Chronic myeloid leukemia with rare e1a3 BCR-ABL transcript

Chronic myelogenous leukemia [CML] results from neoplastic transformation of a hematopoietic stem cell. It is cytogenetically characterized by the presence of Philadelphia chromosome which results from reciprocal translocation t[9;22] that juxtaposes the ABL gene on chromosome 9 with breakpoint cluster region [BCR] on chromosome 22 generating BCR-ABL oncogene. All BCR-ABL fusion proteins display activated tyrosine kinase activity. Their different types are associated with different clinical course and prognosis. We report a rare case of e1a3 BCR-ABL transcript. So far only 4 cases in patients with CML have been reported

Frequency of JAK2 V617F mutation in patients with Philadelphia positive Chronic Myeloid Leukemia in Pakistan

Co-existence of myeloproliferative disorders [MPD] and Janus associated kinase 2 mutation [JAK2 V617F] is a well-established fact. Only few case reports are available showing presence of JAK2 V617F mutation in chronic myeloid leukemia [CML]. Purpose of this study was to determine the frequency of JAK2 V617F mutation in Philadelphia Chromosome positive [Ph [+]] CML patients in Pakistan. The study was conducted from August 2009 to July 2010 at Civil Hospital and Baqai Institute of Hematology [BIH] Karachi. Blood samples from 25 patients with CML were collected. Multiplex reverse transcription polymerase chain reaction [RT-PCR] was performed for Breakpoint Cluster Region - Abelson [BCR-ABL] rearrangement. Conventional PCR was performed for JAK2 V617F mutation on BCR-ABL positive samples. All 25 samples showed BCR-ABL rearrangement. Out of these 11 samples [44%] had JAK2 V617F mutation; the remaining 14 [56%] cases showed JAK2 617V wild type. It is concluded that the co-existence of Ph [+]CML and JAK2 V617F mutation is possible

Single step PCR for the identification of low density lipoprotein receptor [LDL-R] gene mutations

This study was conducted to determine the common mutation of low density lipoprotein receptor in patients with familial hypercholesterolemia [FH] in our population and identify the different point mutation in the LDL-receptor gene. The main aim of this study was to reduce the cost of PCR without extracting DNA and do the diagnosis at single step. This study was carried out in the period of one year, from 2009- 2011. All the patients selected for this study were from Dr. Ziauddin Hospital, National Institute of Cardiovascular Diseases, and Dr. Rubina Ghani's Pathological and Molecular Laboratories. While collecting the blood sample, the patients were in overnight fasting condition. The clinical and biochemical analysis was performed on hyperlipidemic patients [n=120] to determine the frequency of familial hypercholesterolemia in our population. After lipid profile the patients were selected and direct multiplex PCR [Polymerase chain reaction] was performed from whole blood collected in a single tube using forward and reverse primers of exons 3, 4, 9 and 14 of without extracting DNA. Genomic DNA was extracted from blood samples as well as direct whole ETDA blood of healthy control group and hypercholesterolemia patients to detect mutations in exons 3, 4, 9, and 14 of the LDLR gene, with modification in the technique by using type-specific primers. These results for exon 4 mutation were confirmed by DNA sequencing. Screening method based on PCR by using Kappa direct PCR could be a faster and cheaper method with least contamination for screening a large number of FH patients for mutation of LDLR gene

Heterogeneity of breakpoint cluster region-abelson [BCR-ABL] rearrangement in patients with chronic myeloid leukemia

Breakpoint cluster region-Abelson [BCR-ABL] rearrangement or Philadelphia [Ph] chromosome in Chronic Myeloid Leukemia [CML] is derived from a reciprocal chromosomal translocation between ABL gene on chromosome 9 and BCR gene on chromosome 22. This chimeric protein has various sizes and therefore different clinical behaviour. The purpose of this study was to determine the heterogeneity of BCR-ABL rearrangement in patients with Ph+CML in Pakistan. The study was conducted at Civil Hospital and Baqai Institute of Hematology [BIH] Karachi. Blood samples from 25 patients with CML were collected. Multiplex reverse transcription polymerase chain reaction [RT-PCR] was performed to identify various BCR-ABL transcripts. All 25 samples showed BCR-ABL rearrangements. Out of these, 24 [96%] patients expressed p210 BCR-ABL rearrangements i.e. 60% [n=15] had b3a2 and 32% [n=8] had b2a2 rearrangements. Co-expression of b3a2 /b2a2 rearrangement and p190 [e1a3] rearrangement was also identified in two patients. It is apparent that majority of the patients had p210 BCR-ABL rearrangements. Frequency of co-expression and rare fusion transcripts was very low

