review, it is not possible to recommend or refute splint application in neonates. Further well-designed RCTs are needed.

Bacillus subtilis NBRI-W9 simultaneously activates SAR and ISR against Fusarium chlamydosporum NBRI-FOL7 to increase wilt resistance in tomato.

AIMS: The study aimed to determine the pathogenicity of Fusarium species currently prevalent in tomato fields having history of chemical fungicide applications and determine the bio-efficacy of Bacillus subtilis NBRI-W9 as a potent biological control agent. METHODS AND RESULTS: Fusarium was isolated from surface-sterilized infected tomato plants collected from fields. Pathogenicity of 30 Fusarium isolates was determined by in vitro and in vivo assays. Following Koch's postulates, F. chlamydosporum (FOL7) was identified as a virulent pathogen. The biological control of FOL 7 by B. subtilis NBRI-W9 (W9) and the colonization potential of W9 were established using spontaneous rifampicin-resistant mutants. W9 showed 82% inhibition of FOL7 on a dual-culture plate and colonization levels in tomato plants of ∼5.5, ∼3.3, and ∼2.2 log10 CFU/g in root, stem, and leaf tissue, respectively. Antagonistic activity was shown by scanning electron microscopy (SEM) and cell-wall-degradative enzymes. W9 reduced FOL7 infection in net-house and field experiments by 60% and 41%, respectively. Biochemical investigation, defence enzymes, defence gene expression analysis, SEM, and field studies provide evidence of hyperparasitism and induced resistance as the mode of biological control. The study also demonstrates that the potent biocontrol agent W9, isolated from Piper, can colonize tomato plants, control fungal disease by inducing induced systemic resistance (ISR) and systemic acquired resistance (SAR) simultaneously, and increase crop yield by 21.58% under field conditions. CONCLUSIONS: This study concludes that F. chlamydosporum (NBRI-FOL7) is a potent, fungicide-resistant pathogen causing wilt in tomatoes. NBRI-W9 controlled FOL7 through mycoparasitism and simultaneously activated ISR and SAR in plants, providing an attractive tool for disease control that acts at multiple levels.

A critical assessment on scalable technologies using high solids loadings in lignocellulose biorefinery: challenges and solutions.

The pretreatment and the enzymatic saccharification are the key steps in the extraction of fermentable sugars for further valorization of lignocellulosic biomass (LCB) to biofuels and value-added products via biochemical and/or chemical conversion routes. Due to low density and high-water absorption capacity of LCB, the large volume of water is required for its processing. Integration of pretreatment, saccharification, and co-fermentation has succeeded and well-reported in the literature. However, there are only few reports on extraction of fermentable sugars from LCB with high biomass loading (>10% Total solids-TS) feasible to industrial reality. Furthermore, the development of enzymatic cocktails can overcome technology hurdles with high biomass loading. Hence, a better understanding of constraints involved in the development of technology with high biomass loading can result in an economical and efficient yield of fermentable sugars for the production of biofuels and bio-chemicals with viable titer, rate, and yield (TRY) at industrial scale. The present review aims to provide a critical assessment on the production of fermentable sugars from lignocelluloses with high solid biomass loading. The impact of inhibitors produced during both pretreatment and saccharification has been elucidated. Moreover, the limitations imposed by high solid loading on efficient mass transfer during saccharification process have been elaborated.

Endophytic biofungicide Bacillus subtilis (NBRI-W9) reshapes the metabolic homeostasis disrupted by the chemical fungicide, propiconazole in tomato plants to provide sustainable immunity against non-target bacterial pathogens.

