Uncovering Novel Protein Partners of Inducible Nitric Oxide Synthase in Human Testis.

Peroxidative damage to human spermatozoa has been shown to be the primary cause of male infertility. The possible role of nitric oxide (NO) in affecting sperm motility, capacitation, and acrosome reaction has been reported, too. The overproduction of NO by the enzyme inducible nitric oxide synthase (iNOS) could be responsible as it has been implicated in the pathogenesis of many diseases. There have been many studies on regulating iNOS function in various tissues, especially by protein-protein interaction; however, no study has looked for iNOS-interacting proteins in the human testis. Here, we have reported the identification of two proteins that interact with iNOS. We initially undertook a popular yeast two-hybrid assay to screen a human testis cDNA library in yeast using an iNOS-peptide fragment (amino acids 181-335) as bait. We verified our data using the mammalian chemiluminescent co-IP method; first, employing the same peptide and, then, a full-length protein co-expressed in HEK293 cells in addition to the candidate protein. In both cases, these two protein partners of iNOS were revealed: (a) sperm acrosome-associated 7 protein and (b) retinoblastoma tumor-suppressor binding protein.

Evaluation of a disability-inclusive ultra-poor graduation programme in Uganda: study protocol for a cluster-randomised controlled trial with process evaluation.

BACKGROUND: There is little evidence on how to support ultra-poor people with disabilities to adopt sustainable livelihoods. The Disability-Inclusive Graduation (DIG) programme targets ultra-poor people with disabilities and/or women living in rural Uganda. The programme is an adaptation of an ultra-poor graduation model that has been shown to be effective in many contexts but not evaluated for people with disabilities. METHODS: The DIG programme works with project participants over a period of 18 months. Participants receive unconditional cash transfers for 6 months, training, access to savings-and-loans groups, and a capital asset that forms the basis of their new livelihood. The programme is also adapted to address specific barriers that people with disabilities face. Eligible households are clustered by geographical proximity in order to deliver the intervention. Eligibility is based on household screening to identify the 'ultra-poor' based on proxy means testing-both households with and without people with disabilities are included in the programme. Clusters are randomly selected prior to implementation, resulting in 96 intervention and 89 control clusters. The primary outcome of the trial is per-capita household consumption. Before the start of the intervention, a baseline household survey is conducted (November 2020) among project participants and those not offered the programme, a similar endline survey is conducted with participants with disabilities at the end of programme implementation in July 2022, and a second endline survey for all participants in October 2023. These activities are complemented by a process evaluation to understand DIG programme implementation, mechanisms, and context using complementary qualitative and quantitative methods. Ethical approval for the research has been received from Mildmay Uganda Research Ethics Committee and London School of Hygiene and Tropical Medicine. DISCUSSION: DIG is a promising intervention to evaluate for people with disabilities, adapted to be disability inclusive across programme components through extensive consultations and collaboration, and has proven efficacy at reducing poverty in other marginalised groups. However, evaluating a well-evidenced intervention among a new target group poses ethical considerations. TRIAL REGISTRATION: Registry for International Development Impact Evaluations, RIDIE-STUDY-ID-626008898983a (20/04/22). ISRCTN registry, ISRCTN78592382 . Retrospectively registered on 17/08/2023.

Identifying delays in healthcare seeking and provision: The Three Delays-in-Healthcare and mortality among infants and children aged 1-59 months.

Delays in illness recognition, healthcare seeking, and in the provision of appropriate clinical care are common in resource-limited settings. Our objective was to determine the frequency of delays in the "Three Delays-in-Healthcare", and factors associated with delays, among deceased infants and children in seven countries with high childhood mortality. We conducted a retrospective, descriptive study using data from verbal autopsies and medical records for infants and children aged 1-59 months who died between December 2016 and February 2022 in six sites in sub-Saharan Africa and one in South Asia (Bangladesh) and were enrolled in Child Health and Mortality Prevention Surveillance (CHAMPS). Delays in 1) illness recognition in the home/decision to seek care, 2) transportation to healthcare facilities, and 3) the receipt of clinical care in healthcare facilities were categorized according to the "Three Delays-in-Healthcare". Comparisons in factors associated with delays were made using Chi-square testing. Information was available for 1,326 deaths among infants and under 5 children. The majority had at least one identified delay (n = 854, 64%). Waiting >72 hours after illness recognition to seek health care (n = 422, 32%) was the most common delay. Challenges in obtaining transportation occurred infrequently when seeking care (n = 51, 4%). In healthcare facilities, prescribed medications were sometimes unavailable (n = 102, 8%). Deceased children aged 12-59 months experienced more delay than infants aged 1-11 months (68% vs. 61%, P = 0.018). Delays in seeking clinical care were common among deceased infants and children. Additional study to assess the frequency of delays in seeking clinical care and its provision among children who survive is warranted.

