Vitro methods

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  • Bringing together the recent and relevant contributions of over 125 scientists from industry, government, and academia in North America and Western Europe, Alternative Toxicological Methods explores the development and validation of replacement, reduction, and refinement alternatives (the 3Rs) to animal testing. Internationally recognized scientists present what has been accomplished thus far in developing acceptable alternatives to traditional animal toxicological assessment and provide potentially new initiatives.

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  • The molecular definition of tumor antigens, costimulatory signals, and the possibility to genetically engineer tumor cells as well as simple protocols for efficient isolation and preparation of dendritic cells (DC) renew the interest in tumor immunotherapy and vaccination, in particular. Engineering of tumor cells with the gene of a particular cytokine is a way of releasing that cytokine at the tumor site. In contrast to bolus administration, it provides a constant supply of cytokine.

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  • A novel plasmid vector pSELECT-1 is described which can be used for highly efficient site-directed in vitro mutagenesis. The mutagenesis method is based on the use of single-stranded DNA and two primers, one mutagenic primer and a second correction primer which corrects a defect in the ampicillin resistance gene on the vector and reverts the vector to ampicillin resistance. Using T4 DNA polymerase and T4 DNA ligase the two primers are physically linked on the template. The non-mutant DNA strand is selected against by growth in the presence of ampicillin.

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  • The isomeranzin, a coumarin was isolated from the leaves of Clausena heptaphylla. The structure was elucidated by IR, MS, 1H and 13C-NMR. For the first time, the anti-Herpes simplex virus type 1 & 2 in vitro activity of the compound was reported and discussed. I - introduction Clausena heptaphylla (Roxb.) W. & Arn. (Rutaceae) (Vietnamese name: Giæi 7 l¸) is a tropical tree [1]. This species is also widely used in traditional medicine. In Asia (China, India, Vietnam, etc.), it is used for fever.

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  • Cell culture is practiced extensively throughout the world today. The techniques required to allow cells to grow and be maintained outside the body have been developed throughout the 20th century. In the 50 years since the publication of the first human cancer cell line, HeLa (1), thousands of cell lines representing most of the spectrum of human cancer have been derived. These have provided tools to study in depth the biochemistry and molecular biology associated with individual cancer types and have helped enormously in our understanding of normal as well as cancer cell physiology.

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  • The past three decades have witnessed significant advances in the field of assisted human conception. Following the remarkable perseverance and triumph of Robert Edwards, Patrick Steptoe, and Jean Purdy, numerous scientists and physicians from around the world have worked to develop more effective and safer procedures to treat infertile couples.

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  • Tuyển tập các báo cáo nghiên cứu về lâm nghiệp được đăng trên tạp chí lâm nghiệp Original article đề tài: Efficient method of micropropagation and in vitro rooting of teak (Tectona grandis L.) focusing on large-scale industrial plantations...

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  • Meliton et al. Journal of Inflammation 2010, 7:14 http://www.journal-inflammation.com/content/7/1/14 RESEARCH Research Open Access Cytosolic group IVa phospholipase A2 mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro Angelo Y Meliton1, Nilda M Muñoz*1, Lucille N Meliton1, David C Binder1, Christopher M Osan1, Xiangdong Zhu1, Steven M Dudek1 and Alan R Leff1,2 Abstract Background: Cytosolic gIVaPLA2 is a critical enzyme in the generation of arachidonate metabolites and in induction of β2-integrin adhesion in granulocytes.

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  • An efficient in vitro-inoculation method for Tomato yellow leaf curl virus

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  • Tuyển tập các báo cáo nghiên cứu về bệnh thú y được đăng trên tạp chí y học General Psychiatry cung cấp cho các bạn kiến thức về ngành thú y đề tài: No Peri- and Postnatal Effects on Calves Born After Transfer of in Vitro Produced Embryos Vitrified by the Open Pulled Straw (OPS) Method...

