• Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Current Issue in 2019

Volume: 9 Issue: 13

Biochemistry

Adhesive and Cohesive Peel Force Measurement of Human Airway Mucus Authors:  Henry P. Goodell, Siddharth K. Shenoy, Nathan T. Shenkute, Elijah Lackey, Robert G. Dennis and Brian Button, date: 07/05/2019, view: 316, Q&A: 0
In health, the high-speed airflow associated with cough represents a vital backup mechanism for clearing accumulated mucus from our airways. However, alterations in the mucus layer in cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD) leads to the mucus layer adhered to the airway surfaces, representing the nidus of chronic lung ...
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Oxygen Consumption Measurements in Caenorhabditis elegans Using the Seahorse XF24 Authors:  Suraiya Haroon and Marc Vermulst, date: 07/05/2019, view: 170, Q&A: 0
Mitochondria generate 90% of the energy required to sustain life. As a result, loss of mitochondrial function compromises almost every facet of human physiology. Accordingly, most mitochondrial diseases tend to present themselves as complex, multi-systemic disorders that can be difficult to diagnose. Depending on the severity of the mitochondrial ...
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Biophysics

Dynamic and Sequential Protein Reconstitution on Negatively Curved Membranes by Giant Vesicles Fusion Authors:  Nicola de Franceschi, Maryam Alqabandi, Winfried Weissenhorn and Patricia Bassereau, date: 07/05/2019, view: 185, Q&A: 0
In vitro investigation of the interaction between proteins and positively curved membranes can be performed using a classic nanotube pulling method. However, characterizing protein interaction with negatively curved membranes still represents a formidable challenge. Here, we describe our recently developed approach based on ...
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Cell Biology

The Chick Chorioallantoic Membrane (CAM) Assay as a Three-dimensional Model to Study Autophagy in Cancer Cells Authors:  Félice A. Janser, Patricia Ney, Marta Teixeira Pinto, Rupert Langer and Mario P. Tschan, date: 07/05/2019, view: 213, Q&A: 0
The chick chorioallantoic membrane (CAM) is an extra-embryonic organ and thus well accessible for seeding and harvesting 3D cell cultures. Samples from CAM assays are suitable for protein and gene expression analysis as well as for immuno-histochemical studies. Here we present the CAM assay as a possible model to study autophagy in different types ...
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Immunology

Morphological Evaluation of Wound Healing Events in the Excisional Wound Healing Model in Rats Authors:  Lígia Reis de Moura Estevão, Puebla Cassini-Vieira, Ana Greice Borba Leite, Apolônia Agnes Vilar de Carvalho Bulhões, Lucíola da Silva Barcelos and Joaquim Evêncio-Neto, date: 07/05/2019, view: 203, Q&A: 0
Skin wound healing is a complex process involving different events such as blood coagulation, inflammation, new blood vessels formation, and extracellular matrix deposition. These events can be observed by using histology techniques. However, the lack of the standardization of such parameters impacts on the reproducibility of results. Here, we ...
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In vitro Infection of Primary Human Monocytes with HIV-1 Authors:  Patrick O’Connell, Yong-Hui Zheng, Andrea Amalfitano and Yasser A. Aldhamen, date: 07/05/2019, view: 232, Q&A: 0
Monocyte infection by HIV-1 is an important component of chronic HIV pathogenesis. Following infection by HIV-1, monocytes are able to cross the blood brain barrier and set up a viral reservoir in the central nervous system. Additionally, in the setting of chronic HIV-1 infection, monocytes can become activated either directly through HIV-1 ...
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Microbiology