Morpho-molecular investigation of ectoparasitic infestation of companion animals in Sylhet city, Bangladesh.

Ticks (Ixodida) and Fleas (Siphonaptera) are considered among the most important arthropod of public health concern due to their ability to transmit vector-borne pathogens to humans. By sharing a common environment, vector-borne diseases constituted major setbacks to the development of a pet population in Bangladesh. This study aimed to determine companion animal-associated ticks and fleas based on morpho-molecular approaches. Between December 2021 and May 2022, 74 animals (62 cats and 12 dogs) were examined, of which 17 (27.4%) cats and 9 (75.0%) dogs had ectoparasitic infestations, with 35.1% overall prevalence. Morphometrical examination showed the ectoparasites in these animals were Ctenocephalides spp. (flea) and Riphicephalus spp. (tick). Genetic analysis using the mitochondrial markers i.e. Cytochrome c oxidase subunit 1 (cox1) revealed the presence of two flea species i.e., Ctenocephalides canis, Ctenocephalides felis, and one tick species Rhipicephalus sanguineus. Interviews of animal owners indicate that 35.14% of them had no concern about ectoparasitic infestation or ectoparasites-borne diseases. Our results indicated that fleas and ticks were the most common ectoparasites in companion animals of this area. The zoonotic nature of some ectoparasites can be regarded as a public health alert. The findings will assist epidemiologists and policymakers in offering customized guidance for upcoming monitoring and preventive tactics in this area.

Post-mortem investigation of deaths due to pneumonia in children aged 1-59 months in sub-Saharan Africa and South Asia from 2016 to 2022: an observational study.

BACKGROUND: The Child Health and Mortality Prevention Surveillance (CHAMPS) Network programme undertakes post-mortem minimally invasive tissue sampling (MITS), together with collection of ante-mortem clinical information, to investigate causes of childhood deaths across multiple countries. We aimed to evaluate the overall contribution of pneumonia in the causal pathway to death and the causative pathogens of fatal pneumonia in children aged 1-59 months enrolled in the CHAMPS Network. METHODS: In this observational study we analysed deaths occurring between Dec 16, 2016, and Dec 31, 2022, in the CHAMPS Network across six countries in sub-Saharan Africa (Ethiopia, Kenya, Mali, Mozambique, Sierra Leone, and South Africa) and one in South Asia (Bangladesh). A standardised approach of MITS was undertaken on decedents within 24-72 h of death. Diagnostic tests included blood culture, multi-organism targeted nucleic acid amplifications tests (NAATs) of blood and lung tissue, and histopathology examination of various organ tissue samples. An interdisciplinary expert panel at each site reviewed case data to attribute the cause of death and pathogenesis thereof on the basis of WHO-recommended reporting standards. FINDINGS: Pneumonia was attributed in the causal pathway of death in 455 (40·6%) of 1120 decedents, with a median age at death of 9 (IQR 4-19) months. Causative pathogens were identified in 377 (82·9%) of 455 pneumonia deaths, and multiple pathogens were implicated in 218 (57·8%) of 377 deaths. 306 (67·3%) of 455 deaths occurred in the community or within 72 h of hospital admission (presumed to be community-acquired pneumonia), with the leading bacterial pathogens being Streptococcus pneumoniae (108 [35·3%]), Klebsiella pneumoniae (78 [25·5%]), and non-typeable Haemophilus influenzae (37 [12·1%]). 149 (32·7%) deaths occurred 72 h or more after hospital admission (presumed to be hospital-acquired pneumonia), with the most common pathogens being K pneumoniae (64 [43·0%]), Acinetobacter baumannii (19 [12·8%]), S pneumoniae (15 [10·1%]), and Pseudomonas aeruginosa (15 [10·1%]). Overall, viruses were implicated in 145 (31·9%) of 455 pneumonia-related deaths, including 54 (11·9%) of 455 attributed to cytomegalovirus and 29 (6·4%) of 455 attributed to respiratory syncytial virus. INTERPRETATION: Pneumonia contributed to 40·6% of all childhood deaths in this analysis. The use of post-mortem MITS enabled biological ascertainment of the cause of death in the majority (82·9%) of childhood deaths attributed to pneumonia, with more than one pathogen being commonly implicated in the same case. The prominent role of K pneumoniae, non-typable H influenzae, and S pneumoniae highlight the need to review empirical management guidelines for management of very severe pneumonia in low-income and middle-income settings, and the need for research into new or improved vaccines against these pathogens. FUNDING: Bill & Melinda Gates Foundation.

