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J Li, Z Li, S Liu, R Zia, A Liang, L Yang
The normal maturation and ovulation from ovarian follicles is important in ensuring conception and improving fertility of buffalo. The molecular regulation mechanism of buffalo follicles growth, however, remains unknown. This study analyzed the gene expression profiles associated with buffalo ovarian follicle growth. According to the analysis of RNA sequencing, 17,700 unigenes and 13,672 differentially expressed genes (DEGs) were detected. A total of 30 common DEGs were identified during four stages of follicle growth, and the expression patterns are basically synchronized, suggesting the products as a result of expressions of these genes may cooperate to regulate follicular development...
November 6, 2017: Animal Reproduction Science
Charlie J Pyle, Abul K Azad, Audrey C Papp, Wolfgang Sadee, Daren L Knoell, Larry S Schlesinger
Tuberculosis (TB) is a global epidemic caused by the infection of human macrophages with the world's most deadly single bacterial pathogen, Mycobacterium tuberculosis (M.tb). M.tb resides in a phagosomal niche within macrophages, where trace element concentrations impact the immune response, bacterial metal metabolism, and bacterial survival. The manipulation of micronutrients is a critical mechanism of host defense against infection. In particular, the human zinc transporter Zrt-/Irt-like protein 8 (ZIP8), one of 14 ZIP family members, is important in the flux of divalent cations, including zinc, into the cytoplasm of macrophages...
November 9, 2017: International Journal of Molecular Sciences
Arshad Khan, Lovepreet Mann, Ramesha Papanna, Mi-Ae Lyu, Christopher R Singh, Scott Olson, N Tony Eissa, Jeffrey Cirillo, Gobardhan Das, Robert L Hunter, Chinnaswamy Jagannath
Human mesenchymal stem cells (MSCs) express scavenger receptors that internalize lipids, including oxidized low-density lipoprotein (oxLDL). We report that MSCs phagocytose Mycobacterium tuberculosis (Mtb) through two types of scavenger receptors (SRs; MARCO and SR-B1), as blockade of the receptors with antibodies or siRNA knockdown decreased the uptake of Mtb. MSCs also expressed mannose receptor (MR) that was found to endocytose rhodamine-labeled mannosylated BSA (rMBSA), though the receptor was not involved in the uptake of Mtb...
November 8, 2017: Scientific Reports
Eileen Uribe-Querol, Carlos Rosales
Phagocytosis is a fundamental process of cells to capture and ingest foreign particles. Small unicellular organisms such as free-living amoeba use this process to acquire food. In pluricellular organisms, phagocytosis is a universal phenomenon that all cells are able to perform (including epithelial, endothelial, fibroblasts, etc.), but some specialized cells (such as neutrophils and macrophages) perform this very efficiently and were therefore named professional phagocytes by Rabinovitch. Cells use phagocytosis to capture and clear all particles larger than 0...
2017: Frontiers in Immunology
Youhei Egami, Katsuhisa Kawai, Nobukazu Araki
Phagosome formation is a complicated process that requires spatiotemporally regulated actin reorganization. We found that RhoC GTPase is a critical regulator of FcγR-mediated phagocytosis in macrophages. Our live-cell imaging revealed that RhoC, but not RhoA, is recruited to phagocytic cups engulfing IgG-opsonized erythrocytes (IgG-Es). RhoC silencing by RNAi, clustered regularly interspaced short palindromic repeats (CRISPR)/Cas-mediated RhoC knockout and the expression of dominant-negative or dominant-active RhoC mutants suppressed the phagocytosis of IgG-Es...
November 7, 2017: Journal of Cell Science
Eric Cascales
Bacterial contractile injection systems are fascinating particles that use a spring-like mechanism to inject an effector-loaded needle into target cells. A recent study shows that the intracellular bacterium Amoebophilus asiaticus uses arrays of contractile structures to escape from the amoeba phagosome.
November 6, 2017: Current Biology: CB
Rikako Ogawa, Hilo Yen, Kiyoshi Kawasaki, Toru Tobe
During the course of infection, pathogens must overcome a variety of host defense systems. Modulation of lipid A, which is a strong stimulant for host immune systems, is one of the strategies used by microorganisms to evade the host response. The lpxR gene, which encodes a lipid A 3'-O-deacylase, is commonly found in several pathogens and has been shown to reduce the inflammatory response. Here, we demonstrated that the lpxR gene of enterohemorrhagic E. coli (EHEC) was positively regulated by two virulence regulators, Pch and Ler, and that this regulation was coordinated with the LEE genes, which encode major virulence factors for colonization...
