Autosomal recessive non-syndromic hearing loss genes in Pakistan during the previous three decades.

Hearing loss is a clinically and genetically heterogeneous disorder, with over 148 genes and 170 loci associated with its pathogenesis. The spectrum and frequency of causal variants vary across different genetic ancestries and are more prevalent in populations that practice consanguineous marriages. Pakistan has a rich history of autosomal recessive gene discovery related to non-syndromic hearing loss. Since the first linkage analysis with a Pakistani family that led to the mapping of the DFNB1 locus on chromosome 13, 51 genes associated with this disorder have been identified in this population. Among these, 13 of the most prevalent genes, namely CDH23, CIB2, CLDN14, GJB2, HGF, MARVELD2, MYO7A, MYO15A, MSRB3, OTOF, SLC26A4, TMC1 and TMPRSS3, account for more than half of all cases of profound hearing loss, while the prevalence of other genes is less than 2% individually. In this review, we discuss the most common autosomal recessive non-syndromic hearing loss genes in Pakistani individuals as well as the genetic mapping and sequencing approaches used to discover them. Furthermore, we identified enriched gene ontology terms and common pathways involved in these 51 autosomal recessive non-syndromic hearing loss genes to gain a better understanding of the underlying mechanisms. Establishing a molecular understanding of the disorder may aid in reducing its future prevalence by enabling timely diagnostics and genetic counselling, leading to more effective clinical management and treatments of hearing loss.

De novo and bi-allelic variants in AP1G1 cause neurodevelopmental disorder with developmental delay, intellectual disability, and epilepsy.

Adaptor protein (AP) complexes mediate selective intracellular vesicular trafficking and polarized localization of somatodendritic proteins in neurons. Disease-causing alleles of various subunits of AP complexes have been implicated in several heritable human disorders, including intellectual disabilities (IDs). Here, we report two bi-allelic (c.737C>A [p.Pro246His] and c.1105A>G [p.Met369Val]) and eight de novo heterozygous variants (c.44G>A [p.Arg15Gln], c.103C>T [p.Arg35Trp], c.104G>A [p.Arg35Gln], c.229delC [p.Gln77Lys∗11], c.399_400del [p.Glu133Aspfs∗37], c.747G>T [p.Gln249His], c.928-2A>C [p.?], and c.2459C>G [p.Pro820Arg]) in AP1G1, encoding gamma-1 subunit of adaptor-related protein complex 1 (AP1γ1), associated with a neurodevelopmental disorder (NDD) characterized by mild to severe ID, epilepsy, and developmental delay in eleven families from different ethnicities. The AP1γ1-mediated adaptor complex is essential for the formation of clathrin-coated intracellular vesicles. In silico analysis and 3D protein modeling simulation predicted alteration of AP1γ1 protein folding for missense variants, which was consistent with the observed altered AP1γ1 levels in heterologous cells. Functional studies of the recessively inherited missense variants revealed no apparent impact on the interaction of AP1γ1 with other subunits of the AP-1 complex but rather showed to affect the endosome recycling pathway. Knocking out ap1g1 in zebrafish leads to severe morphological defect and lethality, which was significantly rescued by injection of wild-type AP1G1 mRNA and not by transcripts encoding the missense variants. Furthermore, microinjection of mRNAs with de novo missense variants in wild-type zebrafish resulted in severe developmental abnormalities and increased lethality. We conclude that de novo and bi-allelic variants in AP1G1 are associated with neurodevelopmental disorder in diverse populations.

Exome sequencing identifies homozygous variants in MBOAT7 associated with neurodevelopmental disorder.

Intellectual disability (ID) is a large group of neurodevelopmental disorders characterized by a congenital limitation in intellectual functioning (reasoning, learning, and problem solving), adaptive behavior (conceptual, social, and practical skills), originated at birth and manifested before the age of 18. By whole exome sequencing of five consanguineous Pakistani families presenting hallmark features of ID, global developmental delay, aggressive and self-injurious behaviors, microcephaly, febrile seizures and facial dysmorphic features, we identified three novel homozygous missense variants (NM_024298.5: c.588G > T; p.Trp196Cys, c.736 T > C; p.Tyr246His and c.524A > C; p. Asp175Ala) and one rare homozygous in-frame deletion variant (c.758_778del;p.Glu253_Ala259del) in membrane-bound O-acyltransferase family member 7 (MBOAT7) gene previously associated with autosomal recessive neurodevelopmental disorder. The segregation of the variants was validated by Sanger sequencing in all family members. In silico homology modeling of wild-type and mutated proteins revealed substantial changes in the structure of both proteins, indicating a possible effect on function. The identification and validation of new pathogenic MBOAT7 variants in five cases of autosomal recessive ID further highlight the importance of this genes in proper brain function and development.

