Hyperfunction variant rs708035 of interleukin 1 receptor-associated kinases 2 gene involved in the predisposition of leprosy infection.

BACKGROUND: Mycobacterium leprae (slow-growing bacteria) is the etiological agent for leprosy infection, which is a chronic granulomatous disease. Symptoms initiate with the loss of sensation in the affected areas, which can lead to severe injuries, cuts and burns. IRAK2 (interleukin-1 receptor-associated kinases 2) is reported to function in the regulation of the NFκB pathway. The frequency of the IRAK2 polymorphism (rs708035) was unknown in the Pakistani population. Therefore, the study was designed to identify the role of the rs708035 SNP (single nucleotide polymorphism) in susceptibility to leprosy. METHODOLOGY: The case-control study was designed, and participants were selected by Ridley-Jopling Classification. Blood samples from healthy individuals and patients were collected after ethical approval. Genomic DNA was extracted for the amplification of selected polymorphisms by tetra-primer amplification refractory mutation system polymerase chain reaction. The desired products were observed via agarose gel (2.5%) electrophoresis followed by data analysis using bioinformatics tools (SNP Stats and SHEsis) and statistical tests (odds ratio, OR, and chi square). RESULTS: The study revealed that the mutant genotype (TT) was found to be frequent among cases (22.80%) in comparison with the controls (1.66%). The SNP rs708035 was significantly associated with the progression of leprosy (χ2  = 17.62, p < 0.0001). The targeted SNP significantly increases the risk of leprosy 2.3 times (OR = 2.3119, 95% CI 1.2729-4.1989, p < 0.01). The genetic model also confirms the significant association of the A/T genotype with leprosy in the over-dominant model (OR = 2.83, 95% CI 1.16-6.89, p < 0.001). CONCLUSIONS: The study revealed a significant association of the targeted SNP with leprosy and provided baseline data regarding the association of rs708035. The current research could be utilized for the preparation of biomarkers by considering a larger sample size. HIGHLIGHTS: The patients suffering from leprosy faced various comorbidities, including hypertension and diabetes. The study reports for the first time a significant association of interleukin 1 receptor associated kinases 2 (IRAK2) single nucleotide polymorphism (SNP) rs708035 among the Pakistani population (Karachi). The current study provides baseline data to develop diagnostic biomarkers for early detection of leprosy.

Vitamin D receptor gene polymorphism TaqI (rs731236) and its association with the susceptibility to coronary artery disease among Pakistani population.

BACKGROUND: Coronary artery disease (CAD) is a leading cause of mortality in Pakistan and also worldwide. Vitamin D receptor (VDR) regulates the transcription of many genes and has a significant impact on inflammation and the morphology of cardiac cells. Genetic variation in the VDR gene such as the TaqI polymorphism (rs731236) may have an impact that causes adverse effects. Accordingly, it is important to determine possible association of the TaqI polymorphism (rs731236) with CAD. METHODS: The study included blood samples from 1016 subjects: 516 from CAD patients and 500 from age- and gender-matched controls. Genomic DNA was extracted by standard salting out method. Targeted variation was amplified by an allele-specific polymerase chain reaction (PCR). PCR products were examined and genotyped on agarose gel electrophoresis represented by an amplified product size of 148 bp followed by Sanger sequencing to validate variations. RESULTS: Serum vitamin levels, as observed using enzyme-linked immunosorbent assay, were found to be insufficient in both CAD patients (20.52 ± 0.06 ng/ml) and controls (21.6981 ± 0.05 ng/ml). The TaqI polymorphism (rs731236) T>C was found to be significantly associated with CAD (p < 0.0001). The odds ratio showed that the risk increases by 1.8-fold with variant C allele. Dominant, co-dominant and over dominant genetic model analyses suggested that the TC genotype might be a risk factor involved in the possible association with susceptibility to CAD. CONCLUSIONS: The TaqI polymorphism (rs731236) in the coding region may affect the function of the receptor by altering the binding site, which might participate in an inflammatory response and increase the risk for developing susceptibility to CAD.