November 7, 2017: Cellular Microbiology
Qian Shen, Chad A Rappleye
Mammalian body temperature triggers differentiation of the fungal pathogen Histoplasma capsulatum into yeast cells. The Drk1 regulatory kinase and an interdependent network of Ryp transcription factors establish the yeast state. Beyond morphology, the differentiation-dependent expression program equips yeasts for invasion and survival within phagosomes. Yeast cells produce α-glucan and the Eng1 endoglucanase which hide yeasts from immune detection. Secretion of yeast phase-specific Sod3 and CatB detoxify phagocyte-derived reactive oxygen molecules...
October 26, 2017: Current Opinion in Microbiology
Jinchao Liu, Meijiao Li, Lin Li, She Chen, Xiaochen Wang
Apoptotic cells generated by programmed cell death are engulfed by phagocytes and enclosed within membrane-bound phagosomes. Maturation of apoptotic cell-containing phagosomes leads to formation of phagolysosomes where cell corpses are degraded. The class III phosphatidylinositol 3-kinase (PI3-kinase) VPS-34 coordinates with PIKI-1, a class II PI3-kinase, to produce PtdIns3P on phagosomes, thus promoting phagosome closure and maturation. Here, we identified UBC-13, an E2 ubiquitin-conjugating enzyme that functions in the same pathway with VPS-34 but in parallel to PIKI-1 to regulate PtdIns3P generation on phagosomes...
November 1, 2017: Journal of Cell Biology
Sarela Garcia-Santamarina, Marta A Uzarska, Richard A Festa, Roland Lill, Dennis J Thiele
Copper (Cu) ions serve as catalytic cofactors to drive key biochemical processes, and yet Cu levels that exceed cellular homeostatic control capacity are toxic. The underlying mechanisms for Cu toxicity are poorly understood. During pulmonary infection by the fungal pathogen Cryptococcus neoformans, host alveolar macrophages compartmentalize Cu to the phagosome, and the ability to detoxify Cu is critical for its survival and virulence. Here, we report that iron-sulfur (Fe-S) clusters are critical targets of Cu toxicity in both Saccharomyces cerevisiae and C...
October 31, 2017: MBio
Amriya Naufer, Victoria E B Hipolito, Suriakarthiga Ganesan, Akriti Prashar, Vanina Zaremberg, Roberto J Botelho, Mauricio R Terebiznik
Phagocytosis of filamentous bacteria occurs through tubular phagocytic cups (tPCs) and takes many minutes to engulf these filaments into phagosomes. Contravening the canonical phagocytic pathway, tPCs mature by fusing with endosomes. Using this model, we observed the sequential recruitment of early and late endolysosomal markers to the elongating tPCs. Surprisingly, the regulatory early endosomal lipid phosphatidylinositol-3-phosphate (PtdIns(3)P) persists on tPCs as long as their luminal pH remains neutral...
October 31, 2017: Journal of Cell Biology
Stephen Y T Li, Ming Yan, Haiqi Chen, Tito Jesus, Will M Lee, Xiang Xiao, C Yan Cheng
The blood-testis barrier (BTB), conferred by Sertoli cells in the mammalian testis, is an important ultrastructure that supports spermatogenesis. Studies using animal models have shown that a disruption of the BTB leads to meiotic arrest, causing defects in spermatogenesis and male infertility. In order to better understand the regulation of BTB dynamics, we report findings herein to understand the role of rpS6 (ribosomal protein S6), a downstream signaling protein of mTORC1 (mammalian target of rapamycin complex 1), in promoting BTB disruption in the testis in vivo, making the barrier "leaky"...
October 31, 2017: American Journal of Physiology. Endocrinology and Metabolism
Abdullah R Chaudhary, Florian Berger, Christopher L Berger, Adam G Hendricks
Organelles, proteins, and mRNA are transported bidirectionally along microtubules by plus-end directed kinesin and minus-end directed dynein motors. Microtubules are decorated by microtubule-associated proteins (MAPs) that organize the cytoskeleton, regulate microtubule dynamics and modulate the interaction between motor proteins and microtubules to direct intracellular transport. Tau is a neuronal MAP that stabilizes axonal microtubules and crosslinks them into bundles. Dysregulation of tau leads to a range of neurodegenerative diseases known as tauopathies including Alzheimer's disease (AD)...
