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At least 91 records · Page 5Linked to original sources
PubMed2004

AcrAB-TolC directs efflux-mediated multidrug resistance in Salmonella enterica serovar typhimurium DT104.

Multidrug-resistant Salmonella enterica serovar Typhimurium definitive phage type 104 (DT104) strains harbor a genomic island, called Salmonella genomic island 1 (SGI1), which contains an antibiotic resistance gene cluster conferring resistance to ampicillin, chloramphenicol, florfenicol, streptomycin, sulfonamides, and tetracyclines. They may be additionally resistant to quinolones. Among the antibiotic resistance genes there are two, i.e., floR and tet(G), which code for efflux pumps of the major facilitator superfamily with 12 transmembrane segments that confer resistance to chloramphenicol-florfenicol and the tetracyclines, respectively. In the present study we determined, by constructing acrB and tolC mutants, the role of the AcrAB-TolC multidrug efflux system in the multidrug resistance of several DT104 strains displaying additional quinolone resistance or not displaying quinolone resistance. This study shows that the quinolone resistance and the decreased fluoroquinolone susceptibilities of the strains are highly dependent on the AcrAB-TolC efflux system and that single mutations in the quinolone resistance-determining region of gyrA are of little relevance in mediating this resistance. Overproduction of the AcrAB efflux pump, as determined by Western blotting with an anti-AcrA polyclonal antibody, appeared to be the major mechanism of resistance to quinolones. Moreover, chloramphenicol-florfenicol and tetracycline resistance also appeared to be highly dependent on the presence of AcrAB-TolC, since the introduction of mutations in the respective acrB and tolC genes resulted in a susceptible or intermediate resistance phenotype, according to clinical MIC breakpoints, despite the presence of the FloR and Tet(G) efflux pumps. Resistance to other antibiotics, ampicillin, streptomycin, and sulfonamides, was not affected in the acrB and tolC mutants of DT104 strains harboring SGI1. Therefore, AcrAB-TolC appears to direct efflux-mediated resistance to quinolones, chloramphenicol-florfenicol, and tetracyclines in multidrug-resistant S. enterica serovar Typhimurium DT104 strains.

Bacterial Proteins↗
PubMed2002

Induction of protection against the necrotrophic pathogens Phytophthora citrophthora and Alternaria solani in Lycopersicon esculentum Mill. by a novel synthetic glycoside combined with amines.

1,2,3,4-tetra-O-acetyl-6-ethyladipate-beta- D-glucopyranose (TOGE), a glycoside derivative of adipic acid monoethyl ester and 1,2,3,4-tetra-O-acetyl-beta- D-glucopyranose, was synthesized and the resistance-inducing activity shown by TOGE-1 (TOGE plus furfurylamine) and TOGE-2 (TOGE plus 1,3-diaminepropane) was assayed. TOGE-1 and TOGE-2 protected tomato plants against two different fungal pathogens Phytophthora citrophthora and Alternaria solani. Foliar treatments at very low concentrations (2.5 mg l(-1) TOGE, 0.5 mg l(-1) amine) clearly reduced the disease incidence for both pathogens. TOGE-2 application was the most effective on intact plants as well as on detached leaves, reducing fungal growth by more than 46% with respect to control plants. On the other hand TOGE-1 treatment reduced fungal advance by 21%. These results demonstrate a high protective effect against fungal infections for both chemicals. A possible direct antimicrobial effect was discounted due to the weak activity observed in vitro against these pathogens at the low concentrations used in plants. TOGE-2 treatment clearly activates resistance against both pathogens and improves the protective effect previously shown by FGA mixture (adipic acid monoethyl ester, 1,2,3,4-tetra-O-acetyl-beta- D-glucopyranose and furfurylamine) [V. Flors et al. (2001) J Agric Food Chem 49:2569-2575]. The obtained results indicate that TOGE-1 and TOGE-2 act as resistance inducers. Although their mode of action is still unknown, pre-challenge studies have demonstrated the induction of the phenylpropanoid pathway and antioxidant activities. Both chemicals have demonstrated a beneficial effect on plant development, increasing chlorophyll and protein contents, photosynthetic rate and water-use efficiency. The improvement of plant growth and development produced by these treatments suggests crop tolerance to these chemicals, although effective formulations that are safe to humans must be developed before this technology can be used commercially.