Cardiovascular Health and Disease in the Pakistani American Population.

PURPOSE OF REVIEW: This narrative review seeks to elucidate clinical and social factors influencing cardiovascular health, explore the challenges and potential solutions for enhancing cardiovascular health, and identify areas where further research is needed to better understand cardiovascular issues in native and American Pakistani populations. RECENT FINDINGS: The prevalence of cardiometabolic disease is high not only in Pakistan but also among its global diaspora. This situation is further complicated by the inadequacy of current cardiovascular risk assessment tools, which often fall short of accurately gauging the risk among Pakistani individuals, underscoring the urgent need for more tailored and effective assessment methodologies. Moreover, social determinants play a crucial role in shaping cardiovascular health. The burden of cardiovascular disease and upstream risk factors is high among American Pakistani individuals. Future research is needed to better understand the heightened risk of cardiovascular disease among Pakistani individuals.

Investigating the genetic basis of hereditary spastic paraplegia and cerebellar Ataxia in Pakistani families.

BACKGROUND: Hereditary Spastic Paraplegias (HSPs) and Hereditary Cerebellar Ataxias (HCAs) are progressive neurodegenerative disorders encompassing a spectrum of neurogenetic conditions with significant overlaps of clinical features. Spastic ataxias are a group of conditions that have features of both cerebellar ataxia and spasticity, and these conditions are frequently clinically challenging to distinguish. Accurate genetic diagnosis is crucial but challenging, particularly in resource-limited settings. This study aims to investigate the genetic basis of HSPs and HCAs in Pakistani families. METHODS: Families from Khyber Pakhtunkhwa with at least two members showing HSP or HCA phenotypes, and who had not previously been analyzed genetically, were included. Families were referred for genetic analysis by local neurologists based on the proband's clinical features and signs of a potential genetic neurodegenerative disorder. Whole Exome Sequencing (WES) and Sanger sequencing were then used to identify and validate genetic variants, and to analyze variant segregation within families to determine inheritance patterns. The mean age of onset and standard deviation were calculated to assess variability among affected individuals, and the success rate was compared with literature reports using differences in proportions and Cohen's h. RESULTS: Pathogenic variants associated with these conditions were identified in five of eight families, segregating according to autosomal recessive inheritance. These variants included previously reported SACS c.2182 C > T, p.(Arg728*), FA2H c.159_176del, p.(Arg53_Ile58del) and SPG11 c.2146 C > T, p.(Gln716*) variants, and two previously unreported variants in SACS c.2229del, p.(Phe743Leufs*8) and ZFYVE26 c.1926_1941del, p.(Tyr643Metfs*2). Additionally, FA2H and SPG11 variants were found to have recurrent occurrences, suggesting a potential founder effect within the Pakistani population. Onset age among affected individuals ranged from 1 to 14 years (M = 6.23, SD = 3.96). The diagnostic success rate was 62.5%, with moderate effect sizes compared to previous studies. CONCLUSIONS: The findings of this study expand the genotypic and phenotypic spectrum of HSPs and HCAs in Pakistan and emphasize the importance of utilizing exome/genome sequencing for accurate diagnosis or support accurate differential diagnosis. This approach can improve genetic counseling and clinical management, addressing the challenges of diagnosing neurodegenerative disorders in resource-limited settings.

Genetic heterogeneity in epilepsy and comorbidities: insights from Pakistani families.

