scholarly journals Shiga Toxin (Stx)-Binding Glycosphingolipids of Primary Human Renal Cortical Epithelial Cells (pHRCEpiCs) and Stx-Mediated Cytotoxicity

Toxins ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 139
Author(s):  
Johanna Detzner ◽  
Elisabeth Krojnewski ◽  
Gottfried Pohlentz ◽  
Daniel Steil ◽  
Hans-Ulrich Humpf ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Epithelial Cells ◽  
Shiga Toxin ◽  
Saturated Fatty Acids ◽  
Human Kidney ◽  
Enterohemorrhagic Escherichia Coli ◽  
Highly Sensitive ◽  
Liquid Ordered ◽  
Uremic Syndrome

Human kidney epithelial cells are supposed to be directly involved in the pathogenesis of the hemolytic–uremic syndrome (HUS) caused by Shiga toxin (Stx)-producing enterohemorrhagic Escherichia coli (EHEC). The characterization of the major and minor Stx-binding glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer) and globotetraosylceramide (Gb4Cer), respectively, of primary human renal cortical epithelial cells (pHRCEpiCs) revealed GSLs with Cer (d18:1, C16:0), Cer (d18:1, C22:0), and Cer (d18:1, C24:1/C24:0) as the dominant lipoforms. Using detergent-resistant membranes (DRMs) and non-DRMs, Gb3Cer and Gb4Cer prevailed in the DRM fractions, suggesting their association with microdomains in the liquid-ordered membrane phase. A preference of Gb3Cer and Gb4Cer endowed with C24:0 fatty acid accompanied by minor monounsaturated C24:1-harboring counterparts was observed in DRMs, whereas the C24:1 fatty acid increased in relation to the saturated equivalents in non-DRMs. A shift of the dominant phospholipid phosphatidylcholine with saturated fatty acids in the DRM to unsaturated species in the non-DRM fractions correlated with the GSL distribution. Cytotoxicity assays gave a moderate susceptibility of pHRCEpiCs to the Stx1a and Stx2a subtypes when compared to highly sensitive Vero-B4 cells. The results indicate that presence of Stx-binding GSLs per se and preferred occurrence in microdomains do not necessarily lead to a high cellular susceptibility towards Stx.

scholarly journals EnterohemorrhagicEscherichia coliinfection stimulates Shiga toxin 1 macropinocytosis and transcytosis across intestinal epithelial cells

2011 ◽  
Vol 301 (5) ◽  
pp. C1140-C1149 ◽  
Author(s):  
Valeriy Lukyanenko ◽  
Irina Malyukova ◽  
Ann Hubbard ◽  
Michael Delannoy ◽  
Edgar Boedeker ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Shiga Toxin ◽  
Molecular Mechanisms ◽  
Intestinal Epithelial Cells ◽  
Enterohemorrhagic Escherichia Coli ◽  
Intestinal Epithelial ◽  
Systemic Complications ◽  
E Coli ◽  
Systemic Spread ◽  
Uremic Syndrome

Gastrointestinal infection with Shiga toxins producing enterohemorrhagic Escherichia coli causes the spectrum of gastrointestinal and systemic complications, including hemorrhagic colitis and hemolytic uremic syndrome, which is fatal in ∼10% of patients. However, the molecular mechanisms of Stx endocytosis by enterocytes and the toxins cross the intestinal epithelium are largely uncharacterized. We have studied Shiga toxin 1 entry into enterohemorrhagic E. coli-infected intestinal epithelial cells and found that bacteria stimulate Shiga toxin 1 macropinocytosis through actin remodeling. This enterohemorrhagic E. coli-caused macropinocytosis occurs through a nonmuscle myosin II and cell division control 42 (Cdc42)-dependent mechanism. Macropinocytosis of Shiga toxin 1 is followed by its transcytosis to the basolateral environment, a step that is necessary for its systemic spread. Inhibition of Shiga toxin 1 macropinocytosis significantly decreases toxin uptake by intestinal epithelial cells and in this way provides an attractive, antibiotic-independent strategy for prevention of the harmful consequences of enterohemorrhagic E. coli infection.

scholarly journals Primary Human Colon Epithelial Cells (pHCoEpiCs) Do Express the Shiga Toxin (Stx) Receptor Glycosphingolipids Gb3Cer and Gb4Cer and Are Largely Refractory but Not Resistant towards Stx