An integrated coastal ecosystem monitoring strategy: Pilot case in Naf-Saint Martin Peninsula, Bangladesh.

Rapid population growth creating an excessive pressure on the marine environment and thus monitoring of marine ecosystem is essential. However, due to high technical and financial involvement, monitoring of coastal ecosystem is always challenging in developing countries. This study aims to develop an integrated coastal ecosystem monitoring system that combines scientific sampling, numerical model simulation and citizen science observations to monitor the coastal ecosystem of Bangladesh. This concept of integrated monitoring approach was piloted from January 2022 to April 2023 at the South East coastal zone of Bangladesh. Scientific sampling and numerical model simulations were performed for temperature and salinity data collection. Citizen science approach was employed to collect data on environmental conditions, fisheries, plankton, other marine resources, and plastic pollution. Numerical model simulations and citizen scientists observations of temperature and salinity showed good agreement with the scientifically collected data. In addition, citizen scientists observations on fisheries, plankton, other marine resources and plastic pollution were also in line with the existing database and previous studies. The proposed integrated monitoring approach presents a viable technique, creating a new avenue for coastal and marine ecosystem monitoring where infrastructural facilities are limited.

Model organism life extending therapeutics modulate diverse nodes in the drug-gene-microbe tripartite human longevity interactome.

Advances in antiaging drug/lead discovery in animal models constitute a large body of literature on novel senotherapeutics and geroprotectives. However, with little direct evidence or mechanism of action in humans-these drugs are utilized as nutraceuticals or repurposed supplements without proper testing directions, appropriate biomarkers, or consistent in-vivo models. In this study, we take previously identified drug candidates that have significant evidence of prolonging lifespan and promoting healthy aging in model organisms, and simulate them in human metabolic interactome networks. Screening for drug-likeness, toxicity, and KEGG network correlation scores, we generated a library of 285 safe and bioavailable compounds. We interrogated this library to present computational modeling-derived estimations of a tripartite interaction map of animal geroprotective compounds in the human molecular interactome extracted from longevity, senescence, and dietary restriction-associated genes. Our findings reflect previous studies in aging-associated metabolic disorders, and predict 25 best-connected drug interactors including Resveratrol, EGCG, Metformin, Trichostatin A, Caffeic Acid and Quercetin as direct modulators of lifespan and healthspan-associated pathways. We further clustered these compounds and the functionally enriched subnetworks therewith to identify longevity-exclusive, senescence-exclusive, pseudo-omniregulators and omniregulators within the set of interactome hub genes. Additionally, serum markers for drug-interactions, and interactions with potentially geroprotective gut microbial species distinguish the current study and present a holistic depiction of optimum gut microbial alteration by candidate drugs. These findings provide a systems level model of animal life-extending therapeutics in human systems, and act as precursors for expediting the ongoing global effort to find effective antiaging pharmacological interventions.Communicated by Ramaswamy H. Sarma.

Warming influences carbon and nitrogen assimilation between a widespread Ericaceous shrub and root-associated fungi.

High-latitude ecosystems are warming faster than other biomes and are often dominated by a ground layer of Ericaceous shrubs, which can respond positively to warming. The carbon-for-nitrogen (C-for-N) exchange between Ericaceous shrubs and root-associated fungi may underlie shrub responses to warming, but has been understudied. In a glasshouse setting, we examined the effects of warming on the C-for-N exchange between the Ericaceous shrub Empetrum nigrum ssp. hermaphroditum and its root-associated fungi. We applied different 13 C and 15 N isotope labels, including a simple organic N form (glycine) and a complex organic N form (moss litter) and quantified their assimilation into soil, plant biomass, and root fungal biomass pools. We found that warming lowered the amount of 13 C partitioned to root-associated fungi per unit of glycine 15 N assimilated by E. nigrum, but only in the short term. By contrast, warming increased the amount of 13 C partitioned to root-associated fungi per unit of moss 15 N assimilated by E. nigrum. Our study suggests that climate warming affects the short-term exchange of C and N between a widespread Ericaceous shrub and root-associated fungi. Furthermore, while most isotope tracing studies use labile N sources, we demonstrate that a ubiquitous recalcitrant N source may produce contrasting results.

Correction: Prevalence of anemia in diabetes mellitus in South Asia: A systematic review and meta-analysis.

[This corrects the article DOI: 10.1371/journal.pone.0285336.].

Factors contributing to antibiotic use among children younger than five years old with fever, acute respiratory illness, and diarrhea in Bangladesh.