Association analysis and allelic distribution of deletion in CC chemokine receptor 5 gene (CCR5Δ32) among breast cancer patients of Pakistan.

Chemokine CC receptor type 5 (CCR5) is a cell surface receptor that has high affinity for chemotropic cytokines called chemokines. The CCR5 gene contains a 32 base pairs (bp) deletion (CCR5Δ32). This deletion may result in a malformed and nonfunctional receptor, reported to be responsible for the development and dissemination of different cancers. CCR5Δ32 exists in two allelic forms i.e. deletion (D) and wild type (WT). This study aims to detect the role of CCR5Δ32 in breast cancer development. Blood samples were collected from breast cancer patients (330) and controls of same gender (306). Along with this histopathologically diagnosed malignant tissue samples were also excised from breast lesions of 100 patients. Genetic variations within the blood and tissue samples were examined by PCR then observed through gel electrophoresis and confirmed by direct DNA sequencing. Obtained DNA sequences were aligned and analyzed by MEGA6 software. Genotypic and association analyses were done by SPSS software version 17.0. Deletion of 32 bp in CCR5 gene has been analyzed. Genotypic variations of CCR5Δ32 are; homozygous wild type (WT/WT), heterozygous deletion (WT/D) and homozygous deletion (D/D). Statistical analyses of CCR5Δ32 data revealed that WT/D was significantly higher in blood samples of breast cancer patients (7.27% (24/330)) as compare to controls (1.30% (4/306)). In tumor tissue samples WT/WT being the most frequent genotype (99.00% (99/100)) with 1.00 (1/100) of D/D which suggested that it may be acquired. Hence, association analysis showed that CCR5Δ32 is positively associated with breast cancer in Pakistan (p < 0.001). The risk ratio of CCR5Δ32 was 5.6610 (95% confidence interval: 2.0377 to 15.7267) and odds ratio was calculated to be 6.0335 (95% confidence interval: 2.1288 to 17.0999) which signifies that deletion also increases the risk of breast cancer development. Moreover, association analyses also revealed that clinicopathological features do not have any impact on the CCR5Δ32 genotype of breast cancer. This suggests that deletion of 32 bp in CCR5 gene may be associated with breast cancer. CCR5 signals the activation and migration of immune cells at the site of tumor formation. Because of deletion; deformed CCR5 receptor might be unable to express and function properly which may subdue the immunity against cancer hence, leading to its progression.

Addiction of tobacco chewing and smoking in the patients of head and neck squamous cell carcinoma: A descriptive epidemiological study in Pakistan.

OBJECTIVE: The associated risk factors for the majority of patients with head and neck squamous cell carcinoma (HNSCC) are tobacco and betel nut abuse, while there also seems to be a rising proportion of patients who report no history of tobacco or betel nut usage. Therefore, objective of the study was to find out potential risk factors and demographics of HNSCC patients addicted to tobacco and/or betel nut, as well as non-addicted patients. METHODS: This epidemiological study was conducted in Karachi Institute of Biotechnology and Genetic Engineering (KIBGE), University of Karachi, and Jinnah Postgraduate Medical Centre (JPMC) from January to December 2016. All subjects were participants in an epidemiological study of HNSCC. Demographics and clinical characteristics were obtained for 185 addicted and 26 non-addicted patients. RESULTS: Non-addicted patients were more likely to be females (χ2=19.0, p<0.001) and were significantly younger than addicted patients (χ2=21.0, p<0.001). Addicted patients more likely belonged to a lower income group (χ2=14.4, p=0.006) and had a higher proportion of oral cancers (χ2=30.0, p<0.001). Almost all addicted females had oral cancers (97%), whereas more than half of the non-addicted females had oral cancers (53%). CONCLUSIONS: Addicted patients commonly have oral cavity cancers. This might be due to the habit of chewing tobacco and/or betel nut that addicted patients have. Non-addicted patients are commonly young females. It is likely that no single known factor is responsible for HNSCC in non-addicted patients, and several occupational exposure studies in future may be important to the etiology of non-addicted patients.

