The genetic architecture of youth anxiety: a study protocol.

BACKGROUND: Anxiety disorders are the most common psychiatric problems among Canadian youth and typically have an onset in childhood or adolescence. They are characterized by high rates of relapse and chronicity, often resulting in substantial impairment across the lifespan. Genetic factors play an important role in the vulnerability toward anxiety disorders. However, genetic contribution to anxiety in youth is not well understood and can change across developmental stages. Large-scale genetic studies of youth are needed with detailed assessments of symptoms of anxiety disorders and their major comorbidities to inform early intervention or preventative strategies and suggest novel targets for therapeutics and personalization of care. METHODS: The Genetic Architecture of Youth Anxiety (GAYA) study is a Pan-Canadian effort of clinical and genetic experts with specific recruitment sites in Calgary, Halifax, Hamilton, Toronto, and Vancouver. Youth aged 10-19 (n = 13,000) will be recruited from both clinical and community settings and will provide saliva samples, complete online questionnaires on demographics, symptoms of mental health concerns, and behavioural inhibition, and complete neurocognitive tasks. A subset of youth will be offered access to a self-managed Internet-based cognitive behavioral therapy resource. Analyses will focus on the identification of novel genetic risk loci for anxiety disorders in youth and assess how much of the genetic risk for anxiety disorders is unique or shared across the life span. DISCUSSION: Results will substantially inform early intervention or preventative strategies and suggest novel targets for therapeutics and personalization of care. Given that the GAYA study will be the biggest genomic study of anxiety disorders in youth in Canada, this project will further foster collaborations nationally and across the world.

Hydro-chemical characteristics and groundwater quality evaluation in south-western region of Bangladesh: A GIS-based approach and multivariate analyses.

The study focuses on the chemistry of groundwater and if it is suitable for drinking and for use in agriculture using water quality indices, GIS mapping, and multivariate analyses in Sharsa Upazila, Jashore district, Bangladesh. In this study, the concentration of NH4+, K+, Ca2+, EC, Turbidity overstep BDWS drinking standards in 69 %, 14 %, 100 %, 40 % (WHO), 73 % of samples respectively. The value of Water Quality Indices (WQI) results inferred that the maximum specimen was held good quality for drinking uses, and the values distributed central eastern part to the south-eastern part were good quality water in the selected studied area. The study area's PH, EC, SAR, Na (%), TH, and NO3- values were mapped using GIS tools to show their spatial distribution. The cluster and correlation matrix analyses are used to validate for Principle Component Analysis (PCA). The five PCA results exhibited that the presence of EC, turbidity, K+, SO42- and NO3- was significant and was caused by both geogenic (rock weathering and cation exchange) and anthropogenic (agrochemicals, animal feedback) factor. According to the hydro-geochemical data, the maximum number of samples is of the Ca-Mg-HCO3-Cl type and is dominated by rocks. The irrigation water indices like MH, KR, SAR, and %Na indicate show high-quality groundwater for irrigation purposes. Most of the samples were satisfactory and compiled with WHO and Bangladeshi criteria for standard drinking water guideline values.

Human capital as a turnkey resource in resilience to cyclones and storm surges: Empirical evidence from coastal Bangladesh.

What are the best methods to build cyclone and storm resilience in a developing country? We examine the multiple resources that contribute to storm resilience in the highly vulnerable ecological context of coastal Bangladesh, finding that human capital is a critical turnkey resource that enables and facilitates the use of other resources in household responses to floods, storm surges and other cyclone damage. Drawing on a household survey of nine coastal villages in different ecological zones of coastal Bangladesh, we use principal component analysis (PCA) and multiple regression to identify four components of storm resilience and a human capital index that, along with other household resources, predicts resilience to storms and cyclones. We then use this human capital index as a policy tool to map the proportion of highly resilient households in these nine villages and identify additional methods for building a stronger understanding of storm resilience.

Exploring spatial variations in level and predictors of unskilled birth attendant delivery in Bangladesh using spatial analysis techniques: Findings from nationally representative survey data.