Chemical and microbial fungicides (Bio/fungicide) act differentially on plant systems. The present work assessed the metabolic profile of tomato plants vis-a-vis endophytic diversity after spraying of Propiconazole (PCZ) and endophytic biofungicide Bacillus subtilis (W9). Bio/fungicides were sprayed on tomato plants and evaluated for phenotypic, biochemical, and metabolic profiles after one week. In W9 treatment, a significant increase in relative abundance of several metabolites was observed including sugars, sugar alcohols, fatty-acids, organic-acids, and amino-acids. Polysaccharides and fatty acids showed a significant positive correlation with Rhizobiales, Burkholderiales, Bacillales, and Lactobacillales, respectively (p < 0.05). The PCZ and W9 treated plant's metabolic status significantly affected their resistance to non-target, bacterial pathogen P. syringae. Compared to PCZ and control, W9 treatment reduced the ROS deposition and expression of antioxidants gene GPx, PO (~0.1-1.7fold). It enhanced the genes related to the Phenylpropanoid pathway (∼1.6-5.2 fold), PR protein (~1.2-3.4 fold), and JA biosynthesis (~1.7-4.3 fold), resulting in reduced disease incidence. The results provide novel insights into the effects of endophytic biofungicide and chemical fungicides on the plant's metabolic status, its relation to the endophytes, and role in altering the plant's immune system.

Development and content validation of yoga module for the mental health of NICU mothers.

BACKGROUND: Postnatal yoga has been found to be effective for maternal mental health management. But a validated yoga module for the mental health of early postpartum mothers with infants admitted to the Neonatal Intensive Care Unit (NICU) is lacking. AIM: To design and validate a yoga module for the mental health of early postpartum mothers having infants admitted to the NICU. MATERIALS AND METHODS: First phase: A yoga module was designed through a review of published research articles and yogic texts for NICU mothers. Second phase: thirty-eight yoga experts validated the yoga module. Lawshe's formula was used to calculate each item's content validity ratio (CVR). The intra-class correlation coefficient was determined for the validated yoga module. Third phase: The validated yoga module was pilot-tested with a sample size of 20 NICU mothers. RESULTS: Thirty-eight yoga experts validated the yoga module for NICU mothers. Thirteen practices included in the module indicated good content validity (cutoff value: 0.316). The module's content validity index (CVI) and intra-class correlation coefficient were 0.672 and 0.924, respectively. Ten days of practicing the yoga module resulted in a significant reduction in maternal stress levels in the yoga group (p < 0.001) compared to the control group (p = 0.427). CONCLUSION: The present study suggests good content validity of the yoga module for the mental health of NICU mothers. However, future randomized controlled trials must be carried out to determine both the feasibility and clinical efficacy of the Yoga Module for NICU mothers.

Hypoglycemia and anxiolysis mediated by levofloxacin treatment in diabetic rats.

Purpose: The present study was designed to determine the effect of levofloxacin (LVX) treatment on the blood glucose level, insulin sensitivity, anxiety level, nitrite and MDA level of STZ induced diabetic rats. Methods: Wistar rats were used in the present study. The rats were made diabetic by the administration of single dose of STZ (45 mg/kg, i.p.) and NAD (50 mg/kg, i.p.). The rats with the blood glucose level greater than 200 mg/dl were considered as diabetic (confirmed at day-3 of STZ-NAD administration). The non-diabetic rats were considered as control and received saline.Diabetic rats received metformin (50 mg/kg, p.o.) and LVX (20, 25, 30 and 35 mg/kg, i.p.) daily for 14 days (starting from the day at which STZ was injected). Following administration on 14th day,the blood sample was collected and the rats were subjected to behavioral assays for the determination of locomotor activity and anxiety level. Plasma was separated and used for the estimation ofnitrite and malondialdehyde (MDA)level. On 15th day OGTT was performed in the overnight fasted rats for the assessment of insulin sensitivity. Results: The results obtained suggested that the administration of STZ-NAD induced the hyperglycemia at day-3 of administration. Diabetic rats displayed the significant increase in blood glucose, anxiety related behavior, MDA level while significant decrease in the insulin sensitivity and plasma nitrite level. Daily administration of metformin to the diabetic rats decreased the blood glucose level, increased the time spent at the center of open field, reversed the anxiety related behavior in LDT and EPM, did not affect the plasma nitrite level, decreased the plasma MDA level, decreased the fasting glucose level and AUC in OGTT assay. LVX (30 and 35 mg/kg) treatment significantly decreased the blood glucose level of diabetic rats. LVX (20, 25 and 30 mg/kg) treatment significantly decreased the number of square crossing while LVX (20, 25, 30 and 35) treatment significantly increased the time spent at the center of the field by the diabetic rats. LVX (20 and 35 mg/kg) treatment significantly reversed the STZ induced anxiety in LDT while LVX (20, 30 and 35 mg/kg) treatment significantly reversed the STZ induced anxiety in EPM test. LVX (20, 25 and 35 mg/kg) treatment significantly increased the plasma nitrite level and LVX (20-35 mg/kg) treatment significantly decreased the MDA level of diabetic rats. Further only LVX (35 mg/kg) treatment significantly decreased the fasting glucose level and increased the AUC of diabetic rats. Conclusion: In conclusion, STZ-NAD administration increased the blood glucose level, anxiety related behavior, decreased the plasma nitrite and increased the MDA level. LVX administration potentiated the diabetogenic effects of STZ-NAD in rats. Daily administration of LVX decreased the blood glucose level of diabetic rats. LVX administration alleviated the STZ induced anxiety in OFT, LDT and EPM test. LVX administration increased the plasma nitrite level and decreased the lipid peroxidation in diabetic rats. Supplementary Information: The online version contains supplementary material available at 10.1007/s40200-023-01234-0.

