Muhammad Asif

Effect of iron overload on renal functions and oxidative stress in beta thalassemia patients

Objectives: To check the amount of cellular damage caused by serial transfusions of blood in thalassemia patients. Methods: A cross-sectional study was conducted in the University of Lahore, Lahore, Pakistan between August 2012 and December 2012. A total of 43 thalassemia patients underwent at least 10 blood transfusions. Comprehensive biochemical analysis of blood was performed to record the levels of creatinine, urea, uric acid, albumin, liver function tests, malondialdehyde (MDA), and ferritin. Results: Serum creatinine (0.732±0.23mg/dl) and uric acid (6.7±0.94mg/dl, p<0.05) were significantly higher in patient groups as compared with the control. Ferritin levels were significantly higher in patients as compared with the control (3103.9±1747.4, p<0.05). Hemoglobin levels were observed in controls 14±1.3g/dl and in patients 7.1±1.03g/dl. No clear relationship exists between age and hematological parameters of thalassemic patients. Serum ferritin level is positively related with serum alanine transaminase, aspartate aminotransferase, and alkaline phosphatase and MDA (p<0.05). Conclusion: Serum MDA and serum ferritin of patients (r=0.593, p<0.05) reflects that both are crucial parameters estimating the cellular damage in patients suffering from thalassemia.

Presence of novel compound BCR-ABL mutations in late chronic and advanced phase imatinib sensitive CML patients indicates their possible role in CML progression

ABSTRACT BCR-ABL kinase domain (KD) mutations are well known for causing resistance against tyrosine kinase inhibitors (TKIs) and disease progression in chronic myeloid leukemia (CML). In recent years, compound BCR-ABL mutations have emerged as a new threat to CML patients by causing higher degrees of resistance involving multiple TKIs, including ponatinib. However, there are limited reports about association of compound BCR-ABL mutations with disease progression in imatinib (IM) sensitive CML patients. Therefore, we investigated presence of ABL-KD mutations in chronic phase (n = 41), late chronic phase (n = 33) and accelerated phase (n = 16) imatinib responders. Direct sequencing analysis was used for this purpose. Eleven patients (12.22%) in late-CP CML were detected having total 24 types of point mutations, out of which 8 (72.72%) harbored compound mutated sites. SH2 contact site mutations were dominant in our study cohort, with E355G (3.33%) being the most prevalent. Five patients (45%) all having compound mutated sites, progressed to advanced phases of disease during follow up studies. Two novel silent mutations G208G and E292E/E were detected in combination with other mutants, indicating limited tolerance for BCR-ABL1 kinase domain for missense mutations. However, no patient in early CP of disease manifested mutated ABL-KD. Occurrence of mutations was found associated with elevated platelet count (p = 0.037) and patients of male sex (p = 0.049). The median overall survival and event free survival of CML patients (n = 90) was 6.98 and 5.8 y respectively. The compound missense mutations in BCR-ABL kinase domain responsible to elicit disease progression, drug resistance or disease relapse in CML, can be present in yet Imatinib sensitive patients. Disease progression observed here, emphasizes the need of ABL-KD mutation screening in late chronic phase CML patients for improved clinical management of disease.

A Novel Four-Way Complex Variant Translocation Involving Chromosome 46,XY,t(4;9;19;22)(q25:q34;p13.3;q11.2) in a Chronic Myeloid Leukemia Patient

Philadelphia (Ph) chromosome (9;22)(q34;q11) is well established in more than 90% of chronic myeloid leukemia (CML) patients, and the remaining 5–8% of CML patients show variant and complex translocations, with the involvement of third, fourth, or fifth chromosome other than 9;22. However, in very rare cases, the fourth chromosome is involved. Here, we found a novel case of four-way Ph+ chromosome translocation involving 46,XY,t(4;9;19;22)(q25:q34;p13.3;q11.2) with CML in the chronic phase. Complete blood cell count of the CML patient was carried out to obtain total leukocytes count, hemoglobin, and platelets. Fluorescence in situ hybridization technique was used for the identification of BCR–ABL fusion gene, and cytogenetic test for the confirmation of Ph (9;22)(q34;q11) and the mechanism of variant translocation in the bone marrow. The patient is successfully treated with a dose of 400u2009mg/day imatinib mesylate (Gleevec). We observed a significant decrease in white blood cell count of 11.7u2009×u2009109/L after 48-month follow-up. Patient started feeling better generally. There was a reduction in the swelling of the body, fatigue, and anxiety.