BACKGROUND: Epilepsy, a challenging neurological condition, is often present with comorbidities that significantly impact diagnosis and management. In the Pakistani population, where financial limitations and geographical challenges hinder access to advanced diagnostic methods, understanding the genetic underpinnings of epilepsy and its associated conditions becomes crucial. METHODS: This study investigated four distinct Pakistani families, each presenting with epilepsy and a spectrum of comorbidities, using a combination of whole exome sequencing (WES) and Sanger sequencing. The epileptic patients were prescribed multiple antiseizure medications (ASMs), yet their seizures persist, indicating the challenging nature of ASM-resistant epilepsy. RESULTS: Identified genetic variants contributed to a diverse range of clinical phenotypes. In the family 1, which presented with epilepsy, developmental delay (DD), sleep disturbance, and aggressive behavior, a homozygous splice site variant, c.1339-6 C > T, in the COL18A1 gene was detected. The family 2 exhibited epilepsy, intellectual disability (ID), DD, and anxiety phenotypes, a homozygous missense variant, c.344T > A (p. Val115Glu), in the UFSP2 gene was identified. In family 3, which displayed epilepsy, ataxia, ID, DD, and speech impediment, a novel homozygous frameshift variant, c.1926_1941del (p. Tyr643MetfsX2), in the ZFYVE26 gene was found. Lastly, family 4 was presented with epilepsy, ID, DD, deafness, drooling, speech impediment, hypotonia, and a weak cry. A homozygous missense variant, c.1208 C > A (p. Ala403Glu), in the ATP13A2 gene was identified. CONCLUSION: This study highlights the genetic heterogeneity in ASM-resistant epilepsy and comorbidities among Pakistani families, emphasizing the importance of genotype-phenotype correlation and the necessity for expanded genetic testing in complex clinical cases.

Novel and known minor alleles of CNTNAP2 gene variants are associated with comorbidity of intellectual disability and epilepsy phenotypes: a case-control association study reveals potential biomarkers.

BACKGROUND: Neurodevelopmental disorders are heterogeneous due to underlying multiple shared genetic pathways and risk factors. Intellectual disability, epilepsy and autism spectrum disorder phenotypes overlap which indicates the diverse effects of common genes. Recent studies suggested the probable contribution of CNTNAP2 gene polymorphisms to the comorbidity of these neurological conditions. METHODS AND RESULTS: This study was conducted to investigate the role of CNTNAP2 polymorphisms rs147815978 (G>T) and rs2710102 (A>G) as a risk factor for comorbidity of intellectual disability and epilepsy in a group of 345 individuals including 170 patients and 175 healthy controls recruited from various ethnic groups of Pakistani population. Our case-control study group was genotyped by tetra primer ARMS-PCR technique and results were analysed to know the effects of CNTNAP2 rs147815978 (G>T) and rs2710102 (A>G) polymorphisms in the group. The frequency of risk allele T (rs147815978) and risk allele G (rs2710102) for homozygous recessive genotypes (TT/GG) in our study group was 36.47% while odds ratios for risk allele T (rs147815978) was 5.45 (3.90-7.61: 95% CI, P = 0.000) and that for risk allele G (rs2710102) was 2.39 (1.76-3.24: 95% CI, P = 0.0001). Homozygous recessive genotypes (TT/GG) appeared only in cases and not in control group which indicated these as suspected risk genotypes and the significant association (p < 0.05%) of CNTNAP2 gene polymorphisms rs147815978 (G>T) and rs2710102 (A>G) with co-occurrence of intellectual disability and epilepsy phenotypes in our study group which is in HWE (χ2 = 174, P < 0.0001). Logistic regression analysis shows additive (p < 0.0001) and multiplicative (p < 0.001) models which confirms significant association of both the polymorphisms in our data, which are closely located on same haplotype (D' = - 0.168). CONCLUSIONS: We propose that CNTNAP2 rs147815978 (G>T) and rs2710102 (A>G) polymorphisms are possible risk loci for overlapping neurodevelopmental disorders in Pakistani population. We propose the role of a previously reported common SNP rs2710102 (A>G) with a rarely reported novel SNP rs147815978 (G>T) for CNTNAP2 gene association with neurodevelopmental disorders in our data. Our study has expanded the knowledge of CNTNAP2 gene polymorphisms as probable biomarkers for susceptibility of co-occurrence of intellectual disability and epilepsy phenotypes in Pakistani population. We hope that our study will open new horizons of CNTNAP2 gene variants research to cure the neurological conditions in Pakistani population where consanguinity is a tradition and prevalence of neurodevelopmental disorders has increased from 1 to 2% during last 5 years.

Assessment of physical literacy in 8- to 12-year-old Pakistani school children: reliability and cross-validation of the Canadian assessment of physical literacy-2 (CAPL-2) in South Punjab, Pakistan.

