Cationic lipid-conjugated bis-arylidene oxindole derivatives as broad-spectrum breast cancer-selective therapeutics.

Breast cancer is a heterogeneous malignancy with wide-ranging variations in therapeutic responses, overall survival etc. Major challenges for available chemotherapeutic agents in achieving clinical success are in maintaining systemic bio-distribution and avoiding non-specific adverse effects. Bis-arylidene oxindoles are estrogen receptor (ER)-selective bioactive molecules with moderate potency. In here, we have designed, synthesized and evaluated a series of twin aliphatic chain cationic lipid-conjugated bis-arylidene oxindole molecules with variations in nature of linker, lengths of carbon spacer and hydrophobic twin chains. We observed that among the various structural analogues, C8 twin-chain containing molecules, PGC8, S2C8 and S3C8 showed effective cancer cell-selective cytotoxicity in different cancer cell lines with an IC50 ranging from 4 to 7 µM. These molecules selectively induced apoptosis, ROS production and cell cycle inhibition at G1/S phase in ER + breast cancer cells but not in non-cancer cells. Additionally, these molecules formed homogenous self-assemblies exhibiting effective hydrodynamic diameter with positive surface charge. The self-assemblies also showed prominent cancer cell-selective uptake and DNA-binding abilities. Hence, we have shown successful incorporation of dexamethasone to the self-assemblies, and its enhanced cytotoxicity even in ER-negative breast cancer cells. All these results indicate that PGC8, S2C8 and S3C8 molecules, albeit their potent and selective ER-positive anti-breast cancer activity, can be repurposed as targeted delivery systems and hold promise as unique, broader spectrum breast cancer cell-selective therapeutic payloads.

Prevalence, Virulence Gene Profiling, and Characterization of Enteroaggregative Escherichia coli from Children with Acute Diarrhea, Asymptomatic Nourished, and Malnourished Children Younger Than 5 Years of Age in India.

OBJECTIVE: To study the significance of enteroaggregative Escherichia coli (EAEC) as a pathogen causing acute diarrhea and a commensal in healthy nourished and malnourished children younger than five years of age in the Chandigarh region and to address possible traits of EAEC virulence genes, biofilm formation, phylogroups, and antibiotic resistance that would be correlated with diarrhea or carriage. STUDY DESIGN: Stool samples were obtained from children with acute diarrhea (n = 548), as well as nourished (n = 550), and malnourished controls without diarrhea (n = 110). E coli isolates were confirmed as EAEC by pCVD432 polymerase chain reaction. Multiplex polymerase chain reactions were used to identify 22 virulence-related genes and phylogeny. Antibiotic susceptibility, adherence, and biofilm-forming potential also were studied. RESULTS: Overall, 16.6% of children were malnourished. EAEC detection was greater among children with acute diarrhea (16%) than nourished (6%) and malnourished nondiarrheal controls (2.7%). We found an association of EAEC infections with age <2 years (P = .0001) in the diarrheal group. Adhesive variants adhesion fimbriae IV and adhesion fimbriae II were significantly associated with diarrhea. The aggR and aar genes showed a positive and negative association with the severity of disease (P = .0004 and P = .0003). A high degree of multidrug resistance was found (73.8%) in the diarrheal group. Most EAEC strains from the diarrheal group belonged to B2 and D phylogroups, whereas strains from non-diarrheal groups, which belonged to phylogroup B1. CONCLUSIONS: EAEC is a significant contributor to childhood diarrhea, its presence as a commensal, and the significance of the association of various virulence factors among the EAEC isolated from diarrheal and non-diarrheal stools. These data reinforce the importance of aggR and aar as positive and negative regulators and the contribution of AAF/II and AAF/IV fimbria for the pathobiology of EAEC.

Self-Assembling Derivative of Hydrocortisone as Glucocorticoid Receptor-Targeted Nanotherapeutics for Synergistic, Combination Therapy against Colorectal Tumor.

