scholarly journals Intracellular ion concentrations and cation-dependent remodelling of bacterial MreB assemblies

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Dávid Szatmári ◽  
Péter Sárkány ◽  
Béla Kocsis ◽  
Tamás Nagy ◽  
Attila Miseta ◽  
...  

Abstract Here, we measured the concentrations of several ions in cultivated Gram-negative and Gram-positive bacteria, and analyzed their effects on polymer formation by the actin homologue MreB. We measured potassium, sodium, chloride, calcium and magnesium ion concentrations in Leptospira interrogans, Bacillus subtilis and Escherichia coli. Intracellular ionic strength contributed from these ions varied within the 130–273 mM range. The intracellular sodium ion concentration range was between 122 and 296 mM and the potassium ion concentration range was 5 and 38 mM. However, the levels were significantly influenced by extracellular ion levels. L. interrogans, Rickettsia rickettsii and E. coli MreBs were heterologously expressed and purified from E. coli using a novel filtration method to prepare MreB polymers. The structures and stability of Alexa-488 labeled MreB polymers, under varying ionic strength conditions, were investigated by confocal microscopy and MreB polymerization rates were assessed by measuring light scattering. MreB polymerization was fastest in the presence of monovalent cations in the 200–300 mM range. MreB filaments showed high stability in this concentration range and formed large assemblies of tape-like bundles that transformed to extensive sheets at higher ionic strengths. Changing the calcium concentration from 0.2 to 0 mM and then to 2 mM initialized rapid remodelling of MreB polymers.

Water ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 1625
Author(s):  
Rekha Singh ◽  
Woohang Kim ◽  
James A. Smith

This study quantifies the effects of chloride ions on silver and copper release from porous ceramic cubes embedded with silver and copper and its effect on E. coli disinfection in drinking water. Log-reduction of E. coli by silver ions decreased after 4 h of contact time as the chloride ion concentration increased from 0 to 250 mg/L but, it was not changed by copper ions under the same conditions. For silver addition by silver-ceramic cubes, log reductions of E. coli decreased sharply from 7.2 to 1.6 after 12 h as the chloride concentration increased from 0 to 250 mg/L. For the silver-ceramic cube experiments, chloride ion also reduced the total silver concentration in solution. After 24 h, total silver concentrations in solution decreased from 61 µg/L to 20 µg/L for corresponding chloride ion concentrations. According to the MINTEQ equilibrium model analysis, the decrease in disinfection ability with silver embedded ceramic cubes could be the result of precipitation of silver ions as silver chloride. This suggests that AgCl was precipitating within the pore space of the ceramic. These results indicate that, although ionic silver is a highly effective disinfectant for E. coli, the presence of chloride ions can significantly reduce disinfection efficacy. For copper-ceramic cubes, log reductions of E. coli by copper embedded cubes increased from 1.2 to 1.5 when chloride ion concentration increased from 0 to 250 mg/L. Total copper concentrations in solution increased from 4 µg/L to 14 µg/L for corresponding chloride ion concentrations. These results point towards the synergistic effect of chloride ions on copper oxidation as an increased concentration of chloride enhances copper release.


2010 ◽  
Vol 66 ◽  
pp. 142-149 ◽  
Author(s):  
Jang Hsing Hsieh ◽  
T.H. Yeh ◽  
C.S. Chiou ◽  
C.T. Huang ◽  
C. Li

TaN–Cu and TaN-Ag nanocomposite films were deposited by reactive co-sputtering on Si. The films were then annealed using RTA (Rapid Thermal Annealing) at 400 °C for 2, 4, 8, 15 minutes respectively to induce the nucleation and growth of metal particles in TaN matrix and on film surface. The samples were then tested for their anti-bacterial behaviors against Gram-negative and Gram-positive bacteria. The results were compared with the solubility of Ag and Cu in a buffer solution. Under similar ion concentrations, it is concluded that Ag ion or TaN-Ag is more effective against E. Coli (Gram-negative), while Cu ion or TaN-Cu is more effective against Staphylococcus aureus (Gram-positive).


