Muhammad Ayyub

Congenital afibrinogenemia: report of three cases

One female and two male infants, aged 2, 3 and 5 months, respectively, presented with uncontrolled bleeding from the umbilical stump and hematoma formation at intramuscular (vitamin K) injection sites. They were all products of firstcousin marriages and had been born at full term by normal, vaginal delivery. The number of siblings varied from one to three. There was no family history of bleeding tendency in any of the families. On examination, no abnormality was detected except for oozing from the umbilical stump and hematomas at injection sites. Blood counts and liver function tests were normal in all infants. However, prothrombin time (PT), partial thromboplastin time with kaolin (PTTK) and thrombin time (TT) were infinitely prolonged in all three patients but were normal in their parents. Fibrin degradation products were not detected in

Frequency of Gγ-globin promoter −158 (C>T) XmnI polymorphism in patients with homozygous/compound heterozygous beta thalassaemia

BACKGROUND Response to hydroxyurea therapy in homozygous or compound heterozygous beta thalassaemia (BT) has been reported as more favourable in the presence of XmnI polymorphism. The prevalence of XmnI polymorphism may vary with BT phenotypes and genotypes, and differs geographically in distribution. Prevalence of XmnI polymorphism is not known in northern Pakistan. OBJECTIVE To determine the frequency of Gγ-globin promoter -158 (C>T) XmnI polymorphism (XmnI polymorphism) in patients with homozygous or compound heterozygous beta thalassaemia. MATERIALS Polymerase chain reaction (PCR) for common beta thalassaemia mutations and Gγ-globin promoter -158 (C>T) XmnI polymorphism was performed on 107 blood samples of transfusion dependent beta thalassaemia (BT) patients in Pakistan. One hundred samples of unrelated BT traits and 94 samples of healthy subjects as controls were also analysed for BT mutations and XmnI polymorphism. RESULTS Out of 301 DNA samples, XmnI polymorphism was detected in 71(24%); in normal controls, XmnI polymorphism was detected in 34/94 (36%) subjects; while in homozygous/compound heterozygous BT, it was detected in 14/107(13%) patients (Fishers exact test, p=.0002). In heterozygous BT group, XmnI polymorphism was detected in 23/100 subjects (Fishers exact test, p=.03 with normal controls, and p=.049 with homozygous/compound heterozygous BT). The most common BT genotype was Frame Shift (Fr) 8-9/Fr 8-9, and none of the patients with this genotype had XmnI polymorphism. The second most common genotype was IVSI-5/IVSI-5; 4/26 (15%). Cases with this genotype had XmnI polymorphism. CONCLUSION XmnI polymorphism in homozygous/compound heterozygous BT group is 13%. The most common genotype associated with XmnI polymorphism was IVSI-5/IVSI-5.

Outcome of combination antiviral therapy in chronic hepatitis C virus infection during therapy of acute lymphoblastic leukemia.

Chronic hepatitis C virus (HCV) infection is not uncommon in patients with acute leukemia due to frequent blood transfusions. The treatment of HCV in patients with acute leukemia can produce profound immune dysfunction with the risk of severe cytopenia. We report the case of a young man who was treated with combined therapy of peginterferon α 2a and ribavirin for HCV while he was on maintenance anti-leukemic treatment. The patient required reduction in the dose of peginterferon α 2a and the addition of filgrastim due to neutropenia. Therapy for HCV was continued for 72 weeks and at the end of therapy, the patient had undetectable HCV RNA. The patient maintained a sustained viral response two years after the end of therapy and developed complete remission of leukemia, whereupon his anti-leukemic therapy was also discontinued. We recommend conducting further large prospective studies in HCV patients treated for leukemia to determine the safety and efficacy of antiviral therapy in this group of patients.

