Genome-wide identification, characterization and expression profiling of gibberellin metabolism genes in jute.

BACKGROUND: Gibberellin (GA) is one of the most essential phytohormones that modulate plant growth and development. Jute (Corchorus sp.) is the second most important source of bast fiber. Our result has shown that exogenous GA can positively regulate jute height and related characteristics which mean increasing endogenous GA production will help to get a jute variety with improved characteristics. However, genes involved in jute GA biosynthesis have not been analyzed precisely. RESULTS: Genome-wide analysis identified twenty-two candidate genes involved in jute GA biosynthesis pathway. Among them, four genes- CoCPS, CoKS, CoKO and CoKAO work in early steps. Seven CoGA20oxs, three CoGA3oxs, and eight GA2oxs genes work in the later steps. These genes were characterized through phylogenetic, motif, gene structure, and promoter region analysis along with chromosomal localization. Spatial gene expression analysis revealed that 11 GA oxidases were actively related to jute GA production and four of them were marked as key regulators based on their expression level. All the biosynthesis genes both early and later steps showed tissue specificity. GA oxidase genes were under feedback regulation whereas early steps genes were not subject to such regulation. CONCLUSION: Enriched knowledge about jute GA biosynthesis pathway and genes will help to increase endogenous GA production in jute by changing the expression level of key regulator genes. CoGA20ox7, CoGA3ox2, CoGA2ox3, and CoGA2ox5 may be the most important genes for GA production.

An innovative ring-shaped electroeluter for high concentration preparative isolation of protein from polyacrylamide gel.

A ring-shaped electroeluter (RSE) was designed for protein recovery from polyacrylamide gel matrix. The RSE was designed in such a way that a ring-shaped well was used to place gel slices and an enrichment well was used to collect eluted protein samples. With HSA as model protein, the electroelution time was less than 30 min with 80% recovery rate, and the concentration of recovered protein was 50 times higher than that of conventional method. The RSE could be reused at least ten times. The developed device makes great advance towards economic electroelution of biomolecules (such as proteins) from gel matrix.

In-Vial Temperature Gradient Headspace Single Drop Microextraction Designed by Multiphysics Simulation.

Presented herein is a novel headspace single drop microextraction (HS-SDME) based on temperature gradient (TG) for an on-site preconcentration technique of volatile and semivolatile samples. First, an inner vial cap was designed as a cooling device for acceptor droplet in HS-SDME unit to achieve fast and efficient microextraction. Second, for the first time, an in-vial TG was generated between the donor phase in a sample vial at 80 °C and the acceptor droplet under the inner vial cap containing cooling liquid at -20 °C for a TG-HS-SDME. Third, a simple mathematic model and numerical simulations were developed by using heat transfer in fluids, Navier-Stokes and mass balance equations for conditional optimization, and dynamic illumination of the proposed extraction based on COMSOL Multiphysics. Five chlorophenols (CPs) were selected as model analytes to authenticate the proposed method. The comparisons revealed that the simulative results were in good agreement with the quantitative experiments, verifying the design of TG-HS-SDME via the numerical simulation. Under the optimum conditions, the extraction enrichments were improved from 302- to 388-fold within 2 min only, providing 3.5 to 4 times higher enrichment factors as compared to a typical HS-SDME. The simulation indicated that these improvements in the extraction kinetics could be attributed due to the applied temperature gap between the sample matrix and acceptor droplet within the small volume of headspace. Additionally, the experiments demonstrated a good linearity (0.03-100 µg/L, R2 > 0.9986), low limit of detection (7-10 ng/L), and fair repeatability (<5.9% RSD, n = 6). All of the simulative and experimental results indicated the robustness, precision, and usefulness of TG-HS-SDME for trace analyses of analytes in a wide variety of environmental, pharmaceutical, food safety, and forensic samples.

A highly efficient three-phase single drop microextraction technique for sample preconcentration.