Hydrocortisone, a natural glucocorticoid secreted by adrenal and extra-adrenal tissues, locally governs the transcription of genes involved in inflammation, immune response, metabolism, and energy homeostasis via binding to its cognate glucocorticoid receptor (GR). In this study, we show that modified hydrocortisone (HC16), a cancer-selective cytotoxic molecule, showed synergism in combination with drugs like Doxorubicin and docetaxel, self-assembled into vesicles, entrapped docetaxel and complexed with anti-cancer plasmid DNA for enhanced killing of cancer cells. These vesicles exhibited GR-mediated nuclear localization, delivery of the p53 gene, and also inhibited cell viability selectively in RKO, HCT15, and CT26 colon cancer cells but not in normal cells like CHO and HEK293T. Apart from exerting its own anti-cancer activity, the self-assembled HC16 vesicles loaded with docetaxel sensitized the cancer cells to its drug cargo by downregulating the drug metabolizing CYP3A4 gene. This indirectly reduces the risk of nonspecific adverse effects in normal cells, as the viability of sensitized cancer cells could be significantly reduced even in low doses of cytotoxic docetaxel. The near infrared (NIR)-dye-associated self-assemblies accumulated in a colon tumor with higher orders of NIR intensity compared to those in a colon of healthy mice. Thereafter, the treatment of HC16-docetaxel-p53 vesicle/DNA complex led to significant tumor regression, which resulted in a cecum/body weight ratio in tumor-bearing mice similar to that of healthy mice measured at 24 h postcompletion of treatment. There was an up to 2.5-fold enhancement in the overall survivability of colon-tumor-bearing mice treated with HC16-docetaxel-p53 vesicle/DNA complexes when compared against the pristine docetaxel-treated groups. Further, the HC16-docetaxel-p53 vesicle/DNA complex-treated group showed reduced nuclear accumulation of cell proliferation marker Ki67, reduced protein levels of prosurvival and mesenchymal proteins like Bcl-2, PARP, vimentin, and N-cadherin, and increased the levels of pro-apoptotic activated caspases as compared to the pristine docetaxel-treated groups. The therapeutic package described herein is expected to find future use as a rational, multifaceted, GR-targeted approach for inhibiting colon tumor progression.

Mitoapocynin, a mitochondria targeted derivative of apocynin induces mitochondrial ROS generation and apoptosis in multiple cell types including cardiac myoblasts: a potential constraint to its therapeutic use.

Mitoapocynin is a triphenylphosphonium conjugated derivative of apocynin that specifically locates to the mitochondria. It has been developed as a mitochondrially targeted therapeutic antioxidant. We attempted to attenuate the mitochondrial ROS induced in H9c2 cardiac myoblast cells treated with norepinephrine. Mitoapocynin was a poor quencher of total ROS as detected by the fluoroprobe DCFH-DA. Using mitochondrial superoxide specific probe MitoSoxRed, we found that 5-10 µM mitoapocynin itself induces superoxide over and above that is generated by the norepinephrine treatment. A supposedly control molecule to mitoapocynin, the synthetic compound PhC11TPP, having the triphenylphosphonium group and a benzene moiety with C11 aliphatic chain spacer was also found to be a robust inducer of mitochondrial ROS. Subsequent assays with several cell lines viz., NIH3T3, HEK293, Neuro2A, MCF-7 and H9c2, showed that prolonged exposure to mitoapocynin induces cell death by apoptosis that can be partially prevented by the general antioxidant N-acetyl cysteine. Analyses of mitochondrial electron transport complexes by Blue Native Polyacrylamide gel electrophoresis showed that both mitoapocynin and PhC11TPP disrupt the mitochondrial Complex I and V, and in addition, PhC11TPP also damages the Complex IV. Our data thus highlights the limitations of the therapeutic use of mitoapocynin as an antioxidant.

Methoxy-enriched cationic stilbenes as anticancer therapeutics.

