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Course Experiments (course + experiment)

Distribution by Scientific Domains
Medical Sciences28%

Kinds of Course Experiments

  • time course experiment
    		•

    Selected Abstracts

    Cadmium tolerance in the Nile tilapia (Oreochromis niloticus) following acute exposure: Assessment of some ionoregulatory parameters

    ENVIRONMENTAL TOXICOLOGY, Issue 1 2006
    Sofia Garcia-Santos
    Abstract The Nile tilapia (Oreochromis niloticus) can tolerate very high levels of waterborne cadmium. It has one of the highest 96 h LC50 recorded for a freshwater teleost fish (14.8 mg/L Cd; hardness 50 mg/L CaCO3). Cadmium is known to perturb ion balance in teleost fishes. However, in an acute time course experiment, plasma Na+ concentrations were unaffected, and plasma Ca2+ values only decreased after 96 h exposure in a dose-independent manner. Branchial Na+/K+ -ATPase activity and ,-subunit protein level expression in crude gill homogenates were not affected by Cd exposure during this period. Branchial chloride cell numbers, identified as Na+/K+ -ATPase immunoreactive cells using immunohistochemistry, decreased 24 h after exposure but recovered thereafter. Histopathological changes did not follow a consistent pattern of variation with exposure time, and the alterations noted in gill epithelium were basically nonspecific to cadmium. Because of its tolerance, it can be concluded that the tilapia O. niloticus would not be a suitable test organism to evaluate sublethal toxicity of cadmium and the realistic impact of this pollutant in the environment. However, it certainly could contribute significantly to our understanding of the toxic mechanism of cadmium exposure in aquatic organisms. This is the first work to investigate the effect of waterborne pollutants on Na+/K+ -ATPase ,-subunit protein expression in fish gills. © 2006 Wiley Periodicals, Inc. Environ Toxicol 21: 33,46, 2006. [source]

    Gap junctional communication in human osteoclasts in vitro and in vivo

    JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 6a 2008
    A. F. Schilling
    Abstract Bone-forming cells are known to be coupled by gap junctions, formed primarily by connexin43 (Cx43). The role of Cx43 in osteoclasts has so far only been studied in rodents, where Cx43 is important for fusion of mononuclear precursors to osteoclasts. Given the potential importance for human diseases with pathologically altered osteoclasts, we asked whether a similar influence of Cx43 can also be observed in osteoclasts of human origin. For this purpose, Cx43 mRNA expression was studied in a time course experiment of human osteoclast differentiation by RT-PCR. Localization of Cx43 in these cells was determined by immunohistochemistry and confocal microscopy. For the assessment of the effect of gap junction inhibition on cell fusion, gap junctions were blocked with heptanol during differentiation of the cells and the cells were then evaluated for multinuclearity. Paraffin sections of healthy bone and bone from patients with Paget's disease and giant cell tumour of the bone were used to study Cx43 expression in vivo. We found mRNA and protein expression of Cx43 in fully differentiated osteoclasts as well as in precursor cells. This expression decreased in the course of differentiation. Consistently, we found a lower expression of Cx43 in osteoclasts than in bone marrow precursor cells in the histology of healthy human bone. Blockade of gap junctional communication by heptanol led to a dose-dependent decrease in multinuclearity, suggesting that gap junctional communication precedes cell fusion of human osteoclasts. Indeed, we found a particularly strong expression of Cx43 in the giant osteoclasts of patients with Paget's disease and giant cell tumour of the bone. These results show that gap junctional communication is important for fusion of human mononuclear precursor cells to osteoclasts and that gap junctional Cx43 might play a role in the regulation of size and multinuclearity of human osteoclasts in vivo. [source]

    Cytotoxic effects induced by hexachlorobenzene in Squilla mantis (L.) (Crustacea, Stomatopoda)

    ENVIRONMENTAL TOXICOLOGY, Issue 1 2008
    Antonio Dell'Anno
    Abstract Contamination of marine environments by hexachlorobenzene (HCB) represents a serious concern for potential consequences on ecosystem and human health. Despite this, information on cytotoxic effects on marine organisms is still largely lacking. In this study, we investigated cytotoxic effects induced by HCB on gonads and muscular tissue of Squilla mantis by analysing Na+/K+ -ATPase activity and plasma membrane fluidity. This crustacean species was selected as a model for its habitat, trophic level, feeding behavior, and commercial exploitation for human consumption. Time course experiments revealed that low concentrations of HCB (i.e. 50 nM) determine an exponentially decrease of Na+/K+ -ATPase activity and a significant modification of cellular membrane fluidity. Significant negative relationships between Na+/K+ -ATPase activity and membrane fluidity were observed, suggesting that changes in the structure and packing of cellular membranes induced by HCB may be the primary factor affecting the activity of essential bilayer-associated enzymes. Overall these findings suggest that even small concentrations of HCB may determine important changes on cell metabolism with potential cascade effects on recruitment of this commercial species. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2008. [source]

    Gene expression study of Saccharomyces cerevisiae under changing growth conditions

    JOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 8 2009
    Pengcheng Fu
    Abstract BACKGROUND: DNA microarrays technology has been used to obtain expression profiles of thousands of genes at the same time for a given organism at relatively low costs. While gene expression approaches are being developed which allow holistic analysis of complex biological processes, there exist very few illustrative examples on the integration of large scale modeling and high throughput time course experiments to upgrade the information contents on yeast biology. RESULTS:Saccharomyces cerevisiae cell culture experiments with perturbed growth conditions were designed so that the metabolic states would be shifted from one to another. Microarrays were used to explore changes in gene expression across the entire yeast genome during the perturbation experiments. Changes in transcript abundance in these growth periods were investigated to study the cellular response to different glucose and oxygen supply. Computational results and experimental observations representing the three characteristic metabolic states were compared on the S. cerevisiae metabolic pathways, as well as the visualization platform provided by the metabolic phenotypic phase plane (PhPP) for the gene regulation on cell metabolism and adaptation of cells to environmental changes. CONCLUSIONS: The integrated expression study described reveals that S. cerevisiae cells respond to environmental changes mainly by down-regulating a number of genes to alter the cell metabolism so that the cells adapt to the variations in their growth conditions. Copyright © 2009 Society of Chemical Industry [source]

    Hepcidin Regulation in Wild-Type and Hfe Knockout Mice in Response to Alcohol Consumption: Evidence for an Alcohol-Induced Hypoxic Response

    ALCOHOLISM, Issue 8 2009
    Mandy L. Heritage
    Background,/Aims:, Expression of Hamp1, the gene encoding the iron regulatory peptide hepcidin, is inappropriately low in HFE-associated hereditary hemochromatosis and Hfe knockout mice (Hfe,/,). Since chronic alcohol consumption is also associated with disturbances in iron metabolism, we investigated the effects of alcohol consumption on hepcidin mRNA expression in Hfe,/, mice. Methods:,Hfe,/, and C57BL/6 (wild-type) mice were pair-fed either an alcohol liquid diet or control diet for up to 8 weeks. The mRNA levels of hepcidin and ferroportin were measured at the mRNA level by RT-PCR and protein expression of hypoxia inducible factor-1 alpha (HIF-1,) was measured by western blot. Results:,Hamp1 mRNA expression was significantly decreased and duodenal ferroportin expression was increased in alcohol-fed wild-type mice at 8 weeks. Time course experiments showed that the decrease in hepcidin mRNA was not immediate, but was significant by 4 weeks. Consistent with the genetic defect, Hamp1 mRNA was decreased and duodenal ferroportin mRNA expression was increased in Hfe,/, mice fed on the control diet compared with wild-type animals and alcohol further exacerbated these effects. HIF-1, protein levels were elevated in alcohol-fed wild-type animals compared with controls. Conclusion:, Alcohol may decrease Hamp1 gene expression independently of the HFE pathway possibly via alcohol-induced hypoxia. [source]

    On Gene Ranking Using Replicated Microarray Time Course Data

    BIOMETRICS, Issue 1 2009
    Yu Chuan Tai
    Summary Consider the ranking of genes using data from replicated microarray time course experiments, where there are multiple biological conditions, and the genes of interest are those whose temporal profiles differ across conditions. We derive a multisample multivariate empirical Bayes' statistic for ranking genes in the order of differential expression, from both longitudinal and cross-sectional replicated developmental microarray time course data. Our longitudinal multisample model assumes that time course replicates are independent and identically distributed multivariate normal vectors. On the other hand, we construct a cross-sectional model using a normal regression framework with any appropriate basis for the design matrices. In both cases, we use natural conjugate priors in our empirical Bayes' setting which guarantee closed form solutions for the posterior odds. The simulations and two case studies using published worm and mouse microarray time course datasets indicate that the proposed approaches perform satisfactorily. [source]

    A novel microplate-based screening strategy to assess the cellulolytic potential of Trichoderma strains

    BIOTECHNOLOGY & BIOENGINEERING, Issue 3 2010
    Stefano Cianchetta
    Abstract Bioconversion of lignocellulosic biomass to fuel requires a hydrolysis step to obtain fermentable sugars, generally accomplished by fungal enzymes. An assorted library of cellulolytic microbial strains should facilitate the development of optimal enzyme cocktails specific for locally available feedstocks. Only a limited number of strains can be simultaneously assayed in screening based on large volume cultivation methods, as in shake flasks. This study describes a miniaturization strategy aimed at allowing parallel assessment of large numbers of fungal strains. Trichoderma strains were cultivated stationary on microcrystalline cellulose using flat bottom 24-well plates containing an agarized medium. Supernatants obtained by a rapid centrifugation step of the whole culture plates were evaluated for extracellular total cellulase activity, measured as filter paper activity, using a microplate-based assay. The results obtained were consistent with those observed in shake-flask experiments and more than 300 Trichoderma strains were accordingly characterized for cellulase production. Five strains, displaying on shake-flasks at least 80% of the activity shown by the hyper-cellulolytic mutant Trichoderma Rut-C30, were correctly recognized by the screening on 24-well plates, demonstrating the feasibility of this approach. Cellulase activity distribution for the entire Trichoderma collection is also reported. One strain (T. harzianum Ba8/86) displayed the closest profile to the reference strain Rut-C30 in time course experiments. The method is scalable and addresses a major bottleneck in screening programs, allowing small-scale parallel cultivation and rapid supernatant extraction. It can also be easily integrated with high-throughput enzyme assays and could be suitable for automation. Biotechnol. Bioeng. 2010;107: 461,468. © 2010 Wiley Periodicals, Inc. [source]