BACKGROUND: The increasing prevalence of physical inactivity, declining fitness, and rising childhood obesity highlight the importance of physical literacy (PL), as a foundational component for fostering lifelong health and active lifestyle. This recognition necessitates the development of effective tools for PL assessment that are applicable across diverse cultural landscapes. AIM: This study aimed to translate the Canadian Assessment of Physical Literacy-2 (CAPL-2) into Urdu and adapt it for the Pakistani cultural context, to assess PL among children aged 8-12 years in Pakistan. METHOD: The Urdu version of CAPL-2 was administered among 1,360 children aged 8-12 from 87 higher secondary schools across three divisions in South Punjab province, Pakistan. Statistical analysis includes test-retest reliability and construct validity, employing confirmatory factor analysis to evaluate the tool's performance both overall and within specific subdomains. RESULTS: The Urdu version of CAPL-2 demonstrated strong content validity, with a Content Validity Ratio of 0.89. Confirmatory factor analysis supported the four-factor structure proposed by the original developers, evidenced by excellent model fit indices (GFI = 0.984, CFI = 0.979, TLI = 0.969, RMSEA = 0.041). High internal consistency was observed across all domains (α = 0.988 to 0.995), with significant correlations among most, excluding the Knowledge and Understanding domains. Notably, gender and age significantly influenced performance, with boys generally scoring higher than girls, with few exceptions. CONCLUSION: This study marks a significant step in the cross-cultural adaptation of PL assessment tools, successfully validating the CAPL-2 Urdu version for the Pakistani context for the first time. The findings affirm the tool's suitability for assessing PL among Pakistani children, evidencing its validity and reliability across the Pakistani population.

Beyond individual responsibility towards healthy food choices: A qualitative study among Pakistani women in Hong Kong.

While biomedical understandings of food and diet coupled with discourses on individual responsibility towards healthy food choices are nowadays prominent, other social and cultural meanings attached to food and diet are largely devalued. The limits of such a reductionist approach are more evident when related to the experiences of migrant and ethnic populations, whose alternative knowledge(s) and practices about food and health are especially neglected. A multicultural city with a public healthcare system inherited from the British colonial times and largely shaped by biomedical ideas of health, Hong Kong offers a lens into the limits of such a reductionist approach. Due to their vulnerability in the context of Hong Kong as shaped by intersecting social identities, 72 women from Pakistan were recruited to be our community partners in a community-based participatory action research project to investigate their health needs and concerns. 12 focus group discussions were conducted, exploring their experiences of "healthy" food and overweight especially related to their encounters with the Hong Kong public healthcare system, as these issues emerged as key health concerns within the community. Four major themes emerged: unmet expectations of care, health is beyond the individual, constraints to a healthy diet in the context of migration, and beyond health: food as care for diasporic women. This study highlights the limit of a reductionist approach to healthy food as merely based on nutrition and individual responsibility. It stresses the need of a counter-discourse in the field of public health, emphasizing not only alternative cultural ideas of health and food, but also enlarging the field of health in biomedical terms to embrace "care" and acknowledging the structural constraints shaping migrant and ethnic population's vulnerability in making "healthy" food choices.

Perceived barriers and facilitators to healthy eating among Pakistani women participating in the PakCat program in Catalonia: A qualitative approach.

Immigrant women of Pakistani origin are among the most at-risk groups for type 2 diabetes, obesity, and heart failure in Catalonia. As the incidence of these diseases is associated with lifestyle factors, we approached this community with participatory research and conducted six focus groups (N = 36) among Pakistani women participating in the PakCat Program. The research process of this paper adhered to the COREQ checklist. Through the thematic analysis, we identified six main themes: social beliefs and attitudes, family environment, personal factors, dietary acculturation, traditional dietary patterns, and economic factors. We discovered both facilitators and barriers associated with each theme, but the findings indicated that Pakistani women encounter more inhibitors than enablers to following a healthy diet. The determination of these factors can facilitate the reinforcement of the aspects that help Pakistani women to follow a healthy diet and provide adequate tools to overcome the barriers.

Identifying the Health Concerns of Pregnant British Pakistani Women Living in Deprived Areas: A Qualitative Study.