Health Emergency Risk Management in World Health Organization - South-East Asia Region during 2014-2023: synthesis of experiences.

Health Emergency Risk Management (ERM) has become increasingly critical on the global stage, prompted by the escalating frequency and severity of natural disasters and disease outbreaks. This paper offers a comprehensive synthesis of the World Health Organization's (WHO) experiences in the South-East Asia Region during the period 2014-2023, shedding light on its efforts to manage health emergencies and enhance resilience. The South-East Asia Region's unique environmental and economic diversity exposes it to significant health risks, including emerging infectious diseases and their implications for development, particularly in low-income countries. Here we document the transition from reactive emergency responses to proactive preparedness, catalyzed by prioritizing ERM as one of the regional flagship priorities in 2014. Key components of this initiative included capacity-building, the establishment of the South-East Asia Regional Health Emergency Fund (SEARHEF), and the implementation of the International Health Regulations (IHR 2005). This synthesis highlights the region's achievements in event reporting, development of national actions plan, successful Early Warning, Alert, and Response System (EWARS) implementation, and improvements in core capacities under IHR (2005). It also underscores the challenges associated with cross-border data sharing and regional collaboration that could strengthen ERM and enhance readiness for effective synergistic response.

Advancing Universal Health Coverage in the WHO South-East Asia Region with a focus on Human Resources for Health.

The identification in 2014 of Universal Health Coverage, including focus on human resources for health, as a flagship priority for the WHO South-East Asia Region marked critical departure from the prior period of the Millennium Development Goals. The last decade witnessed strong political commitment and action to advance UHC across the Region. At regional level, UHC service coverage index improved from 47 in 2010 to 62 in 2021. Improved availability of human resources for health has been an important contributor, with the regional average of doctors, nurses and midwives increasing by approximately a third between 2014 and 2020. Progress on financial protection has been mixed: proportion of population impoverished declined significantly but catastrophic expenditure did not reduce. Despite important gains, progress is insufficient to achieve UHC targets by 2030. Covid-19 pandemic and subsequent economic challenges have created further urgency to accelerate progress towards UHC, with attention to strengthening primary health care.

T-Piece resuscitator versus self-inflating bag for delivery room resuscitation in preterm neonates: a randomized controlled trial.

