Serum Vitamin D status and VDR (FokI) polymorphism association study in Pakistani females with breast cancer.

Background: Breast cancer is most common in Pakistani women at young age compared to West where it is most common after 60 years of age. Variations in genes controlling vitamin D activity would play a role in determining the risk of breast cancer in women at early age. Purpose: To determine the association of vitamin D receptors (VDR) gene polymorphisms (FokI) with breast cancer risk in Pakistani women. Methods: FokI polymorphisms were studied through the polymerase chain reaction-restriction fragment length polymorphism technique on blood samples of 300 breast cancer and 300 healthy women. Results: This study found that circulating level of 25(OH)D3 was significantly lower among breast cancer patients as well as healthy subjects. Patients with large tumor size had significantly lower vitamin D levels. VDR FokI genotypes were significantly distributed (P ≤ 0.00001) in Pakistani women with newly diagnosed breast cancer. A significant association between different FokI genotypes and circulating levels of 25(OH)D3 was found. Patients with FF genotype was significantly (P < 0.0001) associated with higher risk of breast cancer (OR 8.9, 95% CI 0.17-0.45) compared to Ff and ff genotype. Conclusion: VDR gene FokI polymorphism was associated with plasma vitamin D level and significant differences found in mean serum vitamin D levels between genotype groups of FokI. The study concluded that FokI might be one of the contributors to increase relative risk of breast cancer in Pakistani women.

Phenotypic and Molecular Characterization of CTX-M Type B-Lactamases in Gram Negative Bacterial Strains Isolated from Hospitals, Lahore, Pakistan.

One of the principal mechanisms that contribute resistance to antibiotics is the production of extended spectrum beta lactamase (ESBL) in Gram negative bacteria. In the present study, molecular methods were used to evaluate the prevalence of the extended-spectrum beta-lactamase (ESBL)-encoding CTX-M gene among Gram negative bacterial strains. In total, 148 clinical samples were collected from different tertiary care hospitals of Lahore, Pakistan. Disc synergy diffusion method was used to detect the presence of ESBL production. Moreover, antibiotic resistance patterns and molecular detection of bla CTX-M ESBLs, were also studied. The pathogens isolated from the 148 samples included Escherichia coli (43%) followed by Klebsiella sp. (28%), Proteus sp. (18%) and Pseudomonas sp. (11%). In all 148 strains, 95 (64%) were ESBL producers while 53 (36%) were non ESBL producers. The strains which were phenotypically ESBL producers, bla CTX-M were found in 46% E. coli strains, while 50% Klebsiella sp. were harboring the gene. A high resistance rate was observed against cephalosporins (cefopodoxime 67%, cefoperazone 73%, cephalexin 63% sparaxin 61%). Lower resistance was observed against meropenem among all isolated bacterial strains. Genotypic detection of bla CTX-M genes by PCR revealed 46% of E. coli and 50% of Klebsiella strains harbored bla CTX-M gene. The present study showed that ESBLs producers were resistant to commonly used antibiotics. Similarly, bla CTX-M ESBL production is more prevalent in our clinical isolates.

Molecular characterization of FLT3 mutations in acute leukemia patients in Pakistan.

Fms-like tyrosine kinase 3 (FLT3) performs a vital role in the pathogenesis of hematopoietic malignancies. Therefore in recent times, the focus of several studies was on use of FLT3 as a prognostic marker. The present study investigated the molecular characterization and incidence of FLT3 mutations in acute leukemia patients in Pakistan. A total of 55 patients were studied, of which 25 were suffering from acute lymphoblastic leukemia (ALL) and 30 were suffering from acute myeloid leukemia (AML). The polymerase chain reaction demonstrated FLT3/ ITD mutations in 1 (4%) of 25 ALL patients, a male with the L2 subtype. In AML cases the rate was 4 (13.3%) of 30, three males and one female. The AML-M4 subtype was found in three and the AML M2 subtype in the other. In the AML cases, a statistically significant (p=0.009) relationship was found between WBC (109/L) and FLT3/ ITD positivity. However, no significant relationship was found with other clinical parameters (p>0.05). In acute myeloid leukemia (AML) FLT3/ITD+ mutation was more prevalent in elderly patients 31-40 age groups, 21-30 and 51-60 age groups respectively. In acute lymphoblastic leukemia (ALL) statistically no significant relationship was found between clinical features and FLT3/ITD positivity (p>0.05). However, in acute lymphoblastic leukemia (ALL) FLT3/ITD+ mutation was more commonly found in age groups of 21-30.

High prevalence of BCR-ABL fusion transcripts with poor prognostic impact among adult ALL patients: report from Pakistan.

AIM: Philadelphia (Ph) chromosome positive acute lymphoblastic leukemia (ALL) is a common cytogenetic abnormality associated with poor outcome in adults. This preliminary study, in the absence of substantial evidence, reported the prevalence of the BCR-ABL gene fusion in ALL patients by RT-PCR in Pakistan. Moreover, the prognostic significance of BCR-ABL fusion along with other characteristics was also ascertained. METHODS: One hundred forty six newly diagnosed ALL patients treated at our center between 2005 and 2008 comprised the study group. The patients were treated with a Children and Leukemia Group B induction regimen. RESULTS: Among these patients, BCR-ABL fusion oncogene was present in 43 of 78 patients (55%). A statistically significant difference in BCR-ABL-positivity within three age groups (<20 years, 20-50 years, >50 years) was observed (P= 0.001). The median age was significantly higher in the BCR-ABL+ group (30 vs 19 years; P= 0.001). BCR-ABL+ patients were also characterized by higher median WBC counts (96,000/µL vs 23,000//µL, P= 0.002). Complete remission was achieved in 74% BCR-ABL- patients and 35% BCR-ABL+ patients (P= 0.001). Only 10% of BCR-ABL- patients achieved continued complete remission (CR). None of the BCR-ABL+ patients maintained a CR further to induction therapy. With the available therapeutic protocol, the presence of a BCR-ABL fusion predicted a lower survival (P= 0.001). CONCLUSION: A high prevalence of BCR-ABL gene fusion was observed that appeared as a poor prognostic factor. Identification of this genetic entity at diagnosis is crucial for understanding the nature of adult ALL and for deciding optimal treatment.