INTRODUCTION: Pregnant British Pakistani women have disproportionately poorer health than the wider population. Bradford has a strong Pakistani presence and a wide range of public health problems including high levels of gestational diabetes, high obesity rates and a high infant mortality rate, which is highest for babies of Pakistani origin. For women to be healthy, we need to know what concerns they have about their health so they can be addressed appropriately. The aim of this study, therefore, was to explore the health concerns of pregnant British Pakistani women living in deprived areas. METHODS: Semi-structured qualitative interviews were conducted with 21 pregnant Pakistani women in a hospital setting. Data were analysed using thematic analysis. RESULTS: Pakistani women identified safety issues, barriers to undertaking physical activity in the areas where they live, concerns surrounding exercising during pregnancy and cultural and religious constraints that prevented them from engaging in physical activity. They reported issues around food, concerns around a lack of culturally appropriate diet information, the cost of unhealthy food locally, and the lack of healthy food options in their residences. Women were unsure on where to obtain health promotion information and reported a lack of access in obtaining that information. Language barriers in accessing health promotion information were further reported as a concern. DISCUSSION: Researchers, midwives, health providers, local authority and policy makers interested in improving the health of pregnant Pakistani women may use these findings to develop further research and interventions to improve the poor health of this population.

What is already known on this subject? South Asian women have previously identified issues relating to safety in physical activity and cultural barriers to engaging in physical activity but there has been little investigation into the health concern of pregnant Pakistani women.What this study adds? We now have a clearer understanding of the barriers faced by pregnant Pakistani women living in deprived areas when trying to live a healthy lifestyle. This understanding will contribute to the development of strategies for promoting health and improving the outcomes for this population.

Health literacy in Pakistani migrants in Australia-An emerging and neglected culturally and linguistically diverse community.

ISSUE ADDRESSED: Pakistani migrants are one of the fastest-growing culturally and linguistically diverse (CALD) communities in Australia, but there is currently a lack of information regarding their health literacy. This study aimed to investigate the health literacy of Pakistani migrants residing in Australia. METHODS: Using a cross-sectional study design, health literacy was measured using the Urdu version of Health Literacy Questionnaire (HLQ). Descriptive statistics and linear regression were used to describe the health literacy profile of respondents and to examine its association with their demographic characteristics. RESULTS: The responses of 202 Pakistani migrants were included. The median age of the respondents was 36 years, 61.8% were males and 87.6% had a university education. The majority spoke Urdu at home and almost 80% were Australian permanent residents or citizens. Pakistani respondents scored high on HLQ domains; feeling understood by health providers (Scale 1), social support for health care (Scales 4), engaging with health care providers (Scale 6) and understanding health information (Scale 9). The respondents scored low on HLQ domains; having sufficient information (Scale 2), actively managing health (Scale 3), appraisal of health information (Scale 5), navigating the health care system (Scale 7) and ability to find information (Scale 8). In the regression model, university education and age were significantly associated with health literacy in almost all the domains, but the effect size was small for age. Speaking English at home and being a permanent resident were also associated with better health literacy in two to three HLQ domains. CONCLUSIONS: Health literacy strengths and weaknesses of Pakistani migrants residing in Australia were identified. Health care providers and organisations may use these findings to tailor health information and services to better support health literacy in this community. SO WHAT?: This study will inform future interventions to better support health literacy and reduce health disparities in Pakistani migrants residing in Australia.

A homozygous founder variant in PDE2A causes paroxysmal dyskinesia with intellectual disability.

Intellectual developmental disorder with paroxysmal dyskinesia or seizures (IDDPADS, OMIM#619150) is an ultra-rare childhood-onset autosomal recessive movement disorder manifesting paroxysmal dyskinesia, global developmental delay, impaired cognition, progressive psychomotor deterioration and/or drug-refractory seizures. We investigated three consanguineous Pakistani families with six affected individuals presenting overlapping phenotypes partially consistent with the reported characteristics of IDDPADS. Whole exome sequencing identified a novel missense variant in Phosphodiesterase 2A (PDE2A): NM_002599.4: c.1514T > C p.(Phe505Ser) that segregated with the disease status of individuals in these families. Retrospectively, we performed haplotype analysis that revealed a 3.16 Mb shared haplotype at 11q13.4 among three families suggesting a founder effect in this region. Moreover, we also observed abnormal mitochondrial morphology in patient fibroblasts compared to controls. Belonging to diverse age groups (13 years-60 years), patients presented paroxysmal dyskinesia, developmental delay, cognitive abnormalities, speech impairment, and drug-refractory seizures with variable onset of disease (as early as 3 months of age to 7 years). Together with the previous reports, we observed that intellectual disability, progressive psychomotor deterioration, and drug-refractory seizures are consistent outcomes of the disease. However, permanent choreodystonia showed variability. We also noticed that the later onset of paroxysmal dyskinesia manifests severe attacks in terms of duration. Being the first report from Pakistan, we add to the clinical and mutation spectrum of PDE2A-related recessive disease raising the total number of patients from six to 12 and variants from five to six. Together, with our findings, the role of PDE2A is strengthened in critical physio-neurological processes.

