Altered plasma proteome during an early phase of peritonitis-induced sepsis

2009 ◽  
Vol 116 (9) ◽  
pp. 721-730 ◽  
Author(s):  
Visith Thongboonkerd ◽  
Wararat Chiangjong ◽  
Jan Mares ◽  
Jiri Moravec ◽  
Zdenek Tuma ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Nitrosative Stress ◽  
Inflammatory Responses ◽  
Sepsis Syndrome ◽  
Host Responses ◽  
Plasma Proteome ◽  
Oxidative And Nitrosative Stress ◽  
Two Dimensional Gel Electrophoresis ◽  
Human Sepsis ◽  
Altered Proteins

Sepsis is a systemic response to infection commonly found in critically ill patients and is associated with multi-organ failure and high mortality rate. Its pathophysiology and molecular mechanisms are complicated and remain poorly understood. In the present study, we performed a proteomics investigation to characterize early host responses to sepsis as determined by an altered plasma proteome in a porcine model of peritonitis-induced sepsis, which simulated several clinical characteristics of human sepsis syndrome. Haemodynamics, oxygen exchange, inflammatory responses, oxidative and nitrosative stress, and other laboratory parameters were closely monitored. Plasma samples were obtained from seven pigs before and 12 h after the induction of sepsis, and plasma proteins were resolved with two-dimensional gel electrophoresis (n=7 gels/group; before being compared with during sepsis). The resolved proteins were stained with the SYPRO Ruby fluorescence dye and subjected to quantitative and comparative analyses. From approx. 1500 protein spots visualized in each gel, levels of 36 protein spots were significantly altered in the plasma of animals with sepsis (sepsis/basal ratios or degrees of change ranged from 0.07 to 21.24). Q-TOF (quadrupole–time-of-flight) MS and MS/MS (tandem MS) identified 30 protein forms representing 22 unique proteins whose plasma levels were increased, whereas six forms of five unique proteins were significantly decreased during sepsis. The proteomic results could be related to the clinical features of this animal model, as most of these altered proteins have important roles in inflammatory responses and some of them play roles in oxidative and nitrosative stress. In conclusion, these findings may lead to a better understanding of the pathophysiology and molecular mechanisms underlying the sepsis syndrome.

scholarly journals Intracellular survival ofBurkholderia cepaciacomplex in phagocytic cells

2015 ◽  
Vol 61 (9) ◽  
pp. 607-615 ◽  
Author(s):  
Miguel A. Valvano
Keyword(s):  
Burkholderia Cepacia ◽  
Molecular Mechanisms ◽  
Inflammatory Responses ◽  
Genetic Manipulation ◽  
Intracellular Survival ◽  
Burkholderia Cenocepacia ◽  
Burkholderia Cepacia Complex ◽  
Host Responses ◽  
Current View ◽  
Phagocytic Cells

Burkholderia cepacia complex (Bcc) species are a group of Gram-negative opportunistic pathogens that infect the airways of cystic fibrosis patients, and occasionally they infect other immunocompromised patients. Bcc bacteria display high-level multidrug resistance and chronically persist in the infected host while eliciting robust inflammatory responses. Studies using macrophages, neutrophils, and dendritic cells, combined with advances in the genetic manipulation of these bacteria, have increased our understanding of the molecular mechanisms of virulence in these pathogens and the molecular details of cell-host responses triggering inflammation. This article discusses our current view of the intracellular survival of Burkholderia cenocepacia within macrophages.

scholarly journals Calgranulin C Has Filariacidal and Filariastatic Activity

1999 ◽  
Vol 67 (12) ◽  
pp. 6631-6636 ◽  
Author(s):  
John D. Gottsch ◽  
Steven W. Eisinger ◽  
Sammy H. Liu ◽  
Alan L. Scott
Keyword(s):  
Culture System ◽  
Molecular Mechanisms ◽  
Inflammatory Responses ◽  
Parasite Burden ◽  
Immunohistochemical Analysis ◽  
Host Responses ◽  
Dose Dependent Fashion ◽  
Antifilarial Activity ◽  
Larval Parasites