2021 ◽  
Vol 22 (18) ◽  
pp. 10002
Author(s):  
Johanna Detzner ◽  
Charlotte Püttmann ◽  
Gottfried Pohlentz ◽  
Hans-Ulrich Humpf ◽  
Alexander Mellmann ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Shiga Toxin ◽  
Human Kidney ◽  
Human Colon ◽  
Enterohemorrhagic Escherichia Coli ◽  
Intestinal Lumen ◽  
Brain Endothelial Cells ◽  
Detergent Resistant Membranes ◽  
Globo Series ◽  
Major Target

Shiga toxin (Stx) is released by enterohemorrhagic Escherichia coli (EHEC) into the human intestinal lumen and transferred across the colon epithelium to the circulation. Stx-mediated damage of human kidney and brain endothelial cells and renal epithelial cells is a renowned feature, while the sensitivity of the human colon epithelium towards Stx and the decoration with the Stx receptor glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer, Galα1-4Galβ1-4Glcβ1-1Cer) and globotetraosylceramide (Gb4Cer, GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer) is a matter of debate. Structural analysis of the globo-series GSLs of serum-free cultivated primary human colon epithelial cells (pHCoEpiCs) revealed Gb4Cer as the major neutral GSL with Cer (d18:1, C16:0), Cer (d18:1, C22:1/C22:0) and Cer (d18:1, C24:2/C24:1) accompanied by minor Gb3Cer with Cer (d18:1, C16:0) and Cer (d18:1, C24:1) as the dominant lipoforms. Gb3Cer and Gb4Cer co-distributed with cholesterol and sphingomyelin to detergent-resistant membranes (DRMs) used as microdomain analogs. Exposure to increasing Stx concentrations indicated only a slight cell-damaging effect at the highest toxin concentration of 1 µg/mL for Stx1a and Stx2a, whereas a significant effect was detected for Stx2e. Considerable Stx refractiveness of pHCoEpiCs that correlated with the rather low cellular content of the high-affinity Stx-receptor Gb3Cer renders the human colon epithelium questionable as a major target of Stx1a and Stx2a.

scholarly journals Whole-genome characterization of hemolytic uremic syndrome-causing Shiga toxin-producing Escherichia coli in Sweden

Virulence ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 1296-1305
Author(s):  
Ying Hua ◽  
Milan Chromek ◽  
Anne Frykman ◽  
Cecilia Jernberg ◽  
Valya Georgieva ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Hemolytic Uremic Syndrome ◽  
Shiga Toxin ◽  
Whole Genome ◽  
Genome Characterization ◽  
Uremic Syndrome

scholarly journals Characterization of planktonic and biofilm fatty acid profiles and evaluation of their trophic transfer potential to Hyalella Azteca

2021 ◽  
Author(s):  
Jerry Chien-Yao Chao
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Bacterial Species ◽  
Saturated Fatty Acids ◽  
Hyalella Azteca ◽  
Fatty Acid Profiles ◽  
Fa Composition ◽  
Branched Chain ◽  
Transfer Potential

Fatty acid (FA) composition between biofilms and batch planktonic cultures were compared for two bacterial species Pseudomonas aeruginosa and Staphylococcus aureaus. Biofilm cultures exhibited decrease in saturated fatty acids (SAFA) that potentially conform to a more fluidic biophysical membrane property. The amount of FA in the biofilms' extracellular polymeric substance was not sufficient to consider it having a major contribution to the observed differences between biofilms and batch planktonic cultures. While biofilm grazing by the amphipod Hyalella azteca was evident, only certain bacteria-specific FA appeared to have the potential to be retained (odd-number SAFA and branched-chain FA). H. azteca with diet strictly consisted of bacteria biofilms did not demonstrate significant changes in their nutritional condition in terms of ω-3 and ω-6 polyunsaturated fatty acids (PUFA): combined with the results from fasting trials, H. azteca appears to have the capacity to retain ω-3 and ω-6 PUFAs up to 10 days.

scholarly journals Identification and Characterization of Oleaginous Yeast Isolated from Kefir and Its Ability to Accumulate Intracellular Fats in Deproteinated Potato Wastewater with Different Carbon Sources

2017 ◽  
Vol 2017 ◽  
pp. 1-19 ◽  
Author(s):  
Iwona Gientka ◽  
Marek Kieliszek ◽  
Karolina Jermacz ◽  
Stanisław Błażejak
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Fossil Fuels ◽  
Unsaturated Fatty Acids ◽  
Saturated Fatty Acids ◽  
Carbon Sources ◽  
Debaryomyces Hansenii ◽  
Industry Waste ◽  
Identification And Characterization