Unravelling the genetic basis of retinal dystrophies in Pakistani consanguineous families.

BACKGROUND: Retinitis Pigmentosa (RP) is a clinically and genetically progressive retinal dystrophy associated with severe visual impairments and sometimes blindness, the most common syndromic form of which is Usher syndrome (USH). This study aimed to further increase understanding of the spectrum of RP in the Khyber Pakhtunkhwa region of Pakistan. METHODOLOGY: Four consanguineous families of Pashtun ethnic group were investigated which were referred by the local collaborating ophthalmologists. In total 42 individuals in four families were recruited and investigated using whole exome and dideoxy sequencing. Among them, 20 were affected individuals including 6 in both family 1 and 2, 5 in family 3 and 3 in family 4. RESULT: Pathogenic gene variants were identified in all four families, including two in cone dystrophy and RP genes in the same family (PDE6C; c.480delG, p.Asn161ThrfsTer33 and TULP1; c.238 C > T, p.Gln80Ter) with double-homozygous individuals presenting with more severe disease. Other pathogenic variants were identified in MERTK (c.2194C > T, p.Arg732Ter), RHO (c.448G > A, p.Glu150Lys) associated with non-syndromic RP, and MYO7A (c.487G > A, p.Gly163Arg) associated with USH. In addition, the reported variants were of clinical significance as the PDE6C variant was detected novel, whereas TULP1, MERTK, and MYO7A variants were detected rare and first time found segregating with retinal dystrophies in Pakistani consanguineous families. CONCLUSIONS: This study increases knowledge of the genetic basis of retinal dystrophies in families from Pakistan providing information important for genetic testing and diagnostic provision particularly from the Khyber Pakhtunkhwa region.

Physical activity and TV viewing parenting practices for toddlers among South Asian and white families in the UK: born in Bradford 1000 study.

BACKGROUND: Children of South Asian (SA) origin in the UK have lower levels of physical activity (PA), compared to their White counterparts. Parents play an important role in establishing PA habits among young children. The aim of this study was to compare PA and television (TV) viewing parenting practices for young children between SA British (SAB) and White British (WB) parents living in the UK. METHODS: We conducted a secondary analysis of the Born in Bradford (BiB) 1000 study, using survey data at child ages 24 and 36 months. The study sample included three groups of mothers (n = 1,149): foreign-born SAB (n = 458), UK-born SAB (n = 276), and WB (n = 455). Mothers completed a survey about parenting practices (i.e., PA supports, PA restrictions, TV viewing restrictions) at child age 24 months and child PA and TV viewing behaviors at child ages 24 and 36 months. Parenting practices were compared among the three groups. Multivariable linear regression analyses compared children's weekly walking frequency and daily TV viewing hours by parenting practices in the three groups. RESULTS: The foreign-born SAB group showed the lowest frequencies of PA-supportive parenting practices (verbal encouragement: 3.7 ± 3.1 times/week; logistic support: 1.5 ± 1.8 times/week) and the highest frequencies of PA-restrictive parenting practices (7.8 ± 7.7 times/week) among the three groups (p < 0.01). Children of Foreign-born SAB mothers had the most frequent TV watching during a mealtime (4.0 ± 3.1 times/week) among the three groups (p < 0.01). Less frequent PA-supportive parenting practices and SA ethnicity were associated with lower walking frequency at 24 and 36 months of age among children (p < 0.01). More frequent exposure to TV at mealtimes and SA ethnicity were associated with higher TV viewing time at 24 and 36 months of age among children (p < 0.01). CONCLUSIONS: This study demonstrated that SAB parents, particularly those who are foreign-born, apply parenting practices for their young children that are less supportive of PA and more supportive of TV viewing, and their children have lower PA and higher TV viewing time, compared with their WB counterparts.