INTRODUCTION: Increasing antibiotic usage is a leading health threat that develops antibiotic resistance. The current practice of antibiotic use among under-five children are unavailable in Bangladesh. We aim to identify the factors of antibiotic use among under-five children with infectious diseases. METHODOLOGY: A cross-sectional multiple indicators cluster survey (MICS) was conducted in 2019 across Bangladesh. This survey of 23,099 children under the age of five was randomly selected by using a two-stage stratified sampling method. The first stage involved randomly selecting 32,200 enumeration clusters. In second stage, households where 15-45-years-old women lived were randomly selected from within each cluster. The Poisson regression models were performed to estimate the prevalence ratio (PR). RESULTS: We found 36.7% (8447/23,099) under-five children with infectious diseases. The proportion of antibiotic use was reported as 32.6%. Antibiotic use was associated with wealth (poorest vs. rich adjusted prevalence ratio (APR) = 1.07; 95% CI: 0.94-1.22) and mother's education (pre-primary vs. higher: APR = 1.14; 95% CI: 1.03-1.27). Oral and injectable antibiotics were used in cases of fever (30.5%), diarrhea (4.5%), fever with cough (47.6%). Cotrimoxazole (31.0%) and amoxicillin (29.0%) were consumed for fever with cough while cotrimoxazole (14.0%) and amoxicillin (11.0%) were consumed for fever with diarrhea. They received antibiotics from drug stores (71.9%) without prescription and private healthcare (52.1%). CONCLUSIONS: Overall, one-third of the under-five children in Bangladesh consumed antibiotics to treat infectious diseases. Multiple factors contribute to the prevalence of antibiotic use. The results highlight the need to regulate antibiotic use and prioritize national intervention programs.

Secondhand smoke exposure and associated factors among city residents living in multiunit housing in Bangladesh.

BACKGROUND: Secondhand smoke (SHS) poses a high health risk to those living in multiunit housing (MUH) since it can easily spread from unit to unit and throughout the building's communal areas. MUH residents in Bangladesh are particularly vulnerable to SHS due to the absence of smoking restrictions within a housing complex. Therefore, this study aimed to assess the prevalence of SHS exposure and its associated factors among MUH residents living in seven divisional cities of Bangladesh- Dhaka, Chattogram, Rajshahi, Khulna, Sylhet, Barishal, and Rangpur. METHODS: From April 2019 to November 2019, a cross-sectional survey was conducted with 616 MUH residents aged 18 or older who had been residing in MUH for at least two years in the seven divisional cities of Bangladesh. A multivariable logistic regression model was performed to determine the associated factors of SHS exposure. RESULTS: In MUH complexes, more than half (54.9%) of the 616 respondents were exposed to SHS. The key factors positively associated with SHS exposure were females (aOR: 1.8, 95% CI:1.236-2.681), residents with a low monthly family income (aOR: 1.9, 95% CI: 1.162-3.220), those whose family members smoked (aOR: 2.4, 95% CI: 1.537-3.746), and Dhaka city residents (aOR: 1.9, 95% CI: 1.013-3.440). CONCLUSIONS: This study revealed a high prevalence of SHS exposure among Bangladeshi MUH residents. Therefore, a smoking ban is needed in and around MUH complexes to protect non-smoking residents from SHS exposure.

Phyllanthus emblica (Amla) methanolic extract regulates multiple checkpoints in 15-lipoxygenase mediated inflammopathies: Computational simulation and in vitro evidence.

Amla (Phyllanthus emblica) has long been used in traditional folk medicine to prevent and cure a variety of inflammatory diseases. In this study, the antioxidant activity (DPPH scavenging and reducing power), anti-inflammatory activity (RBC Membrane Stabilization and 15-LOX inhibition), and anticoagulation activity (Serin protease inhibition and Prothrombin Time assays) of the methanolic extract of amla were conducted. Amla exhibited a substantial amount of phenolic content (TPC: 663.53 mg GAE/g) and flavonoid content (TFC: 418.89 mg GAE/g). A strong DPPH scavenging effect was observed with an IC50 of 311.31 µg/ml as compared to standard ascorbic acid with an IC50 of 130.53 µg/ml. In reducing power assay, the EC50 value of the extract was found to be 196.20 µg/ml compared to standard ascorbic acid (EC50 = 33.83 µg/ml). The IC50 value of the RBC membrane stabilization and 15-LOX assays was observed as 101.08 µg/ml (IC50 of 58.62 µg/ml for standard aspirin) and 195.98 µg/ml (IC50 of 19.62 µg/ml for standard quercetin), respectively. The extract also strongly inhibited serine protease (trypsin) activity with an IC50 of 505.81 µg/ml (IC50 of 295.44 µg/ml for standard quercetin). The blood coagulation time (PTT) was found to be 11.91 min for amla extract and 24.11 min for standard Warfarin. Thus, the findings of an in vitro study revealed that the methanolic extract of amla contains significant antioxidant, anti-inflammatory, and anticoagulation activity. Furthermore, in silico docking and simulation of reported phytochemicals of amla with human 15-LOXA and 15-LOXB were carried out to validate the anti-inflammatory activity of amla. In this analysis, epicatechin and catechin showed greater molecular interaction and were considerably stable throughout the 100 ns simulation with 15-lipoxygenase A (15-LOXA) and 15-lipoxygenase B (15-LOXB) respectively.