A highly efficient three-phase single drop microextraction (SDME) method is presented by using an organic-aqueous compound droplet. A coupling microdevice is designed to produce compound droplets in different sizes conveniently. In this way, the volume ratio of organic phase to aqueous phase in a compound droplet can be significantly reduced. Good operability and droplet stability were observed during extraction under vigorous stirring conditions. Five statins were used as model compounds and spiked in river water and human serum samples to evaluate the analytical performance of the proposed method. By using a 1.2 µL toluene-aqueous compound droplet (volume ratio 0.2 : 1), a 350 to 1712 fold enrichment of statins was obtained within 4 minutes. The results indicate that the proposed method is a very rapid and efficient sample pretreatment method, and is promising for automated and high-throughput applications.

A new strategy for highly efficient single-drop microextraction with a liquid-gas compound pendant drop.

Herein, a simple assembly was designed via a capillary and a funnel-like cap to achieve liquid-gas compound pendant drop (CPD) microextraction with great convenience. Due to the increased contact area and adhesion force between the capillary tip and the drop, the proposed method provides considerable flexibility in producing CPDs with different air bubble sizes. Four pesticides were chosen as model analytes to evaluate the proposed method. By using a 1 µL chlorobenzene droplet containing a 1 µL air bubble at a stirring rate of 700 rpm, a 70 to 135-fold enrichment of pesticides was obtained within 3.4 minutes. As compared with a typical SDME, the proposed method showed a 2-fold increase of enrichment factors and a 4-fold decrease of extraction time. Improvement of the extraction efficiency could be ascribed to the increased surface area of the droplet, and the thin film phenomena further improved the extraction kinetics through effective agitation. The results indicate that CPD microextraction could serve as a promising sample pretreatment method for automated high-throughput analyses in a wide variety of research areas.

Captopril challenge test: an underutilized test in the diagnosis of primary aldosteronism.

Primary aldosteronism (PA) is the most common cause of endocrine hypertension and is often underdiagnosed. This condition is associated with increased cardiovascular morbidity and mortality in comparison to age and blood pressure matched individuals with essential hypertension (EH). The diagnostic pathway for PA consists of three phases: screening, confirmatory testing, and subtyping. The lack of specificity in the screening step, which relies on the aldosterone to renin ratio, necessitates confirmatory testing. The Endocrine Society's clinical practice guideline suggests four confirmatory tests, including the fludrocortisone suppression test (FST), saline suppression test (SST), captopril challenge test (CCT), and oral sodium loading test (SLT). There is no universally accepted choice of confirmatory test, with practices varying among centers. The SST and FST are commonly used, but they can be resource-intensive, carry risks such as volume overload or hypokalemia, and are contraindicated in severe/uncontrolled HTN as well as in cardiac and renal impairment. In contrast, CCT is a safe and inexpensive alternative that can be performed in an outpatient setting and can be applied when other tests are contraindicated. Despite its simplicity and convenience, the variability in captopril dose, testing posture, and diagnostic threshold limit its widespread use. This narrative review evaluates the diagnostic accuracy of the CCT across different populations, addresses controversies in its usage, and proposes recommendations for its use in the diagnosis of PA. Furthermore, suggestions for future research aimed at promoting the wider utilization of the CCT as a simpler, safer, and more cost-effective diagnostic test are discussed.

Heavy metals and metalloid contamination and risk evaluation in the surface sediment of the Bakkhali River estuary in Bangladesh.