Stilbene-based compounds are largely described for their antioxidant activity. But their use as anticancer chemotherapeutics is hampered by poor pharmacokinetic properties and non-selectivity towards cancer and non-cancer potency. To overcome these drawbacks, twin chain cationic lipid conjugated, methoxy-enriched stilbene derivatives were designed, synthesized and evaluated for their anticancer potency. Our findings reveal that HMSC16, a molecule with the highest number of methoxy groups and with C16-twin chain lipid, is the most potent as well as the most selective anticancer agent when compared to the other synthesized derivatives and commercially available stilbene-based drug, tamoxifen, and resveratrol. To justify these results, we have conducted a series of mechanistic experiments where we found that HMSC16 induced ROS generation, apoptosis, and autophagy by affecting the mitochondrial, lysosomal and nuclear pathways. Further cell cycle analysis data reveals that HMSC16 not only induces cell death but is also involved in the arrest of the cell cycle at the sub-G1 phase. Moreover, HMSC16 showed self-aggregation property owing to a possibly favorable hydrophilic-lipophilic balance. The self-aggregation property of HMSC16 allowed it to entrap hydrophobic drugs, withaferin. With entrapped withaferin, HMSC16 showed additive if not synergistic cell killing effect in HeLa cells. From the above results, we concluded that HMSC16 can be used not just as a drug but also as a drug delivery agent.

Synthesis and biological evaluation of polyhydroxylated oxindole derivatives as potential antileishmanial agent.

The devastating appearance of numerous drug-unresponsive strains of Leishmania donovani and severe toxic side effects of conventional antileishmanial therapy necessitates the search for novel leads, to treat visceral leishmaniasis efficiently. The current study deals with the synthesis and biological evaluation of a unique C-5 functionalized oxindole based polyphenol to ascertain its activities against L. donovani infection, in vitro. The polyhydroxylated oxindole derivative (1) was generated by coupling styrene derivatives with 5-bromo bis-arylidene oxindole using Heck coupling reaction. The synthesized molecule 1 was tested for its antileishmanial activity using both promastigote and amastigote stages of L. donovani. Molecule 1 showed promising anti-promastigote and anti-amastigote activities with IC50 values 15 µM and 1 µM, respectively, with no cytotoxicity towards host splenocytes. The results revealed that this compound induced parasite death by promoting oxidative stress, thereby triggering apoptosis.

An Offenders-Offenses Shared Component Spatial Model for Identifying Shared and Specific Hotspots of Offenders and Offenses: A Case Study of Juvenile Delinquents and Violent Crimes in the Greater Toronto Area.

Objectives: We attempted to apply the Bayesian shared component spatial modeling (SCSM) for the identification of hotspots from two (offenders and offenses) instead of one (offenders or offenses) variables and developed three risk surfaces for (1) common or shared by both offenders and offenses; (2) specific to offenders, and (3) specific to offenses. Methods: We applied SCSM to examine the joint spatial distributions of juvenile delinquents (offenders) and violent crime (offenses) in the York Region of the Greater Toronto Area at the dissemination area level. The spatial autocorrelation, overdispersion, and latent covariates were adjusted by spatially structured and unstructured random effect terms in the model. We mapped the posterior means of the estimated shared and specific risks for identifying the three risk surfaces and types of hotspots. Results: Results suggest that about 50% and 25% of the relative risks of juvenile delinquents and violent crimes, respectively, could be explained by the shared component of offenders and offenses. The spatially structured terms attributed to 48% and 24% of total variations of the delinquents and violent crimes, respectively. Contrastingly, the unstructured random covariates influenced 3% of total variations of the juvenile delinquents and 51% for violent crimes. Conclusions: The Bayesian SCSM presented in this study identifies shared and specific hotspots of juvenile delinquents and violent crime. The method can be applied to other kinds of offenders and offenses and provide new insights into the clusters of high risks that are due to both offenders and offenses or due to offenders or offenses only.

Assessing the Quality and Coverage of Maternal Postnatal Care in Bangladesh: A Comparative Analysis of Quality Postnatal Care among Home and Facility Births.

BACKGROUND: Bangladesh has achieved remarkable progress in reducing maternal mortality, yet postpartum deaths remain a significant issue. Emphasis on quality postnatal care (qPNC) is crucial, as increased coverage alone has not sufficiently reduced maternal morbidity and mortality. METHODS: This study included data from the Bangladesh Maternal Mortality Survey of 32,106 mothers who delivered within three years prior to the survey. Descriptive statistics were used to report coverage and components of postnatal care stratified by covariates. Log-linear regression models were used to assess the determinants of quality postnatal care among facility and home births. RESULTS: From 2010 to 2016, postnatal care coverage within 48 h of delivery by a qualified provider rose from 23% to 47%. Of the births, 94% were facility births that received timely PNC, contrasted with only 6% for home births. Despite the increased coverage, quality of care remained as low as 1% for home births and 13% for facility births. Key factors affecting qPNC utilization included socio-demographic factors, pregnancy complications, type of birth attendant, delivery method, and financial readiness. CONCLUSION: Importantly, deliveries assisted by skilled birth attendants correlated with higher quality postnatal care. This study reveals a significant gap between the coverage and quality of postnatal care in rural Bangladesh, especially for home births. It underscores the need for targeted interventions to enhance qPNC.

