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Introduction: The human leukocyte antigen (HLA) region on chromosome 6p21 is well known to carry the most important genetic factors in susceptibility to psoriasis. Different HLA alleles and haplotypes have been reported to be associated with psoriasis in different populations. Psoriasis has a variable age of onset and, based on this, it can be classified into two types; type I with age of onset before 40 years of age and type II with age of onset after 40 years of age. The objective of this study was to determine the association of HLA class I and class II alleles and haplotypes with disease and stratification using age of onset in Pakistani psoriatic patients. Methods: A group of 603 individuals (326 cases and 277 controls) were analyzed for HLA class I and II alleles and haplotype association by sequence specific PCR. The association was further analyzed according to the age of onset of the patients. Results: We found that HLA alleles B*57 and Cw*06:02, DQB1*03:03:02 are strongly associated with early onset psoriasis, while alleles B*15, DRB1*13:02 and DQB1*03:03:02 are associated with late-onset psoriasis. Cw*06:02 allele was not associated with late-onset psoriasis patients. Allele DQB1*03:03:02 had the highest odds ratio in all patients. We found a novel association specifically with late-onset psoriasis samples with the haplotype HLA-A*11; B*15; Cw*04; DRB1*15; DQB1*05 (Pc = 3.60 × 10−7). We also found strong association with previously reported extended haplotype EH-57.1: HLA-B*57; Cw*06:02; DRB1*07:01; DQB1*03:03:02 in all our patients (Pc = 8.34 × 10−07). Conclusion: Our results show that different HLA class I and II alleles and haplotypes are associated with psoriasis at different age of onset. In this study, we have reported novel alleles and haplotype association with late-onset psoriasis. Our data confirm the previous strong associations with HLA alleles and haplotypes and also reports novel alleles and haplotype association in Pakistani psoriasis patients. [ABSTRACT FROM AUTHOR]

Asthma, a chronic inflammatory disorder of the lungs and airways, typically results from a combination of multiple environmental and genetic factors. Human leucocyte antigen (HLA) region on chromosome 6p21 encodes the most highly polymorphic loci in the human genome, encoding genes with central roles in the immune function where HLA loci are strongly associated with various immune‐mediated diseases such as autoimmunity, allergies and infection. The alleles of HLA class II genes such as DRB1 and DQB1 are the key genetic markers in the development of asthma and have been extensively studied in different ethnicities of the world population. However, the genetic screening of HLA class II alleles and haplotypes in Pakistani asthmatics has not been studied so far. The aim of the present study was to screen the HLA class II DRB1 and DQB1 alleles in asthma cases and controls in a Pakistani population. Seven hundred and two healthy controls and asthma patients were genotyped for HLA class II by sequence‐specific polymerase chain reaction assays. The HLA‐DRB1 and HLA‐DQB1 allele and haplotype frequencies were calculated, and their risk or protective association with asthma was determined. Two‐locus haplotypes of DRB1 and DQB1 alleles were imputed using Arlequin version 3.1 software. The signals of association with asthma were stronger at the DQB1 locus as compared to DRB1. HLA DQB1*03:03:02 (odds ratio [OR] = 2.42, 95% confidence interval [CI] = 1.34–4.25) was significantly associated with an increased risk of asthma, as was the haplotype comprised allele DRB1*07:01‐DQB1*03:03:02 (OR = 2.40, 95% CI = 1.25–4.62). In contrast, DQB1*06 (OR = 0.39, 95% CI = 0.22–0.70) and DQB1*06:02 (OR = 0.27, 95% CI = 0.10–0.71) emerged as protective alleles for asthma. Our data concludes that the HLA DQB1*03:03:02 allele was a risk allele for asthma, whereas two DQB1 alleles, DQB1*06 and DQB1*06:02, were associated with asthma protection. Our findings highlight a prominent role for HLA‐DQB1 alleles in asthma pathogenesis in studied Pakistani cases. More studies, especially with a larger study cohort are needed to confirm the utility of HLA DQB1*03:03:02 as a predictive marker. [ABSTRACT FROM AUTHOR]