Current state of contamination and subsequent risk of contaminated sediment of a tidal river of Bangladesh was evaluated in the present study. Sediment samples were collected from five locations in the tributary of Bakkhali River estuary during summer (April) and winter (December) season, 2020. Collected samples were processed using standard protocol and the content of heavy metals (Cd, Cr, Cu, Pb and Zn) and metalloid (As), were analyzed by the Flame Atomic Absorption Spectrometer. Sediment contamination was evaluated by pollution load index (PLI), contamination factor (CF), degree of contamination (Cd), potential ecological risk index (PERI), non-carcinogenic and carcinogenic risk (CR) due to the dermal contact of the sediment. Multivariate statistical analysis such as principal component analysis (PCA) and cluster analysis (CA) were also applied to find out the possible sources of the contaminant in the sediment. Results showed the average concentration of As, Cd, Cr, Pb, Cu and Zn was 9.74 ± 3.57, 2.00 ± 0.85, 48.75 ± 8.92, 29.78 ± 8.39, 5.44 ± 2.03 and 56.94 ± 8.57 mg/kg, respectively. Concentration of Cu, Pb and Zn were within the recommended level whereas the concentration of As, Cd and Cr were suppressed the recommended level of WHO and FAO/WHO standards. PLI, CF and Cd revealed considerably low degree of contamination of the sediment. Geo-accumulation index indicated uncontaminated to moderately contaminated condition of the sediment. Although the values of enrichment factor revealed no potential enrichment for most of the metals, Cd showed a minor enrichment during the winter season. Based on the ecological risk assessment, the sediment from all of the sample locations was found to be of moderate to low risk. PCA and CA analysis revealed the origin of contaminants mainly from anthropogenic sources. Although different metals showed non-carcinogenic risk to the inhabitants, cancer risk values for dermal contact (CRderm) were much lower than 10-6 indicating no cancer risk for adult and child. However, the findings also revealed that children were more susceptible to CRderm compared to adults. The present study concluded that long term dermal contact of the sediment of Bakkhali River estuary will be contagious to the people. Therefore, regular monitoring of the estuarine environment is necessary so that contamination does not get worse.

First-Principles Calculations on Electronic, Optical, and Phonon Properties of γ-Bi2MoO6.

The wide band gap γ-Bi2MoO6 (BMO) has tremendous potential in emergent solar harvesting applications. Here we present a combined experimental-first-principles density functional theory (DFT) approach to probe physical properties relevant to the light sensitivity of BMO like dynamic and structural stability, Raman and infrared absorption modes, value and nature of band gap (i.e., direct or indirect), dielectric constant, and optical absorption, etc. We solvothermally synthesized wide band gap Pca21 phase pure BMO (≳3 eV) for two different pH values of 7 and 9. The structural parameters were correlated with the stability of BMO derived from elastic tensor simulations. The desired dynamical stability at T = 0 K was established from the phonon vibrational band structure using a finite difference-based supercell approach. The DFT-based Raman modes and phonon density of states (DOS) reliably reproduced the experimental Raman and infrared absorption. The electronic DOS calculated from Heyd-Scuseria-Ernzerhof HSE06 with van der Waals (vdW) and relativistic spin-orbit coupling (SOC) corrections produced a good agreement with the band gap obtained from diffuse reflectance spectroscopy (DRS). The optical absorption obtained from the complex dielectric constant for the HSE06+SOC+vdW potential closely resembled the DRS-derived absorption of BMO. The BMO shows ∼43% photocatalytic efficiency in degrading methylene blue dye under 75 min optical illumination. This combined DFT-experimental approach may provide a better understanding of the properties of BMO relevant to solar harvesting applications.

Tea (Camellia sinensis) cultivated in three agro-ecological regions of Bangladesh: Unveiling the variability of methylxanthine, bioactive phenolic compound, and antioxidant activity.

Tea (Camellia sinensis) is a widely consumed beverage known for its numerous health benefits, largely attributed to its rich content of quality determining secondary metabolites such as methylxanthine compounds and bioactive phenolic compounds. The goal of this study was to find out variations of the levels of methylxanthines, bioactive phenolic compounds, and antioxidant activity in methanolic and hot water extracts of 129 tea samples grown in three different ecological regions of Bangladesh named Panchagar, Sylhet, and Chattogram. Methylxanthine and bioactive phenolic compounds were determined by using HPLC-DAD, and the antioxidant profile was analysed by UV-vis spectrophotometric methods for methanol and hot water extracts of tea leaves. The IC50 values showed the trend as Panchagar > Sylhet > Chattogram and Sylhet > Chattogram > Panchagar for water and methanol extract, respectively. The results revealed significant (p < 0.05) variations in the levels of methylxanthines content: Panchagar > Chattogram > Sylhet. Caffeine was significantly higher (103.02 ± 5.55 mg/g of dry extract) in the methanolic extract of tea leaves of Panchagar district and lower (53.33 ± 4.30 mg/g of dry extract) in the hot water extract of Sylhet district. Panchagar and Chattogram possessed significantly (p < 0.05) higher catechin content for methanol (57.01 ± 5.50 mg/g dry extract) and hot water (55.23 ± 4.11 mg/g dry extract) extracts, respectively. The utilization of canonical discriminant functions yielded highly favorable outcomes in the classification of tea from three distinct cultivation origins in Bangladesh, relying on their inherent features. This study demonstrated the potential effects of geographical variations on the bioactive compounds and antioxidant properties of tea, emphasizing the importance of regional differences in tea cultivation for optimizing its health benefits as well as dispersing tea cultivation across the country.

