Pakistani Cohort Research Articles

Effects of feeding practices on the diversity of faecal short chain Fatty acids in 1-4 months infants in Pakistani cohort

Introduction: The most important product of fermentation of non-digestible carbohydrates is short chain fatty acids. Formation of these metabolites are mainly dependent on diet and gut microbial colonization. Literature has been supportive of the fact that short chain fatty acids play significant role in chronic inflammation (allergic diseases) and host physiology and metabolic processes including obesity and diabetes. Little is known about the differences in the metabolism between breast-fed and bottle-fed infants in our country. This study was designed to determine the differences in the metabolism between breast-fed and bottle-fed infants in Pakistani cohort. Material & Methods: A cross sectional study was carried out in 50 healthy infants. Enrollment of these children was done from different centers and localities in Peshawar. Stool samples were collected from the included infants and short chain fatty acids were studied by using Gas Chromatography. Results: A total of 50 healthy infants (25 breast-fed and 25 bottle-fed) with mean age (mean ± sd) 78.40 35.88 days (range: 1-4 months) were included in this survey. Breast feeding was associated with lesser absolute concentrations of total short chain fatty acids. Among short chain fatty acids, acetates were more in breast feeding infants (median 651, p= 0.001) compared to formula feeding infants while propionates (median 81.06, p=0.02), butyrates (median 31.86, p=0.007) and iso-butyrates (median 3.76, p=0.003) were dominant in FFI which indicated an important role of short chain fatty acids in human health. Conclusion: It may be concluded that feeding practices strongly influence the gut microbial metabolism and metabolites (short chain fatty acids).

Fatigue assessment by FACIT-F scale in Pakistani cohort with Rheumatoid Arthritis (FAF-RA) study.

Objective:To fine out fatigue frequency and severity by FACIT-F scale in Pakistani cohort with rheumatoid arthritis.Methods:This study was conducted at department of Medicine division of rheumatology CPMC Lahore. After the approval of IRB, 192 patients of RA were recruited. Written, informed consent was taken, demographic details were noted, patients filled the URDU version of FACIT-F (fatigue severity scale). 5-ml of blood was taken for fasting blood sugar, viral markers and ESR by a trained phlebotomist. Each individual’s disease activity was assessed by DAS-28 and FACIT-F score was calculated.Results:The Mean age (39.9±10.5) years, (71.9%) were females. Fatigue frequency was 62% (n=126), age, education, hypertension, DAS-28, exercise levels and HCV gives significant association with fatigue score. Linear regression analysis, results showed one unit increase in DAS-28 will gives 2.71 unit increases in fatigue scores(P <0.05).Conclusions:We have very high frequency of fatigue in RA, increases with disease activity & associated conditions.

Retroperitoneal Soft-Tissue Sarcoma: Retrospective Study from a Cancer Hospital in Pakistan

Objectives: To determine the clinicopathological aspects and factors determining the recurrence and 5-year survival of patients diagnosed with retroperitoneal soft-tissue sarcoma in a Pakistani cohort of patients. Methods: This retrospective descriptive study was conducted at the Department of Surgical Oncology, Shaukat Khanum Memorial Hospital & Research Centre (SKMCH&RC), Lahore, Pakistan. Patients who underwent surgery for retroperitoneal sarcoma from January 2011 to December 2015 were included and followed for 5 years (December 2020). A total of 54 patients was included in this study. Results: Fifty-seven percent (n=31) cases were male with a median age at the time of presentation 43 (30-60) years. Well-differentiated liposarcoma was the most common variant. Non-compartmental resection was done in 40 (74%) of the cases. The overall 5-year survival was 74% with median Disease-Free Survival (DFS) of 14 (6-48) months. Recurrence was seen in 28 cases (52%). Conclusions: In the Pakistani cohort of patients. retroperitoneal soft-tissue sarcoma was more commonly seen in males at the median age of 43 years. The most common presentations were abdominal pain and mass. Well-differentiated liposarcoma was the most common histological variant. The overall 5-year survival was 74% that was affected by histological variant and grade. Recurrence was affected by histological variant and grade, margin positivity, and use of neoadjuvant therapy.