2000 ◽  
Vol 203 (13) ◽  
pp. 2065-2074 ◽  
Author(s):  
J.N. Stinner ◽  
L.K. Hartzler

The aim of this study was to determine the effects of temperature upon pH, protein charge and acid-base-relevant ion exchange in air-breathing ectotherms. Plasma and skeletal muscles in cane toads (Bufo marinus) and bullfrogs (Rana catesbeiana) were examined at 30, 20 and 10 degrees C. In addition, skeletal muscle ion concentrations were examined in black racer snakes (Coluber constrictor) at 30 and 10 degrees C. Cooling the amphibians produced a reduction in most of the plasma ion concentrations (Na(+), K(+), Ca(2+), Cl(−), SO(4)(2)(−)) and in protein concentration because of increased hydration. Between 30 and 10 degrees C, total plasma osmolality fell by 14 % in the toads and by 5 % in the frogs. Plasma protein charge, calculated using the principle of electroneutrality, was unaffected by temperature, except possibly for the toads at 10 degrees C. The in vivo skeletal muscle capdelta pHi/ capdelta T ratio, where pHi is intracellular pH and T is temperature, between 30 and 20 degrees C averaged −0.014 degrees C(−)(1) in the toads and −0.019 degrees C(−)(1) in the frogs. Between 20 and 10 degrees C, there was no change in pHi in the toads and a −0.005 degrees C(−)(1) change in the frogs. The in vitro skeletal muscle capdelta pHi/ capdelta T averaged −0.011 degrees C(−)(1) in both toads and frogs. In all three species, skeletal muscle inulin space declined with cooling. Intracellular ion concentrations were calculated by subtracting extracellular fluid ion concentrations from whole-muscle ion concentrations. In general, temperature had a large effect upon intracellular ion concentrations (Na(+), K(+), Cl(−)) and intracellular CO(2) levels. The relevance of the changes in intracellular ion concentration to skeletal muscle acid-base status and protein charge and the possible mechanisms producing the adjustments in intracellular ion concentration are discussed. It is concluded that ion-exchange mechanisms make an important contribution to adjusting pH with changes in temperature.


2019 ◽  
Author(s):  
Axel Loewe ◽  
Yannick Lutz ◽  
Norbert Nagy ◽  
Alan Fabbri ◽  
Christoph Schweda ◽  
...  

AbstractChanges of serum and extracellular ion concentrations occur regularly in patients with chronic kidney disease (CKD). Recently, hypocalcemia, i.e. a decrease of the extra-cellular calcium concentration [Ca2+]o, has been suggested as potential pathomechanism contributing to the unexplained high rate of sudden cardiac death (SCD) in CKD patients. In particular, there is a hypothesis that hypocalcaemia could slow down natural pacemaking in the human sinus node to fatal degrees. Here, we address the question whether there are inter-species differences in the response of cellular sinus node pacemaking to changes of [Ca2+]o. Towards this end, we employ computational models of mouse, rabbit and human sinus node cells. The Fabbri et al. human model was updated to consider changes of intracellular ion concentrations. We identified crucial inter-species differences in the response of cellular pacemaking in the sinus node to changes of [Ca2+]o with little changes of cycle length in mouse and rabbit models (<83 ms) in contrast to a pronounced bradycardic effect in the human model (up to > 1000 ms). Our results suggest that experiments with human sinus node cells are required to investigate the potential mechanism of hypocalcaemia-induced bradycardic SCD in CKD patients and small animal models are not well suited.


1997 ◽  
Vol 12 (4) ◽  
pp. 225-229
Author(s):  
Cart-in A-S. Gustavsson ◽  
Chritofer T. Lindgren ◽  
Mikael E. Lindström

Abstract The amount of lignin reacting according to the slow residual phase, i.e. the residual phase lignin, is in many perspectives an interesting issue. The purpose of the present investigation was to develop a mathematical model to show how the amount of residual phase lignin in the kraft cooking of spruce chips (Picm ahies) depends on the conditions in the earlier phases of the cook. The variables studied were hydroxide ion concentration, hydrogen sulfide ion concentration and ionic strength. The liquor-to-wood ratio during pulping was very high to maintain approximately constant chemical concentrations throughout each experiment (so called "constant composition" cooks). An increase in hydroxide ion concentration andtor hydrogen sulfide ion concentration leads to a decrease in the amount of residual phase lignin, while an increase in ionic strength, i.e. sodium ion concentration, leads to an increase. A signiticant result is that the hydrogen sulfide ion concentration has a pronounced influence on the amount of residual phase lignin during a cook at a low hydroxide ion concentration. The amount of residual phase lignin expressed as % lignin on wood, L,, can be described by the following equation developed for "constant composition" cooks (when cooking with a constant sodium ion concentration of 2 mol/L): LT=0,55-0.32*[HO-](-1,3)*ln[HS-] This equation is valid for a concentration of HO- in the range from 0.17 to 1.4, and a hydrogen sulfide ion concentration from 0.07 to 0.6 mol/L.