The search for efficient oleaginous microorganisms, which can be an alternative to fossil fuels and biofuels obtained from oilseed crops, has been going on for many years. The suitability of microorganisms in this regard is determined by their ability to biosynthesize lipids with preferred fatty acid profile along with the concurrent utilization of energy-rich industrial waste. In this study, we isolated, characterized, and identified kefir yeast strains using molecular biology techniques. The yeast isolates identified wereCandida inconspicua,Debaryomyces hansenii,Kluyveromyces marxianus,Kazachstania unispora, andZygotorulaspora florentina. We showed that deproteinated potato wastewater, a starch processing industry waste, supplemented with various carbon sources, including lactose and glycerol, is a suitable medium for the growth of yeast, which allows an accumulation of over 20% of lipid substances in its cells. Fatty acid composition primarily depended on the yeast strain and the carbon source used, and, based on our results, most of the strains met the criteria required for the production of biodiesel. In particular, this concerns a significant share of saturated fatty acids, such as C16:0 and C18:0, and unsaturated fatty acids, such as C18:1 and C18:2. The highest efficiency in lipid biosynthesis exceeded 6.3 g L−1.Kazachstania unisporawas able to accumulate the high amount of palmitoleic acid.

scholarly journals Genotypic Characterization of Non-O157 Shiga Toxin–Producing Escherichia coli in Beef Abattoirs of Argentina

2011 ◽  
Vol 74 (12) ◽  
pp. 2008-2017 ◽  
Author(s):  
M. O. MASANA ◽  
B. A. D'ASTEK ◽  
P. M. PALLADINO ◽  
L. GALLI ◽  
L. L. DEL CASTILLO ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Hemolytic Uremic Syndrome ◽  
Shiga Toxin ◽  
Pattern Analysis ◽  
Severe Disease ◽  
Pulsed Field Gel Electrophoresis ◽  
Lower Proportion ◽  
Uremic Syndrome ◽  
Sporadic Cases

The non-O157 Shiga toxin–producing Escherichia coli (STEC) contamination in carcasses and feces of 811 bovines in nine beef abattoirs from Argentina was analyzed during a period of 17 months. The feces of 181 (22.3%) bovines were positive for non-O157 STEC, while 73 (9.0%) of the carcasses showed non-O157 STEC contamination. Non-O157 STEC strains isolated from feces (227) and carcasses (80) were characterized. The main serotypes identified were O178:H19, O8:H19, O130:H11, and O113:H21, all of which have produced sporadic cases of hemolytic-uremic syndrome in Argentina and worldwide. Twenty-two (7.2%) strains carried a fully virulent stx/eae/ehxA genotype. Among them, strains of serotypes O103:[H2], O145:NM, and O111:NM represented 4.8% of the isolates. XbaI pulsed-field gel electrophoresis pattern analysis showed 234 different patterns, with 76 strains grouped in 30 clusters. Nine of the clusters grouped strains isolated from feces and from carcasses of the same or different bovines in a lot, while three clusters were comprised of strains distributed in more than one abattoir. Patterns AREXSX01.0157, AREXBX01.0015, and AREXPX01.0013 were identified as 100% compatible with the patterns of one strain isolated from a hemolytic-uremic syndrome case and two strains previously isolated from beef medallions, included in the Argentine PulseNet Database. In this survey, 4.8% (39 of 811) of the bovine carcasses appeared to be contaminated with non-O157 STEC strains potentially capable of producing sporadic human disease, and a lower proportion (0.25%) with strains able to produce outbreaks of severe disease.

Characterization of a cDNA encoding a human kidney, cytochrome 4A fatty acid ω-hydroxylase and the cognate enzyme expressed in Escherichia coli

1993 ◽  
Vol 1172 (1-2) ◽  
pp. 161-166 ◽  
Author(s):  
Colin N.A. Palmer ◽  
Toby H. Richardson ◽  
Keith J. Griffin ◽  
Mei-Hui Hsu ◽  
A.Scott Muerhoff ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Human Kidney

scholarly journals Shiga toxin remodels the intestinal epithelial transcriptional response to Enterohemorrhagic Escherichia coli

2021 ◽  
Vol 17 (2) ◽  
pp. e1009290
Author(s):  
Alyson R. Warr ◽  
Carole J. Kuehl ◽  
Matthew K. Waldor
Keyword(s):  
Escherichia Coli ◽  
Epithelial Cells ◽  
Diarrheal Disease ◽  
Rabbit Model ◽  
Transcriptional Response ◽  
Enterohemorrhagic Escherichia Coli ◽  
Intestinal Epithelial ◽  
Transcriptional Responses ◽  
Gene Encoding ◽  
Uremic Syndrome

