scholarly journals Biochemical and Functional Analysis of Cyanobacterium Geitlerinema sp. LPS on Human Monocytes

2019 ◽  
Vol 171 (2) ◽  
pp. 421-430 ◽  
Author(s):  
Michelle Swanson-Mungerson ◽  
Philip G Williams ◽  
Joshua R Gurr ◽  
Ryan Incrocci ◽  
Vijay Subramaniam ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Lipid A ◽  
Positive Control ◽  
Cyanobacterial Blooms ◽  
Side Chains ◽  
Human Monocytes ◽  
E Coli ◽  
Proinflammatory Responses ◽  
The Difference ◽  
High Concentrations

Abstract Cyanobacterial blooms are an increasing source of environmental toxins that affect both human and animals. After ingestion of cyanobacteria, such as Geitlerinema sp., toxins and lipopolysaccharide (LPS) from this organism induce fever, gastrointestinal illness, and even death. However, little is known regarding the effects of cyanobacterial LPS on human monocytes after exposure to LPS upon ingestion. Based on our previous data using Geitlerinema sp. LPS (which was previously named Oscillatoria sp., a genus belonging to the same order as Geitlerinema), we hypothesized that Geitlerinema sp. LPS would activate human monocytes to proliferate, phagocytose particles, and produce cytokines that are critical for promoting proinflammatory responses in the gut. Our data demonstrate that Geitlerinema sp. LPS induced monocyte proliferation and TNF-α, IL-1, and IL-6 production at high concentrations. In contrast, Geitlerinema sp. LPS is equally capable of inducing monocyte-mediated phagocytosis of FITC-latex beads when compared with Escherichia coli LPS, which was used as a positive control for our experiments. In order to understand the mechanism responsible for the difference in efficacy between Geitlerinema sp. LPS and E. coli LPS, we performed biochemical analysis and identified that Geitlerinema sp. LPS was composed of significantly different sugars and fatty acid side chains in comparison to E. coli LPS. The lipid A portion of Geitlerinema sp. LPS contained longer fatty acid side chains, such as C15:0, C16:0, and C18:0, instead of C12:0 found in E. coli LPS which may explain the decreased efficacy and toxicity of Geitlerinema sp. LPS in comparison to E. coli LPS.

scholarly journals Uropathogenic Escherichia coli Biofilm-Forming Capabilities are not Predictable from Clinical Details or from Colonial Morphology

Diseases ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 11 ◽  
Author(s):  
Shane Whelan ◽  
Mary Claire O’Grady ◽  
Dan Corcoran ◽  
Karen Finn ◽  
Brigid Lucey
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Culture Medium ◽  
Recurrent Infection ◽  
Positive Control ◽  
Uropathogenic Escherichia Coli ◽  
E Coli ◽  
Recurrent Uti ◽  
Colonial Morphology ◽  
The Difference

Antibiotic resistance is increasing to an extent where efficacy is not guaranteed when treating infection. Biofilm formation has been shown to complicate treatment, whereby the formation of biofilm is associated with higher minimum inhibitory concentration values of antibiotic. The objective of the current paper was to determine whether biofilm formation is variable among uropathogenic Escherichia coli isolates and whether formation is associated with recurrent urinary tract infection (UTI), and whether it can be predicted by phenotypic appearance on culture medium A total of 62 E. coli isolates that were reported as the causative agent of UTI were studied (33 from patients denoted as having recurrent UTI and 29 from patients not specified as having recurrent UTI). The biofilm forming capability was determined using a standard microtitre plate method, using E. coli ATCC 25922 as the positive control. The majority of isolates (93.6%) were found to be biofilm formers, whereby 81% were denoted as strong or very strong producers of biofilm when compared to the positive control. Through the use of a Wilcox test, the difference in biofilm forming propensity between the two patient populations was found to not be statistically significant (p = 0.5). Furthermore, it was noted that colony morphology was not a reliable predictor of biofilm-forming propensity. The findings of this study indicate that biofilm formation is very common among uropathogens, and they suggest that the biofilm-forming capability might be considered when treating UTI. Clinical details indicating a recurrent infection were not predictors of biofilm formation.