Genetic studies in the Pakistani population reveal novel associations with ventricular septal defects (VSDs).

BACKGROUND: With prevalence up to 4%, Ventricular Septal Defect (VSD) is one of the leading causes of neonatal deaths. VSD is a common complex genetic disorder that has been associated with many genetic determinants. Variants from genes for the transcription factors including T-Box TBX5 and NFATc1 (nuclear factor of activated T cells, cytoplasmic 1), Vascular endothelial growth factor (VEGF), ISLET1 (encoded by the ISL1 gene) and enzyme MTHFR, a methylene tetrahydrofolate reductase were selected. Genetic risk score (GRS) is a widely accepted approach used to convert the genetic data into prediction and assessment tool for disease susceptibility. METHODS: A total of 200 participants were recruited for the current study, 100 VSD patients and 100 controls. Genotyping of the ISL1: rs1017, NFATc1: rs7240256, VEGF: rs36208048, TBX5: rs11067075, and MTHFR: rs1801133 variants was performed using tetra primer ARMS PCR and PCR-RFLP. For the statistical analysis, the software SPSS version 23 was used. Genotypic frequencies of cases and controls were calculated using chi-square (χ²) whereas allelic frequencies were calculated by using the SNPStats tool. The association of GRS quartiles with VSD was examined using binary logistic regression. Adjusted p-value 0.01 was used as significance threshold for all analyses. RESULTS: The ISL1 (OD: 0.242, CI: 0.158-0.37, p-value: 2.15 × 10- 4 :), NFATc1 (OD: 2.53, CI: 1.64-3.89, p-value: 2.11 × 10- 5), TBX5 (OD: 2.24, CI: 1.47-3.41, p-value:1.6 × 10- 4) and MTHFR (OD: 10.46, CI: 5.68-19.26, p-value: 2.09 × 10- 9:) variants were found to be in association with VSD. In contrast, the VEGF (OD: 0.952, CI: 0.56-1.62, p-value: 0.8921) variant did not show significance association with the VSD. For cases, the mean GRS score was 3.78 ± 1.285 while in controls it was 2.95 ± 1.290 (p-value: 0.479, CI: 0.474-1.190). Comparison of GRS between cases and control showed that mean GRS of cases was 1.90 ± 0.480 while in controls it was 1.68 ± 0.490 (p-value: 0.001, CI: 0.086-0.354). Higher quartiles were more prevalent in cases whereas lower quartiles were more prevalent in controls. CONCLUSION: GRS of these five loci was strongly associated with VSD. Moreover, genetic risk score can provide better information for the association between variants and disease as compared to a single SNP. We also illustrated that the cumulative power of GRS is greater over the single SNP effect. This is a pilot scale study with a relatively small sample size whose findings should be replicated in a larger sample size for the unique local Pakistani population.

Genome-wide analysis of runs of homozygosity in Pakistani controls with no history of speech or language-related developmental phenotypes.

BACKGROUND: Runs of homozygosity (ROHs) analysis of controls provide a convenient resource to minimize the association of false positive results of disease-associated ROHs and genetic variants for simple and complex disorders in individuals from the same population. Evidence for the value of ROHs to speech or language-related traits is restricted due to the absence of population-matched behaviourally defined controls and limited family-based studies. AIM: This study aims to identify common ROHs in the Pakistani population, focussing on the total length and frequency of ROHs of variable sizes, shared ROHs, and their genomic distribution. SUBJECTS AND METHODS: We performed homozygosity analysis (in PLINK) of 86 individuals (39 males, 47 females) with no history of speech or language-related phenotypes (controls) who had been genotyped with the Illumina Infinium QC Array-24. RESULTS: ROHs of 1-<4 megabases (Mb) were frequent in unrelated individuals. We observed ROHs over 20 Mb among six individuals. Over 30 percent of the identified ROHs were shared among several individuals, indicating consanguinity's effect on the Pakistani population. CONCLUSION: Our findings serve as a foundation for family-based genetic studies of consanguineous families with speech or language-related disorders to ultimately narrow the homozygosity regions of interest to identify pathogenic variants.

