Proteome analysis of mouse brain exposed to chronic hypoxia

Chronic hypoxia exists in many diseases, including cancer. The subject of our study is analysis of molecular pathways affected in the chronically hypoxic mouse brain. Using the emPAl protocol, we performed a quantitative proteomic approach to characterize the global proteome in the mouse brain exposed to 7% O[2] for 48 hours. Utilizing the emPAl protocol to estimate protein abundance and assign molar concentrations to all proteins, we were able to identify 33 proteins with significant changes in their expression. Deregulated proteins were mainly involved in cell metabolism, apoptosis, Ca[2+] signaling, pentose phosphate pathway, 14-3-3 protein mediated signaling cascades and protein degradation. The obtained data will provide some clues for understanding mechanisms with which cells respond and adapt to chronic hypoxia

profile of human sperm proteome; a mini-review

New advances in mass spectrometry-based proteomics technology are having a major impact on our understanding of how human spermatozoa acquire their capacity for fertilization. A complete analysis of the proteins found in the human spermatozoa is essential for understanding the events leading up to, and including, fertilization and early embryo development. In this short review, we have collected the human sperm proteome from the literature and analyzed it by the Database for Annotation, Visualization and Integrated Discovery [DAVID] software. Bioinformatics analysis demonstrated that the collected 1,300 proteins were involved in various metabolic pathways including catabolic processes. Additionally, the majority of the collected human sperm proteome belonged to cytoplasm. Application of the multi-dimensional protein identification technology [MudPIT] for obtaining a better coverage of the hydrophobic and basic proteins of the human sperm proteome is recommended

Comparison of the photobleaching and photostability traits of alexa fluor 568- and fluorescein isothiocyanate- conjugated antibody

Synthetic fluorescent dyes that are conjugated to antibodies are useful tools to probe molecules. Based on dye chemical structures, their photobleaching and photostability indices are quite diverse. It is generally believed that among different fluorescent dyes, Alexa Fluor family has greater photostability than traditional dyes like fluorescein isothiocyanate [FITC] and Cy5. Alexa Fluor 568 is a member of Alexa Fluor family presumed to have superior photostability and photobleahing profiles than FITC. In this experimental study, we conjugated Alexa Fluor 568 and FITC dyes to a mouse anti-human nestin monoclonal antibody [ANM] to acquire their photobleaching profiles and photostability indices. Then, the fluorophore/antibody ratios were calculated using a spectrophotometer. The photobleaching profiles and photostability indices of conjugated antibodies were subsequently studied by immunocytochemistry [ICC]. Samples were continuously illuminated and digital images acquired under a fluorescent microscope. Data were processed by ImageJ software. Alexa Fluor 568 has a brighter fluorescence and higher photostability than FITC. Alexa Fluor 568 is a capable dye to use in photostaining techniques and it has a longer photostability when compared to FITC

Hypoxia: a review

Tissue hypoxia occurs where there is an imbalance between oxygen supply and consumption. Growing evidence from experimental and clinical studies points to the fundamental and pathophysiologic role of hypoxia in cancer, ischemic tolerance, and stroke. Hypoxia-induced changes in ion homeostasis, erythropoiesis, angiogenesis, proliferation and differentiation. This review outlines hypoxia effect at molecular level and describes briefly hypoxia role in the physiological and pathological conditions