Cardiomyocyte-Targeting Peptide to Deliver Amiodarone.

BACKGROUND: Amiodarone is underutilized due to significant off-target toxicities. We hypothesized that targeted delivery to the heart would lead to the lowering of the dose by utilizing a cardiomyocyte-targeting peptide (CTP), a cell-penetrating peptide identified by our prior phage display work. METHODS: CTP was synthesized thiolated at the N-terminus, conjugated to amiodarone via Schiff base chemistry, HPLC purified, and confirmed with MALDI/TOF. The stability of the conjugate was assessed using serial HPLCs. Guinea pigs (GP) were injected intraperitoneally daily with vehicle (7 days), amiodarone (7 days; 80 mg/kg), CTP-amiodarone (5 days; 26.3 mg/kg), or CTP (5 days; 17.8 mg/kg), after which the GPs were euthanized, and the hearts were excised and perfused on a Langendorff apparatus with Tyrode's solution and blebbistatin (5 µM) to minimize the contractions. Voltage (RH237) and Ca2+-indicator dye (Rhod-2/AM) were injected, and fluorescence from the epicardium split and was captured by two cameras at 570-595 nm for the cytosolic Ca2+ and 610-750 nm wavelengths for the voltage. Subsequently, the hearts were paced at 250 ms with programmed stimulation to measure the changes in the conduction velocities (CV), action potential duration (APD), and Ca2+ transient durations at 90% recovery (CaTD90). mRNA was extracted from all hearts, and RNA sequencing was performed with results compared to the control hearts. RESULTS: The CTP-amiodarone remained stable for up to 21 days at 37 °C. At ~1/15th of the dose of amiodarone, the CTP-amiodarone decreased the CV in hearts significantly compared to the control GPs (0.92 ± 0.05 vs. 1.00 ± 0.03 ms, p = 0.0007), equivalent to amiodarone alone (0.87 ± 0.08 ms, p = 0.0003). Amiodarone increased the APD (192 ± 5 ms vs. 175 ± 8 ms for vehicle, p = 0.0025), while CTP-amiodarone decreased it significantly (157 ± 16 ms, p = 0.0136), similar to CTP alone (155 ± 13 ms, p = 0.0039). Both amiodarone and CTP-amiodarone significantly decreased the calcium transients compared to the controls. CTP-amiodarone and CTP decreased the CaTD90 to an extent greater than amiodarone alone (p < 0.001). RNA-seq showed that CTP alone increased the expression of DHPR and SERCA2a, while it decreased the expression of the proinflammatory genes, NF-kappa B, TNF-α, IL-1ß, and IL-6. CONCLUSIONS: Our data suggest that CTP can deliver amiodarone to cardiomyocytes at ~1/15th the total molar dose of the amiodarone needed to produce a comparable slowing of CVs. The ability of CTP to decrease the AP durations and CaTD90 may be related to its increase in the expression of Ca-handling genes, which merits further study.

Conjugation of amiodarone to a novel cardiomyocyte cell penetrating peptide for potential targeted delivery to the heart.

Modern medicine has developed a myriad of therapeutic drugs against a wide range of human diseases leading to increased life expectancy and better quality of life for millions of people. Despite the undeniable benefit of medical advancements in pharmaceutical technology, many of the most effective drugs currently in use have serious limitations such as off target side effects resulting in systemic toxicity. New generations of specialized drug constructs will enhance targeted therapeutic efficacy of existing and new drugs leading to safer and more effective treatment options for a variety of human ailments. As one of the most efficient drugs known for the treatment of cardiac arrhythmia, Amiodarone presents the same conundrum of serious systemic side effects associated with long term treatment. In this article we present the synthesis of a next-generation prodrug construct of amiodarone for the purpose of advanced targeting of cardiac arrhythmias by delivering the drug to cardiomyocytes using a novel cardiac targeting peptide, a cardiomyocyte-specific cell penetrating peptide. Our in vivo studies in guinea pigs indicate that cardiac targeting peptide-amiodarone conjugate is able to have similar effects on calcium handling as amiodarone at 1/15th the total molar dose of amiodarone. Further studies are warranted in animal models of atrial fibrillation to show efficacy of this conjugate.

