Protein expression profiles of intestinal epithelial co-cultures: effect of functionalised carbon nanotube exposure.

To assess the biological effects of low level, water dispersible, functionalised carbon nanotube (f-CNT) exposure in an in vitro model simulating the digestive tract, cellular protein expression was quantified and compared using label-free quantitative mass spectrometry (LFQMS). Co-cultured cells were exposed to well-characterised SWCNT-COOH, MWCNT-COOH, and MWCNT-PVP. The relative expression of 2,282 unique proteins was compared across the dose groups.

Ethanol exposure alters protein expression in a mouse model of fetal alcohol spectrum disorders.

Alcohol exposure during development can result in variable growth retardation and facial dysmorphology known as fetal alcohol spectrum disorders. Although the mechanisms underlying the disorder are not fully understood, recent progress has been made that alcohol induces aberrant changes in gene expression and in the epigenome of embryos. To inform the gene and epigenetic changes in alcohol-induced teratology, we used whole-embryo culture to identify the alcohol-signature protein profile of neurulating C6 mice.

Ethanol-induced changes in the expression of proteins related to neurotransmission and metabolism in different regions of the rat brain.

Despite extensive description of the damaging effects of chronic alcohol exposure on brain structure, mechanistic explanations for the observed changes are just emerging. To investigate regional brain changes in protein expression levels following chronic ethanol treatment, one rat per sibling pair of male Wistar rats was exposed to intermittent (14 h/day) vaporized ethanol, the other to air for 26 weeks. At the end of 24 weeks of vapor exposure, the ethanol group had blood ethanol levels averaging 450 mg%, had not experienced a protracted (> 16 h) withdrawal from ethanol, and revealed only mild evidence of hepatic steatosis.

Differential expression of renal proteins in a rodent model of Meckel syndrome.

Background: Meckel syndrome (MKS) is a fatal autosomal recessive condition with prominent renal cystic pathology. Renal protein misexpression was evaluated in the Wpk rat model of human MKS3 gene disease to identify biomarkers for the staging of renal cystic progression.

Methods: Misexpressed proteins were compared between early and late stages of MKS renal cystic disease using proteomic analysis (two-dimensional gel electrophoresis with LC-MS/MS identification) followed by Western blot analysis.

Protein changes in macrophages induced by plasma from rats exposed to 35 GHz millimeter waves.

A macrophage assay and proteomic screening were used to investigate the biological activity of soluble factors in the plasma of millimeter wave-exposed rats. NR8383 rat macrophages were incubated for 24 h with 10% plasma from male Sprague-Dawley rats that had been exposed to sham conditions, or exposed to 42 °C environmental heat or 35 GHz millimeter waves at 75 mW/cm² until core temperature reached 41.0 °C.

Serum Proteomic Profiles In Subjects with Heavy Alcohol Abuse.

OBJECTIVES: The abuse of alcohol is a major public health problem, and the diagnosis and care of patients with alcohol abuse and dependence is hindered by the lack of tests that can detect dangerous levels of drinking or relapse during therapy. Gastroenterologists and other healthcare providers find it very challenging to obtain an accurate alcohol drinking history. We hypothesized that the effects of ethanol on numerous systems may well be reflected in changes in quantity or qualities of constituent or novel plasma proteins or protein fragments.

A proteomic workflow for discovery of serum carrier protein-bound biomarker candidates of alcohol abuse using LC-MS/MS.

The diagnosis and care of patients with alcohol abuse and dependence is hampered by a lack of sensitive and specific screening and monitoring tests. Proteomics is a good approach to search for biomarkers of alcohol abuse. Serum carrier protein-bound proteins have attracted significant interest because they remain a relatively un-mined region of the proteome.

Sample complexity reduction for two-dimensional electrophoresis using solution isoelectric focusing prefractionation.

Despite its excellent resolving power, 2-DE is of limited use when analyzing cellular proteomes, especially in differential expression studies. Frequently, fewer than 2000 protein spots are detected on a single 2-D gel (a fraction of the total proteome) regardless of the gel platform, sample, or detection method used. This is due to the vast number of proteins expressed and their equally vast dynamic range.

Application of pressure cycling technology to tissue sample preparation for 2-DE.

2-DE is a powerful protein analytical tool whose major strengths include semiglobal quantitation and charge separation of complex protein mixtures, enabling the analysis of differential protein expression, and variable post-translational modification. One of 2-DE's limitations relates to its limited dynamic range and consequently the number of proteins expressed that can be analyzed on a single gel. In an attempt to improve the yield of detectable proteins during sample preparation, we applied a novel extraction technique called pressure cycling technology.

A comparative proteomic study to characterize the vinblastine resistance in human ovarian cancer cells.

Drug resistance is a major impediment to the successful treatment of human cancers, including ovarian cancer. Vinblastine (VLB), an antimicrotubule agent, is one of the chemotherapeutic drugs that exhibit resistance in ovarian cancer patients. To determine the protein factors that are involved in vinblastine resistance in human ovarian cancer cells, a combination of sample pre-fractionation and high-resolution 2-DE proteomic analysis was performed.

A proteomic survey of rat cerebral cortical synaptosomes.

Previous findings from our laboratory and others indicate that two-dimensional gel electrophoresis (2-DE) can be used to study protein expression in defined brain regions, but mainly the proteins which are present in high abundance in glia are readily detected. The current study was undertaken to determine the protein profile in a synaptosomal subcellular fraction isolated from the cerebral cortex of the rat. Both 2-DE and liquid chromatography - tandem mass spectrometry (LC-MS/MS) procedures were used to isolate and identify proteins in the synaptosomal fraction and accordingly >900 proteins were detected using 2-DE; the 167 most intense gel spots were isolated and identified with matrix-assisted laser desorption/ionization - time of flight peptide mass fingerprinting or LC-MS/MS.

Gels and more gels: probing toxicity.

Two-dimensional electrophoresis (2-DE) remains an important technology in the study of protein expression. In many applications, 2-DE has been supplanted by liquid chromatographic and mass spectrometric approaches that overcome some of its limitations and labor intensiveness. Nevertheless, 2-DE has exceptional relevance in toxicology and, despite the challenges, its implementation continues to support toxicologists in understanding the biological effects of chemical exposures in living systems.