Early suppression of the endocannabinoid degrading enzymes FAAH and MAGL alters locomotor development in zebrafish.

The fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) enzymes are the predominant catabolic regulators of the major endocannabinoids (eCBs) anadamide (AEA) and 2-arachidonoylglycerol (2-AG), respectively. The expression and roles of eCBs during early embryogenesis remain to be fully investigated. Here, we inhibited FAAH and MAGL in zebrafish embryos during the first 24 h of life and examined motor neuron and locomotor development at 2 and 5 days post fertilization (dpf). Application of the dual FAAH/MAGL inhibitor, JZL195 (2 µmol l-1), resulted in a reduction in primary and secondary motor neuron axonal branching. JZL195 also reduced nicotinic acetylcholine receptor (nAChR) expression at neuromuscular junctions. Application of URB597 (5 µmol l-1), a specific inhibitor of the FAAH enzyme, also decreased primary motor neuron branching but did not affect secondary motor neuron branching and nAChR expression. Interestingly, JZL184 (5 µmol l-1), a specific inhibitor of MAGL, showed no effects on motor neuron branching or nAChR expression. Co-treatment of the enzyme inhibitors with the CB1R inhibitor AM251 confirmed the involvement of CB1R in motor neuron branching. Disruption of FAAH or MAGL reduced larval swimming activity, and AM251 attenuated the JZL195- and URB597-induced locomotor changes, but not the effects of JZL184. Together, these findings indicate that inhibition of FAAH, or augmentation of AEA acting through CB1R during early development, may be responsible for locomotor deficiencies.

Epidemiology and risk factors for pneumonia severity and mortality in Bangladeshi children <5 years of age before 10-valent pneumococcal conjugate vaccine introduction.

BACKGROUND: Pneumonia is the leading infectious cause of morbidity and mortality in young children in Bangladesh. We present the epidemiology of pneumonia in Bangladeshi children <5 years before 10-valent pneumococcal conjugate vaccine introduction and investigate factors associated with disease severity and mortality. METHODS: Children aged 2-59 months admitted to three Bangladeshi hospitals with pneumonia (i.e., cough or difficulty breathing and age-specific tachypnea without danger signs) or severe pneumonia (i.e., cough or difficulty breathing and ≥1 danger signs) were included. Demographic, clinical, laboratory, and vaccine history data were collected. We assessed associations between characteristics and pneumonia severity and mortality using multivariable logistic regression. RESULTS: Among 3639 Bangladeshi children with pneumonia, 61% had severe disease, and 2% died. Factors independently associated with severe pneumonia included ages 2-5 months (adjusted odds ratio [aOR] 1.60 [95% CI: 1.26-2.01]) and 6-11 months (aOR 1.31 [1.10-1.56]) relative to 12-59 months, low weight for age (aOR 1.22 [1.04-1.42]), unsafe drinking water source (aOR 2.00 [1.50-2.69]), higher paternal education (aOR 1.34 [1.15-1.57]), higher maternal education (aOR 0.74 [0.64-0.87]), and being fully vaccinated for age with pentavalent vaccination (aOR 0.64 [0.51-0.82]). Increased risk of pneumonia mortality was associated with age <12 months, low weight for age, unsafe drinking water source, lower paternal education, disease severity, and having ≥1 co-morbid condition. CONCLUSIONS: Modifiable factors for severe pneumonia and mortality included low weight for age and access to safe drinking water. Improving vaccination status could decrease disease severity.

Follow-up of cases of Haemophilus influenzae type b meningitis to determine its long-term sequelae.