Development and validation of a modified QuEChERS method coupled with LC-MS/MS for simultaneous determination of difenoconazole, dimethoate, pymetrozine, and chlorantraniliprole in brinjal collected from fields and markets places to assess human health risk.

An effective and sensitive analytical method was developed to quantify the most common pesticide residues (difenoconazole, dimethoate, pymetrozine, and chlorantraniliprole) used for brinjal cultivation in Bangladesh. The quantification of the analytes was done using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were extracted using a modified QuEChERS method and followed by purification with dispersive solid phase extraction (d-SPE) sorbents (PSA, GCB, and C18). Matrix-matched calibration with a regression coefficient R2 ≥ 0.9964 were used to minimize the brinjal matrix effect. The method was validated in quintuple (n = 5) at five different spiked levels (8-400 µg/kg) having recoveries in the range of 70.3-113.2% with relative standard deviations RSDs ≤6.8%, limits of detection (LOD) and limits of quantification (LOQ) was in the range of 0.15-0.66 µg/kg and 0.4-2.0 µg/kg, respectively, for the four analytes. A total 100 samples (50 samples directly from fields of Jessore district, Bangladesh and 50 samples from local market of Dhaka, Bangladesh) were collected to analyse the pesticides residue. The result showed that pesticides residue was found in both the field and market collected samples, 54% and 38%, respectively. The overall mean residue levels of four pesticides in field samples were significantly higher than those of market samples. Moreover, 20% of the field samples and 10% of the market samples had dimethoate residues, which were the most abundant among the four analytes and it ranged from 0.017 to 0.252 mg/kg. In terms of health risk assessments, dimethoate showed the highest estimated daily intake (EDI) and hazard quotient (HQ) values that are 3.02 × 10-5 mg/kg/day and 1.51%, respectively, in field samples. Till now, there have been no regulations or guidelines for the maximum admissible pesticide residue in Bangladesh. Therefore, the above findings will be an initial step for the regulatory authorities of Bangladesh to implement regulations and guidelines for pesticide usage.

A high-throughput test enables specific detection of hepatocellular carcinoma.

High-throughput tests for early cancer detection can revolutionize public health and reduce cancer morbidity and mortality. Here we show a DNA methylation signature for hepatocellular carcinoma (HCC) detection in liquid biopsies, distinct from normal tissues and blood profiles. We developed a classifier using four CpG sites, validated in TCGA HCC data. A single F12 gene CpG site effectively differentiates HCC samples from other blood samples, normal tissues, and non-HCC tumors in TCGA and GEO data repositories. The markers were validated in a separate plasma sample dataset from HCC patients and controls. We designed a high-throughput assay using next-generation sequencing and multiplexing techniques, analyzing plasma samples from 554 clinical study participants, including HCC patients, non-HCC cancers, chronic hepatitis B, and healthy controls. HCC detection sensitivity was 84.5% at 95% specificity and 0.94 AUC. Implementing this assay for high-risk individuals could significantly decrease HCC morbidity and mortality.

An assessment of meteorological parameters effects on COVID-19 pandemic in Bangladesh using machine learning models.

Coronavirus (COVID-19) is a highly contagious virus (SARS-CoV-2) that has caused a global pandemic since January 2020. Scientists around the world are doing extensive research to control this disease. They are working tirelessly to find out the origin and causes of the disease. Several studies and experiments mentioned that there are some meteorological parameters which are highly correlated with COVID-19 transmission. In this work, we studied the effects of 11 meteorological parameters on the transmission of COVID-19 in Bangladesh. We first applied statistical analysis and observed that there is no significant effect of these parameters. Therefore, we proposed a novel technique to analyze the insight effects of these parameters by using a combination of Random Forest, CART, and Lasso feature selection techniques. We observed that 4 parameters are highly influential for COVID-19 where [Formula: see text] and Cloud have positive association whereas WS and AQ have negative impact. Among them, Cloud has the highest positive impact which is 0.063 and WS has the highest negative association which is [Formula: see text]. Moreover, we have validated our performance using DLNM technique. The result of this investigation can be used to develop an alert system that will assist the policymakers to know the characteristics of COVID-19 against meteorological parameters and can impose different policies based on the weather conditions.