Structural investigation of frozen-hydrated Omp C specimens prepared by the fatty acid monolayer technique

1983 ◽  
Vol 41 ◽  
pp. 734-735
Author(s):  
Chung-Fu Chang ◽  
Robert M. Glaeser
Keyword(s):  
Stearic Acid ◽  
Outer Membrane ◽  
Lipid A ◽  
Behenic Acid ◽  
Bulk Water ◽  
Structural Investigations ◽  
E Coli ◽  
Periodic Arrays ◽  
Squeeze Out ◽  
High Concentrations

Omp C (M.W. ∽36,000) is one of the major proteins in the outer membrane of E. coli. Trimeric Omp C forms a pore allowing small hydrophilic molecules to diffuse across the membrane. Specimens we have studied are prepared by reconstituting purified Omp C trimers with lipid A (the core structure of the outer membrane 1ipopolysaccharide). These specimens form 2-D periodic arrays with a size of ∽0.5 μm on edge.Initial structural investigations on negatively stained Omp C specimens have been reported by Grano et al. We now present a preliminary structural analysis of frozen-hydrated Omp C, using specimens prepared by a modification of the stearic-acid monolayer technique of Hayward et al. Stearate monolayers can successfully "squeeze out" the bulk water on the surface of the EM grid only at relatively high concentrations of Ca++ and high pH. In the current study, we replaced the stearic acid with behenic acid, CH3(CH2)20COOH, which can adhere to a suitably prepared EM grid from a subphase of distilled water.

252 VITRIFICATION OF BOVINE BLASTOCYSTS PRODUCED AFTER OOCYTE MATURATION IN MEDIA CONTAINING FATTY ACIDS

2008 ◽  
Vol 20 (1) ◽  
pp. 205
Author(s):  
M. A. Shehab-El-Deen ◽  
J. L. M. R. Leroy ◽  
D. Maes ◽  
A. Van Soom
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Stearic Acid ◽  
Dairy Cows ◽  
In Vitro Maturation ◽  
Embryo Quality ◽  
Positive Control ◽  
High Concentrations ◽  
Carry Over

High concentrations of non-esterified fatty acids (NEFA) during negative energy balance (NEB) in high yielding dairy cows have been proven to be partially responsible for reduced fertility. This hypothesis has been tested by the addition of NEFAs to in vitro maturation medium at concentrations present in follicular fluid during NEB. We aimed to evaluate whether high concentrations of palmitic acid (C16:0) (PA), stearic acid (C18:0) (SA), or oleic acid (C18:1) (OA) during oocyte maturation could have a carry-over effect on embryo quality and could subsequently affect embryo cryotolerance. Cumulus–oocyte complexes (n = 4600) were matured in serum-free TCM199 plus epidermal growth factor (EGF, 20 ng mL–1; negative control), supplemented with ethanol alone (positive control) or with 0.133 mmol L–1 PA, 0.067 mmol L–1 SA, or 0.200 mmol L–1 OA (NEFAs dissolved in ethanol). The three NEFAs were tested separately in 4 replicates for PA and 5 replicates for OA or SA. Each fatty acid tested per replicate including a negative and a positive control group. After the embryos were cultured for 7 days in SOF medium, the number of blastocysts was recorded and classified as expanded, hatching, or hatched. Then, blastocysts were cryopreserved by open pulled straw vitrification using the two-step approach described by Vajta et al. (1998 Mol. Reprod. Dev. 51, 53–58). Vitrified warmed embryos were cultured in groups of <25 per 50-μL droplet of modified SOF medium with 5% fetal calf serum (FCS) under mineral oil for 48 h and examined for re-expansion and hatching. The percentages of survival in the different treatment groups were analyzed using logistic regression analyses, including the effect of replicates. Survival or not was included as the dependent variable and group was the independent variable. For every fatty acid a separate model was used. For all analyses, differences were considered to be statistically significant at the P < 0.05 level. Addition of OA to in vitro maturation media had no significant effects on cryotolerance of embryos. However, addition of PA or SA to in vitro maturation media (Table 1) significantly (P < 0.05) decreased the survival of bovine blastocysts from 79% in the positive control to 57% in PA and from 61% to 53% in SA. The results of the present study indicate that maturation of oocytes in the presence of NEB-associated concentrations of PA and SA can have carry-over effects on embryo quality, leading to reduced cryotolerance. We suggest that elevated NEFA concentrations in the follicular fluid may be one of the factors through which NEB exerts its negative effects on fertility in high yielding dairy cows. Table 1. Survival percentage (mean ± SD) of vitrified expanded bovine blastocysts matured in palmitic acid (C16:0) or stearic acid (C18:0) The authors thank J. Mestach and G. Spaepen for their excellent technical support. This research was supported by the Ministry of the Flemish Community, Belgium, in cooperation with the Ministry of Higher Education, Egypt.