Effects of oral isotretinoin subsequent to diamond dermabrasion of post acne scaring

Background: Acne vulgaris is treated by topical and or systemic antibiotics, vitamins and some keratolytics. Micro dermabrasion, a modified technique, useful for scar removal is more natural, gentler and less invasive tool for doing exfoliation

Objective: To observe the comparative effects of diamond dermabrasion on post acne/inflammatory scaring with or without oral Isotretinoin and its probable side effects

Patients and Methods: A total of 232 patients of post acne/inflammatory scaring were randomly enrolled. In this experimental study out of which 159 females were divided into two groups, Fl of 80 and F2 of 79 females. The F1 was prescribed on Isotretinoin 20 mg/day for 45 successive days after diamond dermabrasion performed by NOVA NV60. F2 received no further treatment after the procedure. The males were divided into Ml and M2, of 37 and 36 in each group respectively and were given isotretinoin 20mg/day in Ml group and no treatment in M2 group

Results: In female groups, 82.55% experienced non-inflammatory lesions, erythema in 80%, desquamation in 21.25%, mild to moderate inflammatory lesions in 17.5%, dryness in 88.75%, pruritus in 32.50% and stinging /burning in 13.75% was observed in Group F1. In group F2 these percentages were 74.68%, 77.2%, 17.72%, 25%, 79.74%, 43.03%, and 24.05% of the same parameters, respectively. F1were graded for progress, 02 females were of grade 0, 13 women obtained grade 1, 41 patients of Grade 2 and 24 females were of Grade 3. The progress grades for F2 were, Grade O had 11 females, Grade 1 had 27, Grade 2 had 38 while Grade 3 had only 03 females. In male group Ml, 78.37% experienced non-inflammatory lesions, erythema in 83.78%, 29.72% had desquamation, 21% had mild to moderate inflammatory lesions, dryness in 89.18%, pruritus in 78.37% and stinging/burning in 16.20% in Group Ml. In group M2 these percentage s were 83.33%, 94.44%, 36.11, 25%, 83.33%, 77% and 27.77% of the same parameters, respectively. Progress grades for M1 were; Grade O for 04, Grade 1 for 10, Grade 2 for 16 and Grade 3 for 07. For M2 the same grading system was followed and Grade O had 02, Grade 1 for 19, Grade 2 were 11 and Grade 3 was 04

Conclusion: The suggestion of prescription oflsotretinoin following Diamond Dermabrasion is due to the powerful epithelial generation by the vitamin A analogue. This effect provides good and early healing of the abraded skin. However further studies are suggested with larger sample size

Carotid intima-media thickness correlation with lipid profile in patients with familial hypercholesterolemia versus controls

To determine the variations in carotid intima-media thickness [CIMT] in familial hypercholesterolemia [FH] patients and its use as predictive marker for premature cardiovascular diseases. National Institute of Cardiovascular Diseases and Dr. Ziauddin Hospital, Karachi, from June 2008 to October 2009. Familial hypercholesterolemia was clinically diagnosed by premature coronary diseases, xanthomas, arcus cornealis and family history of premature coronary heart diseases. Controls were age matched normal individuals without hypercholesterolemia. Their lipid profile was tested after overnight fasting. CIMT was measured in mm using B-mode ultrasonography using linear probe. Student t-test was applied to compare mean CIMT of cases and the control. The mean CIMT values of the FH cases were correlated with LDL using Pearson's correlation test. Forty cases with hypercholesterolemia gave consent to participate in the study. These patients had total cholesterol >200 mg/dL and LDL >/= 160 mg/dL as compared to twenty controls of similar age with total cholesterol

Two novel mutations in exon 3 and 4 of low density lipoprotein [LDL] receptor gene in patients with heterozygous familial hypercholesterolemia