The RNA-binding protein Adad1 is necessary for germ cell maintenance and meiosis in zebrafish.

The double stranded RNA binding protein Adad1 (adenosine deaminase domain containing 1) is a member of the adenosine deaminase acting on RNAs (Adar) protein family with germ cell-specific expression. In mice, Adad1 is necessary for sperm differentiation, however its function outside of mammals has not been investigated. Here, through an N-ethyl-N-nitrosourea (ENU) based forward genetic screen, we identified an adad1 mutant zebrafish line that develops as sterile males. Further histological examination revealed complete lack of germ cells in adult mutant fish, however germ cells populated the gonad, proliferated, and entered meiosis in larval and juvenile fish. Although meiosis was initiated in adad1 mutant testes, the spermatocytes failed to progress beyond the zygotene stage. Thus, Adad1 is essential for meiosis and germline maintenance in zebrafish. We tested if spermatogonial stem cells were affected using nanos2 RNA FISH and a label retaining cell (LRC) assay, and found that the mutant testes had fewer LRCs and nanos2-expressing cells compared to wild-type siblings, suggesting that failure to maintain the spermatogonial stem cells resulted in germ cell loss by adulthood. To identify potential molecular processes regulated by Adad1, we sequenced bulk mRNA from mutants and wild-type testes and found mis-regulation of genes involved in RNA stability and modification, pointing to a potential broader role in post-transcriptional regulation. Our findings suggest that the RNA regulatory protein Adad1 is required for fertility through regulation of spermatogonial stem cell maintenance in zebrafish.

Astute exploration of collective mental health events among the residents of elderly care homes.

Background & objective: Developing countries are presently witnessing a great burden of rapid aging followed by losing the social values of older adults due to age-related cognitive impairment as well as rising depression levels. This study was designed to assess the cognitive impairment and depression status combinedly among older adults in elderly care homes. Methods: It was a cross-sectional survey among randomly selected 200 older adults aged between 60 and 80 years residing in some old homes in Dhaka district, Bangladesh. Data were collected through face-to-face interviews while Cognitive function and level of depression were assessed by applying the Standardized Mini-Mental State Examination (MMSE) and Geriatric Depression Scale (GDS). Results: Among all the respondents, the majority (81.5%) were staying in old homes for 1-5 years. The majority (91.0%) had difficulties with vision, 40.7% had difficulties with hearing and 19.6% had difficulties moving around. The MMSE test revealed that 43% had moderate cognitive impairment, 36% had mild and 19.5% were found normal while more than half (56%) had severe depression. Significant cognitive impairment was found among the illiterate respondents who did not have any family care support. In addition, higher-educated respondents were found to have more severe depression (OR/p = 6.33/<0.01; 95% CI: 2.36-16.96). Furthermore, severely depressed respondents had more severe cognitive impairment (COR/p = 3.83/0.01; 95% CI: 1.66-8.83). Functional disabilities were also a greater concern for cognitive impairment and depression. Conclusion: An increasing number of old home residents are suffering from significant mental disorders while there is no mental health support in the elderly care homes in Bangladesh. Finally, there is a great need to develop packages and programs of mental health care for senior citizens and their caregivers residing in old homes, which can be scaled up across the country's mixed healthcare delivery system.

Causes of Death Among Infants and Children in the Child Health and Mortality Prevention Surveillance (CHAMPS) Network.