ABSTRACT The calgranulins are a family of calcium- and zinc-binding proteins produced by neutrophils, monocytes, and other cells. Calgranulins are released during inflammatory responses and have antimicrobial activity. Recently, one of the calgranulins, human calgranulin C (CaGC), has been implicated as an important component of the host responses that limit the parasite burden during filarial nematode infections. The goal of this work was to test the hypothesis that human CaGC has biologic activity against filarial parasites. Brugia malayi microfilariae and adults were exposed in vitro to 0.75 to 100 nM recombinant human CaGC. Recombinant CaGC affected adult and larval parasites in a dose-dependent fashion. Microfilariae were more sensitive to the action of CaGC than were adult parasites. At high levels, CaGC was both macrofilariacidal and microfilariacidal. At lower levels, the percentage of parasites killed was dependent on the level of CaGC in the culture system. The larvae not killed had limited motility. The filariastatic effect of low-level CaGC was reversed when the CaGC was removed from the culture system. Immunohistochemical analysis demonstrated that human CaGC accumulated in the cells of the hypodermis-lateral chord of adult and larval parasites. The antifilarial activity of CaGC was not due to the sequestration of zinc. Thus, the cellular and molecular mechanisms that result in the production and release of CaGC in humans may play a key role in the regulation of filarial parasite numbers.

scholarly journals Reconciling the IPC and Two-Hit Models: Dissecting the Underlying Cellular and Molecular Mechanisms of Two Seemingly Opposing Frameworks

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Carlos F. M. Morris ◽  
Muhammad Tahir ◽  
Samina Arshid ◽  
Mariana S. Castro ◽  
Wagner Fontes
Keyword(s):  
Inflammatory Response ◽  
Clinical Outcomes ◽  
Ischemic Preconditioning ◽  
Molecular Mechanisms ◽  
Inflammatory Responses ◽  
Present Review ◽  
Host Responses ◽  
Ischemia And Reperfusion ◽  
Biological Models

Inflammatory cascades and mechanisms are ubiquitous during host responses to various types of insult. Biological models and interventional strategies have been devised as an effort to better understand and modulate inflammation-driven injuries. Amongst those the two-hit model stands as a plausible and intuitive framework that explains some of the most frequent clinical outcomes seen in injuries like trauma and sepsis. This model states that a first hit serves as a priming event upon which sequential insults can build on, culminating on maladaptive inflammatory responses. On a different front, ischemic preconditioning (IPC) has risen to light as a readily applicable tool for modulating the inflammatory response to ischemia and reperfusion. The idea is that mild ischemic insults, either remote or local, can cause organs and tissues to be more resilient to further ischemic insults. This seemingly contradictory role that the two models attribute to a first inflammatory hit, as priming in the former and protective in the latter, has set these two theories on opposing corners of the literature. The present review tries to reconcile both models by showing that, rather than debunking each other, each framework offers unique insights in understanding and modulating inflammation-related injuries.

scholarly journals The Parasympathetic Nervous System in the Quest for Stroke Therapeutics

2011 ◽  
Vol 31 (5) ◽  
pp. 1187-1195 ◽  
Author(s):  
Cletus Cheyuo ◽  
Asha Jacob ◽  
Rongqian Wu ◽  
Mian Zhou ◽  
Gene F Coppa ◽  
...  
Keyword(s):  
Nervous System ◽  
Molecular Mechanisms ◽  
Nitrosative Stress ◽  
Parasympathetic Nervous System ◽  
Brain Disorders ◽  
Stroke Therapy ◽  
Successful Development ◽  
Oxidative And Nitrosative Stress ◽  
Anatomical Basis ◽  
Further Development

Stroke is a devastating neurovascular disease with limited therapeutic options. The pathogenesis of stroke involves complex interrelated molecular mechanisms including excitotoxicity, oxidative and nitrosative stress, cortical spreading depolarizations, inflammation, necrosis, and apoptosis. Successful development of stroke therapeutics depends on understanding these molecular mechanisms and how to counteract them to limit tissue damage during stroke. Activation of the parasympathetic nervous system (PNS) has been shown to antagonize a multiplicity of pathologic mechanisms. Elements of parasympathetic activation such as vagus nerve stimulation have already been used successfully in treating brain disorders such as epilepsy and depression. This review discusses the anatomical basis and molecular mechanisms involved in activation of the PNS, and assesses the strength of available evidence for the further development of this modality into a stroke therapy.

scholarly journals Antidepressant Like Effect of Sodium Orthovanadate in A Mouse Model of Chronic Unpredictable Mild Stress.