Association of polymorphisms in inflammatory cytokine genes with the development of head and neck cancer in Pakistani population

Head and neck cancer (HNC) is the most common type of carcinoma and represents a major health problem in developing countries. HNC is a multifactorial condition associated with a variety of risk factors comprising genetic, environmental and life style. Genetic polymorphisms of interleukin genes might contribute to the development of HNC. Present study was conducted to investigate a possible association of HNC with single nucleotide polymorphisms (SNPs) in interleukin genes (IL-8 rs4073 A/T, IL-4 rs2070874 C/T, IL-10 rs1800896 T/C, IL-10 rs180072 T/G, IL-6 rs1800796 C/G, IL-6 rs1800795 C/G, and IL-1-α rs17561 A/C) and oral hygiene in Pakistani Population. For this purpose, 231 cases and 219 controls were recruited. The SNPs were assessed through QuantStudio real time PCR system. Analysis of the data showed a significant association of IL-10 rs1800896 with HNC while other SNPs were not associated was HNC pathogenicity. Further, no significant association was observed between hygiene associated factors and HNC in the studied population. This study presents useful aspects of HNC in Pakistani cohort.

Association study of CCR6 rs3093024 with Rheumatoid Arthritis in a Pakistani cohort.

C-C Chemokine receptor 6 (CCR6), an important protein in inflammatory and immunological responses, has been previously reported to be associated with rheumatoid arthritis (RA). Therefore, in order to replicate these findings, a case-control study was conducted on 500 subjects (including 250 RA patients and 250 healthy controls) of Pakistani origin. The aim of this study was to determine the association of CCR6 rs3093024 variant with RA and identify its role in splicing events using bioinformatics tools. The clinical and demographic characteristics of the patients were collected using a well-designed questionnaire. The genotype frequencies of CCR6 rs3093024 variant were determined using tetra-primer ARMS-PCR (amplification of refractory mutation system-polymerase chain reaction) method. A significant difference was found between CCR6 rs3093024 genotype frequencies [P = 0.0016, χ2 = 12.915]. Similarly, a significant difference in the allele frequencies between RA patients and healthy controls was also observed [P = 0.0003 and OR (95% CI) = 0.63 (0.49–0.80)]. The stratification of patients showed that there was a significant increase in AA genotype against AG + GG in patients [P = 0.0014, OR (95% CI) = 2.0 (1.32–3.02)]. Furthermore, using bioinformatics analysis, it was found that CCR6 rs3093024 variant might create a potential splicing enhancer motif (SF2/ASF (IgM-BRCA1) with score of 77.92; Threshold 70.53), which might have important impact on the product of this gene. This study suggests that the A variant of CCR6 rs3093024 variant is significantly associated with RA-risk and its G variant is protective in Pakistani population but a multi-cohort large sized population study is needed to elucidate these results. Moreover, functional studies are needed to highlight the effects of this polymorphism on the function of CCR6 gene.

Predicting emergency interventions in patients with acute ureteral colic using acute renal colic scoring system in a Pakistani cohort.

To evaluate the acute renal colic score (ARC) in predicting the need of emergency intervention (EI) in patients with ureteric colic secondary to a ureteral stone. In an emergency room (ER) of a university hospital, we conducted a prospective cohort study over a period of 6months. ARC score was calculated using four parameters, i.e., serum creatinine, total white cell count (TLC), stone length and level. Primary outcome measure was EI, which was defined as the need of endourological intervention within 48h of presentation. ARC was calculated for each patient against the two possible outcomes, i.e., EI vs. no EI. The need of intervention was based on patient-related clinical factors and the decision of the attending urologist. The study included 132 patients. EI was performed in 85 patients (64.4%). URS was the most common intervention performed in 81 (95.3%) patients, followed by the a insertion of a double J stent in two (2.4%) patients for forniceal rupture and high TLC count and percutaneous nephrostomy in two (2.4%) patients for raised creatinine and TLC. All four variables in ARC score including serum creatinine (p < 0.001), TLC (p < 0.001), stone size (p < 0.001) and stone level (p < 0.001) were found to be significantly associated with need for EI. Using ROC the sensitivity and specificity of the score was 92.9% and 87.5%, respectively, with AUC of 0.93. ARC score is highly sensitive and specific in determining the need of EI in patients with uncomplicated ureteric colic within 48h of initial presentation.

Laparoscopic Curative Resection for Right-Sided Colonic Tumors: Initial Experience From a Specialized Cancer Hospital of a Developing Country.