Spectrum of Superficial and Deep Fungal Isolates in Northern Pakistan

Fungi are an important cause of superficial and deep infections in our population. Lack of expertise in proper identification and inadequate diagnostic facilities often lead to underreporting of these infections and thus underestimation of true disease burden. This study was conducted at Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi, Pakistan, from January 2011 through December 2013. Samples included specimen collected from superficial and deep tissues, respiratory tract specimen, blood, bone marrow and other body fluids. Skin (35.1%) and nail (10.2%) samples were the most common specimens from superficial body sites. Tissue specimens from various body organs and bronchoalveolar lavage fluid were the predominant specimens received for investigation of deep seated fungal infections, contributing 34.9% and 5.9% of the total specimens respectively. Yeasts were isolated from 75(22.6%) samples; different species of Candida accounted for majority of the isolates. Growth of molds was detected in 257(77.4%) samples with Aspergillus spp. accounting for 149 (44.9%) of the isolates. Among dermatophytes, Trichophyton interdigitale 13(3.9%) was the most common isolate. Moulds other than dermatophytes were also isolated from skin, hair and nail samples and Alternaria alternata (4.8%) was the most common non-dermatophyte isolated from these sites. Fungal infections and their spectrum varies considerably in different geographical locations and in all cases not responding to antibiotics and high risk groups, a possibility of fungal cause should be sought.

Prevalence of Prothrombin Gene Mutation (G-A 20210 A) in General Population: A Pilot Study

The objective of this study was to determine the prevalence of prothrombin gene mutation in a sample population from Pakistan. Two hundred apparently healthy unrelated adults (older than 18 years) were included in the study. The sample population comprised 100 Punjabis (male 50, female 50) and 100 Pathans (male 50, female 50). Patients with a history of previous thromboembolism were excluded from the study. Five milliliters (5 mL) of whole blood was drawn in an EDTA bottle. The DNA was extracted by the standard phenol-chloroform method. The DNA was amplified between exon number 14 and the 3’-untranslated region of the prothrombin gene by a polymerase chain reaction in a thermal cycler. Amplified products were digested overnight with HindIII at 37°C. The digested products were electrophoresed on 6% polyacrylamide gel. The fragments were visualized by silver nitrate staining. A heterozygous wild type and an uncut amplified product were included in the electrophoresis strip for quality control. The wild type of DNA ran as a 350-bp fragment and internal control was cut as 550- and 150-bp fragments. The abnormal prothrombin gene was cut into 350-, 322-, and 28-bp fragments. Only two cases of heterozygous prothrombin gene mutation G-A 20210A were found in the sample studied, giving an overall carrier rate of 01% (95% CI 0.4-2.4%) in the target population. Prothrombin gene mutation is present in our population but at a lower frequency than in the white population.

Frequency of Three Different Gene Mutations (TEL-AML1, E2A-PBX1 and MLL-AF4) In Acute Lymphoblastic Leukaemia

Background and Objectives: Acute lymphoblastic leukemia (ALL) is very common in Pakistan. It is a complex genetic disease involving many fusion oncogene (FO) having prognostic significance. Its incidence may not be uniform in different parts of world because of racial, ethnic and environmental variations. These mutations have important implications for prognosis, drug selection and treatment outcome. It shows that studies on fusion oncogenes become very important for risk stratification and planning of treatment at the start of diagnosis. Method: We studied fusion oncogene in 120 pediatric ALL patients from less than one to twenty years of age using RT-PCR and their association with age and gender. It was a cross sectional study carried out in Haematology department, Armed Forces Institute of Pathology, Rawalpindi, Pakistan over a period of one year from 1 Jan 2015 to Dec 2015. Three most common fusion genes i.eE2A-PBX1 t(1; 19), TEL-AML1 t(12; 21), MLL-AF4 t( 4; 11) were studied with relevance to their age and gender. Results: Frequency of MLL-AF4 mutation was found in 5(4.2%) patients. TEL-AML1 was found in 13(10.8%) patients. E2A-PBX1 mutation was found in 12(10%) patients. Conclusions: These mutations are quite frequent in our ALL patients with different age specificities. Frequencies of some of the oncogenes were different from those reported from other areas of Pakistan. These mutations are helpful in risk stratification and have therapeutic and prognostic significance.