Evaluation of 3ß-hydroxysteroid dehydrogenase activity using progesterone and androgen receptors-mediated transactivation.

3ß-Hydroxysteroid dehydrogenases (3ß-HSDs) catalyze the oxidative conversion of delta (5)-ene-3-beta-hydroxy steroids and ketosteroids. Human 3ß-HSD type 2 (HSD3B2) is predominantly expressed in gonadal and adrenal steroidogenic cells for producing all classes of active steroid hormones. Mutations in HSD3B2 gene cause a rare form of congenital adrenal hyperplasia with varying degree of salt wasting and incomplete masculinization, resulting from reduced production of corticoids and androgens. Therefore, evaluation of the HSD3B2 enzymatic activity in both pathways for each steroid hormone production is important for accurately understanding and diagnosing this disorder. Using progesterone receptor (PR)- and androgen receptor (AR)-mediated transactivation, we adapted a method that easily evaluates enzymatic activity of HSD3B2 by quantifying the conversion from substrates [pregnenolone (P5) and dehydroepiandrosterone (DHEA)] to (progesterone and androstenedione). HEK293 cells were transduced to express human HSD3B2, and incubated medium containing P5 or DHEA. Depending on the incubation time with HSD3B2-expressing cells, the culture media progressively increased luciferase activities in CV-1 cells, transfected with the PR/AR expression vector and progesterone-/androgen-responsive reporter. Culture media from human and other mammalian HSD3B1-expressing cells also increased the luciferase activities. HEK293 cells expressing various missense mutations in the HSD3B2 gene revealed the potential of this system to evaluate the relationship between the enzymatic activities of mutant proteins and patient phenotype.

Socio-economic and environmental factors associated with high lymphatic filariasis morbidity prevalence distribution in Bangladesh.

BACKGROUND: Lymphatic filariasis (LF) is a vector-borne parasitic disease which affects 70 million people worldwide and causes life-long disabilities. In Bangladesh, there are an estimated 44,000 people suffering from clinical conditions such as lymphoedema and hydrocoele, with the greatest burden in the northern Rangpur division. To better understand the factors associated with this distribution, this study examined socio-economic and environmental factors at division, district, and sub-district levels. METHODOLOGY: A retrospective ecological study was conducted using key socio-economic (nutrition, poverty, employment, education, house infrastructure) and environmental (temperature, precipitation, elevation, waterway) factors. Characteristics at division level were summarised. Bivariate analysis using Spearman's rank correlation coefficient was conducted at district and sub-district levels, and negative binomial regression analyses were conducted across high endemic sub-districts (n = 132). Maps were produced of high endemic sub-districts to visually illustrate the socio-economic and environmental factors found to be significant. RESULTS: The highest proportion of rural population (86.8%), poverty (42.0%), tube well water (85.4%), and primary employment in agriculture (67.7%) was found in Rangpur division. Spearman's rank correlation coefficient at district and sub-district level show that LF morbidity prevalence was significantly (p<0.05) positively correlated with households without electricity (district rs = 0.818; sub-district rs = 0.559), households with tube well water (sub-district rs = 0.291), households without toilet (district rs = 0.504; sub-district rs = 0.40), mean annual precipitation (district rs = 0.695; sub-district rs = 0.503), mean precipitation of wettest quarter (district rs = 0.707; sub-district rs = 0.528), and significantly negatively correlated with severely stunted children (district rs = -0.723; sub-district rs = -0.370), mean annual temperature (district rs = -0.633.; sub-district rs = 0.353) and mean temperature (wettest quarter) ((district rs = -0.598; sub-district rs = 0.316) Negative binomial regression analyses at sub-district level found severely stunted children (p = <0.001), rural population (p = 0.002), poverty headcount (p = 0.001), primary employment in agriculture (p = 0.018), households without toilet (p = <0.001), households without electricity (p = 0.002) and mean temperature (wettest quarter) (p = 0.045) to be significant. CONCLUSIONS: This study highlights the value of using available data to identify key drivers associated with high LF morbidity prevalence, which may help national LF programmes better identify populations at risk and implement timely and targeted public health messages and intervention strategies.