BACKGROUND: Bangladesh has failed to meet the United Nations goal for reducing maternal mortality in the last decade. The high prevalence of unskilled birth attendant (UBA) delivery (47%) has resulted in negative consequences for the health of mothers and newborn babies in the country. Spatial variations in UBA delivery and its predictors are yet to be explored in Bangladesh, which could be very helpful in formulating cost-effective policies for reducing that. This study examines the spatial variations in UBA delivery and its predictors in Bangladesh. METHODS: This study analyzed the characteristics of 672 clusters extracted from the 2017/18 Bangladesh Demographic and Health Survey, and healthcare facility data from the 2017 Bangladesh Health Facility Survey. These data were analyzed using descriptive and spatial analyses (hot spot analysis, Ordinary Least Squares Regression, and Geographically Weighted Regression) techniques. RESULTS: Statistically significant hot spots of UBA delivery were concentrated in parts of the Mymensingh, Sylhet, Barishal, and Rangpur regions, while Khulna was the safest region. Predictive strengths of the statistically significant predictors of spatial variation in UBA delivery were observed to vary considerably across the regions. Poorest household wealth status and less than four antenatal care contacts emerged as strong predictors of UBA delivery in all the aforementioned hot spot-stricken regions, except Barisal. Additionally, primiparity and all secondary education or higher were strong predictors of lower UBA delivery rates in Mymensingh and Sylhet, while poorer household wealth status was also a strong predictor of UBA delivery in Sylhet. Multiparity was an additional strong predictor of UBA delivery in Rangpur. In Barisal, only poorer household wealth status exerted a strong positive influence on UBA delivery. CONCLUSIONS: The remarkable spatial variations in UBA delivery and its predictors' strengths indicate that geographically-targeted interventions could be a cost-effective method for reducing the UBA delivery prevalence in Bangladesh, thereby improve maternal and child health.

Pharmacogenetic Testing Knowledge and Attitudes among Pediatric Psychiatrists and Pediatricians in Alberta, Canada.

OBJECTIVE: To assess knowledge, attitudes, and barriers as well as ethical, legal and social concerns towards pharmacogenetic (PGx) testing among pediatric psychiatrists and pediatricians in Alberta, Canada. METHOD: An anonymous electronic survey was sent to pediatric psychiatrists (n = 49) and pediatricians (n = 93) in Alberta. RESULTS: A total of 20 surveys were completed (response rate = 14%). Respondents agreed that PGx testing is clinically useful and a majority believed testing had the potential to aid in medication selection, dosing, switching, augmentation, and deprescribing, particularly among children with treatment-resistant conditions. However, most respondents could not identify an appropriate lab to perform testing, did not have the necessary training to interpret PGx results, and did not have access to experts that could assist them in interpreting results. CONCLUSION: The findings suggest additional PGx education and training is required to boost self-efficacy and uptake of PGx testing among pediatric psychiatrists and pediatricians in Alberta, Canada. In addition, local and global efforts to develop clinical practice guidelines, provide clear legal guidance, and ensure equitable access to testing may facilitate the implementation of PGx-informed prescribing.

OBJECTIF: Évaluer les connaissances, les attitudes, et les obstacles ainsi que les préoccupations éthiques, légales et sociales à l'égard des tests pharmacogénétiques (PGx) chez les psychiatres pédiatriques et les pédiatres d'Alberta, Canada. MÉTHODE: Un sondage anonyme électronique a été envoyé à des psychiatres pédiatriques (n = 49) et des pédiatres (n = 93) en Alberta. RÉSULTATS: En tout, 20 sondages ont été remplis (taux de réponse = 14 %). Les répondants convenaient que les tests PGx sont cliniquement utiles et une majorité croyait que les tests avaient le potentiel d'aider à la sélection des médicaments, au dosage, au changement, à l'augmentation et à la déprescription, particulièrement chez les enfants ayant des maladies réfractaires au traitement. Toutefois, la plupart des répondants ne pouvaient pas identifier un laboratoire approprié pour exécuter les tests, n'avaient pas la formation nécessaire pour interpréter les résultats des PGx, et n'avaient pas accès à des experts qui pourraient les aider à interpréter les résultats. CONCLUSION: Les résultats suggèrent qu'il faut plus d'éducation et de formation sur les PGx pour stimuler l'auto-efficacité et l'utilisation des tests PGx chez les psychiatres pédiatriques et les pédiatres de l'Alberta, Canada. En outre, les initiatives locales et mondiales en vue d'élaborer des guides de pratique clinique, d'offrir un guide juridique clair, et d'assurer un accès équitable aux tests peuvent faciliter la mise en œuvre de la prescription éclairée des PGx.