IntroductionLaparoscopic colonic resection is increasingly becoming popular worldwide and aims to provide curative resection in addition to the inherent benefits of laparoscopic surgery. The aim of this study was to evaluate the long-term outcomes of laparoscopic right hemicolectomy in a Pakistani cohort of patients.Methods and proceduresWe retrospectively analyzed the medical records of all patients who presented to our hospital with the diagnosis of right-sided colon carcinoma from January 2010 to December 2018 and underwent laparoscopic right or extended right hemicolectomy. Demographics, operative findings, histopathology report, and follow-up of patients were recorded and the analysis was performed on Statistical Packages for the Social Sciences (SPSS) Version 20 (IBM Corp, Armonk, NY).ResultsSeventy-five patients were included, 56 (74.7%) of whom were males and 19 (25.3%) were females. The median age was 52 years (range 25-82 years). The median hospital stay was five days (Range 3-13 days). The median blood loss was 70 milliliters and the mean operative time was 195.5±77.6 minutes. Laparoscopic extended right hemicolectomy was performed in 23 (16.67%) patients and standard right hemicolectomy in 52 (83.33%) patients. Most (72%) of the patients had a pathological T3 tumor, and the majority (61.3%) of the patients had no nodal involvement (pN0). The mean number of lymph nodes removed was 20+8. The median numbers of involved lymph nodes were 1.14+2.19. All the patients had R0 resection. Postoperatively, two patients had pelvic collection, and there was no 30-day mortality. Local recurrence occurred in four patients and distant metastases were observed in nine patients. The median follow-up in our study was 40.5±18.35 months. The median disease-free survival was 42±2.17 months and the median overall survival was 44±2.16 months.ConclusionOur experience with laparoscopic right colon resections has confirmed the safety and feasibility of the procedure.

Prevalence of EBV, CMV, and HPV in oral squamous cell carcinoma patients in thePakistani population.

Many studies have proposed an important role of viruses in the pathogenesis of oral cancer. The present study aimed to find out the prevalence of Epstein-Barr virus (EBV), cytomegalovirus (CMV), and human papillomavirus (HPV) among patients with oral squamous cell carcinoma (OSCC) in a Pakistani cohort. We investigated tissue samples obtained from 58 patients with OSCC using the polymerase chain reaction assay. No sample was positive for HPV. EBV was identified in 15 patients (25.86%), and CMV in three patients (5.17%). Coinfection with one or more viruses was detected in two cases and was coinfection with EBV and CMV. These results suggest a low prevalence of these viruses in OSCC patients in the Pakistani population compared to most other countries where the prevalence of these viruses has been reported in the past. Nevertheless, further studies are necessary to determine the potential role of EBV and the possible importance of CMV as an infection cofactor in oral cancer.

A novel histological index for evaluation of environmental enteric dysfunction identifies geographic-specific features of enteropathy among children with suboptimal growth.

A major limitation to understanding the etiopathogenesis of environmental enteric dysfunction (EED) is the lack of a comprehensive, reproducible histologic framework for characterizing the small bowel lesions. We hypothesized that the development of such a system will identify unique histology features for EED, and that some features might correlate with clinical severity. Duodenal endoscopic biopsies from two cohorts where EED is prevalent (Pakistan, Zambia) and North American children with and without gluten sensitive enteropathy (GSE) were processed for routine hematoxylin & eosin (H&E) staining, and scanned to produce whole slide images (WSIs) which we shared among study pathologists via a secure web browser-based platform. A semi-quantitative scoring index composed of 11 parameters encompassing tissue injury and response patterns commonly observed in routine clinical practice was constructed by three gastrointestinal pathologists, with input from EED experts. The pathologists then read the WSIs using the EED histology index, and inter-observer reliability was assessed. The histology index was further used to identify within- and between-child variations as well as features common across and unique to each cohort, and those that correlated with host phenotype. Eight of the 11 histologic scoring parameters showed useful degrees of variation. The overall concordance across all parameters was 96% weighted agreement, kappa 0.70, and Gwet's AC 0.93. Zambian and Pakistani tissues shared some histologic features with GSE, but most features were distinct, particularly abundance of intraepithelial lymphocytes in the Pakistani cohort, and marked villous destruction and loss of secretory cell lineages in the Zambian cohort. We propose the first EED histology index for interpreting duodenal biopsies. This index should be useful in future clinical and translational studies of this widespread, poorly understood, and highly consequential disorder, which might be caused by multiple contributing processes, in different regions of the world.