2021 ◽  
Author(s):  
Angel Joshi ◽  
Ansab Akhtar ◽  
Priyanka Saroj ◽  
Anurag Kuhad ◽  
Sangeeta Pilkhwal Sah
Keyword(s):  
Molecular Mechanisms ◽  
Nitrosative Stress ◽  
Tyrosine Phosphatase ◽  
Preference Test ◽  
Tail Suspension Test ◽  
Mild Stress ◽  
Chronic Unpredictable Mild Stress ◽  
Sodium Orthovanadate ◽  
Behavioral Tests ◽  
Oxidative And Nitrosative Stress

Abstract Depression is a psychiatric disorder characterized by low esteem, anhedonia, social deficit, and lack of interest. Decreased BDNF and impaired TrKB signaling be associated with depression. In our study, depressive-like behavior was induced in mice by chronic unpredictable mild stress (CUMS) model. Various behavioral tests like tail suspension test (TST), open field test (OFT), and sucrose preference test (SPT); biochemical analyses for corticosterone, reduced glutathione (GSH), lipid peroxidation (LPO), superoxide dismutase (SOD), nitric oxide (NO) and ELISA for BDNF were performed. Body weight was measured every week. Depressive-like behavior was associated with increased oxidative stress in the brain and subsequent reduction of BDNF. Further, sodium orthovanadate (SOV), a protein tyrosine phosphatase inhibitor was used as a test drug as it is reported to stimulate BDNF levels. Sodium orthovanadate (SOV-5 mg/kg, 10 mg/kg) and fluoxetine (10 mg/kg) was given to mice orally for 21 days before 30 minutes of stress induction. The behavioral tests reflected depressive-like behavior in CUMS, which was attenuated by both SOV and fluoxetine. SOV at 10 mg/kg has demonstrated significant results in our study by decreasing malondialdehyde levels (MDA/LPO), NO levels, and increasing GSH and SOD in both the cortex and hippocampus. Besides, ELISA revealed the elevation of BDNF levels in the treatment groups (SOV-5 mg/kg, 10 mg/kg, and FLX-10 mg/kg) as compared with the disease group (CUMS). Therefore, the treatment with SOV appeared to reverse both oxidative and nitrosative stress. Decreased serum corticosterone levels (SOV-5 mg/kg, 10 mg/kg); FLX (10 mg/kg) + SOV (5 mg/kg); FLX-10 mg/kg and per-se) and elevated BDNF level (SOV-5 mg/kg, 10 mg/kg and FLX-10 mg/kg) were associated with attenuation of depressive-like behavior. The findings of this preliminary study indicate that SOV has the potential to restore antidepressant-like effect or prevention of stress-induced anhedonia and so further molecular mechanisms will be warranted for clinical translation.

scholarly journals DIABETIC NEUROPATHY: MOLECULAR MECHANISMS OF DEVELOPMENT AND POSSIBILITIES FOR PATHOGENETIC THERAPY

2019 ◽  
pp. 8-12
Author(s):  
N.V. Hudiakova ◽  
N.V. Ivanov ◽  
I. Yu. Pchelin ◽  
A.N. Shishkin ◽  
N.V. Vorokhobina ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Diabetic Neuropathy ◽  
Molecular Mechanisms ◽  
Nitrosative Stress ◽  
Treatment Options ◽  
Neurological Complications ◽  
Diabetic Patients ◽  
Oxidative And Nitrosative Stress ◽  
Pathogenetic Therapy ◽  
Mechanisms Of Development

The present review summarizes the results of global studies and assesses contribution of hyperglycemia towards formation of neurologic complications in diabetic patients. Hyperglycemia is believed to play a leading role in the formation of neurological complications in diabetes mellitus. However, the achievement of normalization of glycemia level does not ensure the cessation of their development and progression, which indicates a lack of knowledge about the pathogenetic relationships in diabetic neuropathy. Limited understanding of these issues entails the absence of treatment options that effectively affect the course of this complication. Based on the analysis of experimental and clinical studies of recent years, data on the molecular-biological relationships of hyperglycemia with the formation of neurological complications in diabetes mellitus are summarized. The influence of the oxidative and nitrosative stress, advanced glycation end products, the activation of the polyol and hexosamine pathways on the state of the nerve fiber is analyzed. The data on molecular mechanisms of development of diabetic neuropathy are contradictory. On the basis of recent experimental and clinical data we review possibilities for pathogenetic therapy. The problem of oppositely directed effects of treatment is discussed. Clinical rationale is given for declared direction of further studies.