Enterohemorrhagic Escherichia coli (EHEC) is a food-borne pathogen that causes diarrheal disease and the potentially lethal hemolytic uremic syndrome. We used an infant rabbit model of EHEC infection that recapitulates many aspects of human intestinal disease to comprehensively assess colonic transcriptional responses to this pathogen. Cellular compartment-specific RNA-sequencing of intestinal tissue from animals infected with EHEC strains containing or lacking Shiga toxins (Stx) revealed that EHEC infection elicits a robust response that is dramatically shaped by Stx, particularly in epithelial cells. Many of the differences in the transcriptional responses elicited by these strains were in genes involved in immune signaling pathways, such as IL23A, and coagulation, including F3, the gene encoding Tissue Factor. RNA FISH confirmed that these elevated transcripts were found almost exclusively in epithelial cells. Collectively, these findings suggest that Stx potently remodels the host innate immune response to EHEC.

scholarly journals Cognitive deficits found in a pro-inflammatory state are independent of ERK 1/2 signaling in the murine brain hippocampus treated with Shiga toxin 2 from enterohemorrhagic Escherichia Coli

2020 ◽  
Author(s):  
Clara Berdasco ◽  
Alipio Pinto ◽  
Mariano Blake ◽  
Fernando Correa ◽  
Nadia A. Longo Carbajosa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Central Nervous System ◽  
Nervous System ◽  
Hemolytic Uremic Syndrome ◽  
Signaling Pathway ◽  
Shiga Toxin ◽  
Enterohemorrhagic Escherichia Coli ◽  
Shiga Toxin 2 ◽  
Uremic Syndrome ◽  
The Brain

AbstractShiga toxin 2 (Stx2) from enterohemorrhagic Escherichia coli (EHEC) produces hemorrhagic colitis, hemolytic uremic syndrome (HUS) and acute encephalopathy. The mortality rate in HUS increases significantly when the central nervous system (CNS) is involved. Besides, EHEC also releases lipopolysaccharide (LPS). Many reports have described cognitive dysfunctions in HUS patients, the hippocampus being one of the brain areas targeted by EHEC infection. In this context, a translational murine model of encephalopathy was employed to establish the deleterious effects of Stx2 and the contribution of LPS in the hippocampus. Results demonstrate that systemic administration of a sublethal dose of Stx2 reduced memory index and produced depression like behavior, pro-inflammatory cytokine release and NF-kB activation independent of the ERK 1/2 signaling pathway. On the other hand, LPS activated NF-kB dependent on ERK 1/2 signaling pathway. Cotreatment of Stx2 with LPS aggravated the pathologic state, while dexamethasone treatment succeeded in preventing behavioral alterations. Our present work suggests that the use of drugs such as corticosteroids or NF-kB signaling inhibitors may serve as neuroprotectors from EHEC infection.Author SummaryShiga toxin (Stx) from enterohemorrhagic Escherichia coli (EHEC) is one of the most virulent factors responsible for hemolytic uremic syndrome (HUS). Stx2, the endemic variant targets the brain, among other organs, thus inducing encephalopathies. Central nervous system (CNS) compromise was the main predictor of death in patients with HUS. Stx2 may exert a direct action in the CNS, by disrupting the neurovascular unit. In this context, we investigate the molecular signaling triggered by Stx2 in the murine brain hippocampus involved in inflammatory mechanisms that altered hippocampal-related cognitive behaviors. The present data underscore that the use of drugs such as dexamethasone or those blocking the cascade by preventing NF-kB translocation to the nucleus may serve as effective neuroprotectors with potentially beneficial use in the clinic.

scholarly journals Isolation and Characterization of Sorbitol-Fermenting Shiga Toxin (Verocytotoxin)-Producing Escherichia coli O157:Hßtrains in the Czech Republic

1998 ◽  
Vol 36 (7) ◽  
pp. 2135-2137 ◽  
Author(s):  
Martina Bielaszewska ◽  
Herbert Schmidt ◽  
Mohamed A. Karmali ◽  
Rasik Khakhria ◽  
Jan Janda ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Czech Republic ◽  
Shiga Toxin ◽  
Escherichia Coli O157 ◽  
The Czech Republic ◽  
Isolation And Characterization ◽  
First Finding ◽  
Uremic Syndrome ◽  
Genotypic Characteristics

Two sorbitol-fermenting (SF) Shiga toxin-producingEscherichia coli (STEC) O157:H− strains were isolated from patients with hemolytic-uremic syndrome in the Czech Republic in 1995. Their phenotypic and genotypic characteristics and genomic DNA fingerprints were identical or closely related to those of SF STEC O157:H− strains isolated in Germany in 1988 to 1997. This indicates that the Czech isolates belong to the SF STEC O157 clone which is widespread in Germany. It is the first finding of the clone outside Germany.

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