Age- and Sex-Specific Association Between Vegetation Cover and Mental Health Disorders: Bayesian Spatial Study.

BACKGROUND: Despite growing evidence that reduced vegetation cover could be a putative risk factor for mental health disorders, the age- and the sex-specific association between vegetation and mental health disorder cases in urban areas is poorly understood. However, with rapid urbanization across the globe, there is an urgent need to study this association and understand the potential impact of vegetation loss on the mental well-being of urban residents. OBJECTIVE: This study aims to analyze the spatial association between vegetation cover and the age- and sex-stratified mental health disorder cases in the neighborhoods of Toronto, Canada. METHODS: We used remote sensing to detect urban vegetation and Bayesian spatial hierarchical modeling to analyze the relationship between vegetation cover and mental health disorder cases. Specifically, an Enhanced Vegetation Index was used to detect urban vegetation, and Bayesian Poisson lognormal models were implemented to study the association between vegetation and mental health disorder cases of males and females in the 0-19, 20-44, 45-64, and ≥65 years age groups, after controlling for marginalization and unmeasured (latent) spatial and nonspatial covariates at the neighborhood level. RESULTS: The results suggest that even after adjusting for marginalization, there were significant age- and sex-specific effects of vegetation on the prevalence of mental health disorders in Toronto. Mental health disorders were negatively associated with the vegetation cover for males aged 0-19 years (-7.009; 95% CI -13.130 to -0.980) and for both males (-4.544; 95% CI -8.224 to -0.895) and females (-3.513; 95% CI -6.289 to -0.681) aged 20-44 years. However, for older adults in the 45-64 and ≥65 years age groups, only the marginalization covariates were significantly associated with mental health disorder cases. In addition, a substantial influence of the unmeasured (latent) and spatially structured covariates was detected in each model (relative contributions>0.7), suggesting that the variations in area-specific relative risk were mainly spatial in nature. CONCLUSIONS: As significant and negative associations between vegetation and mental health disorder cases were found for young males and females, investments in urban greenery can help reduce the future burden of mental health disorders in Canada. The findings highlight the urgent need to understand the age-sex dynamics of the interaction between surrounding vegetation and urban dwellers and its subsequent impact on mental well-being.

Job insecurity and mental health related outcomes among the humanitarian workers during COVID-19 pandemic: a cross-sectional study.

BACKGROUND: The COVID-19 remains a public health burden that has caused global economic crises, jeopardizing health, jobs, and livelihoods of millions of people around the globe. Several efforts have been made by several countries by implementing several health strategies to attenuate the spread of the pandemic. Although several studies indicated effects of COVID-19 on mental health and its associated factors, very little is known about the underlying mechanism of job insecurity, depression, anxiety, and stress in Bangladesh. Therefore, this study determined the prevalence of job insecurity and depression, anxiety, stress as well as the association between job insecurity, mental health outcomes also contributing determinants amongst humanitarian workers during the COVID-19 pandemic in Bangladesh. METHODS: We conducted a web-based cross-sectional study among 445 humanitarian workers during the COVID-19 pandemic in six sub-districts of Cox's bazar district of Bangladesh between April and May 2021. The questionnaire was composed of socio-demographic, lifestyle and work related factors. Psychometric instruments like job insecurity scale and depression, anxiety also stress scale (DASS-21) were employed to assess the level of job insecurity and mental health outcomes (depression, anxiety and stress). STATA software version 14 was employed to perform statistical analyses. RESULTS: The prevalence of job insecurity was 42%. The odds of job insecurity was higher in Kutubdia and Pekua (AOR = 3.1, 95% CI 1.36, 7.22) Teknaf (AOR = 2.9, 95% CI 1.33, 6.41), the impact of dissatisfaction on salary (AOR = 2.3, 95% CI 1.49, 3.58) was evident with job insecurity. The prevalence of moderate to severe depression, anxiety and stress among humanitarian worker were (26%, 7%), (25%, 10%) and (15%, 7%) respectively. Further, the region of work, being female, marital status, work environment, and salary dissatisfaction were contributing factors for poor mental health outcomes. Those with job insecurity were almost 3 times more likely to experience depression (AOR = 2.7, 95% CI 1.85, 4.04), anxiety (AOR = 2.6, 95% CI 1.76, 3.71) and stress (AOR: 2.8; 95% CI 1.89, 4.26), respectively. CONCLUSION: Our findings highlight that job security remains essential to help tackle the severity of depression, anxiety and stress in humanitarian workers. The results reflected the critical importance of local and international NGOs addressing poor mental health conditions of their employees to prevent mental health outbreaks.