Adipates↗
PubMed2025

Versatile and Portable Cas12a-mediated Detection of Antibiotic Resistance Markers.

Antibiotic-resistant bacteria are spreading in clinical, industrial, and environmental ecosystems. The spreading dynamics to and from the environment are unknown, largely due to the lack of appropriate (robust, fast, low-cost) analytical assays. In this study, we developed C12a, a versatile molecular toolbox to detect genetic markers of antibiotic resistance using CRISPR/Cas12a. Biochemical characterization show that the C12a toolbox can detect less than 100 attoMolar of pure DNA fragments from the blaCTX-M15 and floR genes, conferring resistance to b-lactams and amphenicols, respectively important for human and veterinary uses. In microbiological assays, C12a detected less than 102 CFU/mL and high concordance was observed if compared to antibiotic susceptibility tests, PCR, or to whole genome sequencing. Additionally, C12a confirmed a high prevalence of the integrase/integron system in E. coli isolates containing multiple antibiotic resistance genes (ARGs). The C12a toolbox shows equivalent detection performance in diverse laboratory settings, results redout (Fluorescence vs FLA) or input sample. Altogether, this work presents a comprehensive proof-of-concept, development description, and biochemical characterization of a collection of molecular tools to detect antibiotic resistance markers in a one health setup.

Antibiotic Resistance Gene↗
PubMed1996

The Phytophthora sojae genome contains tandem repeat sequences which vary from strain to strain.

The oömycete Phytophthora sojae causes root and stem rot of soybean. In P. sojae, 37 cultivarspecific physiological races have been reported with different reactions against 13 single dominant resistance genes in soybean. Recent genetic studies have demonstrated that the P. sojae-soybean interaction follows Flor's gene-for-gene model. Genomic subtraction was carried out to isolate race-specific DNA sequences possibly including avirulence genes. DNA from a race 1 isolate was subtraction-enriched with the DNAs from race 19 and race 22 isolates. The enriched DNAs were used to differentially screen a P. sojae genomic library. Characterization of the clones obtained identified repetitive sequences with variable copy numbers among isolates. Five of these repetitive sequences were tandemly repeated and were localized on single chromosomes. The sixth corresponded to the ribosomal RNA genes. These observations suggest that gene amplification could contribute to the generation of genetic diversity in P. sojae. Enriched sequences corresponding to avirulence genes were not detected.

Base Sequence↗
PubMed1979

[Traumatically induced junctional activity of nevocellular nevi (author's transl)].

Forty papillomatous nevi, mainly with junctional nests, were partially removed surgically in such a way that a small margin was left intact on one side and the deep nevus cells were retained on the flor of the wound. After healing of the wound, a small layer of scar tissue separated the regenerated epidermis from the deep nevus cells, which were left in the dermis. After very differing intervals, as early as 30 days after the partial surgical removal, typical alterations could be observed in the epidermis above the cicatrical tissue. Most frequently, there was a hyperpigmentation with an increase in the number of basal fluorescing dendritic cells, which could also be found along the uppermost parts of epidermal appendages. Moreover, nest-like conglomerations of pigment-producing cells appeared in the form of junctional nevus cell nests. About 2-3 month after the partial surgical removal "Abtropfung" was visible, indicating a regeneration of the nevus. Fluorescence-histochemical and enzyme-histochemical results revealed that marked alterations can be found not only in the newly formed epidermis above the scar, which morphologically show minor alterations.

Adult↗
PubMed1988

Denaturation by heat, sodium dodecyl sulphate and dithiothreitol of globulins and phaseolin from dry bean (Phaseolus vulgaris L.).