Methotrexate induced mitochondrial injury and cytochrome c release in rat liver hepatocytes.

Methotrexate (MTX) is a folic acid antagonist that is widely used to treat a variety of diseases. One of the most serious side effects of MTX therapy is hepatotoxicity. The potential molecular cytotoxic mechanisms of MTX toward isolated rat hepatocytes were investigated using Accelerated Cytotoxicity Mechanism Screening (ACMS) techniques. A concentration and time dependent increase in cytotoxicity and reactive oxygen species (ROS) formation and a decrease in mitochondrial membrane potential (MMP) were observed with MTX. Furthermore, a significant increase in MTX (300 µM)-induced cytotoxicity and ROS formation were observed when glutathione (GSH)-depleted hepatocytes were used whereas addition of N-acetylcysteine (a GSH precursor) decreased cytotoxicity. Catalase inactivation also increased MTX-induced cytotoxicity, while the direct addition of catalase to the hepatocytes decreased cytotoxicity. MTX treatment in isolated rat mitochondria caused swelling and significantly decreased adenosine triphosphate (ATP) and GSH content, and cytochrome c release. Potent antioxidants such as mesna, resveratrol and Trolox decreased MTX-induced cytotoxicity and ROS formation and increased MMP. This study suggests that MTX-induced cytotoxicity caused by ROS formation and GSH oxidation leads to oxidative stress and mitochondrial injury in rat hepatocytes.

(+)-Catechin prevents methylglyoxal-induced mitochondrial dysfunction and apoptosis in EA.hy926 cells.

OBJECTIVE: To investigate whether (+)-catechin, a strong antioxidant, can prevent methylglyoxal (MGO)-induced cytotoxicity and its mechanism. METHODS: Cytotoxicity, apoptosis, reactive oxygen species (ROS) generation, hydrogen peroxide (H2O2) formation, mitochondrial membrane potential (MMP) and mitochondrial morphology were measured in EA.hy926 cells. RESULT: MGO (4 mM)-induced cytotoxicity was markedly inhibited by (+)-catechin (0.1-4 mM) in 24 h. 1 mM MGO-induced apoptotic cell death (44.7%) was significantly inhibited by 4 mM (+)-catechin (to 24.4%), 1 mM aminoguanidine (AG) (to 28.8%) or 4 mM N-acetylcysteine (NAC) (to 24.3%). (+)-Catechin (4 mM) or AG (4 mM) can inhibit the decrease of MMP induced by MGO (2-8 mM) in 3 h. (+)-Catechin (4 mM) or AG (4 mM) can inhibit MGO (4 mM)-induced mitochondrial swelling in 3 h. However, MGO (4 mM)-induced ROS and H2O2 generation was not prevented by (+)-catechin (4 mM). CONCLUSIONS: (+)-Catechin prevents MGO-induced cytotoxicity in EA.Hy926 cells through inhibiting apoptosis and mitochondrial damage.

Flutamide-induced cytotoxicity and oxidative stress in an in vitro rat hepatocyte system.

Flutamide (FLU) is a competitive antagonist of the androgen receptor which has been reported to induce severe liver injury in some patients. Several experimental models suggested that an episode of inflammation during drug treatment predisposes animals to tissue injury. The molecular cytotoxic mechanisms of FLU in isolated rat hepatocytes using an in vitro oxidative stress inflammation system were investigated in this study. When a nontoxic hydrogen peroxide (H2O2) generating system (glucose/glucose oxidase) with peroxidase or iron(II) [Fe(II)] (to partly simulate in vivo inflammation) was added to the hepatocytes prior to the addition of FLU, increases in FLU-induced cytotoxicity and lipid peroxidation (LPO) were observed that were decreased by 6-N-propyl-2-thiouracil or deferoxamine, respectively. N-Acetylcysteine decreased FLU-induced cytotoxicity in this system. Potent antioxidants, for example, Trolox ((±)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), resveratrol (3,5,4'-trihydroxy-trans-stilbene), and DPPD (N,N'-diphenyl-1,4-phenylenediamine) also significantly decreased FLU-induced cytotoxicity and LPO and increased mitochondrial membrane potential (MMP) and glutathione (GSH) levels in the H2O2 generating system with peroxidase. TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl), a known reactive oxygen species (ROS) scavenger and superoxide dismutase mimetic, also significantly decreased toxicity caused by FLU in this system. These results raise the possibility that the presence or absence of inflammation may be another susceptibility factor for drug-induced hepatotoxicity.