scholarly journals Attenuation of Oxidative Stress in HEK 293 Cells by the TCM Constituents Schisanhenol, Baicalein, Resveratrol or Crocetin and Two Defined Mixtures

2015 ◽  
Vol 18 (4) ◽  
pp. 661 ◽  
Author(s):  
John Richard Bend ◽  
Xue Yan (Iris) Xue Yan Xia ◽  
Daofeng Chen ◽  
Abudi Awaysheh ◽  
Andrea Lo ◽  
...  
Keyword(s):  
Real Time ◽  
Molecular Mechanisms ◽  
Nitrosative Stress ◽  
Fluorescent Protein ◽  
Antioxidant Properties ◽  
Oxidative And Nitrosative Stress ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

PURPOSE:  Our working hypothesis is that single bioactive phytochemicals with antioxidant properties that are important constituents of Traditional Chinese Medicine (TCM) and their defined mixtures have potential as chemoprotective agents for chronic conditions characterized by oxidative and nitrosative stress, including Alzheimer’s. Here we evaluate the ability of baicalein, crocetin, trans-resveratrol or schisanhenol and two defined mixtures of these TCM phytochemicals to attenuate the toxicity resulting from exposure to cell permeant t-butyl hydroperoxide (tBPH) in wild-type and bioengineered (to express choline acetyltransferase) HEK 293 cells. METHODS: Endpoints of tBHP-initiated oxidative and nitrosative stress in both types of HEK 293 cells and its attenuation by TCM constituents and mixtures included cytotoxicity (LDH release); depletion of intracellular glutathione (GSH); formation of S-glutathionylated proteins; oxidative changes to the disulfide proteome; and real-time changes in intracellular redox status. RESULTS: At low µM concentrations, each of the TCM constituents and mixtures effectively attenuated intracellular toxicity due to exposure of HEK 293 cells to 50 or 250 µM tBHP for 30 min to 3 h. Confocal microscopy of HEK 293 cells transfected with mutated green fluorescent protein (roGFP2) showed effective attenuation of tBHP oxidation by baicalein in real time. Three redox-regulated proteins prominent in the disulfide proteome of HEK 293 cells were identified by MALDI-TOF mass spectrometry. CONCLUSIONS: We conclude that single TCM chemicals and their simple mixtures have potential for use in adjunct chemoprotective therapy. Advantages of mixtures compared to single TCM constituents include the ability to combine compounds with varying molecular mechanisms of cytoprotection for enhanced biological activity; and to combine chemicals with complementary pharmacokinetic properties to increase half-life and prolong activity in vivo. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

scholarly journals A Putative P-Type ATPase, Apt1, Is Involved in Stress Tolerance and Virulence in Cryptococcus neoformans

2009 ◽  
Vol 9 (1) ◽  
pp. 74-83 ◽  
Author(s):  
Guanggan Hu ◽  
James W. Kronstad
Keyword(s):  
Membrane Fusion ◽  
Cryptococcus Neoformans ◽  
Virulence Factors ◽  
Molecular Mechanisms ◽  
Nitrosative Stress ◽  
Brefeldin A ◽  
Antifungal Drugs ◽  
Oxidative And Nitrosative Stress ◽  
Transport Pathways ◽  
P Type

ABSTRACT The export of virulence factors, such as the capsule polysaccharide, to the cell surface is a critical aspect of the pathogenicity of Cryptococcus neoformans. A view of capsule export via exocytosis and extracellular vesicles is emerging, but the molecular mechanisms underlying virulence factor transport pathways remain to be established. In this study, we characterized the APT1 gene, which encodes a predicted integral membrane P-type ATPase belonging to the type IV, Drs2 family of aminophospholipid translocases (flippases) (APTs). APTs maintain the phospholipid asymmetry that is critical in membrane fusion events for trafficking and in establishing cell polarity. Deletion of the APT1 gene resulted in phenotypes consistent with similar roles in C. neoformans. These included altered actin distribution, increased sensitivity to stress conditions (oxidative and nitrosative stress) and to trafficking inhibitors, such as brefeldin A and monensin, a reduction in exported acid phosphatase activity, and hypersensitivity to the antifungal drugs amphotericin B, fluconazole, and cinnamycin. However, there was no difference in growth, capsule size, or melanin production between the wild type and the apt1 mutant strains at either 30°C or 37°C. Despite the absence of an influence on these major virulence factors, Apt1 was required for survival during interactions with macrophages, and apt1 mutants exhibited attenuated virulence in a mouse inhalation model of cryptococcosis. Therefore, Apt1 contributes to virulence and the stress response in C. neoformans through apparent functions in membrane fusion and trafficking that do not influence the deposition of major virulence factors, such as capsule and melanin, outside the cell.