Fatty liver in Pakistani cohort with rheumatoid arthritis.

Objective:To determine the frequency of fatty liver (non-alcoholic) disease, Framingham 10-year cardiovascular risk score in rheumatoid arthritis patients.Methods:This study was conducted from September 1st to March 19, 2019, at Rheumatology OPD, Central Park Medical College Lahore. One hundred ninety two seropositive rheumatoid arthritis patients were recruited. Demographic details were noted, BP, BMI, smoking habits, and waist circumference were noted, then sent to radiology department for ultrasound scan of abdomen by an expert radiologist. On next day 10 ml blood was taken by phlebotomist for lipid profile and fasting blood sugar levels, after availability of results 10-years Framingham cardiovascular risk score was calculated.Results:Females were (81.3%) mean age of (45.4) years, fatty liver was present in n=39 (20.3%). In positive cases comorbid like metabolic syndrome was present (71.8%), diabetes mellitus (33.3%), hypertension (59%) FRS score (intermediate to high in (33.3%), history of hakeem/desi medication use (51.3%), while on regression analysis all study parameters except DMARD’s had significant association with fatty liver (p<0.05).Conclusion:Nonalcoholic fatty liver disease is very widely prevalent in rheumatoid arthritis patients. As in general population, it is multifactorial in origin and needs careful monitoring and treatment.

Association of VDR Gene Variant (rs1544410) with Type 2 Diabetes in a Pakistani Cohort.

The present study was designed to measure the mean values of body mass index (BMI), random blood sugar/ fast ing blood sugar (RBS/FBS) tests, and Hb A1c and to investigate the role of a genetic variant rs1544410 in the VDR gene in a Pakistani cohort. For this purpose, a total of 917 samples including 469 diabetes mellitus type 2 (T2DM), 145 DM type 1 (T1DM), and 303 healthy control were collected. Out of the total sample set, 500 individuals (250 T2DM cases and 250 controls) were genotyped for rs1544410. It was found that 65 (26.0%) cases and 32 (12.8%) controls had homozygous AA, while 69 (27.6%) cases and 139 (55.6%) controls had heterozygous AG, and 116 (46.4%) cases and 79 (31.6%) controls had homozygous GG (χ2 = 41.81, p = 0.0001). In addition, a similar distribution of allele frequency was determined in cases and controls [p value = 0.866; odds ratio (OR) = 0.967; relative risk (RR) = 1.034]. A significant difference was observed in homozygous dominant [OR = 2.394 (1.501-3.816); RR = 1.46 (1.225-1.740); p = 0.003] and homozygous recessive models [OR = 2.970 (2.086-4.227); RR = 1.798 (1.501-2.154); p = <0.0001] analysis of rs1544410 in the VDR gene. These findings suggest that the VDR gene is associated with T2DM and genotype GG of genetic variant rs1544410 is the susceptible genotype in our Pakistani cohort.

Characterization of the mutation spectrum in a Pakistani cohort of type 3 von Willebrand disease.

Type 3 von Willebrand disease (VWD), a severe autosomal recessive hereditary bleeding disorder, is described by the virtual absence of von Willebrand factor (VWF). In consanguineous populations, for example Pakistan, the disease is reported with a higher incidence rate than the worldwide prevalence. This study aims to characterize molecular pathology and clinical profile of type 3 VWD cohort of Pakistani origin. In total, 48 patients were enrolled in the current study. Initially, the index patients (IPs) were evaluated by a standardized questionnaire for recording bleeding manifestations and by performing conventional coagulation tests. The diagnosis of VWD type 3 was confirmed by VWF antigens less than 5IU/dL. Direct sequencing of VWF gene (VWF) was carried out to identify causative gene variations. We evaluated the potential consequence of novel splice site and missense variations by predictive computational programs and in silico structural analysis. VWF mutations were detected in 46 out of 48 IPs (95.8%), predominantly as homozygous variants. In total, twenty-nine different gene defects were characterized in this cohort from which 10 (34.5%) are novel. The majority of the mutations were null alleles (66%; including gene conversions, nonsense, splice site variations, small deletions and insertions), and 34% of them were missense substitutions. Herein, we reported for the first time, the pattern of gene defects in Pakistani type 3 VWD cohort. We identified a wide heterogeneous mutation spectrum along with variability in the type of bleeding episodes.

