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Cell Apr 2020The recent emergence of the novel, pathogenic SARS-coronavirus 2 (SARS-CoV-2) in China and its rapid national and international spread pose a global health emergency....
The recent emergence of the novel, pathogenic SARS-coronavirus 2 (SARS-CoV-2) in China and its rapid national and international spread pose a global health emergency. Cell entry of coronaviruses depends on binding of the viral spike (S) proteins to cellular receptors and on S protein priming by host cell proteases. Unravelling which cellular factors are used by SARS-CoV-2 for entry might provide insights into viral transmission and reveal therapeutic targets. Here, we demonstrate that SARS-CoV-2 uses the SARS-CoV receptor ACE2 for entry and the serine protease TMPRSS2 for S protein priming. A TMPRSS2 inhibitor approved for clinical use blocked entry and might constitute a treatment option. Finally, we show that the sera from convalescent SARS patients cross-neutralized SARS-2-S-driven entry. Our results reveal important commonalities between SARS-CoV-2 and SARS-CoV infection and identify a potential target for antiviral intervention.
Topics: Ammonium Chloride; Angiotensin-Converting Enzyme 2; Animals; Antibodies, Neutralizing; Antibodies, Viral; Betacoronavirus; COVID-19; Cell Line; Coronavirus; Coronavirus Infections; Drug Development; Esters; Gabexate; Guanidines; Humans; Immunization, Passive; Leucine; Pandemics; Peptidyl-Dipeptidase A; Pneumonia, Viral; Protease Inhibitors; Receptors, Virus; Severe acute respiratory syndrome-related coronavirus; SARS-CoV-2; Serine Endopeptidases; Spike Glycoprotein, Coronavirus; Vesiculovirus; Virus Internalization; COVID-19 Serotherapy
PubMed: 32142651
DOI: 10.1016/j.cell.2020.02.052 -
Current Protocols in Cytometry Jul 2015Human peripheral blood is often studied by flow cytometry in both the research and clinical laboratories. The methods for collection, storage, and preparation of...
Human peripheral blood is often studied by flow cytometry in both the research and clinical laboratories. The methods for collection, storage, and preparation of peripheral blood will vary depending on the cell lineage to be examined as well as the type of assay to be performed. This unit presents protocols for collection of blood, separation of leukocytes from whole blood by lysis of erythrocytes, isolating mononuclear cells by density gradient separation, and assorted non-flow sorting methods, such as magnetic bead separations, for enriching specific cell populations, including monocytes, T lymphocytes, B lymphocytes, neutrophils, and platelets, prior to flow cytometric analysis. A protocol is also offered for cryopreservation of cells, since clinical research often involves retrospective flow cytometric analysis of samples stored over a period of months or years.
Topics: Ammonium Chloride; Antibodies; Anticoagulants; Blood Cells; Blood Platelets; Blood Specimen Collection; Cell Adhesion; Cell Fractionation; Cell Separation; Centrifugation, Density Gradient; Complement System Proteins; Cryopreservation; Erythrocytes; Humans; Indicators and Reagents; Leukocytes; Lymphocytes; Magnetic Phenomena; Microspheres; Monocytes; Plastics; Preservation, Biological; Staining and Labeling
PubMed: 26132177
DOI: 10.1002/0471142956.cy0501s73 -
Clinical Journal of the American... Feb 2018
Topics: Acid-Base Equilibrium; Acidosis, Renal Tubular; Ammonium Compounds; Humans; Kidney Diseases
PubMed: 29311217
DOI: 10.2215/CJN.13791217 -
Molecules (Basel, Switzerland) Feb 2022Quaternary ammonium salt polymers, a kind of polyelectrolyte with a quaternary ammonium group, are widely used in traditional and emerging industries due to their good... (Review)
Review
Quaternary ammonium salt polymers, a kind of polyelectrolyte with a quaternary ammonium group, are widely used in traditional and emerging industries due to their good water-solubility, adjustable cationicity and molecular weight, high efficiency and nontoxicity. In this paper, firstly, the properties and several synthesis methods of typical quaternary ammonium salt monomers were introduced. Secondly, the research progress on the synthesis of polymers was summarized from the perspective of obtaining products with high molecular weight, narrow molecular weight distribution and high monomer conversion, and special functional polymers. Thirdly, the relationships between the structures and properties of the polymer were analyzed from the perspectives of molecular weight, charge density, structural stability, and microstructural regulation of the polymer chain unit. Fourthly, typical examples of quaternary ammonium salt polymers in the application fields of water treatment, daily chemicals, petroleum exploitation, papermaking, and textile printing and dyeing were listed. Finally, constructive suggestions were put forward on developing quaternary ammonium salt polymers with high molecular weights, strengthening the research on the relationships between the structures and their properties and pinpointing relevant application fields.