Understanding the Differential Impact of Vegetation Measures on Modeling the Association between Vegetation and Psychotic and Non-Psychotic Disorders in Toronto, Canada.

Considerable debate exists on whether exposure to vegetation cover is associated with better mental health outcomes. Past studies could not accurately capture people's exposure to surrounding vegetation and heavily relied on non-spatial models, where the spatial autocorrelation and latent covariates could not be adjusted. Therefore, a suite of five different vegetation measures was used to separately analyze the association between vegetation cover and the number of psychotic and non-psychotic disorder cases in the neighborhoods of Toronto, Canada. Three satellite-based and two area-based vegetation measures were used to analyze these associations using Poisson lognormal models under a Bayesian framework. Healthy vegetation cover was found to be negatively associated with both psychotic and non-psychotic disorders. Results suggest that the satellite-based indices, which can measure both the density and health of vegetation cover and are also adjusted for urban and environmental perturbations, could be better alternatives to simple ratio- and area-based measures for understanding the effect of vegetation on mental health. A strong dominance of spatially structured latent covariates was found in the models, highlighting the importance of adopting a spatial approach. This study can provide critical guidelines for selecting appropriate vegetation measures and developing spatial models for future population-based epidemiological research.

Targeting the Trypanothione Reductase of Tissue-Residing Leishmania in Hosts' Reticuloendothelial System: A Flexible Water-Soluble Ferrocenylquinoline-Based Preclinical Drug Candidate.

Since inception, the magic bullets developed against leishmaniasis traveled a certain path and then dropped down due to either toxicity or the emergence of resistance. The route of administration is also an important concern. We developed a series of water-soluble ferrocenylquinoline derivatives, targeting Leishmania donovani, among which CQFC1 showed the highest efficacy even in comparison to other drugs, in use or used, both in oral and intramuscular routes. It did not induce any toxicity to splenocytes and on hematopoiesis, induced protective cytokines, and did not hamper the drug-metabolizing enzymes in hosts. It acts through the reduction and the inhibition of parasites' survival enzyme trypanothione reductase of replicating amastigotes in hosts' reticuloendothelial tissues. Unlike conventional drugs, this molecule did not induce the resistance-conferring genes in laboratory-maintained resistant L. donovani lines. Experimentally, this easily bioavailable preclinical drug candidate overcame all of the limitations causing the discontinuation of the other conventional antileishmanial drugs.

Comparative analysis of virulence determinants, phylogroups, and antibiotic susceptibility patterns of typical versus atypical Enteroaggregative E. coli in India.

Enteroaggregative Escherichia coli (EAEC) is an evolving enteric pathogen that causes acute and chronic diarrhea in developed and industrialized nations in children. EAEC epidemiology and the importance of atypical EAEC (aEAEC) isolation in childhood diarrhea are not well documented in the Indian setting. A comparative analysis was undertaken to evaluate virulence, phylogeny, and antibiotic sensitivity among typical tEAEC versus aEAEC. A total of 171 EAEC isolates were extracted from a broad surveillance sample of diarrheal (N = 1210) and healthy children (N = 550) across North India. Polymerase chain reaction (PCR) for the aggR gene (master regulator gene) was conducted to differentiate tEAEC and aEAEC. For 21 virulence genes, we used multiplex PCR to classify possible virulence factors among these strains. Phylogenetic classes were identified by a multiplex PCR for chuA, yjaA, and a cryptic DNA fragment, TspE4C2. Antibiotic susceptibility was conducted by the disc diffusion method as per CLSI guidelines. EAEC was associated with moderate to severe diarrhea in children. The prevalence of EAEC infection (11.4%) was higher than any other DEC group (p = 0.002). tEAEC occurrence in the diarrheal group was higher than in the control group (p = 0.0001). tEAEC strain harbored more virulence genes than aEAEC. astA, aap, and aggR genes were most frequently found in the EAEC from the diarrheal population. Within tEAEC, this gene combination was present in more than 50% of strains. Also, 75.8% of EAEC strains were multidrug-resistant (MDR). Phylogroup D (43.9%) and B1 (39.4%) were most prevalent in the diarrheal and control group, respectively. Genetic analysis revealed EAEC variability; the comparison of tEAEC and aEAEC allowed us to better understand the EAEC virulence repertoire. Further microbiological and epidemiological research is required to examine the pathogenicity of not only typical but also atypical EAEC.