scholarly journals Acyl Chain Specificity of the Acyltransferases LpxA and LpxD and Substrate Availability Contribute to Lipid A Fatty Acid Heterogeneity in Porphyromonas gingivalis

2008 ◽  
Vol 190 (13) ◽  
pp. 4549-4558 ◽  
Author(s):  
Brian W. Bainbridge ◽  
Lisa Karimi-Naser ◽  
Robert Reife ◽  
Fleur Blethen ◽  
Robert K. Ernst ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Porphyromonas Gingivalis ◽  
Lipid A ◽  
Acyl Chain ◽  
Hydroxy Fatty Acids ◽  
Homologous Gene ◽  
Substrate Availability ◽  
E Coli ◽  
Acyl Chain Length

ABSTRACT Porphyromonas gingivalis lipid A is heterogeneous with regard to the number, type, and placement of fatty acids. Analysis of lipid A by matrix-assisted laser desorption ionization-time of flight mass spectrometry reveals clusters of peaks differing by 14 mass units indicative of an altered distribution of the fatty acids generating different lipid A structures. To examine whether the transfer of hydroxy fatty acids with different chain lengths could account for the clustering of lipid A structures, P. gingivalis lpxA (lpxAPg ) and lpxDPg were cloned and expressed in Escherichia coli strains in which the homologous gene was mutated. Lipid A from strains expressing either of the P. gingivalis transferases was found to contain 16-carbon hydroxy fatty acids in addition to the normal E. coli 14-carbon hydroxy fatty acids, demonstrating that these acyltransferases display a relaxed acyl chain length specificity. Both LpxA and LpxD, from either E. coli or P. gingivalis, were also able to incorporate odd-chain fatty acids into lipid A when grown in the presence of 1% propionic acid. This indicates that E. coli lipid A acyltransferases do not have an absolute specificity for 14-carbon hydroxy fatty acids but can transfer fatty acids differing by one carbon unit if the fatty acid substrates are available. We conclude that the relaxed specificity of the P. gingivalis lipid A acyltransferases and the substrate availability account for the lipid A structural clusters that differ by 14 mass units observed in P. gingivalis lipopolysaccharide preparations.

Contribution of Adrenaline to the Fibrinolytic Activity Evoked by E. Coli Endotoxin in the Rat

1983 ◽  
Vol 50 (02) ◽  
pp. 557-559 ◽  
Author(s):  
J F Fracasso ◽  
A M Rothschild
Keyword(s):  
Fatty Acid ◽  
Adrenal Gland ◽  
Intravenous Injection ◽  
Fibrinolytic Activity ◽  
Blocking Agent ◽  
E Coli ◽  
Adrenergic Blocking

SummaryIntravenous injection of E. coli endotoxin (ETX), of adrenaline (AD) or of carbamylcholine (CBCH), caused fibrinolytic activity (FA), directly detectable on plasminogen-rich fibrin plates, to appear in the plasma of the rat. Adrenodemedul- lation abolished responses to ETX or CBCH, but enhanced those to AD. Rats given ETX exhibited marked hypotension, followed by a compensatory phase of normotension abolished by adrenodemedullation and significantly attenuated by phenoxy- benzamine, an a-adrenergic blocking agent which however failed to block FA caused by either ETX or AD. Aspirin, but not indomethacin, inhibited FA evoked by ETX, AD or CBCH. These results suggest that FA evoked by ETX in the rat is caused by AD released from the adrenal gland and does involve the fatty acid cyclooxygenase system.

