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Anal Biochem. 2003 Jun 15;317(2):186-200.
Modeling of DNA hybridization kinetics for spatially resolved biochips.

Erickson D, Li D, Krull UJ.

Department of Mechanical and Industrial Engineering, University of Toronto, 5 Kings College Road, M5S 3G8, Toronto, Ont., Canada

The marriage of microfluidics with detection technologies that rely on highly selective nucleic acid hybridization will provide improvements in bioanalytical methods for purposes such as detection of pathogens or mutations and drug screening. The capability to deliver samples in a controlled manner across a two-dimensional hybridization detection platform represents a substantial technical challenge in the development of quantitative and reusable biochips. General theoretical and numerical models of heterogeneous hybridization kinetics are required in order to design and optimize such biochips and to develop a quantitative method for online interpretation of experimental results. In this work we propose a general kinetic model of heterogeneous hybridization and develop a technique for estimating the kinetic coefficients for the case of well-spaced, noninteracting surface-bound probes. The experimentally verified model is then incorporated into the BLOCS (biolab-on-a-chip simulation) 3D microfluidics finite element code and used to model the dynamic hybridization on a biochip surface in the presence of a temperature gradient. These simulations demonstrate how such a device can be used to discriminate between fully complementary and single-base-pair mismatched hybridization using fluorescence detection by interpretation of the unique spatially resolved intensity pattern. It is also shown how the dynamic transport of the targets is likely to affect the rate and location of hybridization as well as that, although nonspecific hybridization is present, the change in the concentration of hybridized targets over the sensor platform is sufficiently high to determine if a fully complementary match is present. Practical design information such as the optimum transport speed, target concentration, and channel height is presented. The results presented here will aid in the interpretation of results obtained with such a temperature-gradient biochip.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12758257&dopt=Abstract [PubMed - in process]

Microbes Infect. 2003 May;5(6):481-5.
Vibrio parahaemolyticus infectious for both humans and edible mollusk abalone.

Lee KK, Liu PC, Huang CY.

Department of Aquaculture, National Taiwan Ocean University, 2 Pei-Ning Road, ROC, Keelung, Taiwan

The aims of this study are to report evidence of the first laboratory-acquired infection of Vibrio parahaemolyticus associated with handling experimentally infected abalones and to describe the virulence of the two bacterial strains tested in these animals. Two strains of V. parahaemolyticus, one from the stool of a patient with acute gastroenteritis (strain 880713) and the other from the hemolymph of a diseased small abalone Haliotis diversicolor supertexta (strain 880915), were identified and characterized. Both strains were lethal to small abalone, with similar LD(50) values (8.36-8.41 x 10(4) colony-forming units/g abalone). Laboratory-acquired infection resulted in one individual experiencing two episodes of acute gastroenteritis due to handling virulence tests during a 1-week interval. Our present results suggest that a V. parahaemolyticus strain isolated from the stool of a patient with gastroenteritis was infectious for small abalone, a major species of edible mollusk abalone cultured in Taiwan, while a similar strain isolated from hemolymph of a diseased small abalone was infectious for humans. This is the first report of V. parahaemolyticus virulent to small abalone as a zoonotic pathogen.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12758276&dopt=Abstract [PubMed - in process]

Microbes Infect. 2003 May;5(6):515-26.
The critical role of adenosine A(2A) receptors in downregulation of inflammation and immunity in the pathogenesis of infectious diseases.

Thiel M, Caldwell CC, Sitkovsky MV.

Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 10/11N311, 10 Center Drive, Bethesda, MD 20892-1892, USA

Adenosine can be described as a retaliatory metabolite, the production and release of which is usually enhanced under adverse environmental conditions. Binding via specific receptors, adenosine activates endogenous protective mechanisms aiming at the restoration of tissue homeostasis. While adenosinergic downregulation of tissue damage is beneficial in acute inflammation, chronic suppression of the immune system by adenosine may account for immunoparalysis in long-term septic patients.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12758281&dopt=Abstract [PubMed - in process]

Microbes Infect. 2003 May;5(6):527-34.
Responses to reactive oxygen intermediates and virulence of Salmonella typhimurium.

Janssen R, van der Straaten T, van Diepen A, van Dissel JT.

Department of Infectious Diseases, C5-P, Leiden University Medical Center, P.O. Box 9600, 2300 RC, The, Leiden, Netherlands

Salmonella typhimurium is an intracellular pathogen that can survive and replicate in macrophages. One of the host defense mechanisms that S. typhimurium encounters upon infection is superoxide produced by the phagocytes' NADPH-oxidase. Salmonella has evolved numerous ways of coping with superoxide in the extracellular environment. In addition, Salmonella has to defend itself against superoxide produced as a by-product of aerobic respiration. Over the last decade, research on bacterial mutants has led to the identification of Salmonella strains that differ from their parental strain in susceptibility to superoxide in vitro. However, the consequences of such mutations for bacterial virulence are highly variable, indicating that superoxide sensitivity per se is not a characteristic that renders Salmonella less virulent. By discussing various bacterial mutants classified according to their in vitro sensitivity to superoxide, we will exemplify the complex mechanisms that Salmonella has evolved to cope with superoxide stress.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12758282&dopt=Abstract [PubMed - in process]

J Lipid Res. 2003 Mar;44(3):601-11. Epub 2003 Jan 01.
Different effects of n-6 and n-3 polyunsaturated fatty acids on the activation of rat smooth muscle cells by interleukin-1 beta.

Bousserouel S, Brouillet A, Bereziat G, Raymondjean M, Andreani M.

UMR Physiologie et Physiopathologie, Universite Pierre et Marie Curie, case courrier 256, Batiment A, 5eme etage, 7 quai Saint Bernard, 75252 Paris Cedex 5, France.

There is good evidence that the n-3 polyunsaturated fatty acids (PUFAs) in fish oil have antiinflammatory effects and reduce the pathogenesis of atherosclerosis. However, the mechanisms underlying these actions are largely unknown. This study was designed to investigate the effects of membrane incorporation of two major components of fish oil [eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)], on rat smooth muscle cells (SMCs) activation induced by interleukin-1 beta (IL1 beta). We compared their effects with those of n-6 arachidonic acid (AA). Expression of vascular cell adhesion molecule-1 and monocyte chemoattractant protein-1 adhesion molecules involved in SMCs migration was enhanced by AA, whereas EPA and DHA had no similar effects. We established that AA potentiates IL1 beta-induced expression of the type IIA secreted phospholipase A2 (sPLA2) gene, whereas EPA and DHA reduce this stimulation. EPA and DHA also abolished proinflammatory prostaglandin PGE2 production by inhibiting the IL1 beta-induced production of cyclooxygenase-2 (COX-2) mRNA. Much interest was then focused on three transcriptional factors implicated in inflammation control and especially in modulating rat sPLA2 and COX-2 gene transcription: nuclear factor-kappa B, CCAAT/enhancer binding protein beta, and E26 transformation-specific-1. electrophoretic mobility shift assay revealed that the binding activity of all three factors was increased by AA and reduced (or not affected) by n-3 PUFA. These results indicate that EPA and DHA act in opposition to AA by modulating various steps of the inflammatory process induced by IL1 beta, probably by reducing mitogen-activated protein kinase p42/p44 activity.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12562859&dopt=Abstract

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