Edelfosine-induced metabolic changes in cancer cells that precede the overproduction of reactive oxygen species and apoptosis

Background  

Metabolic flux profiling based on the analysis of distribution of stable isotope tracer in metabolites is an important method widely used in cancer research to understand the regulation of cell metabolism and elaborate new therapeutic strategies. Recently, we developed software Isodyn, which extends the methodology of kinetic modeling to the analysis of isotopic isomer distribution for the evaluation of cellular metabolic flux profile under relevant conditions. This tool can be applied to reveal the metabolic effect of proapoptotic drug edelfosine in leukemia Jurkat cell line, uncovering the mechanisms of induction of apoptosis in cancer cells.     

The use of toluidine blue for in situ detection of micro-organisms in foods

The use of toluidine blue to stain cryosections of food samples to detect sites of microbial growth was examined. The method successfully detected colonies and single cells of both yeasts and bacteria at magnifications of x 400 or below in the majority of foods studied. The method demonstrates great potential for studying the micro-environments in which micro-organisms grow.     

In vitro comparison of agar and microbroth dilution methods for determination of MICs for Mycoplasma hominis

Ten replicates of three Mycoplasma hominis strains were tested by microbroth and agar dilution against levofloxacin, moxifloxacin, gatifloxacin, erythromycin, tetracycline, and clindamycin. Both methods provide reproducible results. Agar dilution tends to yield higher MIC values for some drugs.     

Survival of Theileria parva-infected adult Rhipicephalus appendiculatus under laboratory and quasi-natural conditions

Adult Rhipicephalus appendiculatus Muguga, having high or low intensities of Theileria parva Muguga infection in their salivary glands, were exposed to 20 °C and 85% relative humidity in the laboratory or quasi-natural conditions. Survival of the ticks and T. parva infections in their salivary glands was then monitored over a two year period. Ticks, having an average infection level of 2 infected acini per female, survived for up to 70 or 106 weeks after moulting under the laboratory or quasi-natural conditions respectively. Those having an infection level of 26 infected acini per female, survived for a similar duration except that those under quasi-natural conditions survived for a slightly shorter duration (102 weeks). Similarly, T. parva parasites survived for much longer periods under quasi-natural conditions than under the laboratory conditions. They survived for up to 38 or 78 weeks post salivary gland infection under the laboratory or quasi-natural conditions respectively in both categories of infection levels. There was apparently a density dependent relationship in T. parva survival, with a dramatic fall in infection occurring in ticks with high levels of infection between weeks 10 and 18 or weeks 38 and 46 post salivary gland infection in those exposed to laboratory or quasi-natural conditions before levelling off. This revised version was published online in July 2006 with corrections to the Cover Date.     

Investigation by luminal perfusion of the transfer of compounds into the epididymis of the anaesthetized rat.

Controlled perfusion through the lumen of the distal cauda epididymidis in the anaesthetized rat has been explored as a means of examining physiological exchanges from blood across the epididymal epithelium. The mean length of the perfused, sperm-free, tubule was 14.5 cm (+/- 1.5 s.e.m., n = 9). No cholesterol, protein or sialic acid was detected in the perfusate at flow rates exceeding 10 microliters/min, but at rates of 0.4--1.2 microliters/min, protein appeared at concentrations of 0.21--0.55 mg/ml (i.e. secretion rates of 0.21--0.83 micrograms/min; 3 rats). Glucose was detected at all perfusion rates (3--27 microliters/min) at concentrations of 0.06--0.58 mM (0.8--6.8% blood levels). During intravenous infusions of 3H2O, radioactivity in the perfusate rapidly attained 87% blood plasma concentrations; no radioactivity was detected when carboxy-E114C]dextran or methoxy-[3H]inulin were infused. Radioactivity appeared in the epididymal perfusate to 1--7% of blood levels during intravenous infusions of D-E1U-1RC]glucose or 3-O-methyl[1-3H]glucose. This evidence suggests that the preparation is physiological and could be used to explore the dynamics of exchanges between blood and epididymis.     

Ligand-dependent differences in estrogen receptor beta-interacting proteins identified in lung adenocarcinoma cells corresponds to estrogenic responses

Background

A recent epidemiological study demonstrated a reduced risk of lung cancer mortality in breast cancer patients using antiestrogens. These and other data implicate a role for estrogens in lung cancer, particularly nonsmall cell lung cancer (NSCLC). Approximately 61% of human NSCLC tumors express nuclear estrogen receptor β (ERβ); however, the role of ERβ and estrogens in NSCLC is likely to be multifactorial. Here we tested the hypothesis that proteins interacting with ERβ in human lung adenocarcinoma cells that respond proliferatively to estradiol (E₂) are distinct from those in non-E₂-responsive cells.

Methods

FLAG affinity purification of FLAG-ERβ-interacting proteins was used to isolate ERβ-interacting proteins in whole cell extracts from E₂ proliferative H1793 and non-E₂-proliferative A549 lung adenocarcinoma cell lines. Following trypsin digestion, proteins were identified using liquid chromatography electrospray ionization tandem mass spectrometry (LC-MS/MS). Proteomic data were analyzed using Ingenuity Pathway Analysis. Select results were confirmed by coimmunoprecipitation.

Results

LC-MS/MS identified 27 non-redundant ERβ-interacting proteins. ERβ-interacting proteins included hsp70, hsp60, vimentin, histones and calmodulin. Ingenuity Pathway Analysis of the ERβ-interacting proteins revealed differences in molecular and functional networks between H1793 and A549 lung adenocarcinoma cells. Coimmunoprecipitation experiments in these and other lung adenocarcinoma cells confirmed that ERβ and EGFR interact in a gender-dependent manner and in response to E₂ or EGF. BRCA1 interacted with ERβ in A549 cell lines and in human lung adenocarcinoma tumors, but not normal lung tissue.

Conclusion

Our results identify specific differences in ERβ-interacting proteins in lung adenocarcinoma cells corresponding to ligand-dependent differences in estrogenic responses.