Serum Interleukin-15 and its relationship with adiposity Indices before and after short-term endurance exercise.

OBJECTIVE: The myokine interleukin-15 (IL-15) is capable of modifying the metabolism of both skeletal and adipose tissue. This study compares the change in serum levels of IL-15 in obese and non-obese after a single session of submaximal exercise. METHODS: A cross-sectional study was carried out at Jinnah Medical and Dental College, Karachi, during Aug-Dec 2015, comprising of 133 medical students (aged 17-24 years). Cardiorespiratory fitness was evaluated by Queen's College Step Test. Blood was obtained both before and just after exercise and serum levels of IL-15 determined by enzyme-linked immunosorbent assay. RESULTS: Mean serum level of IL-15 was 3.64±1.59 pg/mL. Higher levels of IL-15 were seen in lean subjects compared to overweight/obese, both before and after three minutes of exercise (all Ptrend<.001). The percent increase in IL-15 upon exercise was 12.7% higher in lean. Significant negative association was seen between interleukin-15 and adiposity, especially visceral fat (r = -.288, p=.001). CONCLUSION: Interleukin-15 correlates negatively with adiposity indices, especially visceral fat. With the proven benefit of IL-15 in terms of adipose tissue stores and skeletal muscle mitochondrial biogenesis, endurance exercises, even of short duration, may possess therapeutic potential towards producing a healthier body.

Association and implications in triple negative and triple positive breast cancer: Relationship with sociodemographic and reproductive factors in Pakistan.

BACKGROUND & OBJECTIVES: Triple negative and triple positive breast cancer have adverse effects than other types of breast cancer. However, triple negative has poor prognosis with short survival as compared with triple positive breast cancer. Good prognosis is one of the key factors for successful treatment trial. This study aimed to find out the association of sociodemographic and reproductive features like parity, menopause, number of child bearing as risk factors in the development and prognosis of triple negative and triple positive breast cancer. METHODS: This study is a part of an ongoing project which is being conducted in Karachi from 2013 to 2020. Informed consent from triple negative breast cancer (n=134) and triple positive breast cancer (n=87) patients were taken prior to their recruitment into the study. Demographic, anthropometric, reproductive and disease history of patients were recorded. Means, frequency distribution, data classification and association analyses were done by SPSS version 17.0. RESULTS: Statistical analyses revealed that delayed first child bearing age and lower number of children are associated with the development of triple negative breast cancer. However, no significant effect of these parameters has been observed on the outcomes of triple positive breast cancer. CONCLUSIONS: Reproductive factors have more pathological implications than sociodemographic factors in both triple positive and triple negative breast cancer development. These findings might prove to be beneficial for effective and better breast cancer management.

Out of pocket (OOP) cost of treating hypertension in Karachi, Pakistan.

Pakistan is categorized to below to middle income countries where two third of the national annual health expenditure is in the form of out of pocket (OOP) cost. A prevalence based study was conducted to determine the OOP cost treatment of hypertension in Karachi by interviewing 350 hypertensive patients aged >30 years through a validated questionnaire. Hypertension (HTN) was classified into stage 1 and stage 2 and was found to be common in females (53.42%) than males (46.57%). The total costs of stage 1and stage 2 HTN were calculated to be 217869.7PKR and17545457.6 PKR respectively. The average treatment cost of stage 1 was observed to be significantly lower (p=0.006) than the cost of stage 2 HTN. Moreover; the cost of antihypertensive drugs, physician fees and laboratory tests were considerably different however; no variation was seen in cost of transport and loss of productivity through absenteeism from work. Overall, the present study indicates that the antihypertensive treatment has imposed a high burden on the pocket of common man and this is a major reason for treatment non-adherence. Consequently, it increases the risks of cardiovascular events, morbidity and mortality. Therefore, effective strategic planning is need of time to reduce OOP cost for better control on hypertension in Pakistan.