Evaluation of azathioprine-induced cytotoxicity in an in vitro rat hepatocyte system.

Azathioprine (AZA) is widely used in clinical practice for preventing graft rejection in organ transplantations and various autoimmune and dermatological diseases with documented unpredictable hepatotoxicity. The potential molecular cytotoxic mechanisms of AZA towards isolated rat hepatocytes were investigated in this study using "Accelerated Cytotoxicity Mechanism Screening" techniques. The concentration of AZA required to cause 50% cytotoxicity in 2 hrs at 37°C was found to be 400 µM. A significant increase in AZA-induced cytotoxicity and reactive oxygen species (ROS) formation was observed when glutathione- (GSH-) depleted hepatocytes were used. The addition of N-acetylcysteine decreased cytotoxicity and ROS formation. Xanthine oxidase inhibition by allopurinol decreased AZA-induced cytotoxicity, ROS, and hydrogen peroxide (H2O2) formation and increased % mitochondrial membrane potential (MMP). Addition of N-acetylcysteine and allopurinol together caused nearly complete cytoprotection against AZA-induced hepatocyte death. TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl), a known ROS scavenger and a superoxide dismutase mimic, and antioxidants, like DPPD (N,N'-diphenyl-p-phenylenediamine), Trolox (a water soluble vitamin E analogue), and mesna (2-mercaptoethanesulfonate), also decreased hepatocyte death and ROS formation. Results from this study suggest that AZA-induced cytotoxicity in isolated rat hepatocytes may be partly due to ROS formation and GSH depletion that resulted in oxidative stress and mitochondrial injury.

N-acetyltransferase 2 (NAT2) genotype as a risk factor for development of drug-induced liver injury relating to antituberculosis drug treatment in a mixed-ethnicity patient group.

PURPOSE: This study aims to assess whether NAT2 genotype affects susceptibility to moderate to severe liver injury in patients undergoing drug treatment for tuberculosis with isoniazid-containing regimens. METHODS: Twenty-six patients of European or South Asian ethnicity, who had suffered liver injury during treatment with isoniazid-containing drug regimens and 101 ethnically matched controls were genotyped for the NAT2*5, NAT2*6, and NAT2*7 alleles. Genotyping for additional polymorphisms in the NAT gene region was also performed on 20 of the 26 cases. NAT2 genotype frequency between cases and controls was compared. RESULTS: NAT2 genotypes predicting a slow acetylator phenotype were found to be associated with an increased risk of isoniazid-related liver injury (odds ratio (OR) = 4.25 (95% confidence interval (CI), 1.36-13.22); p = 0.012) with 85% of the cases being slow acetylators compared with 56% of the controls. There was no evidence for an increased risk for the slow acetylator genotype in patients with the most severe cases of liver injury, who underwent liver transplantation. CONCLUSIONS: The NAT2 slow acetylator genotype appears to be a significant risk factor for moderate and severe drug- induced liver injury. However, the overall effect size is modest and generally in line with effects described previously for this genotype in milder drug-induced liver injury. Additional genetic risk factors may also contribute.

Genotype-phenotype variability in human CYP3A locus in Nepalese people residing in Bangladesh.