2002 ◽  
Vol 2 (3) ◽  
pp. 131-138 ◽  
Author(s):  
D.L. Craig ◽  
H.J. Fallowfield ◽  
N.J. Cromar

A laboratory based microcosm study utilising intact non-sterile sediment cores was undertaken to determine the survival of the faecal indicator organisms Escherichia coli, Enterococcus faecium and somatic coliphage in both recreational coastal water and sediment. Overlying water was inoculated with the test organisms and incubated at 10°C, 20°C or 30°C. E. coli, enterococcus and coliphage were enumerated from the water column and sediment by the membrane filtration method, Enterolert (IDEXX Laboratories) and the double-agar overlay methods respectively on days 0, 1, 2, 7, 14 and 28 following inoculation. It was demonstrated that for all organisms, greater decay (k; d-1) occurred in the water column compared to sediment. Sediment characteristics were found to influence decay, with lowest decay rates observed in sediment consisting of high organic carbon content and small particle size. Decay of E. coli was significantly greater in both the water column and sediment compared with enterococcus and coliphage under all conditions. Decay of enterococcus was found to closely resemble that of coliphage decay. Survival of all organisms was inversely related to temperature, with greatest decay at 30°C. However, increased temperature had a less significant impact on survival of enterococcus and coliphage compared with E. coli. The importance of this study for estimating risk from recreational exposure is great if some pathogenic microorganisms behave similarly to the organisms tested in this study. In particular if survival rates of pathogens are similar to enterococcus and coliphage, then their ability to accumulate in coastal sediment may lead to an increased risk of exposure if these organisms are resuspended into the water column due to natural turbulence or human recreational activity.


1993 ◽  
Vol 27 (3-4) ◽  
pp. 267-270 ◽  
Author(s):  
M. T. Augoustinos ◽  
N. A. Grabow ◽  
B. Genthe ◽  
R. Kfir

A fluorogenic β-glucuronidase assay comprising membrane filtration followed by selective enumeration on m-FC agar at 44.5°C and further confirmation using tlie 4-metliylumbelliferyl-β-D-glucuronide (MUG) containing medium was evaluated for the detection of Escherichia coli in water. A total of 200 typical blue and non-typical blue colonies were isolated from sea and fresh water samples using initial selective enumeration on m-FC agar. Pure cultures of the selected colonies were further tested using the MUG assay and identified using the API 20E method. Of the colonies tested which were shown to be positive using the MUG assay 99.4% were Escherichia coli. The results of this study indicate the combination of the m-FC method followed by the MUG assay to be highly efficient for the selection and confirmation of E. coli from a wide range of environmental waters.


2020 ◽  
Vol 15 (6) ◽  
pp. 665-679
Author(s):  
Alok K. Srivastava ◽  
Lokesh K. Pandey

Background: [1, 3, 4]oxadiazolenone core containing chalcones and nucleosides were synthesized by Claisen-Schmidt condensation of a variety of benzaldehyde derivatives, obtained from oxidation of substituted 5-(3/6 substituted-4-Methylphenyl)-1, 3, 4-oxadiazole-2(3H)-one and various substituted acetophenone. The resultant chalcones were coupled with penta-O-acetylglucopyranose followed by deacetylation to get [1, 3, 4] oxadiazolenone core containing chalcones and nucleosides. Various analytical techniques viz IR, NMR, LC-MS and elemental analysis were used to confirm the structure of the synthesised compounds.The compounds were targeted against Bacillus subtilis, Staphylococcus aureus and Escherichia coli for antibacterial activity and Aspergillus flavus, Aspergillus niger and Fusarium oxysporum for antifungal activity. Methods: A mixture of Acid hydrazides (3.0 mmol) and N, Nʹ- carbonyl diimidazole (3.3 mmol) in 15 mL of dioxane was refluxed to afford substituted [1, 3, 4]-oxadiazole-2(3H)-one. The resulted [1, 3, 4]- oxadiazole-2(3H)-one (1.42 mmol) was oxidized with Chromyl chloride (1.5 mL) in 20 mL of carbon tetra chloride and condensed with acetophenones (1.42 mmol) to get chalcones 4. The equimolar ratio of obtained chalcones 4 and β -D-1,2,3,4,6- penta-O-acetylglucopyranose in presence of iodine was refluxed to get nucleosides 5. The [1, 3, 4] oxadiazolenone core containing chalcones 4 and nucleosides 5 were tested to determined minimum inhibitory concentration (MIC) value with the experimental procedure of Benson using disc-diffusion method. All compounds were tested at concentration of 5 mg/mL, 2.5 mg/mL, 1.25 mg/mL, 0.62 mg/mL, 0.31 mg/mL and 0.15 mg/mL for antifungal activity against three strains of pathogenic fungi Aspergillus flavus (A. flavus), Aspergillus niger (A. niger) and Fusarium oxysporum (F. oxysporum) and for antibacterial activity against Gram-negative bacterium: Escherichia coli (E. coli), and two Gram-positive bacteria: Staphylococcus aureus (S. aureus) and Bacillus subtilis(B. subtilis). Result: The chalcones 4 and nucleosides 5 were screened for antibacterial activity against E. coli, S. aureus and B. subtilis whereas antifungal activity against A. flavus, A. niger and F. oxysporum. Compounds 4a-t showed good antibacterial activity whereas compounds 5a-t containing glucose moiety showed better activity against fungi. The glucose moiety of compounds 5 helps to enter into the cell wall of fungi and control the cell growth. Conclusion: Chalcones 4 and nucleosides 5 incorporating [1, 3, 4] oxadiazolenone core were synthesized and characterized by various spectral techniques and elemental analysis. These compounds were evaluated for their antifungal activity against three fungi; viz. A. flavus, A. niger and F. oxysporum. In addition to this, synthesized compounds were evaluated for their antibacterial activity against gram negative bacteria E. Coli and gram positive bacteria S. aureus, B. subtilis. Compounds 4a-t showed good antibacterial activity whereas 5a-t showed better activity against fungi.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Pedro Seguí ◽  
John J. Aguilera-Correa ◽  
Elena Domínguez-Jurado ◽  
Christian M. Sánchez-López ◽  
Ramón Pérez-Tanoira ◽  
...  