Drug utilization and prescribing pattern of antibiotics in a tertiary care setups; trends and practices.

Irrational, over and misuse of antibiotics arise as global concern in both hospital and community settings and lead to adverse events including antimicrobial resistance, associated health problems, amplified hospitalization stay and cost. Hence, Drug Utilization Evaluation (DUE) studies are designed to evaluate and improve the prescribing, administration and the rational use of medications. The present study was designed to assess the pattern of antimicrobial drug utilization in in-patients cohort of tertiary care setup in Karachi, Pakistan. This cross sectional observational study was conducted in retrospective manner. World health organization (WHO) guidelines and criteria are considered to evaluate the appropriateness of drug use in various disease conditions. ATC/DDD system was applied to determine the study outcome. High frequency of antibiotics utilization found in respiratory tract infections of both lower (LRTI) 16.8% (n=42) and upper (UTI) 13.2% (n=33). The estimated total number of drug units administered per month was greater with cefixime (46) and ciprofloxacin (45) both. DDD/100 bed days drug utilization of antibiotics was higher with ciprofloxacin, cefexime and meropenem (47, 46 and 29.25) correspondingly. In conclusion, the current investigation signifies extensive scope for progress in prescribing trend. Drug adherence to customary guidelines of disease management and constraint policies to endorse judicious drug use may be considered vital in healthcare setup.

CC chemokine receptor 5 Δ32 polymorphism: association analysis and allele distribution among cutaneous leishmaniasis patients from Pakistan.

BACKGROUND: Human immunodeficiency virus (HIV)/leishmaniasis coinfection is a matter of deep concern worldwide. CC chemokine receptor 5 (CCR5) functions as a co-receptor for HIV entry into host immune cells with an elevated expression observed during leishmaniasis, promoting parasite persistence. A 32 bp deletion (Δ32) in the CCR5 gene provides protection against HIV infection and increased resistance to Leishmania infection. METHODS: In this study, CCR5-Δ32 distribution within Pakistani population with cutaneous leishmaniasis was investigated to evaluate genetic susceptibility to HIV infection. CCR5-Δ32 polymorphism was analyzed in 276 leishmaniasis patients and 119 uninfected healthy controls. Genotypic and allelic frequencies were evaluated and tested for Hardy-Weinberg equilibrium (HWE). RESULTS: The overall Δ32 allele frequency was 6.58% of the population (n = 395). There was a significant difference (p < 0.05) in the geographical distribution of Δ32 allele which was higher in the northern region of the country when compared with the south. Five individuals were identified to be homozygous for the Δ32 allele which has not been reported before from Pakistan. However, no significant association was observed between CCR5-Δ32 and cutaneous leishmaniasis. CONCLUSION: The higher frequency of CCR5 wild-type allele among leishmaniasis patients may suggest an increased risk of HIV infection and also support its facilitative role in Leishmania infection.

The association of GSTM1 and GSTT1 polymorphisms with squamous cell carcinoma of cervix in Pakistan.

Gene deletions in GSTM1 and GSTT1 may result in tempering the activation and detoxification of several carcinogens and thereby may increase the risk of cancer pre-disposition. This study aims to investigate the clinical impact of glutathione-S-transferase GSTM1 and GSTT1 polymorphisms on squamous cell carcinoma of cervix (SCCA).The GSTM1 and GSTT1 polymorphisms were analyzed in cervical cancer patients and healthy controls. Touch down multiplex polymerase chain reaction (PCR) strategy was adopted for genotyping of GSTM1 and GSTT1 polymorphisms. The null genotype of GSTM1 exhibited a significantly higher percentage in patients with SCCA (74 %) than in the control group (34.0 %). However, no significant difference was observed in the null genotype of GSTT1 among SCC patients and healthy subjects, respectively. GSTM1 exhibited a significant association with increased risk of squamous cell carcinoma (p < 0.001). The odds ratio for the GSTM1 null genotype was also calculated (odds ratio 3.7484; 95 % confidence interval 1.6562-84834). This suggests that GSTM1 null genotype in cervical cell samples may be associated with more severe precancerous lesions of the cervix.