Chemical composition, antioxidant and antibacterial activity of Piper chaba stem extracts with preservative effects on storage of raw beef patties.

Piper chaba, a traditional South-east Asian medicinal herb and well-known curry spice, was studied to evaluate its suitability as a source of natural preservatives for beef products. Plant extracts that are high in phenolics and have high antimicrobial and antioxidant activities are likely to be useful as a natural preservative. Therefore, the phytochemical composition and the bioactivities of both ethanolic and methanolic extracts of P. chaba stem were examined first. The study revealed a significant antioxidant activities and potential antibacterial activity of P. chaba extracts. Next we investigated the preservation characteristics of P. chaba by using beef patties as a model system. Beef patties were produced and treated with 0.2 % ethanolic extract (mentioned as PEE) of P. chaba and 0.1 % commercial preservative (mentioned as PCP). They were then assessed for various storage quality parameters under refrigerated (4° C ± 1° C) conditions, including free fatty acid, antioxidant contents, and oxidative stability at 0, 6th, 16th, and 33rd days. No significant variations were observed across the products with regard to proximate composition study such as protein, ash and fat contents. In comparison to both PEE and PCP, the control product had higher free fatty acid values throughout the storage period. This indicates that the fat content of the PEE and PCP degraded at a slower rate than the control over the 33-day storage period. Our study also showed that both PCP and PEE had increased antioxidant capacity, implying that lipid oxidation is minimized. In contrast to the control, the oxidative stability of the P. chaba treated products was also higher. Altogether this study revealed that P. chaba could be utilized commercially, particularly in the food industry to preserve muscle foods. Practical Applications: Natural preservatives are becoming more popular as a result of the different carcinogenic and toxic side effects of conventional preservatives. P. chaba, an exquisite culinary herb in Bangladesh, has long been used as a traditional medicine, because of its antimicrobial and antioxidant properties. This study revealed that P. chaba can be utilized as a food preservative, which opens up new possibilities for its development and use in functional foods.

A combined first principles and experimental approach to Bi2WO6.

Here we synthesized Bi2WO6 (BWO) using both solid-state reaction (SBWO) and hydrothermal (HBWO-U and HBWO-S) methods. The orthorhombic Pca21 phase purity in all samples is confirmed from Rietveld refinement of X-ray diffraction data, Raman spectroscopy, and Fourier transform infrared spectroscopy. The HBWO-U and HBWO-S morphology revealed rectangular, spherical, and rod-like features with an average particle size of 55 nm in field emission scanning electron micrographs. A high-resolution transmission electron micrograph showed spherical-shaped particles in the HBWO-U sample with an average diameter of ∼10 nm. The diffuse reflectance-derived indirect electronic band gaps lie within the 2.79-3.23 eV range. The BWO electronic structure is successfully modeled by Hubbard interaction Ud and Up corrected Perdew-Burke-Ernzerhof generalized gradient approximation GGA-PBE+Ud+Up with van der Waals (vdW) force in effect. The optimized (Ud, Up) values are further justified by tuning the Hartree-Fock (HF) exact-exchange mixing parameter αHF from 25% in Heyd-Scuseria-Ernzerhof (HSE06) to 20% in the PBE-HF20% functional. Moreover, no inconsistencies were seen in the GGA-PBE+Ud+Up+vdW simulated crystallographic parameters, and the elastic tensor, phonon, and linear optical properties. Overall, the computationally cheap GGA-PBE+Ud+Up with vdW force may have successfully probed the physical properties of BWO.