PubMed: 35209058
DOI: 10.3390/molecules27041267 -
IUCrData Feb 2023The title compound, CHN·Cl·2HO, crystallizes in the space group 2 with one organic mol-ecule in the asymmetric unit. The compound belongs to a class of benzalkonium...
The title compound, CHN·Cl·2HO, crystallizes in the space group 2 with one organic mol-ecule in the asymmetric unit. The compound belongs to a class of benzalkonium chlorides (BACs) with an alkyl chain length of 16 carbon atoms in an all- conformation.
PubMed: 36911083
DOI: 10.1107/S2414314623000962 -
BioRxiv : the Preprint Server For... May 2023Ammonia is a ubiquitous, toxic by-product of cell metabolism. Its high membrane permeability and proton affinity causes ammonia to accumulate inside acidic lysosomes in...
Ammonia is a ubiquitous, toxic by-product of cell metabolism. Its high membrane permeability and proton affinity causes ammonia to accumulate inside acidic lysosomes in its poorly membrane-permeant form: ammonium (NH ). Ammonium buildup compromises lysosomal function, suggesting the existence of mechanisms that protect cells from ammonium toxicity. Here, we identified SLC12A9 as a lysosomal ammonium exporter that preserves lysosomal homeostasis. SLC12A9 knockout cells showed grossly enlarged lysosomes and elevated ammonium content. These phenotypes were reversed upon removal of the metabolic source of ammonium or dissipation of the lysosomal pH gradient. Lysosomal chloride increased in SLC12A9 knockout cells and chloride binding by SLC12A9 was required for ammonium transport. Our data indicate that SLC12A9 is a chloride-driven ammonium co-transporter that is central in an unappreciated, fundamental mechanism of lysosomal physiology that may have special relevance in tissues with elevated ammonia, such as tumors.
PubMed: 37292735
DOI: 10.1101/2023.05.22.541801 -
Physiological Reports Sep 2022Acute pyelonephritis caused by uropathogenic E. coli (UPEC) can cause renal scarring and lead to development of chronic kidney disease. Prevention of kidney injury...
Acute pyelonephritis caused by uropathogenic E. coli (UPEC) can cause renal scarring and lead to development of chronic kidney disease. Prevention of kidney injury requires an understanding of host factors and/or UPEC adaptive responses that are permissive for UPEC colonization of the urinary tract. Although some studies have suggested urine acidification limits UPEC growth in culture, other studies have described acid-resistance mechanisms (AR) in E. coli such as the CadC/CadBA module that promotes adaptation to acid and nitrosative stress. Herein we confirm and extend our previous study by demonstrating that despite urine acidification, metabolic acidosis induced by dietary ammonium chloride (NH Cl-A) exacerbates cystitis and pyelonephritis in innate immune competent (C3H-HeN) mice characterized by: (1) markedly elevated UPEC burden and increased chemokine/cytokine and NOS2 mRNA expression, (2) accumulation of intravesicular debris noninvasively detected by Power Doppler Ultrasound (PDUS), and (3) collecting duct (CD) dysfunction that manifests as a urine concentration defect. Bladder debris and CD dysfunction were due to the inflammatory response, as neither was observed in Tlr4-deficient (C3H-HeJ) mice. The effect of NH Cl-A was unrelated to acidosis as dietary administration of hydrochloric acid (HCl-A) yielded a comparable acid-base status yet did not increase UPEC burden. NH Cl-A increased polyamines and decreased nitric oxide (NO) metabolites in urine indicating that excess dietary ammonium shifts arginine metabolism toward polyamines at the expense of NO synthesis. Furthermore, despite increased expression of NOS2, NO production post UPEC infection was attenuated in NH Cl-A mice compared to controls. Thus, in addition to induction of metabolic acidosis and urine acidification, excess dietary ammonium alters the polyamine:NO balance and thereby compromises NOS2-mediated innate immune defense.
Topics: Acidosis; Ammonium Chloride; Animals; Arginine; Chemokines; Cystitis; Cytokines; Escherichia coli Infections; Hydrochloric Acid; Mice; Mice, Inbred C3H; Nitric Oxide; Polyamines; Pyelonephritis; RNA, Messenger; Toll-Like Receptor 4; Urinary Tract Infections; Uropathogenic Escherichia coli
PubMed: 36151614
DOI: 10.14814/phy2.15471