Decreased muscle strength in adjuvant-induced rheumatoid arthritis animal model: A relationship to behavioural assessments.

Rheumatoid arthritis (RA) is an autoimmune disorder with unknown aetiology. Patients suffering from RA face persistent pain due to joint inflammation, and tissue destruction. Behavioural phenotyping is an approach to target the role of different behavioural traits associated with disease progression. The study aimed to assess behavioural patterns associated with decreased muscle strength in the adjuvant-induced rheumatoid arthritis animal model. The study was conducted on male Albino Wister rats (n = 30) [Control, Vehicle, and Disease groups]. After taking ethical approvals RA was induced by complete Freund's adjuvant (CFA) intradermally base of tail. The weight of animals, macroscopic analysis of inflammatory signs, and arthritic scores were measured weekly. Grip strength, ganglia-based movement, cataleptic activity, and motor-coordination-related behaviours were assessed among the groups. Radiographs and spleen index assay were performed followed by data analysis using one-way and two-way ANOVA (Analysis of Variance). A significant decrease in weight and an increase in arthritic scores among the diseased group was observed. Behavioural analyses confirmed that diseased animals had significantly decreased grip strength and increased cataleptic activity with less motor coordination. Radiographic images and spleen index assay confirmed the pattern of RA. Therefore, it can be suggested that the development of the disease animal model is an effective approach to identifying the disease progression and associated behavioural changes. Moreover, this prepared laboratory animal model may be utilised for pathway analyses to understand the key role of immune regulators and genetic insight into molecular pathways associated with acute and chronic phases of RA.

In situ electrochemical digital holographic microscopy; a study of metal electrodeposition in deep eutectic solvents.

This study has shown for the first time that digital holographic microscopy (DHM) can be used as a new analytical tool in analysis of kinetic mechanism and growth during electrolytic deposition processes. Unlike many alternative established electrochemical microscopy methods such as probe microscopy, DHM is both the noninvasive and noncontact, the unique holographic imaging allows the observations and measurement to be made remotely. DHM also provides interferometric resolution (nanometer vertical scale) with a very short acquisition time. It is a surface metrology technique that enables the retrieval of information about a 3D structure from the phase contrast of a single hologram acquired using a conventional digital camera. Here DHM has been applied to investigate directly the electro-crystallization of a metal on a substrate in real time (in situ) from two deep eutectic solvent (DES) systems based on mixture of choline chloride and either urea or ethylene glycol. We show, using electrochemical DHM that the nucleation and growth of silver deposits in these systems are quite distinct and influenced strongly by the hydrogen bond donor of the DES.

Ligand exchange in ionic systems and its effect on silver nucleation and growth.

The electrodeposition of metals from ionic solutions is intrinsically linked to the reactivity of the solute ions. When metal salts dissolve, the exchange of the anion with the molecular and ionic components from solution affects the speciation and therefore the characteristics of metal reduction. This study investigates the nucleation mechanism, deposition kinetics, metal speciation and diffusion coefficients of silver salts dissolved in Deep Eutectic Solvents. The electrochemical reduction of AgCl, AgNO3 and Ag2O is studied in 1 : 2 choline chloride : ethylene glycol and 1 : 2 choline chloride : urea. Cyclic voltammetry is used to evaluate electrochemical kinetics. Detailed analysis of chronoamperometric data shows that silver deposits form via multiple 3D nucleation with mass transport controlled hemispherical growth. The nucleation mechanism was found to be potential dependent, varying from progressive to instantaneous as the reduction potential becomes more cathodic. Diffusion coefficients are determined using three different methods. Trends are rationalised in terms of solvent viscosity and silver speciation analysis with EXAFS. The morphology of electroreduced silver is investigated with scanning electron microscopy and shows that deposits from the urea based liquid form more dense morphologies than those from the ethylene glycol based liquid.