Hygienic risk assessment by monitoring pathogens in municipal sewage

2002 ◽  
Vol 2 (3) ◽  
pp. 23-28 ◽  
Author(s):  
C.-H. von Bonsdorff ◽  
L. Maunula ◽  
R.M. Niemi ◽  
R. Rimhanen-Finne ◽  
M.-L. Hänninen ◽  
...  
Keyword(s):  
Enteric Viruses ◽  
Detection Methods ◽  
Municipal Sewage ◽  
Small Community ◽  
E Coli ◽  
Cryptosporidium Oocysts ◽  
High Concentrations ◽  
One Year ◽  
Enteric Protozoa ◽  
Day Care Centre

The purpose of this study was to monitor the levels of human enteric viruses and enteric protozoa and to relate their presence to the microbes used as hygienic quality indicators in domestic sewage from a small community in Finland during a period of one year. Genome-based sensitive detection methods for the pathogens selected (astro- and Norwalk-like viruses, Giardia and Cryptosporidium) have become available only recently and thus no earlier data was available. The effluent sewage is delivered into a river that serves as raw water for a larger town and the pathogens therefore constitute a health risk. The results showed that all the monitored pathogens could be detected, and that enteric viruses were present at considerable concentrations in sewage. High concentrations of astrovirus in raw sewage were observed during a diarrhea epidemic in the local day-care centre. The presence of viruses did not correlate with the monitored bacterial indicators of faecal contamination (coliforms, E. coli and enterococci) or with bacteriophages (somatic coliphages, F-specific RNA phages and B. fragilis phages). Giardia cysts and Cryptosporidium oocysts were detected from one sample (1/10) each.

Identification of N-Substituted Triazolo-azetidines as Novel Antibacterials using pDualrep2 HTS Platform

2019 ◽  
Vol 22 (5) ◽  
pp. 346-354
Author(s):  
Yan A. Ivanenkov ◽  
Renat S. Yamidanov ◽  
Ilya A. Osterman ◽  
Petr V. Sergiev ◽  
Vladimir A. Aladinskiy ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Large Scale ◽  
Underlying Mechanism ◽  
Luciferase Assay ◽  
Reporter System ◽  
E Coli ◽  
Starting Point ◽  
High Concentrations ◽  
Mic Value

Aim and Objective: Antibiotic resistance is a serious constraint to the development of new effective antibacterials. Therefore, the discovery of the new antibacterials remains one of the main challenges in modern medicinal chemistry. This study was undertaken to identify novel molecules with antibacterial activity. Materials and Methods: Using our unique double-reporter system, in-house large-scale HTS campaign was conducted for the identification of antibacterial potency of small-molecule compounds. The construction allows us to visually assess the underlying mechanism of action. After the initial HTS and rescreen procedure, luciferase assay, C14-test, determination of MIC value and PrestoBlue test were carried out. Results: HTS rounds and rescreen campaign have revealed the antibacterial activity of a series of Nsubstituted triazolo-azetidines and their isosteric derivatives that has not been reported previously. Primary hit-molecule demonstrated a MIC value of 12.5 µg/mL against E. coli Δ tolC with signs of translation blockage and no SOS-response. Translation inhibition (26%, luciferase assay) was achieved at high concentrations up to 160 µg/mL, while no activity was found using C14-test. The compound did not demonstrate cytotoxicity in the PrestoBlue assay against a panel of eukaryotic cells. Within a series of direct structural analogues bearing the same or bioisosteric scaffold, compound 2 was found to have an improved antibacterial potency (MIC=6.25 µg/mL) close to Erythromycin (MIC=2.5-5 µg/mL) against the same strain. In contrast to the parent hit, this compound was more active and selective, and provided a robust IP position. Conclusion: N-substituted triazolo-azetidine scaffold may be used as a versatile starting point for the development of novel active and selective antibacterial compounds.

The metachor as a characteristic of the association of electrolytes in aqueous solutions

1984 ◽  
Vol 49 (5) ◽  
pp. 1061-1078 ◽  
Author(s):  
Jiří Čeleda ◽  
Stanislav Škramovský
Keyword(s):  
Aqueous Solutions ◽  
Apparent Volume ◽  
Solutions Of Electrolytes ◽  
Molal Volumes ◽  
The Difference ◽  
High Concentrations ◽  
Experimental Values ◽  
Suitable Characteristic ◽  
Association Of Ions ◽  
Very High

Based on the earlier paper introducing a concept of the apparent parachor of a solute in the solution, we have eliminated in the present work algebraically the effect which is introduced into this quantity by the additivity of the apparent molal volumes. The difference remaining from the apparent parachor after substracting the contribution corresponding to the apparent volume ( for which the present authors suggest the name metachor) was evaluated from the experimental values of the surface tension of aqueous solutions for a set of 1,1-, 1,2- and 2,1-valent electrolytes. This difference showed to be independent of concentration up to the very high values of the order of units mol dm-3 but it was directly proportional to the number of the free charges (with a proportionality factor 5 ± 1 cm3 mol-1 identical for all studied electrolytes). The metachor can be, for this reason, a suitable characteristic for detection of the association of ions and formation of complexes in the solutions of electrolytes, up to high concentrations where other methods are failing.

