l-Amino acid oxidase from Cerastes vipera snake venom: Isolation, characterization and biological effects on bacteria and tumor cell lines.

A homodimeric l-amino acid oxidase enzyme (Cv-LAAOI) was isolated from the venom of Cerastes vipera (Egyptian Sand viper) using gel filtration followed by anion exchange chromatography. The molecular mass of Cv-LAAO is 120 kDa in its native form and 60 kDa in its monomeric form. The optimum enzyme activity was achieved on l-Leucine as a substrate in 50 mM buffer pH 7.5 at 50 °C. The Cv-LAAOI activity was significantly reduced by increasing the temperature over 40 °C, lost 75% of its activity at 60 °C and inhibited completely at 80 °C. The Cv-LAAOI attains the highest substrate specificity towards L-Met. The results have also indicated that Mn2+ enhances the enzyme activity by 10%, while Cu2+, Hg2+, Ni2+, Co2+ have suppressive effects on the Cv-LAAOI activity. On the other hand, EDTA has no significant effect on the enzyme activity. The kinetic parameters of Cv-LAAOI activity (Km, Kcat and Vmax) estimated on l-Leucine at pH 8 and 37 °C were found to be 2 mM, 12 S-1 and 16.7 µmol/min/ml, respectively. In addition, the results have shown that Cv-LAAOI exhibits a significant bactericidal activity against gram-positive and gram-negative bacteria, particularly Staphylococcus aureus and Escherichia coli with MIC values of 20 µg/ml. Moreover, Cv-LAAOI has exhibited a considerable cytotoxic activity against breast cancer cell line (MCF-7) with IC50 value 2.75 ±â€¯0.38 µg/ml compared with different tumor cell lines (liver HepG2, lung A549, colon HCT116 and prostate PC3). Furthermore, Cv-LAAOI has triggered antiproliferative activity via extensive H2O2 generation as indicated by the increase in H2O2 and TBARS levels accompanied by the depletion in the catalase activity (CAT) in MCF-7 treated cells compared to the untreated ones. Thus, these findings clearly indicate that Cv-LAAOI has a selective cytotoxic effect on breast cancer cell line, demonstrating a great prospective for future use in cancer therapy.

Blink reflex in type 2 diabetes mellitus.

PURPOSE: An evaluation of the extent of damage of the central nervous system in diabetes mellitus is of high value in current research. Electrophysiological abnormalities are frequently present in asymptomatic patients with diabetes mellitus. Diabetic cranial neuropathy is one of the complications of the disease. Blink reflex is used to diagnose subclinical cranial neuropathy. The objective is to test the utility of blink reflex in detecting subclinical cranial nerve involvement in patients with type 2 diabetes mellitus. METHODS: Forty patients with type 2 diabetes mellitus, aged from 30 to 60 years examined clinically and neurologically. Blink reflex and nerve conduction studies for the upper and lower limbs were performed and compared with 20 matched normal controls. RESULTS: Diabetic patients with peripheral neuropathy showed significant prolonged distal latency and reduced amplitudes of the R2C response compared with the control, patients without peripheral neuropathy showed insignificant changes. Alteration of R2 correlated with the type of treatment and the duration of the disease. In patients without peripheral neuropathy, ulnar sensory distal latencies showed significant positive correlation with R2I latency, whereas its Conduction Velocity (CV) showed significant positive correlation with R2C amplitudes and negative correlation with R2C latency. CONCLUSIONS: R2C is the most sensitive parameter in the blink reflex, which can help in the diagnosis of subclinical diabetic cranial neuropathy.