OBJECTIVES: The study has been designed to phenotype 200 Nepalese people residing in Bangladesh by measuring urinary ratio of 6ß-hydroxycortisol/cortisol (metabolic ratio) and to genotype all the subjects for the presence of CYP3A4*1B, *2, *4, *5, *6, *10, *18, CYP3A5*3, and *6 alleles. METHODS: Cortisol and 6ß-hydroxycortisol were extracted and quantified from morning spot urine samples (n = 200) by HPLC. Genotyping was carried out using the extracted genomic DNA by amplification of target alleles by PCR. Amplified DNA was digested by appropriate restriction enzymes followed by gel electrophoresis and sequencing to identify the targeted alleles. RESULTS: A wide ratio of 6ß-hydroxycortisol/cortisol was found (0.71 - 10.61) with an average of 4.41. No sample (n = 200) was found positive for CYP3A4*1B, *2, *4, *5, *6, *10, *18, and CYP3A5*6 alleles. CYP3A5*1/*1, *1/*3, and *3/*3 genotype frequency were found to be 20%, 20%, and 60%, respectively. A significantly higher mean metabolic ratio (MR) ± SD (MR = 6.28 ± 3.43) was found for CYP3A5*1/*1 compared to both CYP3A5*1/*3 (MR = 3.68 ± 1.37) and CYP3A5*3/*3 (MR = 3.58 ± 1.95). CONCLUSION: This study demonstrates the absence of common CYP3A4 variant alleles in Nepalese people residing in Bangladesh whereas Nepalese people carrying the CYP3A5*1/*1 genotype appear to show a significantly higher 6ß-hydroxycortisol/cortisol ratios compared to those with CYP3A5*3/*3 genotype.

Comparative pharmacokinetic and relative bioavailability study of coated and uncoated azithromycin powder for suspension in healthy Bangladeshi male volunteers.

Azithromycin (AZT; CAS 83905-01-5) is an efficient antibiotic and is widely prescribed in Bangladesh. The taste of uncoated AZT suspension is bitter. Although several taste masked oral suspensions of AZT are available in Bangladesh, information regarding the bioavailability of these formulations in Bangladeshi population is unavailable. The purpose of this study was to compare the relative bioavailability and other pharmacokinetic properties of two oral formulation of AZT (200 mg/5 ml) suspensions, the uncoated reference product and coated test product (Tridosil 200 mg/5 ml) and to evaluate whether these formulations meet the FDA criteria to assume bioequivalence in Bangladeshi male volunteers. A randomized, single-dose, two-way cross-over, open-label pharmacokinetic study was conducted in 24 healthy male volunteers after administration of a single dose of 500 mg AZT suspension under fasting condition following a washout period of three weeks. Blood samples were collected in different time intervals and analyzed for serum AZT concentration using a validated LC/MS/MS method. The pharmacokinetic parameters were determined by the non-compartmental method. From serum data, the obtained values for test and reference products were 383.21 +/- 11.59 and 432.28 +/- 7.22 ng/ ml for Cmax; 5677.47 +/- 1229.53 and 6144.56 +/- 1098.70 h x ng/ml for AUC(0-120); and 6085.29 +/- 1267.53 and 6694.15 +/- 1222.50 h x ng/ml for AUC(0-infinity), respectively. On analysis of variance, no period or sequence effects were observed for any pharmacokinetic property; however, a significant formulation effect was observed for Cmax and AUMC(0-infinity). The 90% confidence intervals of the test formulation/reference mean ratios of the Intransformed Cmax, AUC(0-120) and AUC(0-infinity) mean values were found to be 87.89% to 89.36%, 87.96% to 95.71% and 86.77% to 94.29% respectively. In this single-dose study of AZT, it was found that the test formulation met the regulatory criteria for bioequivalence to the reference suspension formulation.

Relative bioavailability and pharmacokinetic study of two trimetazidine modified release formulations in healthy Bangladeshi male volunteers.

Trimetazidine (CAS 5011-34-7) is an effective and well-tolerated antianginal drug that possesses protective properties against ischemia-induced heart injury. The relative bioavailability and pharmacokinetic characteristics of two modified release formulations of 35 mg trimetazidine, one as the test product (Metacard MR) and one as the reference product, were compared in healthy Bangladeshi male volunteers. The randomized, two-way crossover study was conducted in 24 healthy male volunteers after administration of a single 35 mg dose of each modified release formulation after 12-h overnight fasting, with a washout period of two weeks. Blood samples were collected at various time intervals following oral administration and analyzed for trimetazidine concentrations using a validated HPLC method. The pharmacokinetic parameters were determined by a non-compartmental method. After administering a single dose of 35 mg of each trimetazidine formulation, the obtained mean (SD) values for the test and reference products were 104.78 (29.3) and 98.57 (28.7) ng/ml for Cmax; 4.00 (1.1) and 3.54 (1.32) h for t(max); 423.81 (173.9) and 410.01 (195.87) ng x h/ml for AUC0-12; and 472.51 (195.2) and 462.78 (225.13) ng x h/ml for AUC0-infinity respectively. The mean t1/2 was found 3.69 (1.1) h and 3.45 (0.72) h for test and reference products respectively. From paired t-test, no significant differences were observed (p > 0.05) for any pharmacokinetic parameters. The 90% confidence intervals of the test/reference mean ratios of the In-transformed AUC0-12, AUC0-infinity, and Cmax mean values were 106.19% (97.16%-116.06%), 104.74% (95.04%-115.42%) and 106.30% (95.23%-118.66%), respectively. The two formulations demonstrated similar bioavailability with respect to both the rate and extent of trimetazidine absorption.

