scholarly journals Examining the Osmotic Response of Acidihalobacter aeolianus after Exposure to Salt Stress

2021 ◽  
Vol 10 (1) ◽  
pp. 22
Author(s):  
Melissa K. Corbett ◽  
Liam Anstiss ◽  
April Gifford ◽  
Ross M. Graham ◽  
Elizabeth L. J. Watkin
Keyword(s):  
Osmotic Stress ◽  
Compatible Solutes ◽  
Organic Osmolytes ◽  
Osmotic Response ◽  
Environmental Instability ◽  
Acidic Environments ◽  
High Concentrations ◽  
Cellular Stresses ◽  
The Impact ◽  
Halophilic Species

Acidihalobacter aeolianus is an acidophilic, halo-tolerant organism isolated from a marine environment near a hydrothermal vent, an ecosystem whereby levels of salinity and total dissolved salts are constantly fluctuating creating ongoing cellular stresses. In order to survive these continuing changes, the synthesis of compatible solutes—also known as organic osmolytes—is suspected to occur, aiding in minimising the overall impact of environmental instability. Previous studies on A. aeolianus identified genes necessary for the accumulation of proline, betaine and ectoine, which are known to act as compatible solutes in other halophilic species. In this study, the impact of increasing the osmotic stress as well as the toxic ion effect was investigated by subjecting A. aeolianus to concentrations of NaCl and MgSO4 up to 1.27 M. Exposure to high concentrations of Cl- resulted in the increase of ectC expression in log-phase cells with a corresponding accumulation of ectoine at stationary phase. Osmotic stress via MgSO4 exposure did not trigger the same up-regulation of ectC or accumulation of ectoine, indicating the transcriptionally regulated response against osmotic stress was induced by chloride toxicity. These findings have highlighted how the adaptive properties of halo-tolerant organisms in acidic environments are likely to differ and are dependent on the initial stressor.

Membrane protein stability depends on the concentration of compatible solutes – a single molecule force spectroscopic study

2013 ◽  
Vol 394 (11) ◽  
pp. 1465-1474 ◽  
Author(s):  
Arpita Roychoudhury ◽  
Adeline Bieker ◽  
Dieter Häussinger ◽  
Filipp Oesterhelt
Keyword(s):  
Membrane Protein ◽  
Protein Stability ◽  
Single Molecule ◽  
Protein Unfolding ◽  
Compatible Solutes ◽  
Protein Stabilization ◽  
Intramolecular Interactions ◽  
Organic Osmolytes ◽  
Dynamic Relationship ◽  
The Impact

Abstract Compatible solutes are small, uncharged, zwitter ionic, osmotically active molecules produced and accumulated by microorganisms inside their cell to counteract different kinds of environmental stress. They enhance protein stability without interfering with the metabolic pathways even at molar concentrations. In this paper, we report the stabilizing effects of compatible solutes, ectoine, betaine and taurine on membrane protein bacteriorhodopsin at different concentrations. Using atomic force microscopy based single molecule force spectroscopy the impact of the osmolytes was quantified by measuring the forces required to pull the protein out of the membrane and the change in the persistence lengths of the unfolded polypeptide chain. Increase in unfolding forces were observed, indicating the strengthening of intramolecular interactions, which are vital for protein stability. The decrease in persistence lengths was recorded and showed increasing tendencies of the polypeptide strand to coil up. Interestingly, it was revealed that these molecules have different stabilizing effects on protein unfolding at different concentrations. The results show that the unfolding of single protein provides insight to the structure-dynamic relationship between the protein and compatible solute molecules at sub-nanometer scale. This also helps to understand the molecular mechanism involved in protein stabilization by organic osmolytes.

scholarly journals Compatible-Solute-Supported Periplasmic Expression of Functional Recombinant Proteins under Stress Conditions

