An in vitro cytotoxicity of glufosfamide in HepG2 cells relative to its nonconjugated counterpart.

BACKGROUND: Glufosfamide (ß-D-glucosylisophosphoramide mustard, GLU) is an alkylating cytotoxic agent in which ifosforamide mustard (IPM) is glycosidically linked to the ß-D-glucose molecule. GLU exerted its cytotoxic effect as a targeted chemotherapy. Although, its cytotoxic efficacy in a number of cell lines, there were no experimental or clinical data available on the oncolytic effect of oxazaphosphorine drugs in hepatocellular carcinoma. Therefore, the main objective of the current study is to assess the cytotoxic potential of GLU for the first time in the hepatocellular carcinoma HepG2 cell line model. METHODS: Cytotoxicity was assayed by the MTT method, and half-maximal inhibitory concentration (IC50) was calculated. Flow cytometric analysis of apoptosis frequencies was measured by using Annexin V/PI double stain, an immunocytochemical assay of caspase-9, visualization of caspase-3, and Bcl2 gene expression were undertaken as apoptotic markers. Mitochondrial membrane potential was measured using the potentiometric dye; JC-1, as a clue for early apoptosis as well as ATP production, was measured by the luciferase-chemiluminescence assay. RESULTS: Glufosfamide induced cytotoxicity in HepG2 cells in a concentration- and time-dependent manner. The IC50 values for glufosfamide were significantly lower compared to ifosfamide. The frequency of apoptosis was much higher for glufosfamide than that of ifosfamide. The contents of caspase-9 and caspase-3 were elevated following exposure to GLU more than IFO. The anti-apoptotic Bcl2 gene expression, the mitochondrial membrane potential, and the cellular ATP levels were significantly decreased than in case of ifosfamide. CONCLUSIONS: The current study reported for the first time cytotoxicity activity of glufosfamide in HepG2 cells in vitro. The obtained results confirmed the higher oncolytic activity of glufosfamide than its aglycone ifosfamide. The generated data warrants further elucidations by in vivo study.

Impact of Foliar Application of Chitosan Dissolved in Different Organic Acids on Isozymes, Protein Patterns and Physio-Biochemical Characteristics of Tomato Grown under Salinity Stress.

In this study, the anti-stress capabilities of the foliar application of chitosan, dissolved in four different organic acids (acetic acid, ascorbic acid, citric acid and malic acid) have been investigated on tomato (Solanum lycopersicum L.) plants under salinity stress (100 mM NaCl). Morphological traits, photosynthetic pigments, osmolytes, secondary metabolites, oxidative stress, minerals, antioxidant enzymes activity, isozymes and protein patterns were tested for potential tolerance of tomato plants growing under salinity stress. Salinity stress was caused a reduction in growth parameters, photosynthetic pigments, soluble sugars, soluble proteins and potassium (K+) content. However, the contents of proline, ascorbic acid, total phenol, malondialdehyde (MDA), hydrogen peroxide (H2O2), sodium (Na+) and antioxidant enzyme activity were increased in tomato plants grown under saline conditions. Chitosan treatments in any of the non-stressed plants showed improvements in morphological traits, photosynthetic pigments, osmolytes, total phenol and antioxidant enzymes activity. Besides, the harmful impacts of salinity on tomato plants have also been reduced by lowering MDA, H2O2 and Na+ levels. Chitosan treatments in either non-stressed or stressed plants showed different responses in number and density of peroxidase (POD), polyphenol oxidase (PPO) and superoxide dismutase (SOD) isozymes. NaCl stress led to the diminishing of protein bands with different molecular weights, while they were produced again in response to chitosan foliar application. These responses were varied according to the type of solvent acid. It could be suggested that foliar application of chitosan, especially that dissolved in ascorbic or citric acid, could be commercially used for the stimulation of tomato plants grown under salinity stress.

Complete Genome Sequence of Chikungunya Virus Isolated from an Aedes aegypti Mosquito during an Outbreak in Yemen, 2011.

Chikungunya virus is recognized as a serious public health problem. The complete genome was sequenced for a chikungunya virus isolated from the mosquito Aedes aegypti during a 2011 outbreak in Al Hodayda, Yemen, which resulted in significant human fatalities. Phylogenetic analysis demonstrated that this Yemeni isolate is most closely related to Indian Ocean strains of the east/central/south African genotype.

Effect of thalidomide dithiocarbamate analogs on the intercellular adhesion molecule-1 expression.

Thalidomide has been reported to have anti-angiogenic and antimetastatic effects. Intercellular adhesion molecule-1 (ICAM-1) was shown to be involved in monocyte adherence to epithelial cells and cancer cell invasion. Novel thalidomide dithiocarbamate analogs (containing 2 sulfur atoms) were designed and synthesized as potential anti-tumor agents. The aim of this work is to investigate their anti-tumor effect against transplantable experimental tumor, Ehrlich ascites carcinoma (EAC), in mice by studying the changes in cell's biochemical profile, the expression of ICAM-1 and nitric oxide (NO) and their association with tumor burden. As shown in our results, treatment of solid tumor-bearing mice with thalidomide 1 resulted in a significant reduction in tumor volume with 75.4% inhibition, a significant decrease in lactate dehydrogenase (LDH), ICAM-1 expression and NO. Thalidomide dithiocarbamate analogs 2 and 3 exhibited a potent effect to reduce the volume of solid tumor with 96.7% and 96.5% inhibition, respectively, a significant ability to increase the albumin, alanine aminotransferase (ALT) and glucose levels and to diminish LDH, ICAM-1 expression and NO. Thalidomide dithiocarbamate analog 3 has more potent anti-tumor activity as compared with thalidomide 1 or its dithiocarbamate analog 2. Taken together, our study improved that the dithiocarbamate analogs 2 and 3 are more potent anti-tumor agents with more pronounced effect than thalidomide 1 itself.

Production of synthetic methionine-free and synthetic methionine-limited alpha casein: protein foodstuff for patients with homocystinuria due to cystathionine beta-synthase deficiency.

The aim of this study was to evaluate the possibility that synthetic forms of methionine-free alpha-casein and methionine-limited alpha casein could be produced by recombinant means to form the basis for developing an industrial-scale process for the provision of a foodstuff suitable for patients with homocystinuria due to cystathionine beta-synthase (CBS) deficiency. As a first step, two forms of alpha casein gene, encoding methionine-free alpha casein (Fcas) or a methionine-limited alpha casein (Mcas), were synthesised and expressed in Escherichia coli. Using the overexpression vector pET28a, both genes were highly expressed in E. coli in soluble form as well as in inclusion bodies. The two recombinant proteins were purified by the one step methods using the fused His-tag and the Ni(2+)column and validated by Western blot analysis. This work paves the way for industrial-scale production of proteins suitable for patients with homocystinuria due to CBS deficiency.