Circadian rhythms are endogenous and self-sustained oscillations of multiple biological processes with approximately 24-h rhythmicity. Circadian genes and their protein products constitute the molecular components of the circadian oscillator that form positive/negative feedback loops and generate circadian rhythms. The circadian regulation extends from core clock genes to various clock-controlled genes that include various cell cycle genes. Aberrant expression of circadian clock genes, therefore, may lead to genomic instability and accelerated cellular proliferation potentially promoting carcinogenesis. The current study encompasses the investigation of simultaneous expression of four circadian clock genes (Bmal1, Clock, Per1 and Per2) and three clock-controlled cell cycle genes (Myc, Cyclin D1 and Wee1) at mRNA level and determination of serum melatonin levels in peripheral blood samples of 37 CLL (chronic lymphocytic leukemia) patients and equal number of age- and sex-matched healthy controls in order to indicate association between deregulated circadian clock and manifestation of CLL. Results showed significantly down-regulated expression of Bmal1, Per1, Per2 and Wee1 and significantly up-regulated expression of Myc and Cyclin D1 (P < 0.0001) in CLL patients as compared to healthy controls. When expression of these genes was compared between shift-workers and non-shift-workers within the CLL group, the expression was found more aberrant in shift-workers as compared to non-shift-workers. However, this difference was found statistically significant for Myc and Cyclin D1 only (P < 0.05). Serum melatonin levels were found significantly low (P < 0.0001) in CLL subjects as compared to healthy controls whereas melatonin levels were found still lower in shift-workers as compared to non-shift-workers within CLL group (P < 0.01). Our results suggest that aberrant expression of circadian clock genes can lead to aberrant expression of their downstream targets that are involved in cell proliferation and apoptosis and hence may result in manifestation of CLL. Moreover, shift-work and low melatonin levels may also contribute in etiology of CLL by further perturbing of circadian clock.

Aging is associated with increased levels of coagulation factors and decrease in natural anticoagulant factors. This strongly supports that age-related hypercoagulable state occurs in elderly. This study aimed to measure the plasma levels of coagulation factors and anticoagulant levels in young and elderly to observe the effect of age on haemostatic system. Ninety healthy individuals, both men and women were divided into two groups on the basis of age. Group I included participants of less than 40 years of age, whereas, group II comprised of participants more than 60 years of age. Fibrinogen activity was assessed by using Clauss technique. Coagulation factor VII, and factor VIII activity by corresponding one stage assay based on prothrombin time and activated partial thromboplastin time. Antithrombin III was measured by the chromogenic method. Our results showed that significantly increased levels of fibrinogen (P = 0.001) were observed in the elderly population as compared to young. Higher fibrinogen levels in younger women than men and comparatively higher level than other races was observed in our local population. Increase in factor VII levels (P = 0.05) was also observed in the elderly group. This increase was statistically significant with age in women (P = 0.03). Factor VIII rose with age in both sexes (P = 0.001). Higher antithrombin activity was observed in the younger group whereas the older group demonstrated significantly lower antithrombin activity (P = 0.001). We conclude that considerable effects of age and sex are observed on coagulation factors and naturally occurring inhibitors.

Abstract. Dengue illness has been a major health concern in Pakistan during the last decade. Dengue infection can result in a spectrum of clinically distinct outcomes, ranging from asymptomatic infection to potentially life-threatening forms of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). A single-nucleotide polymorphism in FcγRIIa (rs1801274) results in altered affinity of the receptor for different subclasses of immunoglobulin G, and is a key player in determining the susceptibility to or protection from severe clinical infection of dengue. In this study, we analyzed the allelic and genotypic distribution of rs1801274 in subjects of Pakistani origin with subclinical dengue infection (n = 40), dengue fever (DF) (n = 40), and DHF/DSS (n = 30). We found that HH homozygotes and heterozygotes were significantly more likely to develop clinical dengue (odds ratio [OR] = 3.21, 95% confidence interval [CI] = 1.29-7.97, P = 0.009), either DF (OR = 2.82, 95% CI = 1.00-7.97, P = 0.045) or DHF/DSS (OR = 3.90, 95% CI = 1.13-13.07, P = 0.024) than the asymptomatic dengue infection. Results of allelic distribution comparisons and logistic regression analysis also supported the same relationship. The results suggest complex nature of interacting factors in determining the course for severe dengue illness.

Background/aim: Fanconi anemia (FA) is an autosomal recessive disease determined by mutations in at least 16 genes, with distinct distributions in different populations. To the best of our knowledge, there are no reports regarding the molecular basis of the disease in FA patients in Pakistan. The current study aimed to determine the frequency of FANCC gene mutations, i.e. IVS4+4A>T, del322G, and R548X, in FA patients.Materials and methods: Genomic DNA was obtained from 36 FA patients. All samples were analyzed by polymerase chain reaction and restriction fragment length polymorphism techniques.Results: Mutation IVS4+4A>T was identified in 26 (72.2%) patients. It was homozygous in 6 and heterozygous in 20 patients. Del322G and R548X were found with the following prevalences: del322G, 5.6%, and R548X, 5.6%. Patients with these two mutations were compound heterozygotes having concomitant IVS4+4A>T mutation. Conclusion:These results suggest that mutation IVS4+4A>T is the most prevalent mutation in our group of patients. This analysis of Pakistani patients also suggests that there is no significant difference between IVS4+4A>T homozygotes and the rest of the patients with regard to severity of clinical phenotype.

Aim: To determine impact of platelet rich plasma therapy for the management of knee osteoarthritis. Study design: Prospective double-blinded, add –on randomized, placebo-controlled trial Place and duration of study: Department of Orthopedics, Shaikh Zayed Hospital Lahore from 1st January 2019 to 30th June 2019 Methodology: Fifty volunteer participants fulfilling inclusion criteria were enrolled. One hundred knees of patients were randomly allocated into two groups. Knees were assigned either of the two groups. Platelet rich plasma group which was assigned to receive 5ml of platelet rich plasma and normal saline (NS) group which was assigned to receive 5ml of NS labeled as control group. All patients given 3 successive intra-articular injections of 5 mL of autologous platelet rich plasma or 5ml of normal saline was given at weekly intervals. Patients were blinded and subjected to a standardized injection protocol and the intensity of pain was assessed on visual analog scale (VAS) for pain. Results: The VAS scores decreased from 50.9±14.7 at baseline assessment to 43.6±16.2 at 1-month follow-up after completion of therapy for PRP group (p<0.0203). The improvement was maintained from the end of the therapy to 3 and 6 months’ follow-up, as measure of VAS score as 30.54±11.8 and 20.2±8.6 respectively (p<0.0001). In normal saline group, there was very slight decrease in VAS score from baseline i.e. 49.8±19.5 to 48±22.7, 44±16.6, and 42±21.7 at 1, 3 and 6 months interval. Conclusion: Platelet rich plasma therapy can provide effective pain control up to 6 months post injection in knee osteoarthritis. Key words: Platelet rich plasma (PRP), Knee osteoarthritis (OA), Pain score