Importance: The number of deaths of children younger than 5 years has been steadily decreasing worldwide, from more than 17 million annual deaths in the 1970s to an estimated 5.3 million in 2019 (with 2.8 million deaths occurring in those aged 1-59 months [53% of all deaths in children aged <5 years]). More detailed characterization of childhood deaths could inform interventions to improve child survival. Objective: To describe causes of postneonatal child deaths across 7 mortality surveillance sentinel sites in Africa and Asia. Design, Setting, and Participants: The Child Health and Mortality Prevention Surveillance (CHAMPS) Network conducts childhood mortality surveillance in sub-Saharan Africa and South Asia using innovative postmortem minimally invasive tissue sampling (MITS). In this cross-sectional study, MITS was conducted in deceased children aged 1 to 59 months at 7 sites in sub-Saharan Africa and South Asia from December 3, 2016, to December 3, 2020. Data analysis was conducted between October and November 2021. Main Outcomes and Measures: The expert panel attributed underlying, intermediate, and immediate conditions in the chain of events leading to death, based on histopathologic analysis, microbiological diagnostics, clinical data, and verbal autopsies. Results: In this study, MITS was performed in 632 deceased children (mean [SD] age at death, 1.3 [0.3] years; 342 [54.1%] male). The 6 most common underlying causes of death were malnutrition (104 [16.5%]), HIV (75 [11.9%]), malaria (71 [11.2%]), congenital birth defects (64 [10.1%]), lower respiratory tract infections (LRTIs; 53 [8.4%]), and diarrheal diseases (46 [7.2%]). When considering immediate causes only, sepsis (191 [36.7%]) and LRTI (129 [24.8%]) were the 2 dominant causes. An infection was present in the causal chain in 549 of 632 deaths (86.9%); pathogens most frequently contributing to infectious deaths included Klebsiella pneumoniae (155 of 549 infectious deaths [28.2%]; 127 [81.9%] considered nosocomial), Plasmodium falciparum (122 of 549 [22.2%]), and Streptococcus pneumoniae (109 of 549 [19.9%]). Other organisms, such as cytomegalovirus (57 [10.4%]) and Acinetobacter baumannii (39 [7.1%]; 35 of 39 [89.7%] considered nosocomial), also played important roles. For the top underlying causes of death, the median number of conditions in the chain of events leading to death was 3 for malnutrition, 3 for HIV, 1 for malaria, 3 for congenital birth defects, and 1 for LRTI. Expert panels considered 494 of 632 deaths (78.2%) preventable and 26 of 632 deaths (4.1%) preventable under certain conditions. Conclusions and Relevance: In this cross-sectional study investigating causes of child mortality in the CHAMPS Network, results indicate that, in these high-mortality settings, infectious diseases continue to cause most deaths in infants and children, often in conjunction with malnutrition. These results also highlight opportunities for action to prevent deaths and reveal common interaction of various causes in the path toward death.

Neural tube defects as a cause of death among stillbirths, infants, and children younger than 5 years in sub-Saharan Africa and southeast Asia: an analysis of the CHAMPS network.

BACKGROUND: Neural tube defects are common birth defects resulting in severe morbidity and mortality; they can largely be prevented with periconceptional maternal intake of folic acid. Understanding the occurrence of neural tube defects and their contribution to mortality in settings where their burden is highest could inform prevention and health-care policy. We aimed to estimate the mortality attributed to neural tube defects in seven countries in sub-Saharan Africa and southeast Asia. METHODS: This analysis used data from the Child Health and Mortality Prevention Surveillance (CHAMPS) network and health and demographic surveillance systems from South Africa, Mozambique, Bangladesh, Kenya, Mali, Ethiopia, and Sierra Leone. All stillbirths and infants and children younger than 5 years who died, who were enrolled in CHAMPS, whose families consented to post-mortem minimally invasive tissue sampling (MITS) between Jan 1, 2017, and Dec 31, 2021, and who were assigned a cause of death by a determination of cause of death panel as of May 24, 2022, were included in this analysis, regardless the cause of death. MITS and advanced diagnostic methods were used to describe the frequency and characteristics of neural tube defects among eligible deaths, identify risk factors, and estimate the mortality fraction and mortality rate (per 10 000 births) by CHAMPS site. FINDINGS: Causes of death were determined for 3232 stillbirths, infants, and children younger than 5 years, of whom 69 (2%) died with a neural tube defect. Most deaths with a neural tube defect were stillbirths (51 [74%]); 46 (67%) were neural tube defects incompatible with life (ie, anencephaly, craniorachischisis, or iniencephaly) and 22 (32%) were spina bifida. Deaths with a neural tube defect were more common in Ethiopia (adjusted odds ratio 8·09 [95% CI 2·84-23·02]), among female individuals (4·40 [2·44-7·93]), and among those whose mothers had no antenatal care (2·48 [1·12-5·51]). Ethiopia had the highest adjusted mortality fraction of deaths with neural tube defects (7·5% [6·7-8·4]) and the highest adjusted mortality rate attributed to neural tube defects (104·0 per 10 000 births [92·9-116·4]), 4-23 times greater than in any other site. INTERPRETATION: CHAMPS identified neural tube defects, a largely preventable condition, as a common cause of death among stillbirths and neonatal deaths, especially in Ethiopia. Implementing interventions such as mandatory folic acid fortification could reduce mortality due to neural tube defects. FUNDING: Bill & Melinda Gates Foundation.

Prevalence of anemia in diabetes mellitus in South Asia: A systematic review and meta-analysis.