The electrophoretic pattern of the untreated common bean globulin cv. Flor de Mayo had 7 protein fractions ranging from 620 to 120 kilodaltons (kd). The last molecular weight corresponds to the monomeric form. One of the objectives of the present work was to establish a comparison among denaturation by heat, sodium dodecyl sulphate (SDS) and dithiothreitol (DTT). At pH 6.0, two bands were resistant to heat treatment, after SDS treatment of the untreated globulin several bands disappeared and two new bands with 26 and 15 kd appeared. DTT did not change the electrophoretic pattern, due to the small quantity of free SH groups in the globulin. At pH 6.0, phaseolin is partly resistant to heat denaturation due to aggregation in an acidic environment. Isoelectrofocusing (IEF) and SDS polyacrylamide gel electrophoresis (SDS-PAGE) separated phaseolin into 10 protein fractions. The hypothesis is that phaseolin or globulin aggregation is due to the charge difference of fractions.

Dithiothreitol↗
PubMed2002

Correlation between acetaldehyde and ethanol resistance and expression of HSP genes in yeast strains isolated during the biological aging of sherry wines.

In the production of sherry wines, the process of biological aging is essential for the development of their organoleptic properties. This process involves velum formation by "flor" yeasts. Several of these yeast strains have been isolated and characterized with regard to their genetic, physiological and metabolic properties. In this work, we studied their resistance to cold-, osmotic-, oxidative-, ethanol- and acetaldehyde-stress, and found, in most cases, a correlation between resistance to acetaldehyde stress and ethanol stress and isolation from "soleras." Moreover, gene expression analysis revealed induction of the heat shock protein (HSP) genes HSP12, HSP82, and especially HSP26 and HSP104, under acetaldehyde stress in most of the strains. In strain C, there was a clear correlation between resistance to ethanol and acetaldehyde, the high induction of HSP genes by these compounds and its presence as the predominant strain in most levels of several soleras.

Acetaldehyde↗
PubMed2025

Genomic insights into Shigella species isolated from small ruminants and manure in the North West Province, South Africa.

This study investigated Shigella species' antibiotic resistance patterns and genomic characteristics from small ruminants and manure collected in Potchefstroom, North West, South Africa. Whole genome sequencing was used to determine resistome profiles of Shigella flexneri isolates from small ruminants' manure and Shigella boydii from sheep faeces. Comparative genomics was employed on the South African 261 S. flexneri strains available from GenBank, including the sequenced strains in this study, by investigating the serovars, antibiotic resistance genes (ARGs), and plasmid replicon types. The S. flexneri strains could not be assigned to known sequence types, suggesting novel or uncharacterized lineages. S. boydii R7-1A was assigned to sequence type 202 (ST202). Serovar 2A was the most common among South African S. flexneri strains, found in 96% of the 250 compared human-derived isolates. The shared mdf(A) was the most prevalent gene, identified in 99% of 261 S. flexneri genomes, including plasmid replicon types ColRNAI_1 (99%) and IncFII_1 (98%). Both species share a core set of resistance determinants mainly involving β-lactams (ampC1, ampC, ampH), macrolides (mphB), polymyxins (eptA, pmrF), multidrug efflux pumps (AcrAB-TolC, Mdt, Emr, Kpn families), and regulatory systems (marA, hns, crp, baeRS, evgAS, cpxA, gadX). However, S. boydii possesses additional resistance genes conferring resistance to tetracyclines (tet(A)), phenicols (floR), sulphonamides (sul2), and aminoglycosides (APH(3'')-Ib, APH(6)-Id), along with the acrEF efflux pump components (acrE, acrF). In contrast, S. flexneri harboured unique genes linked to polymyxin resistance (ugd) and regulatory functions (sdiA, gadW) that were absent in S. boydii. These findings highlight Shigella strains' genomic diversity and antimicrobial resistance potential in livestock-associated environments. Moreover, S. boydii highlights the potential risk of multidrug-resistant bacteria in farming and environmental routes. KEY POINTS: • First whole genome study of Shigella from manure and small ruminants in South Africa. • Shigella boydii strain carried multiple resistance genes to β-lactams and tetracycline. • Multidrug efflux pump gene mdf(A) was detected in 99% of South African Shigella flexneri strains.