2000 ◽  
Vol 66 (4) ◽  
pp. 1572-1579 ◽  
Author(s):  
S. Barth ◽  
M. Huhn ◽  
B. Matthey ◽  
A. Klimka ◽  
E. A. Galinski ◽  
...  
Keyword(s):  
Osmotic Stress ◽  
Recombinant Proteins ◽  
Glycine Betaine ◽  
Metal Ion ◽  
Halophilic Bacteria ◽  
Compatible Solutes ◽  
Compatible Solute ◽  
Size Exclusion ◽  
High Yield ◽  
High Concentrations

ABSTRACT The standard method of producing recombinant proteins such as immunotoxins (rITs) in large quantities is to transform gram-negative bacteria and subsequently recover the desired protein from inclusion bodies by intensive de- and renaturing procedures. The major disadvantage of this technique is the low yield of active protein. Here we report the development of a novel strategy for the expression of functional rIT directed to the periplasmic space of Escherichia coli. rITs were recovered by freeze-thawing of pellets from shaking cultures of bacteria grown under osmotic stress (4% NaCl plus 0.5 M sorbitol) in the presence of compatible solutes. Compatible solutes, such as glycine betaine and hydroxyectoine, are low-molecular-weight osmolytes that occur naturally in halophilic bacteria and are known to protect proteins at high salt concentrations. Adding 10 mM glycine betaine for the cultivation of E. coliunder osmotic stress not only allowed the bacteria to grow under these otherwise inhibitory conditions but also produced a periplasmic microenvironment for the generation of high concentrations of correctly folded rITs. Protein purified by combinations of metal ion affinity and size exclusion chromatography was substantially stabilized in the presence of 1 M hydroxyecotine after several rounds of freeze-thawing, even at very low protein concentrations. The binding properties and cytotoxic potency of the rITs were confirmed by competitive experiments. This novel compatible-solute-guided expression and purification strategy might also be applicable for high-yield periplasmic production of recombinant proteins in different expression systems.

Guardians in a stressful world: the Opu family of compatible solute transporters from Bacillus subtilis

2017 ◽  
Vol 398 (2) ◽  
pp. 193-214 ◽  
Author(s):  
Tamara Hoffmann ◽  
Erhard Bremer
Keyword(s):  
Bacillus Subtilis ◽  
Osmotic Stress ◽  
Cytoplasmic Membrane ◽  
Compatible Solutes ◽  
Transport Systems ◽  
Osmotic Gradient ◽  
Organic Osmolytes ◽  
Water Fluxes ◽  
Water Efflux ◽  
Response Systems

AbstractThe development of a semi-permeable cytoplasmic membrane was a key event in the evolution of microbial proto-cells. As a result, changes in the external osmolarity will inevitably trigger water fluxes along the osmotic gradient. The ensuing osmotic stress has consequences for the magnitude of turgor and will negatively impact cell growth and integrity. No microorganism can actively pump water across the cytoplasmic membrane; hence, microorganisms have to actively adjust the osmotic potential of their cytoplasm to scale and direct water fluxes in order to prevent dehydration or rupture. They will accumulate ions and physiologically compliant organic osmolytes, the compatible solutes, when they face hyperosmotic conditions to retain cell water, and they rapidly expel these compounds through the transient opening of mechanosensitive channels to curb water efflux when exposed to hypo-osmotic circumstances. Here, we provide an overview on the salient features of the osmostress response systems of the ubiquitously distributed bacteriumBacillus subtiliswith a special emphasis on the transport systems and channels mediating regulation of cellular hydration and turgor under fluctuating osmotic conditions. The uptake of osmostress protectants via the Opu family of transporters, systems of central importance for the management of osmotic stress byB. subtilis, will be particularly highlighted.

