Yasar Mehmood Yousafzai

Background: Thalidomide induces fetal hemoglobin and renders most thalassemia patients transfusion-independent. Some patients, however, do not respond. Underlying genetic variations responsible for variable responses to thalidomide are unexplored. Aims and objectives: To discover genetic variations that influence response to thalidomide in transfusion-dependent beta-thalassemia patients. Methods: Twenty beta-thalassemia patients (14 excellent responders and six non-responders) who had received thalidomide were included in the study by a non-probability purposive sampling technique. Patients who showed a rise of >2 mg/dl in hemoglobin level and/or whose hemoglobin levels reached 9 gm/dl without blood transfusions were designated as excellent responders. Patients whose hemoglobin levels did not show an increment rise of >2 and/or whose hemoglobin levels did not rise above 5.9 gm/dl and needed blood transfusions to maintain optimal hemoglobin levels were designated as non-responders. DNA was extracted, and whole-exome sequencing was performed on an Illumina HiSeq System. Aligning and variant calling were done by the Sentieon software. Annotation was done by Annovar. Results: The age of study participants ranged from 1–12 years, with a mean of 5.45 ± 3.81 years. There were 17 (85%) males and three (15%) females. A total of 222,180 germline variants were identified across 20 subjects, from which 24 candidate variants across 24 genes were identified. The three most common polymorphisms in the excellent responder group were found in the exon region of CHI3L1 (rs880633), NPNT (rs35132891), and ZNF 208 (rs10425763), which were found in 92%, 85%, and 71% cases, respectively. The commonest polymorphisms in the non-responder group were found in the PM20D1 gene (rs7518979), LGR6 (rs75658797), MYH15 (rs4299484), and RESF1 (rs3207618), each of which was found in 66.6% cases. Conclusion: This study shows a significant association of single-nucleotide polymorphisms rs880633, rs35132891, and rs10425763 with excellent response status, while rs7518979, rs75658797, rs4299484, and rs3207618 are associated with non-response status.

Objective: To assess empathy scores among postgraduate students enrolled in the M. Phil programs of various Pathology disciplines at Khyber Medical University (KMU) Peshawar, Pakistan. Methods: This cross-sectional study was conducted at Institute of Pathology and Diagnostic Medicine, KMU, Peshawar, Pakistan, between March and October 2023. A total of 48 students from M. Phil Histopathology, Hematology, Oral Pathology, and Microbiology were enrolled through consecutive sampling. Participants completed the Jefferson Scale of Empathy – Health Professions Student (JSE-HPS) questionnaire. Demographic and clinical variables were recorded, and data were analyzed using SPSS v.26. Results: Out of 48 students, 43 completed the questionnaire (response rate: 89.5%). Of these, 15 (34.9%) were males and 28 (65.1%) females; 51.2% held MBBS/BDS degrees and 48.8% were from Allied Health Sciences. The mean JSE score was 92.9 ± 9.2, with a median of 93.0. Subscale means for perspective-taking, compassionate care, and walking in patients’ shoes were 54.1±6.4, 35.5±6.3, and 7.4±2.5, respectively. Empathy scores showed no statistically significant association with gender, age group, academic qualification, M. Phil specialization, prior clinical experience, or level of seniority. Median scores reflected a similar pattern, with slightly higher empathy noted among males, Allied Health graduates, and those without prior clinical exposure, though none reached statistical significance. Conclusion: Empathy scores among M. Phil students in pathology disciplines were modest and showed no significant association with demographic or educational variables. Compared to international literature, the overall empathy levels appear lower, highlighting the need for enhanced focus on empathy development in postgraduate pathology education