To determine the common mutation of low density lipoprotein receptor in hypercholesterolemja patients requiring screening for heterozygous familial hypercholesterolemia [HeFH] in Karachi. Case-series. Dr. Ziauddin Hospital Laboratory and Dr. Rubina Ghani's Pathological and Molecular Laboratories, Karachi, for the PCR bench work from June 2008 to October 2009. All the patients selected for this study were from Dr. Ziauddin Hospital and National Institute of Cardiovascular Diseases. All the patients having high total cholesterol and LDL-cholesterol were included in this study with premature coronary artery diseases or a family history of hypercholesterolemia. Exclusion criteria included Diabetes mellitus, hypertension, renal disease, hypothyroidism and steroid therapy. After lipid profile with overnight fasting, DMA was extracted from whole blood collected in EDTA [ethylenediamine tetra acetic acid] tube and multiplex PCR [polymerase chain reaction] using forward and reverse primers of exons 3, 4, 9 and 14 of base pairs 162, 431, 550 and 496 respectively. Out of total of 120 hypercholesterolemia cases, 42 patients were classical cases of HeFH [heterozygous familial hypercholesterolemia] with xanthomas, xanthelasmas and LDL-C > 160 mg/dl. The total cholesterol [260 +/- 57 mg/dL] and LDL-C [192 +/- 39 mg/dL] of cases was significantly high as compared to, controls having total cholesterol [184 +/- 27 mg/dL] and LDL-C [105 +/- 22 mg/dL], p > 0.001. Two novel point mutations were noted in exon 3 and exon 4. The other 78 cases were probable with raised LDL-C [low density lipoprotein cholesterol] and family history of premature coronary heart diseases. The frequency of HeFH was 35% classical and 65% probable cases out of total 120 hypercholesterolemia patients from two tertiary care hospitals in Karachi. The point mutation on exon 3 and exon 4 of LDLR gene was the most common. PCR is useful for the detection of large re-arrangements in the LDL-receptor gene and is a rapid and reliable method for diagnosis of familial hypercholesterolemia

Screening of five common beta thalassemia mutations in the Pakistani population: a basis for prenatal diagnosis

Thalassemia is one of the most common autosomal single-gene disorder worldwide. The highest prevalence of the disease is in the "thalassemia belt" which includes the Mediterranean region, parts of the Middle East, the Indian subcontinent, the southern parts of the Far East, Pakistan and South-East Asia. This study aimed to detect the common molecular abnormalities of the beta thalassemia syndrome in Pakistan. The study was conducted at the Institute of Hematology, Baqai Medical University, Karachi, Pakistan from August 2004 to November 2007. Blood samples of patients with beta thalassemia major [n=400] were collected from hospital transfusion centres and diagnostic laboratories in different districts of Karachi representing five major ethnic groups including Punjabi, Pathan, Sindhi, Baluchi and Urdu speaking. All the samples were analysed for five common mutations by using the polymerase chain reaction technique ARMS [amplification of refractory mutation system]. Results: The data revealed five common mutations including IVS 1-5[G-C], Fr 41/42[-CTTT], Fr 8/9 [+G], IVS 1-1 and Del 619. These accounted for 90% of the total beta thalassemia genes in Pakistan. The IVS 1-5[G-C] was found to be the most common beta thalassemia gene in the Pakistani population with a frequency of 44.4% present in all major ethnic groups. The results of this study will be helpful in the establishment of a large scale prenatal diagnosis programme in Pakistan

Clozapine induced neutrophil cytotoxicity in rats

The study was carried out to understand the pathogenesis of hematological dyscrasias and cytotoxicity following administration of both purified and commercially available form of Clozapine in an animal model. The Albino Sprague Dawley rats [n=30] with an average weight of 180g were taken and divided into three groups. Haematological parameters including haemoglobin, haematocrit, RBC and differential counts, absolute indices, Red cell Distribution Width [RDW] and morphological features of RBCs by peripheral blood smear were performed by standard laboratory methods. Additionally Serum Iron Concentration [SIC], Total Iron Binding Capacity [TIBC] [Roche Ltd.] and the serum ferritin level [Randox Ltd.] were also determined in each group. All statistical analysis was performed using graph pad prism. Clozapine induced neutrophil toxicity was manifested in both experimental groups, with condensation and subsequent breakdown of chromatin material. Our data, raised concerns about haematological safety and the potential mechanisms of neutrophil cytotoxicity related to the use of this drug