Apoprotein C effect on triglyceride transport in tandem-perfused rat hind end and liver

1984 ◽  
Vol 247 (3) ◽  
pp. G305-G310
Author(s):  
W. J. Kortz ◽  
J. R. Nashold ◽  
M. R. Greenfield ◽  
H. Hilderman ◽  
S. H. Quarfordt
Keyword(s):  
Fatty Acid ◽  
Free Fatty Acid ◽  
Fatty Acyl ◽  
Liver Fat ◽  
Molar Ratio ◽  
Perfusion System ◽  
In Vitro Perfusion ◽  
Arterial Inflow ◽  
High Concentrations

The metabolism of double-labeled triglyceride in a synthetic emulsion was defined in an in vitro perfusion system of rat hind end and liver described previously [Am. J. Physiol. 245 (Gastrointest. Liver Physiol. 8): G106-G112, 1983]. The metabolism of [3H]glycerol-[14C]triolein was defined in the absence of added apoproteins and with additions of human CII and both CII and CIII. Without apoprotein, a pronounced lipolysis of the triglyceride was recognized by high concentrations of radiolabeled glycerol and free fatty acid in the perfusate. The removal of an aliquot of hind-end venous effluent 5 min after adding the labeled triglyceride emulsion to the arterial inflow demonstrated a brisk lipolysis of the substrate when incubated outside the perfusion system. The addition of CII protein to the emulsion before its introduction into the tandem system eliminated perfusate lipolysis, both within the perfusion system and in incubations of aliquots withdrawn from the system. Intravascular lipolysis was not seen with triglyceride emulsions containing both CII and CIH or when an aliquot of hind-end venous effluent was incubated with triglycerides that had not been exposed to the perfusion system. The intravascular lipolysis observed for the [14C]triglyceride added to the tandem system without apoproteins was associated with relatively greater recoveries of 14C-fatty acyl in liver, fat, and muscle and relatively greater recoveries of 14CO2 than when CII alone or both CII and CIII were added with the triglyceride. The addition of CIII to CII in a 1:1 molar ratio increased the recovery of 14C-fatty acyl in muscle and the recovery as 14CO2.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Effects of Amino Acid Side-Chain Length and Chemical Structure on Anionic Polyglutamic and Polyaspartic Acid Cellulose-Based Polyelectrolyte Brushes

Polymers ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1789
Author(s):  
Dmitry Tolmachev ◽  
George Mamistvalov ◽  
Natalia Lukasheva ◽  
Sergey Larin ◽  
Mikko Karttunen
Keyword(s):  
Glutamic Acid ◽  
Chain Length ◽  
Chemical Structure ◽  
Side Chain ◽  
Side Chains ◽  
Side Chain Length ◽  
Polyelectrolyte Brushes ◽  
Grafting Density ◽  
The Difference ◽  
Cellulose Surface

We used atomistic molecular dynamics (MD) simulations to study polyelectrolyte brushes based on anionic α,L-glutamic acid and α,L-aspartic acid grafted on cellulose in the presence of divalent CaCl2 salt at different concentrations. The motivation is to search for ways to control properties such as sorption capacity and the structural response of the brush to multivalent salts. For this detailed understanding of the role of side-chain length, the chemical structure and their interplay are required. It was found that in the case of glutamic acid oligomers, the longer side chains facilitate attractive interactions with the cellulose surface, which forces the grafted chains to lie down on the surface. The additional methylene group in the side chain enables side-chain rotation, enhancing this effect. On the other hand, the shorter and more restricted side chains of aspartic acid oligomers prevent attractive interactions to a large degree and push the grafted chains away from the surface. The difference in side-chain length also leads to differences in other properties of the brush in divalent salt solutions. At a low grafting density, the longer side chains of glutamic acid allow the adsorbed cations to be spatially distributed inside the brush resulting in a charge inversion. With an increase in grafting density, the difference in the total charge of the aspartic and glutamine brushes disappears, but new structural features appear. The longer sides allow for ion bridging between the grafted chains and the cellulose surface without a significant change in main-chain conformation. This leads to the brush structure being less sensitive to changes in salt concentration.

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