Risk assessment of FLT3 and PAX5 variants in B-acute lymphoblastic leukemia: a case–control study in a Pakistani cohort

AIMSB-cell acute lymphoblastic leukemia (B-ALL) is amongst the most prevalent cancers of children in Pakistan. Genetic variations in FLT3 are associated with auto-phosphorylation of kinase domain that leads to increased proliferation of blast cells. Paired box family of transcription factor (PAX5) plays a critical role in commitment and differentiation of B-cells. Variations in PAX5 are associated with the risk of B-ALL. We aimed to analyze the association of FLT3 and PAX5 polymorphisms with B cell leukemia in Pakistani cohort.METHODSWe collected 155 B-ALL subject and 155 control blood samples. For analysis, genotyping was done by tetra ARMS-PCR. SPSS was used to check the association of demographic factors of SNPs present in the population with the risk of B-ALL.RESULTSRisk allele frequency A at locus 13q12.2 (rs35958982, FLT3) was conspicuous and showed positive association (OR = 2.30, CI [1.20–4.50], P = 0.005) but genotype frequency (OR = 3.67, CI [0.75–18.10], P = 0.088) failed to show any association with the disease. At locus 9p13.2 (rs3780135, PAX5), the risk allele frequency was significantly higher in B-ALL subjects than ancestral allele frequency (OR = 2.17, CI [1.37–3.43], P = 0.000). Genotype frequency analysis of rs3780135 polymorphism exhibited the protective effect (OR = 0.55, CI [0.72–1.83], P = 0.029). At locus 13q12.2 (rs12430881, FLT3), the minor allele frequency G (OR = 1.15, CI [1.37–3.43], P = 0.043) and genotype frequency (OR = 2.52, P = 0.006) reached significance as showed p < 0.05.CONCLUSIONIn the present study, a strong risk of B-cell acute lymphoblastic leukemia was associated with rs35958982 and rs12430881 polymorphisms. However, rs3780135 polymorphism showed the protective effect. Additionally, other demographic factors like family history, smoking and consanguinity were also found to be important in risk assessment. We anticipate that the information from genetic variations in this study can aid in therapeutic approach in the future.

GPT2 mutations in autosomal recessive developmental disability: extending the clinical phenotype and population prevalence estimates.

The glutamate pyruvate transaminase 2 (GPT2) gene produces a nuclear-encoded mitochondrial enzyme that catalyzes the reversible transfer of an amino group from glutamate to pyruvate, generating alanine and alpha-ketoglutarate. Recessive mutations in GPT2 have been recently identified in a new syndrome involving intellectual and developmental disability (IDD), postnatal microcephaly, and spastic paraplegia. We have identified additional families with recessive GPT2 mutations and expanded the phenotype to include small stature. GPT2 loss-of-function mutations were identified in four families, nine patients total, including: a homozygous mutation in one child [c.775T>C (p.C259R)]; compound heterozygous mutations in two siblings [c.812A>C (p.N271T)/c.1432_1433delGT (p.V478Rfs*73)]; a novel homozygous, putative splicing mutation [c.1035C>T (p.G345=)]; and finally, a recurrent mutation, previously identified in a distinct family [c.1210C>T (p.R404*)]. All patients were diagnosed with IDD. A majority of patients had remarkably small stature throughout development, many < 1st percentile for height and weight. Given the potential biological function of GPT2 in cellular growth, this phenotype is strongly suggestive of a newly identified clinical susceptibility. Further, homozygous GPT2 mutations manifested in at least 2 of 176 families with IDD (approximately 1.1%) in a Pakistani cohort, thereby representing a relatively common cause of recessive IDD in this population, with recurrence of the p.R404* mutation in this population. Based on variants in the ExAC database, we estimated that approximately 1 in 248 individuals are carriers of moderately or severely deleterious variants in GPT2.

Association of MIR146A rs2910164 variation with a predisposition to sporadic breast cancer in a Pakistani cohort.