Background: Fanconi anemia (FA) is an inherited bone marrow failure syndrome associated with characteristic dysmorphology primarily caused by biallelic pathogenic germline variants in any of 22 different DNA repair genes. There are limited data on the specific molecular causes of FA in different ethnic groups. Methods: We performed exome sequencing and copy number variant analyses on 19 patients with FA from 17 families undergoing hematopoietic cell transplantation evaluation in Pakistan. The scientific literature was reviewed, and we curated germline variants reported in patients with FA from South Asia and the Middle East. Results: The genetic causes of FA were identified in 14 of the 17 families: seven FANCA, two FANCC, one FANCF, two FANCG, and two FANCL. Homozygous and compound heterozygous variants were present in 12 and two families, respectively. Nine families carried variants previously reported as pathogenic, including two families with the South Asian FANCL founder variant. We also identified five novel likely deleterious variants in FANCA, FANCF, and FANCG in affected patients. Conclusions: Our study supports the importance of determining the genomic landscape of FA in diverse populations, in order to improve understanding of FA etiology and assist in the counseling of families. [ABSTRACT FROM AUTHOR]

Typhoid fever caused by Salmonella enterica serovar Typhi has become a serious public health issue due to the development of resistance worldwide especially in developing countries including Pakistan. The aim of this study was to assess the antimicrobial susceptibility pattern of Salmonella Typhi isolated from human blood samples and detect genes by using PCR technique. Total 400 blood samples of typhoid confirm patients were cultured on Salmonella-Shigella agar to isolate Salmonella Typhi. The bacterium was identified through biochemical tests and susceptibility was checked against ten different antibiotics by using disk diffusion method. PCR technique was used to detect different genes after extraction of DNA using phenol-chloroform method. Salmonella typhi (27%) was isolated from blood samples of typhoid patients. Among these, 41% were resistant to ciprofloxacin followed by 34% of ceftriaxone and 25% of streptomycin. Isolated strains were positive for the presence of gyrase A, CMY-2, spvC and InvA genes detected through PCR using specific primers. This study concluded that Salmonella typhi is developing resistance to antimicrobial agents due to the transfer of virulence and resistant genes which may lead to the failure in treatment of typhoid patients in Pakistan. [ABSTRACT FROM AUTHOR]