Genotypes and phenotypes of CYP3A in Bangladeshi population.

BACKGROUND: To investigate whether interindividual variation in CYP3A levels can partly be explained by genetic polymorphisms, this study was designed to phenotype 200 healthy Bangladeshi subjects by measuring urinary ratio of 6ß-hydroxy-cortisol/cortisol and to genotype all the subjects for the presence of CYP3A4*1B, *2, *4, *5, *6, *10 and *18 and CYP3A5*3 alleles. METHODS: For phenotyping, cortisol and 6ß-hydroxy-cortisol were extracted and quantified by HPLC from morning spot urine samples (n=200). Genotyping was done using the extracted genomic DNA from all the subjects followed by amplification of target alleles by PCR. Amplified DNA was digested by restriction enzymes (MboII, XcmI, BsmAI, ClaI, HinfI, HpyCH4III, HpaII and RsaI) followed by gel electrophoresis and sequencing to identify the targeted alleles. RESULTS: The ratio of 6ß-hydroxy-cortisol/cortisol ranged from 0.01 to 31.98 with an average of 3.91. No sample (n=200) was positive for CYP3A4*2, *4, *5, *6, *10 and *18 alleles. Two samples heterozygous for CYP3A4*1B (1.0%) and twenty six samples with the genotype CYP3A5*1/*1 (13.0%) were found to have relatively high 6ß-hydroxy-cortisol/cortisol ratios. CONCLUSION: CYP3A4 variant alleles are present at a low frequency in the Bangladeshi population whereas 50% of the Bangladeshi population carrying a CYP3A5*3/*3 genotype appear to show lower 6ß-hydroxy-cortisol/cortisol ratios compared with those with a CYP3A5*1/*1 genotype.

Investigation of serum trace element, malondialdehyde and immune status in drug abuser patients undergoing detoxification.

Drug abuser patients (n=104), age ranging from 19 to 42 years, were randomly recruited to investigate the serum levels of trace elements (Cu, Zn, Fe, and Mg), malondialdehyde (MDA), and immunoglobulin (IgG, IgA, and IgM) before and after clinical intervention. Control group also included 104 healthy individuals. Blood samples were analyzed for determining trace elements, MDA, and immunoglobulin using atomic absorption spectroscopy, Ultraviolet-Visible (UV-VIS) spectroscopy, and turbidimetry method, respectively. For serum level of Zn and Fe, the differences between the groups (before intervention, after intervention, and control) were not significant (p>0.05). However, significant differences were found in serum copper levels between control group, drug abuser patients, and before and after intervention (p<0.05). The concentration of Mg was found to be significantly higher (p=0.007) in drug abuser patients than the controls, and after intervention, the level was restored to control value. A displacement of elemental homeostasis was observed in drug abuser patients compared to control, and it was improved after intervention. An increase in serum concentration of MDA was found in drug abuser patients compared to control subjects (p>0.05) but was not statistically significant. After intervention, the concentration was restored to control value (p>0.05). The serum concentrations of IgA and IgM were found to be significantly higher (p<0.05) in drug abuser patients before intervention than the controls, and the level tended to be restored to control level after clinical intervention. Serum IgG level was found to be lower in drug abuser patients compared to controls and further declined significantly (p<0.05) after intervention. These findings may suggest a possible imbalance in the levels of micronutrients, antioxidants, and immunoglobulin in drug abuser patients, which tend to be restored to control values after detoxification.