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Past Issues in 2014

Volume: 4 Issue: 19

Cancer Biology

Hypoxia Studies with Pimonidazole in vivo Authors: Kristina Y. Aguilera and Rolf A. Brekken, date: 10/05/2014, view: 16067, Q&A: 0

Therapy-induced hypoxia drives changes in the tumor microenvironment that contribute to the poor response to therapy. Hypoxia is capable of driving the expression and/or activation of specific signaling cascades (e.g., c-Met, Axl, CTGF), the recruitment of tumor promoting immune cells, and the induction of cell survival pathways including ...

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Immunology

Coagulation Assay Authors: Praveen Papareddy, Martina Kalle and Artur Schmidtchen, date: 10/05/2014, view: 8443, Q&A: 0

Clotting times can be measured by using citrate plasma. The intrinsic pathway of coagulation is measured by the activated partial thromboplastin time (aPTT), the extrinsic pathway of coagulation, monitored by measuring the prothrombin time (PT), and thrombin-induced fibrin-network formation (thrombin clotting time; TCT).

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Isolation and in vivo Transfer of Antigen Presenting Cells Authors: Pooja Arora, Shalu Sharma Kharkwal and Steven A. Porcelli, date: 10/05/2014, view: 7801, Q&A: 0

Transfer of antigen presenting cells in vivo is a method used by immunologists to examine the potency of antigen presentation by a selected population of cells. This method is most commonly used to analyze presentation of protein antigens to MHC class I or II restricted T cells, but it can also be used for studies of nonconventional ...

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Microbiology

Chromogenic Substrate Assay for Determining the Activity of Plasma Kallikrein Authors: Praveen Papareddy, Martina Kalle and Artur Schmidtchen, date: 10/05/2014, view: 5800, Q&A: 0

The activation of the intrinsic pathway takes place at negatively charged surfaces, such as bacteria, and involves activation of cogulation Factor XII, which then leads to the activation of plasma kallikrein (PK) and coagulation Factor XI. To determine the PK activity on bacterial surfaces, bacteria were pre-incubated with peptides, followed by ...

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Construction of Glycine Oxidase Mutant Libraries by Random Mutagenesis, Site Directed Mutagenesis and DNA Shuffling Author: Tao Zhan, date: 10/05/2014, view: 6421, Q&A: 0

Glyphosate, a broad spectrum herbicide widely used in agriculture all over the world, inhibits 5-enolpyruvylshikimate-3-phosphate synthase in the shikimate pathway, and glycine oxidase (GO) has been reported to be able to catalyze the oxidative deamination of various amines and cleave the C-N bond in glyphosate (Pedotti et al., 2009). ...

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Expression and Purification of the Thermus thermophilus Argonaute Protein Authors: Daan C. Swarts, Matthijs M. Jore and John van der Oost, date: 10/05/2014, view: 6507, Q&A: 0

The Argonaute protein of Thermus thermophilus (TtAgo) has recently been studied in detail. For its in vitro characterization, TtAgo was purified after heterologous expression in Escherichia coli (E. coli). As TtAgo expression is toxic, a tightly controlled system was used for protein ...

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Neuroscience

Novel Object Recognition for Studying Memory in Mice Authors: Tzyy-Nan Huang and Yi-Ping Hsueh, date: 10/05/2014, view: 22094, Q&A: 0

Memory tests are important indexes of the brain functions for rodents behavior assay. Many memory tasks require external forces (e.g. electric shocks) or intrinsic forces (e.g. hunger and thirsty) to trigger the responses. Under those conditions, rodents are under stresses, such as pain, tired, malnutrition or dehydration, which ...

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Two-choice Digging Task in Mouse for Studying the Cognitive Flexibility Authors: Hsiu-Chun Chuang, Tzyy-Nan Huang and Yi-Ping Hsueh, date: 10/05/2014, view: 7715, Q&A: 1

Cognitive flexibility, the higher-order cognition involving reversal learning, has been defined as having the ability to shift one’s previous thoughts or actions to new situations depending on situational demands. Studies of neuropsychiatric disorders such as autism spectrum disorder (ASD) showed that restricted and repetitive patterns of ...