Bioaccumulation of Chloropyrifos Organo-pesticide and Its Toxicogenic Association with Antioxidant GSTP1 in Pakistani Pest Control Workers.

Synthetic pesticides are employed to enhance agricultural production. Chronic exposure to organophosphate (OP) pesticides may be a source of health problems. The present study was designed to examine an association of GSTP1 (rs1695) polymorphism with OP pesticide chronic exposure. A case-control study was recruited with 250 subjects comprising exposed (n = 100) and controls (n = 150). A survey was conducted to determine the pesticide type to which workers had exposed. According to recorded survey assessment, two compounds of OP pesticides chloropyrifos and malathion were investigated in the blood samples of exposed study subjects using high-performance liquid chromatography (HPLC). For screening of genetic polymorphism in GSTP1 (rs1695) polymerase chain reaction, restriction length polymorphism (PCR-RFLP) and agarose gel electrophoresis were performed. Statistically, data were analyzed using SPSS v. 20.0 and MedCal© software. Total chrom© navigator programmer was used for detection of OP residues in serum and local pesticide solution. chloropyrifos-OP pesticide residues were detected in serum of estimated chronically exposed subjects at 206 nm HPLC optimal conditions. The pattern of GSTP1 (rs1695) genotypic frequencies depicted that heterozygous genotype was higher in Chloropyrifos exposed subjects (0.56) when compared with controls (0.44). Statistical outcomes showed an insignificant association with GSTP1 (rs1695) polymorphism and chloropyrifos-OP pesticide toxicity (Fisher's exact test 1.0, p = 0.25). An insignificant allelic investigation reflected a protective effect of mutant allele G against chloropyrifos-OP pesticide toxicity in exposed subjects. Findings may be helpful in identifying bioaccumulated pesticide residues, but in studied Pakistani exposed workers, no significant association of GSTP1 (rs1695) variant with chloropyrifos-OPs was demonstrated.

Single nucleotide polymorphisms (rs3736228 and rs4988321) in low-density lipoprotein receptor-related protein-5 gene with predisposition to rheumatoid arthritis.

BACKGROUND: LRP5 (Lipoprotein Receptor 5) is one of the representatives of the low-density lipoprotein receptors family that play a crucial role in the process of bone homeostasis and bone remodeling. The role of LRP5 single nucleotide polymorphisms (SNPs) rs3736228 and rs4988321 has been associated with the susceptibility to osteoporosis and bone fracture. The frequency of mentioned LRP5 SNPs is unknown among RA (Rheumatoid Arthritis) patients. The case-control study was designed to determine the association of targeted SNPs among RA patients. METHODOLOGY: Patients were selected by ACR/EULAR 2010 criteria. After ethical approval blood samples of patients and healthy individuals were collected. DNA was extracted from the whole blood followed by amplification of the targeted region by T-ARMS PCR (Tetra-primer Amplification Refractory Mutation System) obtained product was observed on agarose gel electrophoresis. The data were analyzed by statistical and bioinformatic tools. RESULTS: It was observed that the SNPs rs3736228 and rs4988321 showed significant association with the risk of RA [χ2 = 44, p =< 0.001, O.R 95 % CI = 2.495, (1.865 âˆ¼ 3.337), p =< 0.001] and [χ2 = 85, p =< 0.001, O.R 95 % CI = 2.05, (1.571 âˆ¼ 2.69), p =< 0.001] respectively. Haplotypes AT, GC, and GT were found to be significantly associated with the risk of RA. Furthermore, both SNPs were in 40 % LD (Linkage Disequilibrium). CONCLUSIONS: The study revealed that SNPs rs3736228 and rs4988321 were significantly associated with the increased susceptibility to RA. The study serves as the baseline data considering targeted SNPs and their association with the progression of the disease. The study might be utilized for the development of potential biomarker for diagnostic purposes and in the precision medicine approach.

Mutations in PGRN gene associated with the risk of psoriasis in Pakistan: a case control study.