OBJECTIVE: Anemia and Diabetes Mellitus (DM) are amongst major clinical and public health challenges in South Asia that influence the progression of chronic health problems in this population. Despite a growing body of research on these problems, there is a lack synthesized evidence on the burden of anemia among people with DM in this region. This meta-analytic review was conducted to estimate the prevalence of anemia among people with DM in South Asia. METHODS: A systematic search of the literature was conducted in five primary databases and additional sources up to July 29, 2022, that reported the prevalence of anemia among DM patients in any of the eight South Asian countries. Observational studies that met pre-determined eligibility criteria according to the protocol registered in PROSPERO (CRD42022348433) were included in this meta-analysis. Random effect models were used to estimate pooled prevalence. RESULTS: Of the 40 eligible studies, 38 underwent meta-analysis representing 14,194 participants with DM. The pooled prevalence of anemia was 45% (95% CI: 37.0-54.0, I2 = 99.28%, p = 0.00) among diabetic people in South Asia. In sub-group analysis, the pooled prevalence of anemia was higher in females (48%, 95% CI: 37.0-60.0, I2 = 98.86%, p = 0.00) compared to males (39%, 95% CI: 29.0-48.0, I2 = 98.18%, p = 0.00). Diabetic patients with older age (≥ 50 years) reported higher pooled estimates of anemia (48%, 95% CI: 38.0-58.0, I2 = 99.07%) than younger age group (< 50 years) (34%, 95% CI: 21.0-47.0, I2 = 98.83%). In addition, we found variation in pooled prevalence estimates of anemia considering the type of DM, such as type 1 reported 2% (95% CI: 0.00-4.00), type-2 reported 48% (95% CI: 40.0-56.0, I2 = 98.94%), and Gestational diabetes mellitus (GDM) reported 6% (95% CI: 3.00-12.0). CONCLUSION: High pooled estimates of anemia among diabetic patients in South Asia, including publication bias, warrants further clinical and public health research following standard research methods to understand the more context-specific epidemiological insights and evidence.

Cardiomyocyte Targeting Peptide to Deliver Amiodarone.

Background: Amiodarone is underutilized due to significant off-target toxicities. We hypothesized that targeted delivery to the heart would lead to lowering of dose by utilizing a cardiomyocyte targeting peptide (CTP), a cell penetrating peptide identified by our prior phage display work. Methods: CTP was synthesized thiolated at the N-terminus, conjugated to amiodarone via Schiff base chemistry, HPLC purified and confirmed with MALDI/TOF. Stability of the conjugate was assessed using serial HPLCs. Guinea pigs (GP) were injected intraperitoneally daily with vehicle (7 days), amiodarone (7 days; 80mg/Kg), CTP-amiodarone (5 days;26.3mg/Kg), or CTP (5 days; 17.8mg/Kg), after which GPs were euthanized, hearts excised, perfused on a Langendorff apparatus with Tyrode's solution and blebbistatin (5µM) to minimize contractions. Voltage (RH237) and Ca 2+ -indicator dye (Rhod-2/AM) were injected, fluorescence from the epicardium split and focused on two cameras capturing at 570-595nm for cytosolic Ca 2+ and 610-750nm wavelengths for voltage. Subsequently, hearts were paced at 250ms with programmed stimulation to measure changes in conduction velocities (CV), action potential duration (APD) and Ca 2+ transient durations at 90% recovery (CaTD 90 ). mRNA was extracted from all hearts and RNA sequencing performed with results compared to control hearts. Results: CTP-amiodarone remained stable for up to 21 days at 37°C. At ∼1/15 th of the dose of amiodarone, CTP-amiodarone decreased CV in hearts significantly compared to control GPs (0.92±0.05 vs. 1.00±0.03m/s, p=0.0007), equivalent to amiodarone alone (0.87±0.08ms, p=0.0003). Amiodarone increased APD (192±5ms vs. 175±8ms for vehicle, p=0.0025), while CTP-amiodarone decreased it significantly (157±16ms, p=0.0136) similar to CTP alone (155±13ms, p=0.0039). Both amiodarone and CTP-amiodarone significantly decreased calcium transients compared to controls. CTP-amiodarone and CTP decreased CaTD 90 to an extent greater than amiodarone alone (p<0.001). RNA-seq showed that CTP alone increased the expression of DHPR and SERCA2a, while decreasing expression of proinflammatory genes NF-kappa B, TNF-α, IL-1ß, and IL-6. Conclusions: Our data suggests that CTP can deliver amiodarone to cardiomyocytes at ∼1/15 th the total molar dose of amiodarone needed to produce comparable slowing of CVs. The ability of CTP to decrease AP durations and CaTD 90 may be related to its increase in expression of Ca-handling genes, and merits further study.