Osmoregulation in Bacteria: Compatible Solute Accumulation and Osmosensing

2006 ◽  
Vol 3 (2) ◽  
pp. 94 ◽  
Author(s):  
Hans Jörg Kunte
Keyword(s):  
Osmotic Stress ◽  
Surrounding Medium ◽  
Compatible Solutes ◽  
Compatible Solute ◽  
Water Soluble ◽  
Fine Tuning ◽  
Transport Systems ◽  
Organic Osmolytes ◽  
Organic Osmolyte ◽  
Solute Accumulation

Environmental Context.Bacteria and Archaea have developed two basic mechanisms to cope with osmotic stress. The ‘salt-in-cytoplasm mechanism’ involves adjusting the salt concentration in the cytoplasm according to the environmental osmolarity and the ‘organic-osmolyte mechanism’ involves accumulating uncharged, highly water-soluble organic compounds in order to maintain an osmotic equilibrium with the surrounding medium. This highlight gives an overview of the osmoadaptation of prokaryotes employing the organic-osmolyte strategy and introduces a model explaining the fine-tuning of osmoregulatory osmolyte synthesis. Abstract.Bacteria and Archaea have developed two basic mechanisms to cope with osmotic stress, the salt-in-cytoplasm mechanism, and the organic-osmolyte mechanism. Organic osmolytes or so-called compatible solutes can be accumulated in molar concentration in the cytoplasm and allow for the adaptation of bacterial cells to varying salt concentrations. The biosynthetic pathways of compatible solutes and different compatible solute transport systems are described. A model for osmoregulatory compatible solute accumulation is introduced.

scholarly journals Spatial Variation of Water Chemistry in Aries River Catchment Western Romania

2021 ◽  
Vol 11 (14) ◽  
pp. 6592
Author(s):  
Ana Moldovan ◽  
Maria-Alexandra Hoaghia ◽  
Anamaria Iulia Török ◽  
Marius Roman ◽  
Ionut Cornel Mirea ◽  
...  
Keyword(s):  
Heavy Metals ◽  
Water Quality ◽  
Water Chemistry ◽  
Water Samples ◽  
Mine Drainage ◽  
Pollution Indices ◽  
Mining Activities ◽  
River Catchment ◽  
High Concentrations ◽  
The Impact

This study aims to investigate the quality and vulnerability of surface water (Aries River catchment) in order to identify the impact of past mining activities. For this purpose, the pollution and water quality indices, Piper and Durov plots, as well vulnerability modeling maps were used. The obtained results indicate that the water samples were contaminated with As, Fe, Mn, Pb and have relatively high concentrations of SO42−, HCO3−, TDS, Ca, K, Mg and high values for the electrical conductivity. Possible sources of the high content of chemicals could be the natural processes or the inputs of the mine drainage. Generally, according to the pollution indices, which were correlated to high concentrations of heavy metals, especially with Pb, Fe and Mn, the water samples were characterized by heavy metals pollution. The water quality index classified the studied water samples into five different classes of quality, namely: unsuitable for drinking, poor, medium, good and excellent quality. Similarly, medium, high and very high vulnerability classes were observed. The Durov and Piper plots classified the waters into Mg-HCO3− and Ca-Cl− types. The past and present mining activities clearly change the water chemistry and alter the quality of the Aries River, with the water requiring specific treatments before use.

scholarly journals Effect of Maritime Traffic on Water Quality Parameters in Santa Marta, Colombia

2021 ◽  
Vol 9 (5) ◽  
pp. 474
Author(s):  
René Rodríguez-Grimón ◽  
Nestor Hernando Campos ◽  
Ítalo Braga Castro
Keyword(s):  
Water Quality ◽  
Petroleum Hydrocarbons ◽  
Quality Parameters ◽  
Water Quality Parameters ◽  
Maritime Traffic ◽  
Natural Park ◽  
High Concentrations ◽  
The Impact ◽  
The Relationship ◽  
Multiple Chemical

Since 2013, there has been an increase (>23%) in naval traffic using maritime routes and ports on the coastal fringe of Santa Marta, Colombia. Of major concern, and described by several studies, is the relationship between maritime traffic and coastal contamination. This study proposed a maritime traffic indicator considering the simultaneous effects of several relevant measurements of water quality parameters to estimate the impact of naval activity. The approach involved developing a model including the number of vessels, hull length, and permanence time in berths. In addition, water quality variables, considering climatic seasons, were used to verify association with maritime traffic and touristic activities. The high concentrations of total coliforms (TC) and dissolved/dispersed petroleum hydrocarbons in chrysene equivalents (DDPH) reported by the International Marina of Santa Marta (SM) were affected by the local anthropic activities, including tourism, naval traffic, and urban wastewater discharges. Moreover, our results suggest the occurrence of multiple chemical impacts within Tayrona National Natural Park (PNNT) affecting conservation goals. The estimation of the maritime traffic indicator proposed in this study may be an easy and more complete tool for future studies evaluating the impact of naval activities on environmental quality.