BACKGROUND: Psoriasis is a chronic, autoimmune, papulosquamous skin disorder, characterized by the formation of drop-like papules and silvery-white plaques surrounded by reddened or inflamed skin, existing predominantly on the scalp, knees and elbows. The characteristic inflammation and hyperproliferation of keratinocytes in psoriasis is regulated by progranulin (PGRN), which suppresses the expression and release of inflammatory cytokines, such as TNF-α. METHODOLOGY: In this study mutation analysis of the PGRN gene was performed by extracting the genomic DNA from blood samples of 171 diagnosed psoriasis patients and controls through standard salting-out method, followed by amplification and sequencing of the targeted region of exon 5-7 of PGRN gene. RESULTS: Three single nucleotide polymorphisms, rs25646, rs850713 and a novel point mutation 805A/G were identified in the PGRN gene with significant association with the disease. The variant alleles of the polymorphisms were significantly distributed among cases and controls, and statistical analysis suggested that the mutant genotypes conferred a higher risk of psoriasis development and progression. Multi-SNP haplotype analysis indicated that the CAA (OR = 8.085, 95% CI = 5.16-12.66) and the CAG (OR = 3.204, 95% CI = 1.97-5.21) haplotypes were significantly associated with psoriasis pathogenesis. CONCLUSIONS: These findings demonstrate that polymorphisms in PGRN might act as potential molecular targets for early diagnosis of psoriasis in susceptible individuals.

Association of serum vitamin D levels and TaqIrs731236 among patients with hypertensive coronary heart disease.

The development of cardiovascular diseases (CVD) is influenced through multiple risk factor and hypertension. It may increase the risk of cardiac events, and has a significant impact when combined with other risk factors including low levels of vitamin D and genetic variations like single nucleotide variations (SNV) (TaqIrs731236) in vitamin D receptor (VDR) gene. Blood samples from 500 study participants gathered including 250 hypertensive coronary heart disease patients, 250 age and gender matched healthy controls. To isolate genomic DNA, conventional salting out procedure used followed by amplification of targeted variations through Amplification Refractory Mutation System- Polymerase Chain Reaction (ARMS-PCR) Assay. The amplicon consists of 148 base pairs which was visualized on 2 % agarose gel electrophoresis and confirmed by DNA sequencing. The compared clinical parameters including systolic blood pressure (SBP), diastolic blood pressure (DBP), body mass index (BMI), high density lipoproteins (HDL), low density lipoproteins (LDL), cholesterol, triglycerides found significantly different among patients when compared with controls (P < 0.001). The Vitamin D exhibited insufficient levels at different stages of hypertension which were statistically, found significantly associated among patients with hypertensive coronary heart disease showing compared to controls (P < 0.001). The genotype association SNV (TaqIrs731236) T > C showed significant association with hypertensive coronary heart disease compared to healthy controls (Chi-Square χ2 = 60.75 and P < 0.00001). Further, the odds ratio of allelic association for risk allele (C) showed the strength of association with risk of disease, which increases by 2.02 times(P = 0.01). The results suggest that (TaqIrs731236) T > C as genetic predisposition factor, may contribute to develop the risk of hypertensive coronary heart disease. Hypertension as a risk factor along with insufficient levels of vitamin D and SNV (TaqIrs731236) as genetic variations may have been an important contributor to disease risk of hypertensive coronary heart disease.

Effect of soft and prolonged Graeco-Arabic massage in low back pain - A randomized controlled clinical trial.