OBJECTIVE: To measure physical and neurologic impact of Haemophilus influenzae type b (Hib) meningitis on surviving children through short- and long-term follow-up. STUDY DESIGN: Cases of Hib meningitis, diagnosed at a tertiary level pediatric hospital, were subjected to short- and long-term follow-up and compared with age, sex, and area of residence matched healthy controls. Follow-up assessments included thorough physical and neurodevelopmental assessments using a standardized protocol by a multidisciplinary team. RESULTS: Assessments of short-term follow-up cohort (n = 64) revealed hearing, vision, mental, and psychomotor deficits in 7.8%, 3%, 20%, and 25% of the cases, respectively. Deficits were 10%, 1.4%, 21%, and 25% in long-term follow-up cohort (n = 71), in that order. Mental and psychomotor deficits were found in 2% of the controls, none of whom had vision or hearing deficits. CONCLUSIONS: In addition to risk of death, Hib meningitis in children causes severe disabilities in survivors. These data facilitated a comprehensive understanding of the burden of Hib meningitis, specifically in developing countries where disabled children remain incapacitated because of lack of resources and facilities. The evidence generated from this study is expected to provide a compelling argument in favor of introduction and continuation of Hib conjugate vaccine in the national immunization program for children.

Ciprofloxacin-resistant Escherichia coli in hospital wastewater of Bangladesh and prediction of its mechanism of resistance.

Hospital and agriculture wastewater is mostly responsible for causing environmental pollution by spreading un-metabolized antibiotics and resistant bacteria, especially in Bangladesh. Here, we studied the influence of the most frequently prescribed antibiotic, fluoroquinolone (~72%), on the development of antibiotic resistance in Escherichia coli. Out of 300, 24 ciprofloxacin resistant E. coli isolates were selected for the study that showed the MBC(100) higher than expected (600 µg/mL). Here, we profiled plasmid, sequenced gyr genes, screened mutations and analyzed the effect of mutation on drug-protein interaction through molecular docking approach. We found that (1) out of 10, most of them (n = 7) had large plasmid(s); (2) all ciprofloxacin-resistant isolates had gyrA double mutations (S83L and D87Y); (3) no isolate had qnr gene; and (4) docking of ciprofloxacin with DNA gyrase A subunit suggests that acquisition of double mutation leads to alteration of the ciprofloxacin binding pocket.

Coding-Complete Genome Sequence of SARS-CoV-2 Isolate from Bangladesh by Sanger Sequencing.

A coding-complete genome sequence of a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) isolate was revealed. The sample for the virus was isolated from a female patient from Dhaka, Bangladesh, suffering from coronavirus disease-2019 (COVID-19).

Neurodevelopmental sequelae in pneumococcal meningitis cases in Bangladesh: a comprehensive follow-up study.

BACKGROUND: Evaluation of the long-term impact of pneumococcal meningitis on surviving children and their families is critical to fully comprehending the burden of pneumococcal disease and to facilitating an evidence-based decision for the introduction of pneumococcal vaccine. This study was an investigation of the short- and long-term impacts of pneumococcal meningitis among Bangladeshi children. METHODS: Case patients with pneumococcal meningitis who were hospitalized between January 2006 and March 2007 were subjected to short-term follow-up within 30-40 days of discharge. Case patients discharged prior to January 2005 were selected for long-term follow-up at 6-24 months after the date of discharge. Both cohorts were enrolled from Dhaka Shishu Hospital, a pediatric hospital in Bangladesh. Healthy children matched for age, sex, socioeconomic status, and area of residence were recruited from the community as control subjects. During follow-up visits, case patients and control subjects were assessed for their physical and neurodevelopmental status by use of a standardized protocol. The impact of pneumococcal meningitis on siblings and the family as a whole was assessed by means of qualitative interviews. RESULTS: Neurodevelopmental assessments of the short-term follow-up cohort (n=51) revealed hearing, vision, mental, and psychomotor deficits in 33%, 8%, 41%, and 49% of the case patients, respectively. These deficits were 18%, 4%, 41%, and 35% in the long-term follow-up cohort (n=51), respectively. Such deficits were seen in only 2% of the control subjects, none of whom had vision or hearing deficits. CONCLUSIONS: In addition to the risk of death, pneumococcal meningitis in children causes severe disabilities among survivors, as well as disruption of the life of other siblings and family members. This study demonstrated that high rates of sequelae are associated with pneumococcal meningitis. Neurodevelopmental assessment during follow-up of patients with meningitis is critical to our understanding of the burden of the adverse consequences of pneumococcal disease. These data, along with the fact of poor access to health care, provide a compelling argument in favor of the introduction of pneumococcal vaccine, specifically in a setting where access to health care is poor and disabled children remain incapacitated because of a lack of resources and facilities.