Seromonitoring of Peste des Petits Ruminants in goats and molecular characterization of PPR virus from field cases.

OBJECTIVES: The study was undertaken with the objectives to perform seromonitoring of Peste des Petits Ruminants (PPR) antibodies in goats vaccinated with PPR vaccine and molecular characterization of PPR virus (PPRV) from field cases in Bangladesh. MATERIALS AND METHODS: Seromonitoring work was conducted in Char Kalibari, Mymensingh Sadar, Mymensingh. For this, a total of 50 goats were randomly selected and were divided into two groups; vaccinated (Group A; n = 25) and non-vaccinated (Group B; n = 25). The goats of both groups were again sub-divided into four age groups; (i) 0-6 months (n = 5), (ii) 6-12 months (n = 5), (iii) 12-24 months (n = 10), and (iv) >24 months (n = 5). Blood samples were collected on Day-0 and after 21 days of post-vaccination (DPV), and the sera were prepared. The sera were examined for the presence of antibodies against PPRV by competitive enzyme-linked immunosorbent assay. For molecular characterization, nasal swabs (n = 10) were collected from PPR infected goats in Jessore during PPR outbreak (February 2016). The causative agent, PPRV isolated from field cases were confirmed by N gene based on reverse transcription polymerase chain reaction (RT-PCR), followed by sequencing, phylogenetic analysis, and multiple sequence alignment analyses. RESULTS: In the case of seromonitoring, the results revealed that before vaccination (at Day-0), overall, 44% (n = 22/50) goats were seropositive for PPRV. In Group A, 48% (n = 12/25) goats were seropositive, but after 21 DPV, 96% (n = 24/25) goats become seropositive. On the other hand, in Group B, 40% (n = 10/25) and 16% (n = 04/25) seropositive goats found at Day-0 and after 21 DPV, respectively, indicating that the antibody titer was increasing after vaccination and decreasing in convalescent goats. Out of 10 nasal swab samples, 40% (n = 4/10) was confirmed by RT-PCR targeting nucleocapsid (N gene). Phylogenetically, our isolate (KY039156/PPRV/BDG/Jes/2016) was similar to the other strains of PPRV under lineage IV. However, there was a unique amino acid substitution, where glycine (G) was recorded in place of arginine (R). The strain is closely related with other Chinese or Indian strains. The nucleotide sequence homology by NCBI BLAST search of the isolated strain ranged from 95% to 99% with other strains circulating in Bangladesh. CONCLUSION: The PPRV is prevailing in the Mymensingh and Jessore regions of Bangladesh. Effective control of PPR in goats may depend on vaccination with PPR vaccine. Molecular characterization of PPRV in Jessore reveals that the virus is differing from the strain prevalent in other regions of Bangladesh and the world.

Development and validation of BLRI Mastitis Test Kit at Bangladesh Livestock Research Institute Regional Station, Sirajganj.