Are neutrophil extracellular traps the link for the cross-talk between periodontitis and rheumatoid arthritis physiopathology?

Rheumatology ◽  
2021 ◽  
Author(s):  
Sicília Rezende Oliveira ◽  
José Alcides A de Arruda ◽  
Ayda Henriques Schneider ◽  
Valessa Florindo Carvalho ◽  
Caio Machado ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Cross Talk ◽  
Neutrophil Extracellular Traps ◽  
Periodontal Therapy ◽  
Clinical Parameters ◽  
Extracellular Traps ◽  
Early Ra ◽  
The Cross ◽  
High Concentrations ◽  
The Impact

Abstract Objectives Neutrophil extracellular traps (NETs) play a role in the pathogenesis of periodontitis and rheumatoid arthritis (RA). However, it remains poorly understood whether NETs participate in the cross-talk between periodontitis and RA. Herein, we investigated the production of NETs in individuals with periodontitis and RA and its association with clinical parameters. The impact of periodontal therapy on RA and NET release was also assessed. Methods The concentration of NETs and cytokines was determined in the saliva and plasma of individuals with early RA (n = 24), established RA (n = 64), and individuals without RA (n = 76). The influence of periodontitis on the production of NETs and cytokines was also evaluated. Results Individuals with early RA had a higher concentration of NETs in saliva and plasma than individuals with established RA or without RA. Periodontitis resulted in an increase in the concentration of NETs of groups of individuals without RA and with early RA. The proportion of individuals with high concentrations of IL-6, IL-10 and GM-CSF was higher among individuals with periodontitis than among individuals without periodontitis. The concentrations of TNF-α, IL-6, IL-17/IL-25, and IL-28A were particularly high in individuals with early RA. Worse periodontal clinical parameters, RA onset and RA activity were significantly associated with circulating NETs. Periodontal therapy was associated with a reduction in the concentration of NETs and inflammatory cytokines and amelioration in periodontitis and RA. Conclusion This study reveals that NETs are a possible link between periodontitis and RA, with periodontal therapy resulting in a dramatic switch in circulating NET levels.

scholarly journals Photocrosslinking and photopatterning of magneto-optical nanocomposite sol–gel thin film under deep-UV irradiation

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
C. Bidaud ◽  
D. Berling ◽  
D. Jamon ◽  
E. Gamet ◽  
S. Neveu ◽  
...  
Keyword(s):  
Uv Irradiation ◽  
Cobalt Ferrite ◽  
Sol Gel ◽  
Optical Devices ◽  
High Concentration ◽  
Nanocomposite Thin Films ◽  
Deep Uv ◽  
High Concentrations ◽  
193 Nm ◽  
The Impact

AbstractThis paper is aimed at investigating the process of photocrosslinking under Deep-UV irradiation of nanocomposite thin films doped with cobalt ferrite magnetic nanoparticles (MNPs). This material is composed of a hybrid sol–gel matrix in which MNP can be introduced with high concentrations up to 20 vol%. Deep-UV (193 nm) is not only interesting for high-resolution patterning but we also show an efficient photopolymerization pathway even in the presence of high concentration of MNPs. In this study, we demonstrate that the photocrosslinking is based on the free radical polymerization of the methacrylate functions of the hybrid precursor. This process is initiated by Titanium-oxo clusters. The impact of the nanoparticles on the photopolymerization kinetic and photopatterning is investigated. We finally show that the photosensitive nanocomposite is suitable to obtain micropatterns with sub-micron resolution, with a simple and versatile process, which opens many opportunities for fabrication of miniaturized magneto-optical devices for photonic applications.