The establishment of adequate ventilation is the cornerstone of neonatal resuscitation in the delivery room (DR). This parallel-group, accessor-blinded randomized controlled trial compared the changes in peripheral oxygen saturation (SpO2), heart rate (HR), and cerebral regional oxygen saturation (crSO2) with the use of a T-piece resuscitator (TPR) versus self-inflating bag (SIB) as a mode of providing positive pressure ventilation (PPV) during DR resuscitation in preterm neonates. Seventy-two preterm neonates were randomly allocated to receive PPV with TPR (n = 36) or SIB (n = 36). The primary outcome was SpO2 (%) at 5 min. The secondary outcomes included the time to achieve a SpO2 ≥ 80% and > 85%, HR > 100/min, fractional-inspired oxygen (FiO2) requirement, minute-specific SpO2, HR and FiO2 trends for the first 5 min of life, need for DR-intubation, crSO2, need and duration of respiratory support, and other in-hospital morbidities. Mean SpO2 at 5 min was 74.5 ± 17.8% and 69.4 ± 22.4%, in TPR and SIB groups, respectively [Mean difference, 95% Confidence Interval 5.08 (-4.41, 14.58); p = 0.289]. No difference was observed in the time to achieve a SpO2 ≥ 80% and > 85%, HR > 100/min, the requirement of FiO2, DR-intubation, and the need and duration of respiratory support. There was no significant difference in the minute-specific SpO2, HR, and FiO2 requirements for the first 5 min. CrSO2 (%) at one hour was lower by 5% in the TPR group compared to SIB; p = 0.03. Other complications were comparable. CONCLUSIONS: TPR and SIB resulted in comparable SpO2 at 5 min along with similar minute-specific SpO2, HR, and FiO2 trends. CLINICAL TRIAL REGISTRATION: Clinical trial registry of India, Registration no: CTRI/2021/10/037384, Registered prospectively on: 20/10/2021, https://ctri.icmr.org.in/ . WHAT IS KNOWN: • Compared to self-inflating bags (SIB), T-piece resuscitators (TPR) provide more consistent inflation pressure and tidal volume as shown in animal and bench studies. • There is no strong recommendation for one device over the other in view of low certainty evidence. WHAT IS NEW: • TPR and SIB resulted in comparable peripheral oxygen saturation (SpO2) at 5 min along with similar minute-specific SpO2, heart rate, and fractional-inspired oxygen requirement trends. • Short-term complications and mortality rates were comparable with both devices.

Selection of Highly Specific DNA Aptamer for the Development of QCM-Based Arsenic Sensor.

Heavy metal arsenic is a water pollutant that affects millions of lives worldwide. A novel aptamer candidate for specific and sensitive arsenic detection was identified using Graphene Oxide-SELEX (GO-SELEX). Eleven rounds of GO-SELEX were performed to screen As(III) specific sequences. The selected aptamer sequences were evaluated for their binding affinity. The dissociation constant of the best aptamer candidate, As-06 was estimated by fluorescence recovery upon target addition, and it was found to be 8.15 nM. A QCM-based biosensing platform was designed based on the target-triggered release of aptamer from the QCM electrode. An rGO-SWCNT nanocomposite was adsorbed on the gold surface, and the single-stranded probe was stacked on the rGO-CNT layer. Upon addition of the target to the solution, a concentration-dependent release of the ssDNA probe was observed and recorded as the change in the electrode frequency. The developed QCM sensor showed a dynamic linear range from 10 nM to 100 nM and a low detection limit of 8.6 nM. The sensor exhibited excellent selectivity when challenged with common interfering anions and cations.

Pain Assessment and Management Practices via Education & Reinforcement (PAMPER): A Quality Improvement Initiative.

OBJECTIVES: To establish neonatal pain management practices as an essential developmental supportive measure at a tertiary care unit. METHODS: This quality improvement initiative was conducted as per Point-of-Care-Quality-Improvement Model over 6 mo, followed by 2 y of sustenance phase at a Neonatal Intensive Care Unit (NICU) in northern India. Infants of birth weight ≤1800 g were included and assessed for pain. Pain Assessment and Management Practices via Education and Reinforcement (PAMPER) group was created by resident doctors and nursing staff. The Premature Infant Pain Profile score was used for the assessment of pain. Limiting factors were analyzed using a fishbone diagram and interventions were done in multiple Plan-Do-Study-Act cycles. RESULTS: At the end of interventions, 100% of procedures were assessed for pain. The mean (standard deviation) documented pain score for the first seven days was reduced from 12.8 (0.3) in the baseline phase to 7 (2.5). These interventions helped to sustain the practice in >70% of infants in the next 2 y. CONCLUSIONS: Low-cost interventions improved the pain assessment and management policy of authors' NICU with the establishment of a standard protocol. Audits and reinforcement at regular intervals helped in its long-term sustenance.