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In utero Electroporation of the Embryonic Mouse Retina Authors: Francisco Nieto-Lopez and Luisa Sanchez-Arrones, date: 10/05/2014, view: 9740, Q&A: 0

This protocol is useful to manipulate gene expression in the embryonic retina and compare the result with the contralateral non electroporated retina. In addition, the electroporation of a membrane or cytoplasmic tagged GFP allows to determine the effects of gene manipulation on the outgrowth of retinal ganglion cell axons (Garcia-Frigola et al. ...

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Plant Science

Analysis of Total Se Content in Rice Authors: Lianhe Zhang, Feiyan Yu, Kun Deng, Bin Hu and Chengcai Chu, date: 10/05/2014, view: 4882, Q&A: 0

Total Se content in rice is normally low and it is difficult to determine it exactly because of Se volatilization and pollution during the digestion process. In this method, rice sample is digested thoroughly and Se volatilization is reduced greatly by designing a specific digestion tube, increasing digestion temperature by three steps, ...

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Pea Aphid Survival Assays on Arabidopsis thaliana Authors: David C. Prince, Sam T. Mugford, Thomas R. Vincent and Saskia A. Hogenhout, date: 10/05/2014, view: 6012, Q&A: 0

Aphids are phloem-feeding insects that successfully colonize specific host plant species. Aphid performance on a given plant is commonly measured by assessing fecundity of an aphid species that is adapted to the host. However, this approach may not reveal roles for plant genes in defense pathways that adapted aphids suppress. The host range of the ...

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Carotenoid Extraction and Quantification from Capsicum annuum Authors: Richard D. Richins, James Kilcrease, Laura Rodgriguez-Uribe and Mary A. O’Connell, date: 10/05/2014, view: 8250, Q&A: 0

Carotenoids are ubiquitous pigments that play key roles in photosynthesis and also accumulate to high levels in fruit and flowers. Specific carotenoids play essential roles in human health as these compounds are precursors for Vitamin A; other specific carotenoids are important sources of macular pigments and all carotenoids are important ...

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Endogenous ABA Extraction and Measurement from Arabidopsis Leaves Authors: Ning Liu, Yong Ding, Michael Fromm and Zoya Avramova, date: 10/05/2014, view: 7497, Q&A: 0

The endogenous messenger, the phytohormone abscisic acid (ABA) plays a major role in plant’s adaption to drought, salinity, cold and other abiotic stresses. In addition to abiotic stress signaling, ABA is involved also in developmental regulation and in responses to diverse biotic stresses. Dehydration stress results in a strong increase in ...

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Agrobacterium tumefaciens-mediated Transformation of Walnut (Juglans regia) Authors: Sriema L. Walawage, Charles A. Leslie, Matthew A. Escobar and Abhaya M. Dandekar, date: 10/05/2014, view: 7785, Q&A: 0

Like many woody plant species, walnut (Juglans regia) can be difficult to genetically transform and regenerate. However, somatic embryos have been used successfully for over two decades as a target tissue for transformation and regeneration of transgenic walnut plants. Walnut somatic embryos, initiated originally from developing zygotic ...

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Localisation and Quantification of Reactive Oxygen Species and Nitric Oxide in Arabidopsis Roots in Response to Fungal Infection Authors: Kapuganti J. Gupta, Yariv Brotman and Luis A. J. Mur, date: 10/05/2014, view: 7881, Q&A: 0

Nitric oxide and reactive oxygen species have emerged as important signalling molecules in plants. The half-lives of NO and ROS are very short therefore rapid and precise measurements are required for the understanding biological roles of these redox active species. Various organelles and compartments generate NO and ROS thus it is important to ...

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