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Surveillance of genetic diversity of the SARS‐CoV‐2 is extremely important to detect the emergence of more infectious and deadly strains of the virus. In this study, we evaluated mutational events in the SARS‐CoV‐2 genomes through whole genome sequencing. The samples were collected from COVID‐19 patients in different major cities of Pakistan during the four waves of the pandemic (May 2020 to July 2021) and subjected to whole genome sequencing. Using in silico and machine learning tools, the viral mutational events were analyzed, and variants of concern and of interest were identified during each of the four waves. The overall mutation frequency (mutations per genome) increased during the course of the pandemic from 12.19 to 23.63, 31.03, and 41.22 in the first, second, third, and fourth waves, respectively. We determined that the viral strains rose to higher frequencies in local transmission. The first wave had three most common strains B.1.36, B.1.160, and B.1.255, the second wave comprised B.1.36 and B.1.247 strains, the third wave had B.1.1.7 (Alpha variant) and B.1.36 strains, and the fourth waves comprised B.1.617.2 (Delta). Intriguingly, the B.1.36 variants were found in all the waves of the infection indicating their survival fitness. Through phylogenetic analysis, the probable routes of transmission of various strains in the country were determined. Collectively, our study provided an insight into the evolution of SARS‐CoV‐2 lineages in the spatiotemporal local transmission during different waves of the pandemic, which aided the state institutions in implementing adequate preventive measures. [ABSTRACT FROM AUTHOR]

Objective: To compare the mean pain score and range of motion in chronic knee osteoarthritis patients treated with intraarticular Steroid injection versus ultrasound therapy. Study Design: Quasi-experimental study. Place and Duration of Study: Department of Rehabilitation Medicine, Combined Military Hospital, Pano Aqil Pakistan, from Oct 2020 to Apr 2021. Methodology: Patients were divided into two groups (50 patients in each group). One group received ultrasound therapy for six weeks, with three sessions per week. The second group received a single 80mg intraarticular Triamcinolone Acetonide. Patients in both the groups were monitored for six weeks to assess the pain score and range of motion. Results: A total of one hundred (100) patients with knee pain due to chronic osteoarthritis were included in the study. The mean age in the ultrasound group was 54 ± 4.8 years and in the intra-articular group, the mean age was 54 ± 4.7 years. The mean reduction in pain score and mean improvement in the range of motion were compared between two groups and statistically significant difference was observed (p-value <0.001). Conclusion: According to our findings, intra-articular Corticosteroid injection is more effective in comparison to ultrasound therapy. However, there were statistically significant reductions in pain scores and improvements in the range of motion in both groups. [ABSTRACT FROM AUTHOR]

Objective: To report the seasonal trends of Guillain-Barré Syndrome in a cohort of Pakistani population from southern Punjab, Pakistan. Study Design: A retrospective observational study. Place and Duration of Study: Department of Physical Medicine & Rehabilitation, Combined Military Hospital Bahawalpur, from Jan 2016 to Oct 2018. Methodology: One-hundred and fifty-two individuals fulfilling the electrophysiological criteria of Guillain-Barré Syndrome were included from the records of the department and subdivided into four main subtypes on the basis of electrodiagnostic studies including Acute Inflammatory Demyelinating Polyneuropathy, Acute Motor Axonal Neuropathy, Acute Motor and Sensory Axonal Neuropathy, and Miller Fisher Syndrome. The seasons were divided as: summer; May to August, autumn; September and October, winter; November to February, and spring; March and April. Demographic characteristics, type of Guillain-Barré Syndrome, and month and season of presentation were investigated. Data were analyzed with SPSS version 20. Results: The sample (mean age: 39 ± 21 years) consisted of 114 (75%) males and 38 (25%) females. The highest reporting (63, 41.4%) was in winter followed by (53, 34.9%) summer (p<0.001). May was the peak reporting (n=24) month with January to follow (n=20) (p=0.002). Acute Inflammatory Demyelinating Polyneuropathy was the most frequent sub-type 88 (57.9%). Acute Inflammatory Demyelinating Polyneuropathy and Acute Motor Axonal Neuropathy were significantly more frequent in winter season (p<0.001 and p=0.008 respectively) while Acute Motor and Sensory Axonal Neuropathy was in summer season (p=0.01). Regarding the association with a particular month, Acute Inflammatory Demyelinating Polyneuropathy was significantly more common in May (p=0.025). Acute Motor Axonal Neuropathy and Acute Motor and Sensory Axonal Neuropathy could not find any association. Statistics for Miller-Fisher Syndrome were not computable. Conclusion: Clustering of Guillain-Barré Syndrome cases was found in winter. Acute Inflammatory Demyelinating Polyneuropathy and Acute Motor Axonal Neuropathy were significantly more frequent in winter while Acute Motor and Sensory Axonal Neuropathy was in summer. [ABSTRACT FROM AUTHOR]