A novel mutation in RDH5 gene causes retinitis pigmentosa in consanguineous Pakistani family

Retinitis pigmentosa (RP) is the most frequent genetically and clinically heterogeneous inherited retinal degeneration. To date, more than 80 genes have been identified that cause autosomal dominant, autosomal recessive and X linked RP. However, locus and allelic heterogeneity of RP has not been fully captured yet. This heterogeneity and lack of an accurate genotype phenotype correlation makes molecular dissection of the disease more difficult. The present study was designed to characterize the underlying pathogenic variants of RP in Pakistan. For this purpose, a large consanguineous family with RP phenotype showing autosomal recessive mode of inheritance was selected after a complete ophthalmological examination. Next generation sequencing was used for the identification of molecular determinant followed by Sanger-sequencing for confirmation. After sequence analysis a novel homozygous missense mutation, (c.602 Cu2009>u2009T) in exon 4 of the RDH5 gene (MIM: 601617) was identified. This mutation resulted in substitution of phenyl alanine for serine at amino acid 201 (p.Ser201Phe) of the RDH5 gene. The same mutation was not detected in the 200 ethnically-matched control samples by Sanger sequencing. The identified mutant allele segregated in homozygous fashion in all the affected individuals of pedigree. Identification of this mutation reveals the allelic heterogeneity of RDH5 in patients with RP phenotype. The findings of this study demonstrate the clinical significance of next generation sequencing to understand the molecular basis of diseases and would help to reveal new proteins and their function in visual cycle will pave the way for early diagnosis, genetic counseling and better therapeutic inventions.

Serologic autoantibodies against tumor associated antigens as potential biomarkers in patients suffering from Hepatocellular carcinoma and Cholangiocarcinoma

Background: Hepatocellular carcinoma (HCC) and Cholangiocarcinoma (CC) are primary liver cancers. HCC develops in hepatocytes while CC originates from bile duct epithelium. Due to the lack of early diagnosis, the incidence of HCC and CC are ever increasing and providing effective therapy to the patients suffering from these cancers have increasingly become difficult. The significance of clinical diagnosis of HCC by the detection of tumor markers has not been well documented. So far, none of the studies that indicate the presence of specific autoantibodies in CC, on the contrary to HCC, has been reported. The aim of our current study is to identify new immunological tumor markers for the diagnosis of HCC and CC.

Assessment of Circulating Biochemical Markers in Mice Receiving Cinnamon and Glycyrrhizin Under Carbon Tetrachloride Induced Hepatic Injury

The study evaluated the hepatoprotective activity of plant extracts of cinnamon and glycyrrhizin in distinct dosage ways to minimize the oxidative stress induced by carbon tetrachloride (CCl4) in BALB/cJ inbred albino mice. Fifteen albino mice were divided into five groups, each group containing three mice. Group A was referred as positive control while group B, C, D and E were injected intraperitoneally with 1xa0mL/kg body weight of CCl4 twice a week for 1xa0month. Group C and D were treated orally with isolated extracts of cinnamon @50xa0mg/kg and glycyrrhizin @50xa0mg/kg respectively on daily basis for 1xa0month. However, group E was treated orally with combination dose of cinnamon @50xa0mg/kgxa0+xa0glycyrrhizin @50xa0mg/kg body weight. The increase in the levels of alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP), triglyceride (TG), malondialdehyde (MDA) and glucose were recorded in CCl4 induced liver injury in mice while there is decrease in the levels of total protein (TP), reduced glutathione (GSH), Superoxide dismutase (SOD) and catalase (CAT) in CCL4 intoxicated mice. Isolated therapy of plant extracts of cinnamon and glycyrrhizin decreased the levels of ALT, AST, ALP, MDA, TG and glucose whereas increase in TP, GSH, SOD and CAT was observed in plant extracts treated mice. The best restoration of all the above said parameters near to control was observed in group of mice treated with combination dose of cinnamon and glycyrrhizin @50xa0mg/kg. Therefore, the present study declared the antioxidative, anti-inflammatory and hepatoprotective activity of standardized extracts of cinnamon and glycyrrhizin and their potent defensive property.

A rare case of three-way complex variant translocation in chronic myeloid leukemia t(6;9;22)(p21;q34;q11): A case report

Philadelphia (Ph)-positive chromosome or Ph translocation has been recognized in 90-95 chronic myeloid leukemia (CML) cases worldwide. However, only 5-8% CML patients show complex variant translocations. In the present study, hematological tests for a 47-year-old female CML patient were performed to determine the hemoglobin, platelets and total leukocyte values. A FISH test was carried out to recognize the BCR/ABL gene fusion, and a cytogenetic analysis was performed. The hematological results showed an increase in WBC (414000/mm3) and a decrease in hemoglobin (8.9 mg/dl), indicating the anemic condition in the CML patient. Furthermore, cytogenetic karyotyping results showed 46,XX,t(6;9;22)(p21;q34;q11) and positive for Ph chromosome. In conclusion, in the present study, we report a rare three-way complex variant translocation in a CML patient.