BACKGROUND: Low back pain (waja-uz-zahr) due to lumbar spondylosis affects approximately 60-80% of the population at any point of life. The present study was aimed to evaluate the efficacy of soft and prolonged massage with roghan-i-suranjan on low back pain. MATERIAL AND METHODS: Clinically and radiologically diagnosed patients of low back pain were randomized into test and control groups consisting 20 patients in each group. The patients in test group received soft and prolonged massage with roghan-i-suranjan; while, in control group, short wave diathermy was given on every alternate day up to 20 min for 3 weeks. The patients were assessed before and after the treatment using the VAS and ODI (OLBP) scales. RESULTS: VAS score was reduced to 42.14%, (p < 0.001) in massage and 13.94%, (p < 0.008) in control group after the treatment [Percentual Change (PC) between the groups = 54.35%, p < 0.001]. OLBP score was improved to 37.16% (p < 0.001) in massage and 5.93% (p < 0.0012) in control group after the treatment [PC between the groups = 29.57%, p < 0.02]. CONCLUSION: Apart from the muhallil-i-awram (anti-inflammatory), mulattif (attenuative), daf-i-alam (analgesic), and musakkin (sedative) activities of the tested-drugs, the specific strokes soft and prolonged massage further enhances the evacuative and rejuvenative process through its own action of tahleel (dissolution), talteef (attenuation), taskeen (sedation), and taskheen (diaphoresis) induced by the various manipulations used in this massage. Soft and prolonged massage with tested oil was found effective in reducing the pain and improving the quality of life in the patients of low back pain.

The role of institutional factors and cognitive absorption on students' satisfaction and performance in online learning during COVID 19.

With the rise of the Covid-19 pandemic, there has been a severe negative impact on all aspects of life, whether it be a job, business, health, education, etc. As a result, institutions, schools, colleges and universities are being shut down globally to control the spread of Covid-19. Due to this reason, the mode of education has a dramatic shift from on-campus to online learning with virtual teaching using digital technologies. This sudden shift has elevated the stress level among the students because they were not mentally prepared for it, and hence their academic performance has been adversely affected. So, there needs to figure out the underlying process to make online learning more productive. Thus, to obtain this objective, the present study has integrated the modified Technology Acceptance Model (TAM), Task Technology Fit Model (TTF), DeLone and McLean Model of Information Systems Success (DMISM) and Unified Theory of Acceptance and Use of Technology (UTAUT) model. A sample of 404 students was obtained, where 202 students were from the top ten public sector universities, and 202 were from the top ten private sector universities of Punjab. Structural Equation Modelling (SEM) was used to analyze the hypothesized framework using AMOS. The results reveal that institutional factors positively impact students' performance mediated by user satisfaction and task technology fit. Similarly, institutional factors affect performance through mediation by user satisfaction and actual usage in sequence. Cognitive absorption was used as a moderator between institutional factors and user satisfaction. In the end, theoretical and practical inferences have also been discussed.

Insights of OPs and PYR cytotoxic potential Invitro and genotoxic impact on PON1 genetic variant among exposed workers in Pakistan.

Different pesticide chemicals are used to enhance crop yield by protecting from pests. Organophosphate (OPs) and Pyrethroid (PYR) are used in fields of Sanghar, Sindh Pakistan. PON1 an antioxidant enzyme implicated in OPs detoxification may predispose by OPs chronic exposure. This study was conducted to evaluate the toxic potential of active pesticide chemicals at cellular and genetic levels. To examine toxic potential, locally consumed pesticide n = 2 and reference pesticide compounds organophosphate (OPs): Chloropyrifos, Malathion and Pyrethroid (PYR): Cyprmethrin, Cyhalothrin n = 4 were tested against NIH 3T3 cells using MTS assay. Local consumer pesticides demonstrated relevance for half-maximum inhibitory concentration (IC50) 0.00035 mg/mL with selected compound. Malathion IC50 exhibited the highest cytotoxicity among four compounds at 0.0005 mg/mL. On genotoxicity analysis in exposed subjects n = 100 genotypes and alleles n = 200 exhibited significant differences in genotypic and allelic frequencies of pesticide exposed subjects and controls n = 150 (X2 = 22.9, p = 0.001). Screening of genotypes were performed by PCR- RFLP. Statistical assessment carried out using online software and tools. Results suggested that higher heterozygous genotype A/G (74%) may confer low PON1 metabolic activity towards pesticides in exposed subjects. Findings could be helpful to establish health plans by avoiding toxic chemicals that harming exposed population.