Single-nucleotide polymorphisms (SNPs) in genes coding for microRNAs (miRNAs) play a pivotal role in the progression of breast cancer (BC). We investigated the association of miR-146a rs2910164 GC polymorphism with the risk of BC in the Pakistani population. The miR-146a rs2910164 polymorphism was genotyped in 300 BC cases and 300 age- and gender-matched healthy controls using T-ARMS-PCR. Genotype and allele frequencies were calculated and the association between genotypes and the risk of BC was calculated by odds ratio (OR) and confidence interval (95%). A significant difference in genotypic frequencies (χ2 =63.10; P=<0.0001) and allelic frequencies (OR=0.3955 (0.3132-0.4993); P=<0.0001) was observed between cases and controls. Furthermore, we also found that miR-146 rs2910164 CC homozygote increased the risk of BC in the dominant (OR=0.2397 (0.1629-0.3526); P=0.0001; GG vs. GC + CC) and recessive (OR=2.803 (1.865-4.213); P=<0.0001; CC vs. GC + GG) inheritance models. In summary, miR-146a rs2910164 GC is significantly associated with BC in the Pakistani population. To our knowledge, this is the first study that assessed MIR146a rs2910164 G>C SNP in Pakistani population. By analyzing the secondary structure of MIR146A variant, a significant structural modification was noted. Study with a larger sample size is needed to further confirm of these findings.

Association of rs10490924 in ARMS2/HTRA1 with age-related macular degeneration in the Pakistani population.

Age-related macular degeneration (AMD) is a disease of the elderly in which central vision is lost because of degenerative changes of the macula. The current study investigated the association of single-nucleotide polymorphisms (SNPs) with AMD in the Pakistani population. Four SNPs were analyzed in this study: rs1061170 in the CFH, rs429608 near CFB, rs2230199 in the C3, and rs10490924 in ARMS2/HTRA1. This case-control association study was conducted on 300 AMD patients (125 wet AMD and 175 dry AMD) and 200 unaffected age- and gender-matched control individuals. The association of the SNP genotypes and allele frequency distributions were compared between patients and healthy controls, keeping age, gender, and smoking status as covariates. A significant genotype and variant allele association was found of rs10490924 in ARMS2/HTRA1 with wet AMD, while the SNPs in CFH, CFB, and C3 were not associated with AMD in the current Pakistani cohort. The lack of association of CFH, CFB, and C3 may be attributed to limited sample size. This study demonstrates that genetic causative factors of AMD differ among populations and supports the need for genetic association studies among cohorts from various populations to increase our global understanding of the disease pathogenesis.

Clinicopathological Features of Young Versus Older Patients With Breast Cancer at a Single Pakistani Institution and a Comparison With a National US Database.

PURPOSEThe age at which women present with breast cancer varies widely among nations, and breast cancer may behave differently in younger women. Differences in clinicopathological characteristics based on age have not been well characterized in Pakistani patients with breast cancer.METHODSWe conducted a retrospective review of patients with symptoms of breast cancer presenting to Aga Khan University Hospital (AKUH), a large tertiary care center in Pakistan, between 2001 and 2010; we compared young (≤ 40 years) versus older (> 40 years) patients in terms of their clinicopathological characteristics. We also compared this Pakistani cohort with the US population using data from the National Cancer Database (NCDB).RESULTSA total of 1,334 patients with breast cancer presented to our center over the 10-year review period. The median age at diagnosis was 50 years, compared with 60 years for patients in the NCDB. In the AKUH cohort, younger patients were significantly more likely than their older counterparts to present with metastatic disease (13.1% v 10.8%; P < .01). They also were more likely to present with higher-grade tumors (grade 3: 40.1% v 28.3%; P < .001), have triple hormone receptor–negative phenotype (25.4% v 14.1%, P < .001), and have positive axillary lymph node involvement (70.9% v 57.5%; P < .001) compared with older women. Younger and older patients in the AKUH cohort tended to present with higher-stage disease (P < .001) and were more likely to have triple hormone receptor–negative disease (P < .001), compared with all patients in the NCDB and with those of Indo-Pakistani origin.CONCLUSIONYoung Pakistani women, similar to their Western counterparts, present with more advanced disease and more aggressive tumor biology than their older counterparts.

Functional polymorphism within miR-23a∼27a∼24-2 cluster confers clinical outcome of breast cancer in Pakistani cohort.