Reshaping Insanity in Pakistani Law: The Case of Safia Bano.

This Article analyzes the 2021 judgment of the Supreme Court of Pakistan in the case of Mst. Safia Bano v. Home Department, Government of Punjab. The case has garnered significant local and international attention due to the Court's ruling that a death sentence may not be carried out on a defendant who has a mental illness. Setting the case against the backdrop of Pakistan's Islamic and colonial contexts, this article argues that the Supreme Court has reshaped the insanity defense in Pakistani law by placing the determination of a defendant's mental state mainly in the hands of medical professionals. However, the Court's reliance on medical professionals and the subsequent downplaying of the "moral capacity" element of the insanity defense-a determination of law made by courts-has created an obstacle for courts to punish offenders more stringently in future cases due to the popular belief that mental health professionals are ill-equipped to answer broader questions of justice for victims and society. The article recommends that this issue can be remedied by establishing an objective legal test for insanity that considers Islamic law, Pakistani precedent, and advances in medical science.

Expanding OBSL1 Mutation Phenotype: Disproportionate Short Stature, Barrel Chest, Thoracic Kyphoscoliosis, Hypogonadism, and Hypospadias.

We present a Pakistani kinship afflicted with a syndrome with features including short stature, reduced sitting height, orofacial symptoms including prominent forehead and thick eyebrows, short and broad thorax, and variable features such as long philtrum, short broad neck, barrel chest, thoracic kyphoscoliosis, hypogonadism, and hypospadias. Phenotypic variation even within different sibships was considerable. The unique combination of the phenotypic characteristics prompted us to determine the shared homozygosity regions in patient genomes and the pathogenic variants by next generation technologies like single nucleotide polymorphism (SNP) genotyping and whole exome sequencing (WES). Through these analyses, we detected homozygous OBSL1 c.848delG (p.Gly283AlafsTer54) as the causal variant. Biallelic variants in OBSL1 are known to cause Three M Syndrome 2 (3M2), a rare disorder of growth retardation with characteristic facial dysmorphism and musculoskeletal abnormalities. Affected members of the family do not have the 3M2 hallmark features of dolichocephaly, hypoplastic midface, anteverted nares, low nasal bridge, pectus excavatum, sacral hyperlordosis, spina bifida occulta, anterior wedging of thoracic vertebrae, prominent heels, and prominent talus. Moreover, they have some variable features not typical for the syndrome such as round face, disproportionate short stature, barrel chest, thoracic kyphoscoliosis, hypogonadism, and hypospadias. Our study facilitated genetic diagnosis in the family, expanded the clinical phenotype for 3M2, and unraveled the considerable clinical variation within the same kinship. We conclude that unbiased molecular analyses such as WES should be more integrated into healthcare, particularly in populations with high parental consanguinity, given the potential of such analyses to facilitate diagnosis.

Surveillance of aetiologies, clinical presentation, and most common types of epilepsy among paediatric patients at a tertiary care hospital in Pakistan.

Epilepsy is the third most common neurological disease in the world associated with a high frequency in the paediatric age group. This study aims to evaluate the prevalence, types and aetiologies of epilepsy within the Pakistani population. A retrospective review of the charts of all patients, below the age of 18 years, presenting with epilepsy to the Department of Neurology at The Children's Hospital and Institute of Child Health, Lahore, from January 2016 to December 2020, was carried out. Analysis was performed using SPSS Version 26. A p value of <0.05 was considered statistically significant. A total of 1,097 patients were studied, of which 644 (58.8%) were males and 451 (41.2%) females. a vast majority, i.e. 1,021 (96.1%), of the study participants, belonged to the province of Punjab. Afebrile seizures [n=798 (72.7%)] were more commonly reported than febrile seizures [n=299 (27.3%)]. Among seizure types, generalised seizures were the most common type of seizure reported in 520 (49.8%) patients. Refractory seizures were the least common type reported in 3 (0.3%) patients. Aetiology was mostly idiopathic [n=540 (49.2)], followed by congenital [n=228 (20.8%)]. The most frequently reported duration of seizure was between one and three minutes [n=116 (42.3%)]. The most common ictal features seen were a combination of up-rolling of eyes and frothing from the mouth [n=206 (34.9%)]. Results from this study can be used by health care providers to better formulate therapeutic interventions for a timely diagnosis and effective treatment of epilepsy.