FAS/FASL signaling system plays a vital role in the regulation of apoptosis, envisaged as a death process required for immune surveillance to prevent autoimmunity and tumorigenesis along with several other biological activities. Several single-nucleotide polymorphisms (SNPs) of FAS/FASL system can result in aberrant apoptosis, which can cause different cancers and autoimmune diseases. Aplastic anemia (AA) is an autoimmune dysfunction characterized by peripheral blood pancytopenia associated with hypoplasia of bone marrow. The aim of this study was to screen Pakistani AA patients and controls for two Fas SNPs rs2234767 and rs1800682 and two FASLG SNPs rs763110 and rs5030772. Genotyping of 392 DNA samples was done by Tetra-ARMS polymerase chain reaction. Genotypic frequencies of Fas rs1800682 and FASLG rs5030772 showed significance difference in their distribution in both controls and patients, while Fas rs2234767 and FASLG rs763110 SNPs had no such difference. Carriers of rs1800682 AG+GG had a very odd ratio of 4.63, with 95% confidence interval (CI) of 3.01-7.11, while individuals with FASLG rs5030772 AG+GG were more common in controls than patients with OR 0.53 and 95% CI of 0.34-0.83. Cumulative effects of these SNPs were analyzed, and they showed almost similar trends; however, Fas rs2234767 and FASLG rs763110 genotypes in combination with Fas rs1800682 and FASLG rs5030772 demonstrated significant association. This study provided information that endorsed the involvement of FAS/FASL system SNPs in the pathogenesis of AA; further studies should be designed to understand the exact role of SNPs that can help in early diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Objective:: To understand the role of three single nucleotide polymorphisms (SNP) of vitamin D receptor (VDR) gene rs7975232, rs731236 and rs2228570 in Pakistani vitiligo patients and ethnically matched controls. Methods: 196 vitiligo patients and 262 controls were included in this study. Genomic DNA was extracted and polymerase chain reaction, restriction length polymorphism (PCR-RFLP) was done. Results: Genotype CA (rs7975232) was found to be associated with susceptibility (Odd ratio=1.46, 95% CI=1.01-2.13, p=0.046) and TT genotype (rs731236) with protection (Odd ratio=0.48, 95% CI=0.25-0.92, p=0.003) to vitiligo. TT genotype for rs731236 was significantly high in controls and absent in undetermined/unclassified group of patients. Vitiligo patients and controls were also compared on the basis of gender. Genotype CA (OR=2.12, 95% CI=1.18-3.79, p=0.013) and CC (OR =0.21 95% CI=0.06-0.73, p=0.007) for rs7975232 were significantly high in male patients and controls, respectively. For rs2228570 CC genotype was significantly high in controls (OR=0.52, 95% CI=0.2-0.97), p=0.04) while TC genotype showed significant difference between patient and controls (OR=1.93, 95% CI=1.02-3.6), p=0.05). In combined effect haplotype A-T-C was found significantly high in controls as compared with the patients (Odd ratio=0.57, 95% CI=0.34-0.97, p=0.04). Conclusion: VDR polymorphisms may be involved in etiology of vitiligo and future studies should be design to screen large sample size for more VDR polymorphism to get more precise picture. [ABSTRACT FROM AUTHOR]

Asthma is a chronic inflammatory and remodeling disorder of the airways, in which many cells, cellular elements, and cytokines play important roles. The role of tumor necrosis factor-α (TNF-α) in asthma is unclear in Pakistani population. The aim of this study was to assess the relationship between TNF-α-308 polymorphism and asthma. Polymorphism of TNF-α (G-308-A locus; rs 1800629) in 329 asthmatic patients and 151 healthy controls was evaluated. DNA was prepared from blood samples of cases and controls. Samples were genotyped for TNF-α 308 G/A polymorphism. There was no significant difference in the frequency of GG (OR 1.049 with 95% CI 0.68-1.63) and GA (OR 0.987 with 95% CI 0.64-1.53) genotypes of TNF-α-308. The AA genotype was absent in cases and only one AA genotype was observed in the controls. The genetic polymorphism of TNF-α does not seem to be associated with asthma in Pakistani population. [ABSTRACT FROM AUTHOR]

Hematological disorders like Aplastic anemia are quite frequent in Pakistan. Human leukocyte antigen (HLA) system, have been implicated in the development of Aplastic anemia in various population-based studies, The aim of this study was to determine the role of the HLA Class I and Class II alleles in genetic susceptibility to Aplastic anemia in Pakistani patients. HLA A*, B* and DRB1* alleles were analyzed, in 61 Pakistani patients (Females n = 22, Males n = 39) and the control group consisted of 200 ethnically matched individuals (Females n = 89, Males n = 111). The allele frequency of DRB1*15 was found significantly higher in patients 0.36 (p = 0.001 with odds ratio = 1.97), as compared to the controls 0.212. Although DRB1*03 percent frequency was significantly higher in controls 0.175 (p = 0.023) with odds ratio = 0.514, as compared to patients 0.106.Therefore DRB1*15 emerged as a susceptible allele and DRB1*03 as a protective allele in Pakistani Aplastic anemia patients and control samples. No significant difference was found in allele frequencies of other HLA class I and HLA class II alleles for both patients and controls. Three haplotypes A*02 B*40 DRB1*15 (p = 0.021), A*31 B*51 DRB1*13 (p = 0.12) and A*33 B*58 DRB1*15 (p = 0.000) showed significant variations in the two groups. [ABSTRACT FROM AUTHOR]