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  • RNA interference (RNAi) is a powerful tool to silence gene expression post-transcriptionally. Delivering sequences of RNAi in vivo remains a problem. The aim of this study was to use JC virus (JCV) virus-like particles (VLPs) as a vector for delivering RNAi in silencing the cytokine gene of IL-10. Methods: JCV VLPs were generated by recombinant JCV VP1 protein in yeast expression system. DNA fragment containing IL-10 shRNA was packaged into VLPs by osmotic shock. Results: In RAW 264.7 cells, IL-10 shRNA was found to reduce IL-10 expression by 85 to 89%, as compared with VLPs alone.

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  • Previously, we demonstrated apoptotic cell death in the chorion laeve trophoblast layer of human fetal membrane tissues during the late stages of pregnancy, the progression of apoptosis during incubation in vitro, and its suppression by a low concentration of glucocorticoid hormones.

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  • A novel method, based upon primer extension, has been developed for measuring the reopening temperature of a single type of DNA hairpin structure. Two DNA oligo-nucleotides have been utilized and designated as primers 1 and2.Primer1,withits5- and3¢-termini fully comple-mentary to the hairpin flanking sequences, was used to evaluate primer extension conditions, and primer 2, with its 3¢-end competing with the DNA hairpin stem, was used to detect the DNA hairpin reopening temperature.

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  • The (ba)8-barrel is the most common enzyme fold and it is capable of cata-lyzing an enormous diversity of reactions. It follows that this scaffold should be an ideal starting point for engineering novel enzymes by directed evolution. However, experiments to date have utilized in vivo screens or selections and the compatibility of (ba)8-barrels with in vitro selection methods remains largely untested.

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  • Great efficiencies have been achieved in the drug discovery process as a result of technological advances in target identification, high-throughput screening, high-throughput organic synthesis, just-in-time in vitro ADME (absorption, distribution, metabolism, and excretion), and early pharmacokinetic screening of drug leads. These advances, spanning target selection all the way through to clinical candidate selection, have placed greater and greater demands on the analytical community to develop robust high-throughput methods.

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  • This book describes many aspects of tissue culture models in an extensive manner.

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  • SPB2 2/27/2003 4:06 PM Page 35 2 Plant tissue culture Introduction Most methods of plant transformation applied to GM crops require that a whole plant is regenerated from isolated plant cells or tissue which have been genetically transformed. This regeneration is conducted in vitro so that the environment and growth medium can be manipulated to ensure a high frequency of regeneration. In addition to a high frequency of regeneration, the regenerable cells must be accessible to gene transfer by whatever technique is chosen (gene transfer methods are described in Chapter 3).

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  • In this study, structural features of Tricholoma matsutake polysaccharide (TMP-A) were investigated by a combination of infrared (IR) spectra, gas chromatography–mass spectrometry (GC–MS), nuclear magnetic resonance (NMR) spectroscopy. The results indicated that TMP-A had a backbone of 1,4-- d-glucopyranose residue which branches at O-6 based on the experimental results. The branches were mainly composed of an (1→3)--d-galactopyranose residue, and terminated with -d-xylopyranose residue.

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  • Toll-like receptor 4 (TLR4) plays an important role in innate immunity. The role of TLR4 in stretched cardiomyocytes is not known. We sought to investigate whether mechanical stretch could regulate TLR4 expression, as well as the possible molecular mechanisms and signal pathways mediating the expression of TLR4 by cyclic mechanical stretch in cardiomyocytes. Methods: Neonatal Wistar rat cardiomyocytes grown on a flexible membrane base were stretched by vacuum to 20% of maximum elongation at 60 cycles/min.

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  • Sindbis virus (SV) is the prototype of alphaviruses which are a group of widely distributed human and animal pathogens. Heterogeneous nuclear ribonucleoprotein (hnRNP) A1 is an RNA-binding protein that shuttles between the nucleus and the cytoplasm. Our recent studies found that hnRNP A1 relocates from nucleus to cytoplasm in Sindbis virus (SV)-infected cells. hnRNP A1 binds to the 5′ UTR of SV RNA and facilitates the viral RNA replication and translation.

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