Animals↗
PubMed2006

Enological functions of parietal yeast mannoproteins.

Parietal yeast mannoproteins play a very important role in the overall vinification process. Their production and release, both during winemaking and aging on lees, depends on the specific yeast strain and the nutritional conditions. The following enological functions of parietal yeast mannoproteins have been described: (a) adsorption of ochratoxin A; (b) combination with phenolic compounds; (c) increased growth of malolactic bacteria; (d) inhibition of tartrate salt crystallization; (e) interaction with flor wines; (f) prevention of haze; (g) reinforcement of aromatic components; (h) wine enrichment during aging on fine lees; (i) yeast flocculation and autolysis in sparkling wines. Further discoveries related to their enological functions are foreseeable. Yeast-derived mannoproteins may well induce chemical, sensorial and health benefits, thus greatly improving wine quality.

Fermentation↗
PubMed2006

Relationship among antimutagenic, antioxidant and enzymatic activities of methanolic extract from common beans (Phaseolus vulgaris L).

Common beans are rich in phenolic compounds, which can provide health benefits to the consumer. The objective of this work was to study the relationship among antimutagenicity, antioxidant and enzymatic activities of methanolic extract and trolox by principal components multivariate analysis. Antimutagenicity of phenolic compounds present in methanolic extract from the seed coat of common beans (P. vulgaris, Flor de Mayo Bajío cultivar) and trolox against AFB1 mutagenicity were evaluated in the Salmonella typhimurium microsuspension assay. Antioxidant capacity of methanolic extract and trolox were evaluated using beta-carotene and 1,1-diphenyl-2-picryhydrazyl (DPPH) in vitro model assays. Cythrome P450 activity was measured by fluorometric assay. For phenolic extract, trolox and phenolic extract+trolox, the inhibition on AFB1 mutagenicity in tester strain TA100 was 47, 59 and 69%, respectively. While in TA98 was 39, 48 and 68%. The inhibition of phenolic compounds, trolox and phenolic compounds+trolox on cytochrome P450 (CYP450) activity was 48, 59 and 88%, respectively. Correlation analysis showed that phenolic extract and trolox have high antimutagenic and antioxidant activity and also inhibited enzymatic activity. The results suggest that the primary mechanism of action of phenolic compounds in beans against AFB1 mutagenicity may be extra-cellular in the microsuspension assay.

Aflatoxin B1↗
PubMed2006

Relationship between ethanol tolerance, H+ -ATPase activity and the lipid composition of the plasma membrane in different wine yeast strains.

Ethanol tolerance, ATPase activity and the lipid composition of the plasma membrane to study potential relationship among them were examined in five different wine yeast strains. Yeast cells were subjected to ethanol stress (4% v/v). Principal component analysis of the results revealed that the wine yeasts studied can be distinguished in terms of ATPase activity and oleic acid (C18:1), and palmitoleic acid (C16:1), in plasma membrane. Multiple regression analysis was used to identify a potential influence of some components of the plasma membrane on ethanol tolerance and ATPase activity. Based on the results, the ergosterol, oleic acid and palmitoleic acid are highly correlated with ATPase activity and ethanol tolerance. Ethanol tolerance and the ATPase activity of the plasma membrane were correlated at the 96.64% level with the oleic acid and ergosterol in plasma membrane. The Saccharomyces cerevisiae var. capensis flor yeast strain, which exhibited the highest ergosterol concentration in plasma membrane when grown in the presence of 4% v/v ethanol, was found to be the most ethanol-tolerant.

Adenosine Triphosphatases↗
PubMed2004

International dissemination of antibiotic resistant strains of bacterial pathogens.