MicroRNAs (miRNAs) are small regulatory RNA molecules that control gene activity by base pairing with target messenger RNA leading to their cleavage or translational repression. Previous studies show an involvement of miRNAs in various diseases including cancer. Members of the Mir-23a cluster (MIR23A, MIR24-2and MIR27A) are involved in breast cancer (BC). In the present study, miR-23a/24-2/27a cluster was screened for genetic mutation in BC patients. Heterozygous (A/G allele) as well as homozygous (G/G allele) variants were found in mir-27a gene in screened BC patients. RNA structural analysis revealed that the single nucleotide polymorphism (SNP) effects the size of the terminal loop in the precursor miRNA (pre-miRNA). The altered (G allele) hairpin structure observed was two bases longer than the reference (A allele) hairpin.

Impaired renal functions in Pakistani cohort of rheumatoid arthritis.

Objective:To determine the frequency of impaired renal functions and hypertension in rheumatoid arthritis.Methods:This study was conducted between May 1st 2018 to February 1st 2019 at Rheumatology Division, Department of Medicine Central Park Medical College Lahore, total 260 study participants were selected, demographic detail were asked in detail, disease duration of RA and hypertension, DMARD’s, self-use NSAID’s,/hakeem medications, smoking were asked in detail, BMI and blood pressure were measured,5 ml of blood was taken by trained phlebotomist, and sent for the estimation of serum urea and creatinine on (COBAS-III) machine, after availability of results each individuals eGFR (creatinine clearance) was calculated by Cockroft Gualt(CG) and Modification in diet in renal disease method (MDRD).Results:In this study the mean age of study participants was 42.4 (± 9.5) years with disease duration of 7.7(±4.8) years, prevalence of Impaired renal functions of 14.6% (n=38) and hypertension in 53.5% (n=139).Regression analysis shows there is significant association between hypertension, smoking and self/hakeem medications with impaired renal functions (p-0.5). Kappa analysis shows both (MDRD & CG methods) had uniformity in picking up cases of impaired renal functions 75.6% (p-0.05).Conclusion:In RA decline in renal functions is seen with self-use NSAID’s/hakeem medications along with other modifiable factors like smoking and hypertension, while conventional DMARD’s don’t show association with decline. There is very high prevalence of hypertension in rheumatoid arthritis.

Genome-wide implicated risk variants of TCF7L2 gene contribute to type 2 diabetes susceptibility by modulating serum lipids in Pakistani population

Diabetes is a socioeconomic burden worldwide as well as in Pakistan. International Diabetes Federation reported 7 million cases of diabetes and 86,384 deaths due to diabetes in Pakistan in 2015. Transcription factor 7 like 2 is a transcription factor encoded by TCF7L2 gene and regulates several physiological processes in the cell. The aim of the current study was to determine the prevalence of two TCF7L2 gene polymorphisms and analyze its effect on various anthropometric and biochemical parameters in a Pakistani cohort. We collected 926 samples, 500 healthy controls (fasting blood sugar 99 mg/dL, random blood sugar > 126 mg/dL). The genotyping was done by RFLP-PCR and serum biochemical parameters were determined. The genotyping results of rs7903146 by RLFP-PCR showed the allelic frequency of C = 0.64 and T = 0.36 in controls while C = 0.59 and T = 0.41 in cases and genotypic frequency CC = 43.2%, CT = 41.6%, and TT = 15.2% in controls while CC = 34.03%, CT = 49.06%, and TT = 16.91% in cases. The minor T allele appeared to be a risk allele (OR = 1.25, CI = 1.03–1.51, p = 0.016), i.e., it increased the risk of diabetes in the selected study subjects. The genotyping results of rs12255372 showed allelic frequency G = 0.75 and T = 0.25 in controls while G = 0.63 and T = 0.37 in cases and genotypic frequency GG = 54.2%, GT = 42.2%, and TT = 3.6% in controls, while GG = 40.14%, GT = 46.24%, and TT = 13.62% in cases. The minor T allele appeared to be a risk allele for this SNP as well (OR = 1.77, CI = 1.44–2.16, p = 1.9 × 10−8). The biochemical parameters and general characteristics were checked for association with the TCF7L2 variant rs7903146, showed no association with all tested parameters. While rs12255372 had significant association with fasting plasma glucose, triglycerides, HDLC, LDLC, and leptin, but no association with total cholesterol, age, height, weight, and BMI. However, the multivariate analysis showed significant association of lipid parameters only when homozygous risk genotype of rs12255372 co-existed with all genotypes of rs7903146. In conclusion, the polymorphisms in TCF7L2 gene can increase the susceptibility to diabetes and the effect may involve modulating certain serum traits.