Molecular diagnosis of duchennemuscular dystrophy by multiplexpolymerase chain reaction

Duchenne or Becker muscular dystrophy [DMD/BMD] is one of the most common X-linked lethal genetic diseases with a worldwide frequency of one in 3500 live male births. The Dmd locus is, the largest human gene known, span 2.4 Mb in Xp21. It consists of 79 exons that encode a14 Kb transcript. We have investigated the frequency of deletion in the Dmd gene in 37 unrelated Duchenne muscular dystrophy [DMD] Egyptian patients. All patients were screened for 21 exonic deletion which pro-posed by Multicenter Study Group. The screening of deletion was done using single strand conformation polymorphism [SSCP] after multiplex PCR. The most common exonic deletion in our study are exon 48 [46 %],exons-19, 45 [37.8 % of each] followed by exons-43 [ 29.7 %], PM, 44, 47,and 50 [27 %]. The overall deletion in our study about 83.8% in DMD. Deletions were clustered at two spots: 76 % at the hot spot in the distal region of the gene and 24 % at the hot spot in the proximal region of the gene. The multiplex PCR is simple, reliable and rapid technique for detection of carrier in families with DMD

Lymphocyte apoptosis and proliferation related gene products in childhood acute lymphoblastic leukemia

The aim of the study is to characterize markers of apoptosis in children with ALL in relation to treatment outcome of the disease. The study was performed on 34 children with ALL and 60 healthy children as a control group. Apoptosis was assessed by cell morphology; DNA fragmentation; ELISA and RT-PCR for CD95, CD95L, BcL2 and NF-KB; and flowcytometry for CD95, CD40, CD49d, and CD11a. Apoptosis was significantly lower in cases than controls. Apoptosis detected by CD95 ligand was significantly lower in cases with no remission after treatment than those with remission. Antiapoptotic factors: CD40, BcL2, and NF-KB were all found to be higher in cases than controls and in cases with no remission than those with remission, CD49d was significantly lower in cases than controls, and significantly lower in cases with no remission. CD11a levels were not different among various groups. Delayed apoptosis of ALL cells is genetically controlled either directly or indirectly by a network of oncogenes and tumor suppressor genes. CD40 appeared to stimulate both T and lineage and is considered the most potent influencer and predictor to resistance to therapy. Inhibitors for the activity of CD40, 8c/2 and NF-kB as well as stimulants to CD95 could have a potential therapeutic benefit