A topographic analysis of the proliferating tumor cells in an autopsied brain with infiltrative thalamic glioma.

Deep-seated gliomas, including thalamic gliomas, have a poor prognosis because of difficulty of accessibility for surgery. In addition, an infiltrative pattern of the tumor is related to a poor prognosis. In this study, the infiltrative/invasive profile of the proliferating tumor cells of a right thalamic glioma was evaluated in an autopsied brain. A 71-year-old man died from extensive infiltration of a right thalamic glioma. The distribution of the proliferating tumor cells at the right thalamic tumor level was represented by the topographic map of MIB-1 labeling indices (LI) on the whole-brain coronal slice, and this map was analyzed with pathological findings and postmortem T2-weighted magnetic resonance imaging (MRI). The highest MIB-1 LI was 24% for the whole autopsy brain at the thalamic tumor level, whereas the MIB-1 LI was 21% for the biopsy sample of the right thalamic glioma. Because this patient survived only 9 months after diagnosis of the tumor as anaplastic astrocytoma, it was confirmed that 21% MIB-1 LI of the biopsy sample was relevant to his prognosis. The topographic map of MIB-1 LI showed that the proliferating tumor cells of the right thalamic glioma invaded the ventricular walls and the contralateral thalamus by the periventricular route, but there was no exophytic extension to the cortex. In conclusion, topographic analysis of the proliferative potential detected by MIB-1 immunostaining provides information on the growth pattern of human glioma.

Antimicrobial and cytotoxic activity of three bitter plants enhydra fluctuans, andrographis peniculata and clerodendrum viscosum.

PURPOSE: In this study, three important medicinal plants (Enhydra fluctuans Lour, Clerodendrum viscosum Vent and Andrographis peniculata Wall) of Bangladesh were investigated to analyze their antimicrobial and cytotoxic activities against some pathogenic microorganisms and Artemia salina (brine shrimp nauplii). METHODS: The coarse powder material of leaves of each plant was extracted separately with methanol and acetone to yield methanol extracts of leaves of Enhydra fluctuans (MLE), Clerodendrum viscosum (MLC) and Andrographis peniculata (MLA), and acetone extracts of leaves of Enhydra fluctuans (ALE), Clerodendrum viscosum (ALC) and Andrographis peniculata (ALA). The disc diffusion method and the method described by Meyer were used to determine the antimicrobial and cytotoxic activities of each plant extract. RESULTS: Among the test samples, MLE and ALE showed comparatively better antimicrobial activity against a number of bacteria and fungi with inhibition zones in the range of 06-15 mm and according to the intensity of activity, the efficacy against microorganisms were found in the order of Enhydra fluctuans > Andrographis peniculata > Clerodendrum viscosum. In cytotoxicity assay, all samples were found to be active against brine shrimp nauplii (Artemia salina) and ALA produced lowest LC50 value (7.03 µg/ml). CONCLUSION: Enhydra fluctuans and Andrographis peniculata possesses significant antimicrobial and cytotoxic activities.

B- and T-cell epitope mapping of human sapovirus capsid protein: an immunomics approach.

Human sapovirus is one of the major causes of viral gastroenteritis. Although the capsid protein (VP1) confers antigenic cross-reactivity, immunity against sapovirus is still unclear. Using immunoinformatics approach, we defined putative T- and B-cell epitopes of VP1 and mapped on to its predicted three-dimensional structure. Identified five putative T-cell epitopes also occupied the putative B-cell epitope region. These putative epitopes were conserved in all existing serotypes. Predicted epitopes can be generated through proteasome cleavage and may be useful in designing peptide-based subunit vaccine to confer both humoral and cell-mediated immunity.