Relative evaluation of in-vitro antioxidant potential and phenolic constituents by HPLC-DAD of Brassica vegetables extracted in different solvents.

Cabbage, cauliflower and broccoli are well-known vegetables from the Brassica family having functional effects on human health. This study was carried out to identify different antioxidant properties and to quantify phenolic compounds by HPLC-DAD in different extracts (methanol, ethanol and water: acetic acid: acetone) of these vegetables. The results showed that, the methanolic dry extract of cabbage possessed the highest antioxidant activity (549 ± 7.30 µg/g) and IC50 was 90 ± 2.52 µg/mL than others. Whereas the ethanolic dry extract of cauliflower had 348 ± 5.20 µg/g of flavonoid, which was the highest among all. The maximum levels of total tannin (414 ± 5.20 µg/g) and total phenolic content (465 ± 3.25 µg/g) was found in broccoli dry extract. Several polyphenolic compounds were identified in different extracts of the vegetables and they were Cauliflower (8) > Cabbage (10) > Broccoli (9) in total. Therefore, the use of total vegetables rather than extracts in the food industry would be more appropriate to get greater health benefit.

CDKAL1 gene rs7756992 A/G and rs7754840 G/C polymorphisms are associated with gestational diabetes mellitus in a sample of Bangladeshi population: implication for future T2DM prophylaxis.

BACKGROUND: Association of single nucleotide polymorphisms (SNP) rs7756992 A/G and rs7754840 G/C of cyclin-dependent kinase 5 regulatory subunit-associated protein 1-like 1 (CDKAL1) gene with the susceptibility of gestational diabetes mellitus (GDM) has been studied in a group of Bangladeshi women. METHODS: In this case-control study, 212 GDM patients and 256 control subjects were genotyped for rs7756992 and rs7754840 by PCR-RFLP and TaqMan™ allelic discrimination assay method respectively. Genotyping results were confirmed by DNA sequencing and replicated TaqMan™ assay. The odds ratios and their 95% confidence interval were calculated by logistic regression to determine the associations between genotypes and GDM. RESULTS: The genotype frequencies of rs7756992-AA/AG/GG in the GDM group and the control group were 37%/48%, 53%/45%, 10%/7% and those of rs7754840-CC/CG/GG were 51%/55%, 40.1%/39.8%, 9%/5% respectively. Under dominant and log additive models rs7756992 was revealed significantly associated with GDM after being adjusted for family history of diabetes (FHD) and gravidity. Conversely, rs7754840 was significantly associated (P = 0.047) with GDM only under the recessive model after the same adjustment. The risk allele frequency of both SNPs was higher in the GDM group but significantly (P = 0.029) increased prevalence was observed in the rs7756992 G allele. When positive FHD and risk alleles of these SNPs were synergistically present in any pregnant woman, the chance of developing GDM was augmented by many folds. The codominant model revealed 2.5 and 2.1 folds increase in odds by AG (rs7756992) and GC (rs7754840) genotypes and 3.7 and 4.5 folds by GG (rs7756992) and CC (rs7754840) genotypes respectively. A significant 2.7 folds (P = 0.038) increase in odds of GDM resulted from the interaction of rs7756992 and family history of diabetes under the dominant model. The cumulative effect of multigravidity and risk alleles of these SNPs increased the odds of GDM more than 1.5 folds in different genotypes. CONCLUSION: This study not only revealed a significant association between rs7756992 and rs7754840 with GDM but also provided the possibility as potential markers for foretelling about GDM and type 2 diabetes mellitus in Bangladeshi women.