Accumulation of natural and synthetic betaines by a mammalian renal cell line

1996 ◽  
Vol 74 (2) ◽  
pp. 283-287 ◽  
Author(s):  
K. Randall ◽  
M. Lever ◽  
B. A. Peddie ◽  
S. T. Chambers
Keyword(s):  
Osmotic Stress ◽  
Glycine Betaine ◽  
Mdck Cells ◽  
Intracellular Accumulation ◽  
Madin Darby Canine Kidney ◽  
Renal Cell Line ◽  
High Concentrations ◽  
Inhibition Studies ◽  
Darby Canine Kidney ◽  
Canine Kidney

Intracellular accumulation of different betaines was compared in osmotically stressed Madin Darby canine kidney (MDCK) cells to model the betaine accumulation specificity of the mammalian inner medulla and to show how this accumulation differed from that of bacteria. All betaines accumulated less than glycine betaine. Arsenobetaine (the arsenic analogue of glycine betaine) accumulated to 12% of the glycine betaine levels and the sulphur analogue dimethylthetin accumulated to >80%. Most substituted glycine betaine analogues accumulated to 2–5% of intracellular glycine betaine concentrations, however, serine betaine accumulated to <0.5% of glycine betaine levels. Inhibition studies to distinguish the betaine ports were performed by the addition of proline. Butyrobetaine and carnitine accumulation was not proline sensitive, whereas that of omer betaines was. As with glycine betaine, the accumulation of propionobetaine and dimethylthetin was proline sensitive and osmoregulated. Pyridinium betaine was accumulated by both proline-sensitive and -insensitive systems, with a small increase under osmotic stress. High concentrations (10 times that of glycine betaine) of the dietary betaines proline betaine and trigonelline inhibited total betaine accumulation. Because α-substituted betaines are accumulated by bacteria and not by MDCK cells, these betaines may be the basis for design of antimicrobial agents.Key words: MDCK cells, betaine accumulation, osmolytes, betaine analogues.

scholarly journals Defects in the Secretory Pathway and High Ca2+ Induce Multiple P-bodies

2010 ◽  
Vol 21 (15) ◽  
pp. 2624-2638 ◽  
Author(s):  
Cornelia Kilchert ◽  
Julie Weidner ◽  
Cristina Prescianotto-Baschong ◽  
Anne Spang
Keyword(s):  
Osmotic Stress ◽  
Secretory Pathway ◽  
Small Gtpase ◽  
Processing Bodies ◽  
P Bodies ◽  
Intracellular Signals ◽  
Intracellular Ca ◽  
Response To Stress ◽  
Cellular Stresses ◽  
Translational Block

mRNA is sequestered and turned over in cytoplasmic processing bodies (PBs), which are induced by various cellular stresses. Unexpectedly, in Saccharomyces cerevisiae, mutants of the small GTPase Arf1 and various secretory pathway mutants induced a significant increase in PB number, compared with PB induction by starvation or oxidative stress. Exposure of wild-type cells to osmotic stress or high extracellular Ca2+ mimicked this increase in PB number. Conversely, intracellular Ca2+-depletion strongly reduced PB formation in the secretory mutants. In contrast to PB induction through starvation or osmotic stress, PB formation in secretory mutants and by Ca2+ required the PB components Pat1 and Scd6, and calmodulin, indicating that different stressors act through distinct pathways. Consistent with this hypothesis, when stresses were combined, PB number did not correlate with the strength of the translational block, but rather with the type of stress encountered. Interestingly, independent of the stressor, PBs appear as spheres of ∼40–100 nm connected to the endoplasmic reticulum (ER), consistent with the idea that translation and silencing/degradation occur in a spatially coordinated manner at the ER. We propose that PB assembly in response to stress occurs at the ER and depends on intracellular signals that regulate PB number.

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