scholarly journals The bacillithiol pathway is required for biofilm formation in Staphylococcus aureus

2018 ◽  
Author(s):  
Megha Gulati ◽  
Jason M. Thomas ◽  
Mamta Rawat ◽  
Clarissa J. Nobile
Keyword(s):  
Nitric Oxide ◽  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Antimicrobial Agents ◽  
Nitrosative Stress ◽  
Host Responses ◽  
Host Immune System ◽  
Oxidative And Nitrosative Stress ◽  
Biofilm Thickness ◽  
Superficial Skin

Staphylococcus aureus is a major human pathogen that can cause infections that range from superficial skin and mucosal infections to life threatening disseminated infections. S. aureus can attach to medical devices and host tissues and form biofilms that allow the bacteria to evade the host immune system and provide protection from antimicrobial agents. To counter host-generated oxidative and nitrosative stress mechanisms that are part of the normal host responses to invading pathogens, S. aureus utilizes low molecular weight (LMW) thiols, such as bacillithiol (BSH). Additionally, S. aureus synthesizes its own nitric oxide (NO), which combined with its downstream metabolites may also protect the bacteria against specific host responses. We have previously shown that LMW thiols are required for biofilm formation in Mycobacterium smegmatis and Pseudomonas aeruginosa. Our data show that the bshC mutant, which is defective in the last step of the bacillithiol pathway and lacks BSH, is impaired in biofilm formation. We also identify a putative S-nitrosobacillithiol reductase (BSNOR), similar to a S-nitrosomycothiol reductase found in M. smegmatis, and show that the BSNOR mutant has reduced levels of BSH and decreased biofilm formation. Our studies also show that NO plays an important role in biofilm formation and that acidified sodium nitrite severely reduces biofilm thickness. These studies provide insight into the roles of oxidative and nitrosative stress mechanisms on biofilm formation and indicate that bacillithiol and nitric oxide are key players in normal biofilm formation in S. aureus.

scholarly journals The Role of S-Nitrosylation and S-Glutathionylation of Protein Disulphide Isomerase in Protein Misfolding and Neurodegeneration

2013 ◽  
Vol 2013 ◽  
pp. 1-15 ◽  
Author(s):  
M. Halloran ◽  
S. Parakh ◽  
J. D. Atkin
Keyword(s):  
Neurodegenerative Diseases ◽  
Er Stress ◽  
Protein Misfolding ◽  
Molecular Mechanisms ◽  
Nitrosative Stress ◽  
Therapeutic Interventions ◽  
Oxidative And Nitrosative Stress ◽  
Protective Function ◽  
Protein Disulphide Isomerase

Neurodegenerative diseases involve the progressive loss of neurons, and a pathological hallmark is the presence of abnormal inclusions containing misfolded proteins. Although the precise molecular mechanisms triggering neurodegeneration remain unclear, endoplasmic reticulum (ER) stress, elevated oxidative and nitrosative stress, and protein misfolding are important features in pathogenesis. Protein disulphide isomerase (PDI) is the prototype of a family of molecular chaperones and foldases upregulated during ER stress that are increasingly implicated in neurodegenerative diseases. PDI catalyzes the rearrangement and formation of disulphide bonds, thus facilitating protein folding, and in neurodegeneration may act to ameliorate the burden of protein misfolding. However, an aberrant posttranslational modification of PDI, S-nitrosylation, inhibits its protective function in these conditions. S-nitrosylation is a redox-mediated modification that regulates protein function by covalent addition of nitric oxide- (NO-) containing groups to cysteine residues. Here, we discuss the evidence for abnormal S-nitrosylation of PDI (SNO-PDI) in neurodegeneration and how this may be linked to another aberrant modification of PDI, S-glutathionylation. Understanding the role of aberrant S-nitrosylation/S-glutathionylation of PDI in the pathogenesis of neurodegenerative diseases may provide insights into novel therapeutic interventions in the future.

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