Formulation of nutrient enriched germinated wheat and mung-bean based weaning food compare to locally available similar products in Bangladesh.

Poor weaning practice and malnutrition among under 5 (yrs) children are still major public health issues in Bangladesh. This study aimed to develop a cheap and nutritious weaning food for the children of Bangladesh. For this purpose, three weaning formulations of Q1, Q2, and Q3 with different ratios of germinated wheat, germinated mung-bean, and soya-bean, and a constant amount of sweet potato, sugar, salt, and milk flavor were processed and evaluated. The prepared formulations were investigated for proximate composition and sensory evaluation and compared with six commercial weaning food products. The proximate composition values indicated that the fat content of formulated foods ranged between 09.29% and 11.40%. The carbohydrate content was ranged between 52.80% and 61.20%, which was low compared with commercial ones. The protein content of the formulated foods was 20.33%-27.70%, and that was approximately two times more than available commercial foods. The energy content was also more than locally available commercial weaning foods, which were 411.40 ± 1.51 kcal to 419.30 ± 1.12 kcal. Sample Q2 had an 8.4 acceptance score in sensory analysis of a 9-point hedonic scale scorecard, which made it more acceptable than the other two samples. The values of mineral elements (Na, K, Fe) were similar to all analyzed varieties of commercial weaning foods. This nutrient-enriched weaning food will easily be affordable for the people of developing countries like Bangladesh. The results showed that the formulated weaning food had the desired characteristics of a weaning food; hence, it could decrease malnutrition in children.

Whole exome sequencing uncovered highly penetrant recessive mutations for a spectrum of rare genetic pediatric diseases in Bangladesh.

Collectively, rare genetic diseases affect a significant number of individuals worldwide. In this study, we have conducted whole-exome sequencing (WES) and identified underlying pathogenic or likely pathogenic variants in five children with rare genetic diseases. We present evidence for disease-causing autosomal recessive variants in a range of disease-associated genes such as DHH-associated 46,XY gonadal dysgenesis (GD) or 46,XY sex reversal 7, GNPTAB-associated mucolipidosis II alpha/beta (ML II), BBS1-associated Bardet-Biedl Syndrome (BBS), SURF1-associated Leigh Syndrome (LS) and AP4B1-associated spastic paraplegia-47 (SPG47) in unrelated affected members from Bangladesh. Our analysis pipeline detected three homozygous mutations, including a novel c. 863 G > C (p.Pro288Arg) variant in DHH, and two compound heterozygous variants, including two novel variants: c.2972dupT (p.Met991Ilefs*) in GNPTAB and c.229 G > C (p.Gly77Arg) in SURF1. All mutations were validated by Sanger sequencing. Collectively, this study adds to the genetic heterogeneity of rare genetic diseases and is the first report elucidating the genetic profile of (consanguineous and nonconsanguineous) rare genetic diseases in the Bangladesh population.

LEYDIG CELL HYPOPLASIA: A UNIQUE PARADOX IN THE DIAGNOSIS OF 46,XY DISORDERS OF SEX DEVELOPMENT.

OBJECTIVE: Disorders of sex development (DSD) are defined as conditions in which chromosomal sex is inconsistent with phenotypic sex, or in which the phenotype is not classifiable as either male or female. Mutations in genes present in X, Y or autosomal chromosomes can cause abnormalities of testis determination or 46,XY DSD. Leydig cell hypoplasia (LCH), also known as Leydig cell agenesis, is a rare autosomal recessive endocrine syndrome of 46,XY DSD. Our objective here is to present the case of a 27-year-old, phenotypic female who presented with primary amenorrhea and later found to have LCH. METHODS: We used formatted history and clinical examination followed by necessary hormonal investigations. The diagnosis was confirmed by histopathology of resected testes and genetic mutation analysis. RESULTS: The patient's physical examination was unremarkable except 2 ovoid lumps present in the inguinovulvar region. There were no müllerian structures on sonography. Estrogen and both basal and stimulated testosterone levels were low whereas luteinizing hormone and follicle-stimulating hormone were high. Her chromosomal sex was found to be 46,XY. The histopathology of the resected inguinal lumps showed atrophic testicular change lacking Leydig cells with relative preservation of Sertoli cells. Genetic mutation analysis failed to reveal any significant aberration in the LHCGR gene. At present she is on estrogen replacement therapy having undergone bilateral orchidectomy and vaginoplasty. CONCLUSION: LCH represents a unique example of diagnostic dilemma in gender identification. It requires a multidisciplinary approach for optimum outcome.