Preparation and Purification of Active Recombinant Human Pancreatic Lipase in Escherichia coli Authors:  Nanami Kawaguchi, Haruko Ogawa and Kimie Date, date: 07/05/2019, view: 186, Q&A: 0
Human pancreatic lipase (HPL) is the main lipolytic enzyme involved in the digestion of dietary fat. An active recombinant human pancreatic lipase (recHPL) was successfully prepared for the first time in an Escherichia coli (E. coli) expression system using a short Strep-tag II (ST II). The recHPL-ST II was solubilized with 8 M urea from ...
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In vitro and in vivo Assessment of Protein Acetylation Status in Mycobacteria Authors:  Krishna K. Singh, Devendra P. Singh, Rambir Singh and Deepak K. Saini, date: 07/05/2019, view: 196, Q&A: 0
Protein acetylation is one of the standard post-translational modifications found in proteins across all organisms, along with phosphorylation which regulates diverse cellular processes. Acetylation of proteins can be enzymatically catalyzed through acetyltransferases, acetyl CoA synthetases or non-enzymatically through acyl carrier metabolic ...
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Gentamicin Protection Assay to Determine the Number of Intracellular Bacteria during Infection of Human TC7 Intestinal Epithelial Cells by Shigella flexneri Authors:  Atin Sharma and Andrea Puhar, date: 07/05/2019, view: 252, Q&A: 0
Shigella flexneri is an intracellular bacterial pathogen that gains access to the gut epithelium using a specialized Type III Secretion System (T3SS). Various determinants mediating this invasive infection have been experimentally verified using the classical gentamicin protection assay presented here. In this assay epithelial cell lines ...
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Plaque Assay to Determine Invasion and Intercellular Dissemination of Shigella flexneri in TC7 Human Intestinal Epithelial Cells Authors:  Atin Sharma and Andrea Puhar, date: 07/05/2019, view: 216, Q&A: 0
Shigella flexneri invades the epithelial cells lining the gut lumen and replicates intracellularly. The specialized Type III Secretion System (T3SS) and its effector proteins, encoded on a large virulence plasmid, assist the bacterium to gain access to the cytosol. Thereafter Shigella disseminates to neighboring cells in an ...
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Fluorescence Microscopy Assay to Measure HIV-1 Capsid Uncoating Kinetics in vitro Authors:  Chantal L. Márquez, Derrick Lau, James Walsh, K. M. Rifat Faysal, Michael W. Parker, Stuart G. Turville and Till Böcking, date: 07/05/2019, view: 413, Q&A: 0
The stability of the HIV-1 capsid and the spatiotemporal control of its disassembly, a process called uncoating, need to be finely tuned for infection to proceed. Biochemical methods for measuring capsid lattice disassembly in bulk are unable to resolve intermediates in the uncoating reaction. We have developed a single-particle fluorescence ...
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Molecular Biology

Detection of Heteroplasmic Variants in the Mitochondrial Genome through Massive Parallel Sequencing Authors:  Joke Mertens, Filippo Zambelli, Dorien Daneels, Ben Caljon, Karen Sermon and Claudia Spits, date: 07/05/2019, view: 175, Q&A: 0
Detecting heteroplasmies in the mitochondrial DNA (mtDNA) has been a challenge for many years. In the past, Sanger sequencing was the main option to perform this analysis, however, this method could not detect low frequency heteroplasmies. Massive Parallel Sequencing (MPS) provides the opportunity to study the mtDNA in depth, but a controlled ...
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An Efficient Approach for RNA Extraction from Boar Sperm and Seminal Plasma Authors:  Vanmathy R. Kasimanickam and Ramanathan K. Kasimanickam, date: 07/05/2019, view: 202, Q&A: 0
Despite transcriptional silencing in mature sperm and cytoplasmic expulsion of RNA during the final sperm maturation process, thousands of RNAs have been successfully identified in ejaculated sperm. Although most of RNAs’ function is still unknown, it is suggested that sperm RNAs have a vital biological role in fertilization and post-fertilization ...
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Optogenetic Inactivation of Transcription Factors in the Early Embryo of Drosophila Authors:  Stephen L. McDaniel and Melissa M. Harrison, date: 07/05/2019, view: 265, Q&A: 0
The early embryo of Drosophila melanogaster exists as a rapidly dividing syncytium of nuclei that are transcriptionally silent. Maternally deposited factors are required to awaken the genome and assist in the transition from maternal to zygotic control of development. Because many of these essential factors are maternally deposited and ...
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Stem Cell

CRISPR/Cas9 + AAV-mediated Intra-embryonic Gene Knocking in Mice Authors:  Naoaki Mizuno, Eiji Mizutani, Hideyuki Sato, Mariko Kasai, Hiromitsu Nakauchi and Tomoyuki Yamaguchi, date: 07/05/2019, view: 239, Q&A: 0
Intra-embryo genome editing by CRISPR/Cas9 has enabled rapid generation of gene knockout animals. However, large fragment knock-in directly into embryos’ genome is still difficult, especially without microinjection of donor DNA. Viral vectors are good transporters of knock-in donor DNA for cell lines, but seemed unsuitable for pre-implantation ...
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