AbstractThis study was designed to propose alternative therapeutic compounds to fight against bacterial pathogens. Thus, a library of nitrogen-based compounds bis(triazolyl)methane (1T–7T) and bis(pyrazolyl)methane (1P–11P) was synthesised following previously reported methodologies and their antibacterial activity was tested using the collection strains of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa. Moreover, the novel compound 2P was fully characterized by IR, UV–Vis and NMR spectroscopy. To evaluate antibacterial activity, minimal inhibitory concentrations (MICs), minimal bactericidal concentrations (MBCs), minimum biofilm inhibitory concentrations (MBICs), and minimum biofilm eradication concentrations (MBECs) assays were carried out at different concentrations (2–2000 µg/mL). The MTT assay and Resazurin viability assays were performed in both human liver carcinoma HepG2 and human colorectal adenocarcinoma Caco-2 cell lines at 48 h. Of all the synthesised compounds, 2P had an inhibitory effect on Gram-positive strains, especially against S. aureus. The MIC and MBC of 2P were 62.5 and 2000 µg/mL against S. aureus, and 250 and 2000 µg/mL against E. faecalis, respectively. However, these values were > 2000 µg/mL against E. coli and P. aeruginosa. In addition, the MBICs and MBECs of 2P against S. aureus were 125 and > 2000 µg/mL, respectively, whereas these values were > 2000 µg/mL against E. faecalis, E. coli, and P. aeruginosa. On the other hand, concentrations up to 250 µg/mL of 2P were non-toxic doses for eukaryotic cell cultures. Thus, according to the obtained results, the 2P nitrogen-based compound showed a promising anti-Gram-positive effect (especially against S. aureus) both on planktonic state and biofilm, at non-toxic concentrations.


Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 295
Author(s):  
Sebastián Candelaria-Dueñas ◽  
Rocío Serrano-Parrales ◽  
Marisol Ávila-Romero ◽  
Samuel Meraz-Martínez ◽  
Julieta Orozco-Martínez ◽  
...  

In Tehuacán-Cuicatlán valley (Mexico), studies have been carried out on the essential oils of medicinal plants with antimicrobial activity and it was found that they present compounds in common such as: α-pinene, β-pinene, carvacrol, eugenol, limonene, myrcene, ocimene, cineole, methyl salicylate, farnesene, and thymol. The goal of this study was to assess the antimicrobial activity of essential oils’ compounds. The qualitative evaluation was carried out by the Kirby Baüer agar diffusion technique in Gram-positive bacteria (11 strains), Gram-negative bacteria (18 strains), and yeasts (8 strains). For the determination of the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), the agar dilution method was used. All the evaluated compounds presented antimicrobial activity. The compounds eugenol and carvacrol showed the largest inhibition zones. Regarding yeasts, the compounds ocimene, cineole, and farnesene did not show any activity. The compounds eugenol, carvacrol, and thymol presented the lowest MIC; bactericidal effect was observed at MIC level for S. aureus 75MR, E. coli 128 MR, and C albicans CUSI, for different compounds, eugenol, carvacrol, and thymol. Finally, this study shows that the essential oils of plants used by the population of Tehuacán-Cuicatlán valley share compounds and some of them have antibacterial and fungicidal activity.


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