Methanol soluble fraction of fruits of Annona muricata possesses significant antidiarrheal activities.

Medicinal plants are the major sources of traditional treatment of disease in Indian subcontinent due to abundant presence of plants and vast side effects of synthetic drug. The present study was subjected to observe in vitro thrombolytic, antibacterial, and in vivo antidiarrheal activities of methanol soluble fraction of fruits of Annona muricata. In thrombolytic activity assay, various concentrations (2 ─ 10 mg/ml) of methanol soluble fraction was used and dose dependently less potent activity was found. The maximum clot lysis 18.33% (p* < 0.05) was achieved at 10 mg/ml of methanolic fruit extract, whereas standard drug streptokinase showed 55.50% (p*** < 0.001) clot lysis. In antibacterial assay, disc diffusion method was used comprising two gram positive (S. aureus and Micrococcus luteus) and two gram negative (E. coli and P. aeruginosa) bacteria. None of four (0.25, 0.5, 1, and 5 mg/disc) concentration of fruit extract showed antibacterial potentiality, whereas standard amikacin (3 mg/disc) revealed strong antibacterial activities (=~ 23 ─ 24 mm of MIC). To evaluate antidiarrheal activity, castor oil induced diarrhea was created in Swiss albino mice and different doses (100, 200, and 400 mg/kgbw) of fruit extract was introduced post orally. All of three different doses of fruit extract showed significant (p < 0.05 ─ 0.001) antidiarrheal activities. Notably, the percent inhibition of diarrhea by methanolic extract of fruits of Annona muricata was found to be 58.38% at a dose of 400 mg/kgbw. The effect of vehicle saline (10 ml/kgbw) was considered as control and loperamide (5 mg/kgbw) as standard that provided 67.01% inhibition of diarrhea. The results suggest that, the fruits of Annona muricata possess potent antidiarrheal properties, providing scientific basis of using the plant parts in the treatment of diarrheal disease.

Postoperative expression of Cushing disease in a young male: metamorphosis of silent corticotroph adenoma?

SUMMARY: Silent corticotroph adenoma (SCA) is an unusual type of nonfunctioning pituitary adenoma (NFA) that is silent both clinically and biochemically and can only be recognized by positive immunostaining for ACTH. Under rare circumstances, it can transform into hormonally active disease presenting with severe Cushing syndrome. It might often produce diagnostic dilemma with difficult management issue if not thoroughly investigated and subtyped accordingly following surgery. Here, we present a 21-year-old male who initially underwent pituitary adenomectomy for presumed NFA with compressive symptoms. However, he developed recurrent and invasive macroadenoma with severe clinical as well as biochemical hypercortisolism during post-surgical follow-up. Repeat pituitary surgery was carried out urgently as there was significant optic chiasmal compression. Immunohistochemical analysis of the tumor tissue obtained on repeat surgery proved it to be an aggressive corticotroph adenoma. Though not cured, he showed marked clinical and biochemical improvement in the immediate postoperative period. Anticipating recurrence from the residual tumor, we referred him for cyber knife radio surgery. LEARNING POINTS: Pituitary NFA commonly present with compressive symptoms such as headache and blurred vision. Post-surgical development of Cushing syndrome in such a case could be either drug induced or endogenous. In the presence of recurrent pituitary tumor, ACTH-dependent Cushing syndrome indicates CD. Rarely a SCA presenting initially as NFA can transform into an active corticotroph adenoma. Immunohistochemical marker for ACTH in the resected tumor confirms the diagnosis.