Objective: To assess clinical presentations and subtypes of Guillain-Barré Syndrome (GBS) in Pakistan. Study Design: Retrospective study. Place and Duration of Study: CMH Lahore, Abbotabad, Quetta & Armed Forces Institute of Rehabilitation Medicine form Jan 2007 to Feb 2015. Material and Methods: The relevant history, demographic features, clinical presentations and subtypes of GBS in 211 patients fulfilling the clinical and electrodiagnostic criteria were investigated. Results: The average age of the patients was 37.36 years (62.7% M, 37.3% F). Clinically 66.8%, 17.8% and 15.4% presented as ascending paralysis, simultaneous quadriparesis and paraparesis respectively. About 38.1% of the patients presented with cranial nerves involvement, 87.4% presented with areflexia while 59.5% reported pain. The GBS subtypes identified were acute inflammatory demyelinating polyneuropathy (21.9%), acute motor axonal neuropathy (38.9%), acute motor sensory axonal neuropathy (35.6%) and fisher syndrome (3.6%). Conclusion: The GBS presents in young, as an ascending paralysis, main subtypes of GBS in Pakistan are axonal and this highlights the importance of local management guidelines and preventive medicine. [ABSTRACT FROM AUTHOR]

Objectives: To assess factors for the non-compliance of BCG administration at appropriate time, primarily. This was also to find level of awareness of parents regarding the EPI schedule in District Jhelum; to assess timely vaccination with special emphasis on BCG; and to discuss the tangible factors for delay and their remedial measures. Methods: A cross-sectional study was conducted in District Jhelum, over a period of six months, with the target population of all parents having children > 2 years of age. A representative sample of 450 parents having children of less than two years of age was selected. Data was collected through a questionnaire by teams of two members-one male and one female. Data was entered in MS Excel and analysed using SPSS, Version 20. For qualitative analysis, interviews of the key persons of Dist. Jhelum in EPI and their suggestions and recommendations were gathered for the remedy. Results: Out of 450 children under 2 years, 164, i.e. 36.4%, were vaccinated with BCG during the first seven days after their birth. Regarding the awareness of parents about EPI 389, i.e. 86.4%, are aware of EPI. Monthly incomes greater than 20,000 Rupees, 90.8 had their child vaccinated within the first week. This shows that only 36.4% were vaccinated during the first 7 days, while the remaining was vaccinated after the first week of their life signifying the noteworthy delay in the administration of the vaccine. Regarding the qualitative part of the study, it was revealed that the factors for delayed BCG vaccination were 20-dose vile, inadequate resources including logistic, and manpower and inefficient supervising and monitoring. Conclusions: This study concludes that there is a significant delay in timely vaccination in children even with generally omniscient parents regarding the EPI schedule. Increase in the respective household incomes, educated parents- especially mothers, deliveries in hospitals, outreach vaccination, punctuality of the vaccinator and the mother's 'more than' two antenatal visits are factors that help raise BCG-vaccination rates. Thus the introduction of the single-dose vial, the provision of resources and strict monitoring can help alleviate these concerns. [ABSTRACT FROM AUTHOR]

The article focuses on a study which aimed to determine the frequency of at risk couples for beta thalassaemia minor through pre-marital screening, and to observe the acceptability of this approach among masses in a suburban area of Karachi, Pakistan. As mentioned, the study concluded that these is a need for the implementation of a premarital screening programme in Pakistan with the help of mass education and the involvement of health care providers and religious leaders.