October 27, 2017: Traffic
Jessie N Green, Anna L P Chapman, Cynthia J Bishop, Christine C Winterbourn, Anthony J Kettle
The neutrophil enzyme, myeloperoxidase, by converting hydrogen peroxide (H2O2) and chloride to hypochlorous acid (HOCl), provides important defense against ingested micro-organisms. However, there is debate about how efficiently HOCl is produced within the phagosome and whether its reactions with phagosomal constituents influence the killing mechanism. The phagosome is a small space surrounding the ingested organism, into which superoxide, H2O2 and high concentrations of proteins from cytoplasmic granules are released...
October 18, 2017: Free Radical Biology & Medicine
Sónia Castanheira, Francisco García-Del Portillo
Bacteria of the Salmonella genus cause diseases ranging from gastroenteritis to life-threatening typhoid fever and are among the most successful intracellular pathogens known. After the invasion of the eukaryotic cell, Salmonella exhibits contrasting lifestyles with different replication rates and subcellular locations. Although Salmonella hyper-replicates in the cytosol of certain host cell types, most invading bacteria remain within vacuoles in which the pathogen proliferates at moderate rates or persists in a dormant-like state...
2017: Frontiers in Cellular and Infection Microbiology
María M López de Armentia, María C Gauron, María I Colombo
Staphylococcus aureus is a pathogen that causes severe infectious diseases that eventually lead to septic and toxic shock. S. aureus infection is characterized by the production of virulence factors, including enzymes and toxins. After internalization S. aureus resides in a phagosome labeled with Rab7 protein. Here, we show that S. aureus generates tubular structures marked with the small GTPases Rab1b and Rab7 and by the autophagic protein LC3 at early times post-infection. As shown by live cell imaging these tubular structures are highly dynamic, extend, branch and grow in length...
2017: Frontiers in Cellular and Infection Microbiology
Birgitta Ehrnström, Kai Sandvold Beckwith, Mariia Yurchenko, Siv Helen Moen, June Frengen Kojen, Germana Lentini, Giuseppe Teti, Jan Kristian Damås, Terje Espevik, Jørgen Stenvik
TLR8 is the major endosomal sensor of degraded RNA in human monocytes and macrophages. It has been implicated in the sensing of viruses and more recently also bacteria. We previously identified a TLR8-IFN regulatory factor 5 (IRF5) signaling pathway that mediates IFNβ and interleukin-12 (IL-12) induction by Staphylococcus aureus and is antagonized by TLR2. The relative importance of TLR8 for the sensing of various bacterial species is however still unclear. We here compared the role of TLR8 and IRF5 for the sensing of Group B Streptococcus (GBS), S...
2017: Frontiers in Immunology
Timothy M Potter, Sarah L Skoczen, Jamie C Rodriguez, Barry W Neun, Anna N Ilinskaya, Edward Cedrone, Marina A Dobrovolskaia
This chapter provides a protocol for analysis of nanoparticle effects on the function of phagocytic cells. The protocol relies on luminol chemiluminescence to detect zymosan uptake. Zymosan is an yeast particle which is typically eliminated by phagocytic cells via the complement receptor pathway. The luminol, co-internalized with zymosan, is processed inside the phagosome to generate a chemiluminescent signal. If a test nanoparticle affects the phagocytic function of the cell, the amount of phagocytosed zymosan and, proportionally, the level of generated chemiluminescent signal change...
2018: Methods in Molecular Biology
Nicolás Cuenca, Isabel Ortuño-Lizarán, Isabel Pinilla
PURPOSE: Optical coherence tomography has been a technological breakthrough in the diagnosis, treatment, and follow-up of many ocular diseases, especially retinal and neuro-ophthalmologic pathologic conditions. Until now, several controversies have arisen over the specific cell types that the bands observed in the OCT represent, especially over the 4 outer retinal bands. DESIGN: To correlate the 4 outer hyperreflective bands observed in the OCT with the histologic structures using human retinal sections and immunocytochemistry at the fovea level...
October 13, 2017: Ophthalmology
Ankur Gupta-Wright, Dumizulu Tembo, Kondwani C Jambo, Elizabeth Chimbayo, Leonard Mvaya, Shannon Caldwell, David G Russell, Henry C Mwandumba
The accurate assessment of immune competence through ex vivo analysis is paramount to our understanding of those immune mechanisms that lead to protection or susceptibility against a broad range of human pathogens. We have developed a flow cytometry-based, whole blood phagocyte functional assay that utilizes the inflammatory inducer zymosan, coupled to OxyBURST-SE, a fluorescent reporter of phagosomal oxidase activity. The assay measures both phagocytic uptake and the superoxide burst in the phagocyte populations in whole blood...
2017: Frontiers in Immunology
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