Pattern of gynaecological malignancies in south western region of Pakistan: An overview of 12 years

Gynaecological malignancies contribute significantly to cancer burden and have a higher rate of mortality and morbidity. The aim of this retrospective study was to determine the pattern of gynaecological malignancies identified between January, 2000 and December, 2011, at the Centre for Nuclear Medicine and Radiotherapy (CENAR). At CENAR 5,072 female patients were registered with different malignancies, of which 632 cases were gynaecological malignancies. Ovarian cancer (47%) was the most common gynaecological malignancy, followed by cervical cancer (29%), uterine cancer (14%), vulvar and vaginal cancer (6%), and gestational trophoblastic neoplasm (4%). Of the ovarian cancer cases, 72.5% had epithelial while 26.5% had non-epithelial cancer. Squamous cell carcinoma was 75.9% in cervix and 87.8% in vulva and vagina while endometrial carcinoma (75.9%) was more frequent in uterus. For gestational trophoblastic neoplasm, 69.2% of patients had choriocarcinoma. Ovarian cancer was the most common type for the age range of 50-59 years. In the case of cervical and gestational trophoblastic neoplasm the majority of patients presented at the ages of 40-49 and 30-39 years while uterus, vulvar and vaginal tumor presented in the elderly (>60 years). Thus, ovarian cancer is the leading gynecological malignancy in Pakistan.

Ethnomedicinal Cichorium intybus Seed Extracts: An Impending Preparation against Multidrug Resistant Bacterial Pathogens

Background The present study was undertaken to analyze the phytochemical content and biological activity of Cichorium intybus seeds traditionally used in Charsadda, Pakistan against multidrug resistant (MDR) bacterial pathogens. Objectives This study explored the qualitative and quantitative antibacterial potential of C. intybus. Further qualitative analysis of phytochemical content was performed. Methods Cichorium intybus seed extracts were prepared in aqueous, chloroform, ethanol, and hexane separately. Results All the extracts of C. intybus seeds were screened for antibacterial activity and phytochemical content. Cichorium intybus seed extract showed considerable activity against MDR pathogenic bacteria. In the well diffusion method, aqueous extracts showed a higher zone of inhibition against Pseudomonas aeruginosa (16 mm ± 0.7 mm) and Acinetobacter baumannii (13 mm ± 0.5 mm), whereas chloroform, ethanol, and hexane extracts showed activity against P. aeruginosa (11 mm ± 0.3 mm, 12 mm ± 0.5 mm, and 11 mm ± 0 mm, respectively) as compared to Imipenem, a broad spectrum antibiotic. Minimum inhibitory concentration and minimum bactericidal concentration values for aqueous and ethanol extracts indicate that they were more effective against MDR bacteria. Phytochemical analysis revealed that aqueous and ethanol extracts were rich in alkaloids, carbohydrates, gallotannins, and triterpenoids, whereas chloroform and hexane extracts were more concentrated with phenolics, pseudotannins, saponins, and tannins. Cichorium intybus seed extract demonstrated potential activity against MDR human pathogenic bacteria. Conclusions The undertaken study has for the first time reported the effects of C. intybus seed extracts against MDR bacterial pathogens. Findings of the current study will be helpful for further elucidation of bioactive molecules for therapeutic use against MDR bacterial pathogens.

Influence of One Week Education Program on the Knowledge and Approach of Pharmacy Students towards Diabetes mellitus

The objective of this study was to assess the improvement in knowledge and attitude of Pharmacy student towards diabetes by implementing one week long education program. Study design: Study was conducted between April 25, 2011 to April 29, 2011 which engaged the 3rd professional year Pharm. D. students comprising two groups: experimental and control groups. Lectures and visual presentations on diabetic education and practice were conducted for five days. Three surveys were performed on two groups i.e. non intervened control, non- intervened experimental and intervened experimental surveys. Questionnaire was used as survey tool comprising of 46 questions under ten categories. Results: Scores in all diabetic knowledge aspect was increased after education intervention and was significantly (P<0.05) different from that of control group. Conclusion: The marked increase in knowledge about diabetes among pharmacy students illustrates the importance of educational intervention. Educational programs may markedly increase the educational skills, efficiency and confidence of pharmacy students as well as professionals.