Background: β-thalassemia is a significant public health problem in Pakistan with higher carrier rate and disease frequencies. Its mutation prevalence is highly geographical and ethnicity dependent. Objective: To investigate the prevalence of genetic mutations and demographics in transfusion-dependent β-thalassemia (TDT) patients in Pakistani communities. Methodology: Samples were gathered from TDT patients who had been diagnosed and met the inclusion criteria. The SPPS 27 was applied to analyze comprehensive patient demographics, clinical history, and prognostic data. For detection of genetic mutations in TDT patients’ DNA extraction and polymerase chain reaction (PCR) were applied using the relevant screening protocols. Results: A total of 384 patients of TDT were screened from almost 79 various ethnic castes of Pakistan. The Cd 8-9 (+G) mutation was discovered to have the highest number of mutations followed by IVS 1-5. In demographics, Dera Ghazi Khan led in the disease prevalence, followed by Peshawar, Malakand, and Kohat divisions. The areas that have a remarkably high Excellent and Good (Ex+G) responders were Rawalpindi, Multan, Faisalabad, Gujrat, Sukkur, Shaheed Benazirabad, and Merged Districts (Fata). The areas that have comparatively poor Ex+G responders (≤50%) were Sargodha, Sahiwal, Loralai, Pasheen, and Gilgit-Baltistan. Conclusion: Our findings determined the prevalence of genetic mutations and demographics of TDT patients in the diverse community of Pakistan with significant response to low-dose thalidomide in most of the Pakistani communities. For precision medicine and further ensuring the safety and efficacy of low-dose thalidomide in TDT patients, multicenter and large scale clinical trials with Bayesian approach are recommended.

Background: β-thalassemia is an autosomal recessive hereditary hemoglobin production disorder characterized by abnormal hemoglobin synthesis. The efficient and safe therapies for transfusion-dependent β-thalassemia (TDT) patients are desperately required to minimize the requirement for blood transfusion. Objective: To establish the safety and efficacy of low-dose thalidomide in TDT patients. Methodology: The retrospective cohort study was conducted in Khyber Medical University, Peshawar and Blood Diseases Clinic, Peshawar to investigate the safety and efficacy of low-dose thalidomide therapy in TDT patients. Samples were collected from the diagnosed cases of TDT patients whose ages were ≥3 years and were on thalidomide treatment for a period of ≥6 months. Complete blood count CBC was performed on Sysmex XP-100, Japan and biochemical tests were performed on Cobas 6000 analyzer series. Comprehensive patients’ demographics, clinical history and prognosis data were recorded and analyzed using the SPPS 27. Results: The patient characteristics, biochemical parameters and hematological response were assessed and statistically tested for 384 TDT patients using low-dose thalidomide therapy. Applying the criteria for the Hb level achievement, we marked the significant results (<0.001) for Excellent, Good, and Partial responders that were 184 (47.9%), 96 (25%), and 60 (15.6%), respectively. The common side effects observed with the drug included abdominal discomfort, nausea, vomiting, headache, constipation, dizziness, fatigue, anxiety, repeated infections, and skin rash, that were controlled with symptomatic treatment and/or dose adjustment. Conclusion: Collectively, thalidomide is safe and effective in increasing Hb levels thus reducing the requirements of blood transfusion in TDT patients. Future clinical trials are suggested to strengthen the efficacy and safety of low-dose thalidomide and to establish clinical guidelines for its rational use in TDT patients.

Myeloproliferative neoplasms (MPNs) are blood cell disorders, characterized by overproduction of abnormal cells in bone marrow due to stem cell mutation. The proliferations of blood cell are controlled by many genes particularly MPL gene which encodes thrombopoietin receptor, a hematopoietic growth factor involved in the production and regulation of the platelets and multipotent hematopoietic progenitor cells. Acquired mutations including (W515L and W515K) in this gene have been observed in patients with primary myelofibrosis or essential thrombocythemia lacking JAK2 (V617F) mutations. MPL mutation detection is important for MPNs diagnosis, but due to low frequency of mutant allele burden (< 15%) may be missed by already available common assays such as Sanger sequencing. Furthermore, these techniques are costly, time-consuming, and less sensitive. In present study, we aimed to develop sensitive, less time-consuming, and cost-effective real-time PCR assay for the detection of MPL mutations that is based on TaqMan fluorescent probes. DNA was extracted from blood sample of 128 MPNs patients collected and further analysis was performed on TaqMan RT-PCR. Reference curve was obtained for amplified product of MPL gene containing mutated sequence. The predicted sensitivity level was at least 5% mutant allele burden by our developed assay that is much higher than sequencing output. Out of 128, 2 (1.56%) patients harbored W515L mutation and 1 (0.78%) harbored W515K mutation. It was concluded that TaqMan qRT-PCR assay is an efficient, sensitive, cost-effective, and less time-consuming method capable of detecting MPL mutation in MPNs patients. We suggested that this assay might be helpful in investigating mutant allele load in MPNs patients.