Genotoxicity of the Kishon River,Israel: the application of an in vitro cellular assay

The alkaline comet assay, a sensitive method for DNA strand breaks and alkali labile site detection in individual cells, was employed here as an ecotoxicological monitoring tool for evaluation of genotoxicity in the Kishon River, Israel. This river, the most polluted river in Israel, has recently elicited major public concern with regard to cancer incidences in people who have dived there over many years. Five water samples were collected every odd month throughout the year 2001, from four localities. The comet assay was employed on fish hepatoma cell line RTH-149. Cells were exposed for 2h, in triplicate, to Kishon water: medium (1:1) samples that were pre-adjusted for pH and salinity percentage levels. Three DNA damage parameters (comet percentages, score of damage, and cumulative tail lengths of the comet), revealed significantly higher genotoxic values in Kishon water-treated cells as compared with the controls (up to 2.4, 3.2, and 3.6-fold, respectively). Part of the sampling sites revealed higher genotoxicity than other polluted sites. The results of this study demonstrate that the comet assay with RTH-149 cells can be successfully applied to a variety of aquatic samples revealing freshwater, marine and estuary conditions. The method found to be fast, sensitive, and suitable for monitoring programs.     

CADM1 Is a Key Receptor Mediating Human Mast Cell Adhesion to Human Lung Fibroblasts and Airway Smooth Muscle Cells

Background

Mast cells (MCs) play a central role in the development of many diseases including asthma and pulmonary fibrosis. Interactions of human lung mast cells (HLMCs) with human airway smooth muscle cells (HASMCs) are partially dependent on adhesion mediated by cell adhesion molecule-1 (CADM1), but the adhesion mechanism through which HLMCs interact with human lung fibroblasts (HLFs) is not known. CADM1 is expressed as several isoforms (SP4, SP1, SP6) in HLMCs, with SP4 dominant. These isoforms differentially regulate HLMC homotypic adhesion and survival.

Objective

In this study we have investigated the role of CADM1 isoforms in the adhesion of HLMCs and HMC-1 cells to primary HASMCs and HLFs.

Methods

CADM1 overexpression or downregulation was achieved using adenoviral delivery of CADM1 short hairpin RNAs or isoform-specific cDNAs respectively.

Results

Downregulation of CADM1 attenuated both HLMC and HMC-1 adhesion to both primary HASMCs and HLFs. Overexpression of either SP1 or SP4 isoforms did not alter MC adhesion to HASMCs, whereas overexpression of SP4, but not SP1, significantly increased both HMC-1 cell and HLMC adhesion to HLFs. The expression level of CADM1 SP4 strongly predicted the extent of MC adhesion; linear regression indicated that CADM1 accounts for up to 67% and 32% of adhesion to HLFs for HMC-1 cells and HLMCs, respectively. HLFs supported HLMC proliferation and survival through a CADM1-dependent mechanism. With respect to CADM1 counter-receptor expression, HLFs expressed both CADM1 and nectin-3, whereas HASMCs expressed only nectin-3.

Conclusion and Clinical Relevance

Collectively these data indicate that the CADM1 SP4 isoform is a key receptor mediating human MC adhesion to HASMCs and HLFs. The differential expression of CADM1 counter-receptors on HLFs compared to HASMCs may allow the specific targeting of either HLMC-HLF or HLMC-HASMC interactions in the lung parenchyma and airways.     

Interaction of the isolated DNA of lambda phage with spheroplasts of E. coli treated with sturine]

The method of centrifugation in sucrose density gradient (30-55%) of the spheroplast membrane preparations treated and untreated with sturine and infected with phage lambda DNA demonstrated that sturine, treatment increased the phage lambda DNA absorption three-fold. About 50% of the lambda DNA molecules adsorbed by spheroplasts are bound with the cytoplasmic membrane of spheroplasts treated with sturine; 50% of the lambda DNA molecules are bound with the cell wall membrane on the sturine-untreated spheroplasts. The data obtained allow to conclude that the stimulating effect of sturine in E. coli spheroplasts transfection by lambda DNA is connected with redistribution of phage DNA absorbed on spheroplasts from the cell wall to the cytoplasmic membrane facilitating the penetration of DNA and its fastening on the membrane.     

Structural and functional properties of irradiated nuclear membranes at the air-water phase interface

The monolayer technique, the methods of electron microscopy and IR-spectroscopy were used to study isolated nuclear membranes of calf thymus cells. The data obtained permitted to study the relationship between structural disorders induced by irradiation of membranes, the changes in their functional status, and the role played by a lipid component of membranes in these processes.     

Does epigenetic polymorphism contribute to phenotypic variances in <Emphasis Type="Italic">Jatropha curcas</Emphasis> L.?

Background  

There is a growing interest in Jatropha curcas L. (jatropha) as a biodiesel feedstock plant. Variations in its morphology and seed productivity have been well documented. However, there is the lack of systematic comparative evaluation of distinct collections under same climate and agronomic practices. With the several reports on low genetic diversity in jatropha collections, there is uncertainty on genetic contribution to jatropha morphology.