Study of Isocitrate Dehydrogenase 1and 2 Mutations in Adult Egyptian Patients with Denovo Acute Myeloblastic Leukemia, Their Relation to Clinical Characteristics, FLT3/ITD and Nucleophosmin 1 Mutations and Impact on Treatment Outcome

Background: Acute myeloid leukaemia (AML) is a malignancy that is heterogeneous in nature characterized by genetic abnormalities some of which are established in the diagnosis and prognosis of the disease. An additional role for alterations in epigenetic mechanisms has been also highlighted in the pathogenesis of the disease. This may have a role in determining the disease outcome, impact the treatment decision and provide options for targeted therapies especially in patients who lack genetic aberrations. One of the modes of epigenetic dysregulation is mutation in genes encoding isocitrate dehydrogenase 1 and 2 that has been observed in AML with a higher incidence in patients with normal karyotype (NK). Aim of the work: The aim of this work was to study the frequency of IDH1 (R132) and IDH2 (R140Q, R172K) mutations in adult Egyptian patients with de novo acute myeloblastic leukemia (AML), their relation with clinical characteristics, other molecular markers (the internal tandem duplication (ITD)) mutation of FLT3 gene and NPM1 gene mutation) and impact on treatment outcome. Methods: Peripheral blood samples from 50 adult patients with denovo acute myeloid leukemia, admitted to the haematology unit at Alexandria Main University Hospital from February 2015 to February 2017, were used. The polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP) was used for detection of IDH1 codon R132 and IDH2 codons (R140, R172) mutations on genomic DNA. PCR was used for detection of FLT3-ITD mutation on genomic DNA. PCR was used for detection of NPM1 mutation on RNA. Results: IDH 1and 2 mutations occurred in 30% of newly diagnosed AML patients and 47.6% of NK patients. Both mutations did not co-occur except in one case. IDH positive patients were significantly older than IDH negative patients (p=0.003). There was no statistically significant correlation between any of the clinical parameters and the IDH mutations. FAB-M2 was the most common FAB subtype among IDH positive patients. No correlation between IDH mutations and NPM1 or FLT3 could be demonstrated. IDH positive patients had significantly lower CR rates after induction chemotherapy than IDH negative patients (p=0.021). Conclusion: IDH1, 2 mutations are recurring genetic alterations in AML with a higher incidence in patients with normal karyotype and they may have an unfavorable impact on clinical outcome in adult AML patients.

Plasma and ascitic fluid level of calprotectin in chronic liver disease: malignant and non malignant

Calprotectin was widely investigated in alcoholic liver disease and proved to be a new prognostic marker of survival independent of the severity of liver disease as well as marker of malignancy. However it was not widely investigated in other causes of liver cirrhosis. Of the present work was to study the level of calprotectin both in plasma and ascitic fluid in patients with hepatitis C [HCV] related chronic liver disease with and without malignancy, and to find out whether one or both of them correlate with the severity of liver damage and presence of malignancy. This study was conducted at the Faculty of Medicine, Alexandria University and the National Liver Institute, Menoufiya University. Thirty patients with Hepatitis C related liver cirrhosis were recruited. Fifteen of these patients suffered from concomitant hepatocellular carcinoma [HCC] diagnosed by elevated alpha foeto-protein [AFP] and one imaging technique OR by two imaging techniques in the case of normal AFP. Calprotectin was significantly elevated in patients with cirrhosis and cirrhosis/HCC than in controls [p=<0.01]. However there was no significant difference in the levels of plasma or ascitic calprotectin between the cirrhotic group and the group with HCC. There was no correlation between plasma and ascitic calprotectin levels. Ascitic calprotectin correlated significantly with bilirubin, and markers of synthetic liver function [p=<0.05], but plasma calprotectin correlated only with prothombin activity [p=<0.05]. In patients with spontaneous bacterial peritonitis, ascitic calprotectin was significantly higher in patients having this complication [879.8 +/- 67.5] than patients without SBP [534.2 +/- 59.3 [p<0.01] and a highly significant correlation was found between ascitic calprotectin and total leucocytic count in ascitic fluid [p=<0.01]. Calprotectin is elevated in HCV-related cirrhosis but not further elevation with the occurrence of hepatocellular carcinoma. Ascitic calprotectin correlated with the degree of hepatocellular injury and was significantly higher in patients with SBP. Further studies are warranted to establish a role of plasma calprotectin for the risk assessment of infectious complications secondary to bacterial translocation in patients with HCV- related liver cirrhosis

Transforming growth factor- beta 1 in patients with angiographically proven coronary heart disease

Transforming growth factor- beta 1 [TGF- beta 1] is a multifunctional cytokine that exhibits vasculoprotective properties. Production and plasma levels of TGF- beta 1 are influenced by polymorphisms in the TGF- beta 1 gene. A T-C transition at nucleotide 29 of the TGF- beta 1 gene results in a Leu-Pro substitution at amino acid 10 of the signal peptide. We investigated whether the T29-C polymorphism of TGF- beta 1 is associated with myocardial infarction among a sample of Egyptian coronary artery diseased males. The study population consisted of 90 patients with angiographically proven myocardial infarction and 30 control individuals without signs or symptoms of myocardial infarction. Polymorphism-related genotypes were determined with allele-specific PCR [polymerase chain reaction]. In this study; we found no differences between patients with MI and healthy subjects regarding the genotype and allele frequencies of codon 10 TGF- beta 1 polymorphism. Thus, our results indicate that this polymorphism does not appear to predispose to the development of MI. There was an agreement between genotypes observed and those predicted by the Hardy-Weinberg equilibrium in the control group [Codon 10, x[2] goodness of fit was not significant = 0.777, p; 0.87]. Negative association findings in this study did not exclude that TGF- beta 1 is a susceptibility locus for myocardial infarction, but further study on a larger scale is needed