Objective: To determine the antibody levels (IgM and IgG), using ELISA in suspected patients of COVID-19., Study Design: Descriptive cross-sectional study. Place & Duration of Study: Real Time PCR Diagnostic and Research Laboratory, Peshawar, Khyber Pakhtunkhwa, Pakistan, from May to July 2020., Methodology: A total of 94 blood specimens were collected from suspected COVID-19 patients. The antibody levels (IgG and IgM) were determined, using a COVID-19 ELISA IgG and IgM kit., Results: Out of a total 94 serum specimens, specimens were predominantly collected from males (70.2%, n=66) as compared to females (29.8%, n=28). Amongst six different age groups, the majority of the samples were found in the 31-45 years, 16-30 years, and 46-60 years groups, 42.6% (n=40), 23.4% (n=22) and 22.3% (n=21), respectively. Of the 94 suspected COVID-19 patients' serum specimens, IgG and IgM were detected in 29.8% (n=28) and 39.4% (n=37), specimens, respectively. The IgG antibodies were detected more in males (60.0%, n=18) than females (40.0%, n=12) samples. Similarly, IgM antibodies were also found more frequently in males (61.1%, n=22) as compared to females (38.9%, n=14)., Conclusion: Detection of antibodies in COVID-19 infected patients provides vital clinical information for clinicians and could be used for the identification of suspected cases. Moreover, males were more prone to disease compared to females, and the 31-45 years age group was also more affected. Key Words: Serological assays, IgG, IgM, Peshawar, Pakistan.

Unlabelled: Prostaglandins produced by Cyclooxygenase-2 enzyme have been implicated to have a role in breast carcinogenesis. Several single nucleotide polymorphisms (SNPs) linked to COX-2 enzyme are reported to modulate its expression. The aim of the present study was to examine association of these SNPs to breast cancer risk in Pakistani patients., Methods: In this case-control study, three sequence variants rs689465, rs689466, rs20417 in the promoter region of COX-2 were screened to evaluate the association with breast cancer risk. A total of 150 breast cancer patients and 101 healthy control genomic DNA were genotyped for rs689456, rs689466, rs20417 and their genotypes distribution in cases and control were compared using Pearson chi square test. Risk association was analyzed through odd ratio calculated by logistic regression., Results: A screening analysis of COX-2 SNPs in 101 healthy controls showed distribution of Minor allelic frequency distribution of SNPs as follows : rs689465 (0.12), rs689466 (0.15), rs20417 (0.23). Further analyses revealed that their observed genotype frequencies were consistent with Hardy Weinberg equilibrium and strong linkage disequilibrium was identified between rs20417, rs689465 and rs689466. The Combined allele variants analysis showed that Haplotype rs68965G- 689466A-20417C (OR 2.909; CI 95 %1.3776.327; P = 0.007) was significantly associated with breast cancer., Conclusions: Our results indicate no strong association between three most frequent COX-2 SNPs rs689465 rs689466, rs20417 studied with breast cancer risk in the single locus analysis. However, our data suggested that combined COX-2 SNP haplotype have a role in breast cancer associated risk in Pakistani patients.

HER2 gene amplification in invasive breast cancer is a robust predictive marker for response to transtuzumab therapy. This study was undertaken to measure concordance between immunohistochemistry (IHC) and FISH for HER2 gene amplification in invasive breast tumors, as well as the presence of polysomy 17 and possible correlation with demographics and histopathological variables, including ER and PR positivity. A total of 425 cases of infiltrating carcinoma of breast (99% IDC-NOS) were studied. HER2 over expression was tested by IHC and FISH methods. Association between IHC and FISH in both subsets was calculated by amplification ratio including polysomy 17. Out of 425 specimens, 128 (30%) were positive for HER2 amplification by FISH test, whereas only 78 (24%) tumors with 2+ expression showed amplification. In contrast, 39 (74%) demonstrated 3+ IHC score and HER2 gene amplification. The histological variables including tumor size, tumor type, and lymph node involvement did not influence the outcome of FISH analysis. The ER and PR status showed significantly greater positivity in patients negative for HER2 amplification. Polysomy 17 was detected in 23.7% patients and was positively associated with ER and PR expression (P= <0.05). Our study showed a concordance of 24% between 2+ IHC and FISH amplification, while in 3+ IHC cases the concordance was 74%. Significant links of HER2 amplification was seen with ER andPR negativity and higher tumor grade. In addition, non-significant correlations were noted with other variables like tumor type, size and lymph node status.