OBJECTIVE: The objective of this study was to develop a low-cost kit for the detection of subclinical mastitis (SCM) and to check its validity, reproducibility, and efficacy at the field level. MATERIALS AND METHODS: A total of 550 quarter milk samples from crossbred dairy cows were collected, of which 400 milk samples were used to validate the newly developed BLRI mastitis test (BMT) kit to justify its efficacy as an individual test kit in detecting SCM based on somatic cell count (SCC) by direct microscopic count (DMC). The efficacy of the newly developed BMT was compared with the California Mastitis Test (CMT) kit. Another 150 milk samples were subjected to SCC determined by DMC and DCC (De Laval cell counter®) categorized by CMT and BMT scores. RESULTS: A SCM test kit, namely, BMT kit was successfully developed in this study. The percentage accuracy of CMT and BMT were 76.75% and 75.75%; sensitivity 69.36% and 67.56%; specificity 85.95% and 85.85%; positive predictive value 86.03% and 85.71%; negative predictive value 69.23% and 68%, respectively. A p value of 0.001 was found for both CMT and BMT. However, CMT and BMT had no significant difference in sensitivity (p = 0.778). Average SCCs (cells/ml) determined by DCC and DMC, respectively, were mostly corresponded to the SCC ranges of both CMT and BMT scores. CONCLUSION: The newly developed BMT kit is an independent, cheap, farmer-friendly, first country made, and reliable SCM diagnostic test kit that can be used at field condition.

Analyzing spatial and space-time clustering of facility-based deliveries in Bangladesh.

BACKGROUND: A spatial and temporal study of the distribution of facility-based deliveries can identify areas of low and high facility usage and help devise more targeted interventions to improve delivery outcomes. Developing countries like Bangladesh face considerable challenges in reducing the maternal mortality ratio to the targets set by the Sustainable Development Goals. Recent studies have already identified that the progress of reducing maternal mortality has stalled. Giving birth in a health facility is one way to reduce maternal mortality. METHODS: Facility delivery data from a demographic surveillance site was analyzed at both village and Bari (comprising several households with same paternal origins) level to understand spatial and temporal heterogeneity. Global spatial autocorrelation was detected using Moran's I index while local spatial clusters were detected using the local Getis G i * statistics. In addition, space-time scanning using a discrete Poisson approach facilitated the identification of space-time clusters. The likelihood of delivering at a facility when located inside a cluster was calculated using log-likelihood ratios. RESULTS: The three cluster detection approaches detected significant spatial and temporal heterogeneity in the distribution of facility deliveries in the study area. The hot and cold spots indicated contiguous and relocation type diffusion and increased in number over the years. Space-time scanning revealed that when a parturient woman is located in a Bari inside the cluster, the likelihood of delivering at a health facility increases by twenty-seven times. CONCLUSIONS: Spatiotemporal studies to understand delivery patterns are quite rare. However, in resource constraint countries like Bangladesh, detecting hot and cold spot areas can aid in the detection of diffusion centers, which can be targeted to expand regions with high facility deliveries. Places and periods with reduced health facility usages can be identified using various cluster detection techniques, to assess the barriers and facilitators in promoting health facility deliveries.

Using geospatial techniques to develop an emergency referral transport system for suspected sepsis patients in Bangladesh.

BACKGROUND: A geographic information system (GIS)-based transport network within an emergency referral system can be the key to reducing health system delays and increasing the chances of survival, especially during an emergency. We employed a GIS to design an emergency transport system for the rapid transfer of pregnant or early post-partum women, newborns, and children under 5 years of age with suspected sepsis under the Interrupting Pathways to Sepsis Initiative (IPSI) project. METHODS: A GIS database was developed by mapping the villages, roads, and relevant physical features of the study area. A travel-time algorithm was developed to incorporate the time taken by different modes of local transport to reach the health complexes. These were used in a network analysis to identify the shortest routes to the hospitals from the villages, which were categorized into green, yellow, and red zones based on their proximity to the nearest hospitals to provide transport facilities. An emergency call-in centre established for the project managed the transport system, and its data was used to assess the uptake of this transport system amongst distant communities. RESULTS: Fifteen pre-existing and two new routes were identified as the shortest routes to the health complexes. The call-in centre personnel used this route information to direct both patients and transport drivers to the nearest transport hubs or pick-up points. Adherence with referral advice was high in areas where the IPSI transport operated. Over the study period, the utilisation of the project's transport doubled and referral compliance from distant zones similarly increased. CONCLUSIONS: The GIS system created for this study facilitated rapid referral of patients in emergency from distant zones, using locally available transport and resources. The methodology described in this study to develop and implement an emergency transport system can be applied in similar, rural, low-income country settings.