The increasing incidence of methicillin and multiple resistant Staphylococcus aureus (MRSA) in nosocomial infections is mainly associated with a wide, international dissemination of well defined clonal lineages (epidemic MRSA) which are clearly different from community acquired MRSA by molecular typing patterns and structure of the staphylococcal cassette chromosome containing the mecA gene. Although belonging to a definite subpopulation within the species Enterococcus faecium, hospital associated vancomycin resistant isolates also containing the esp gene have very likely evolved by acquisition of glycopeptide resistance gene clusters at different occasions and at different times by a susceptible already disseminated clonal lineage. There is obviously a continuous selection of new types of extended spectrum beta-lactamases in enterobacteriaceae and also horizontal spread of bla-genes. Intrahospital dissemination of particular strains has often been observed, however, an international dissemination until now has been described only for Salmonella enterica serovar Typhimurium producing the CTX-M-3 enzyme. Multiresistant isolates of Salmonella enterica serovar Typhimurium phage type DT104 harbour a multiresistant gene cluster with resistance genes from taxonomically more unrelated species (tetG, floR, bla(PSE1)). Although in vitro transduction has been demonstrated, this gene cluster has only rarely been reported from isolates exhibiting other phage patterns of the same serovar or from other serovars of S. enterica.

Animals↗
PubMed1996

Characterization of the binding properties and physiological action of divalinal-angiotensin IV, a putative AT4 receptor antagonist.

Divalinal-Ang IV [V psi (CH2-NH2)YV psi (CH2-NH2)HPF] is being employed increasingly as a specific AT4 antagonist. This use, which necessitates a comprehensive physiological and pharmacological evaluation of Divalinal-Ang IV's functional and receptor binding characteristics in order to ensure its efficacy and specificity, was the stimulus for this study using bovine adrenal membranes. [125I]Ang IV and [125I]Divalinal-Ang IV were shown to bind with high affinity to a similar number of binding sites, suggesting that both bound the same receptor. This notion was verified by competition curves using [125I]Ang IV and [125I]Divalinal-Ang IV that indicated identical rank order affinities for several angiotensin-related peptides and 100% cross-displacement by Ang IV and Divalinal-Ang IV. Furthermore, an autoradiographic comparison of [125I]Ang IV and [125I]Divalinal-Ang IV in 20 microns sections of bovine adrenals revealed near identical binding distributions characterized by heavy binding in the glomerulosa layer and the medulla. Physiological studies in which test compounds were injected into the internal carotid of the rat and cerebral blood flor (CBF) was measured by laser Doppler flowmetry indicated that pretreatment with Divalinal-Ang IV, but not DuP 753 or PD123177, blocked the increased flow observed with Ang IV infusion. Conversely, DuP 753, but not Divalinal-Ang IV or PD123177, inhibited the decrease in flow witnessed with Ang II. Metabolic stability studies utilizing rat kidney homogenates as a peptidase source, demonstrated that the structural changes present in Divalinal-Ang IV greatly increased its resistance to metabolism as compared to Ang IV. Together, these studies show that Divalinal-Ang IV is a stable, efficacious and specific inhibitor of AT4 receptors.

Angiotensin II↗
PubMed1998

Group III human metabotropic glutamate receptors 4, 7 and 8: molecular cloning, functional expression, and comparison of pharmacological properties in RGT cells.

Cloning and expression in a stable mammalian cell line co-transfected with a glutamate transporter (RGT cells) were used as tools for studying the functions and pharmacological properties of group III metabotropic glutamate receptors (mGluRs). Complementary DNAs (cDNAs) encoding the human mGluR4, human mGluR7, and human mGluR8 were isolated from human cerebellum, fetal brain or retinal cDNA libraries. The human mGluR4, mGluR7 and mGluR8 receptors were 912, 915 and 908 amino acid residues long and share 67-70% amino acid similarity with each other and 42-45% similarity with the members of mGluR subgroups I and II. The human mGluR4 and mGluR7 had amino acid identity of 96% and 99.5% with rat mGluR4 and 7, respectively, whereas the human mGluR8 has 98.8% amino acid identity with the mouse mGluR8. The nucleotide and amino acid sequences in the coding region of human mGluR4 and mGluR7 were found to be identical to the previously published sequences by Flor et al. and Makoff et al. Following stable expression in RGT cells, highly significant inhibitions of forskolin stimulation of cAMP production by group III agonists were found for each receptor. The relative potencies of the group III agonist L-AP4 varied greatly between the group III clones, being mGluR8>mGluR4 >> mGluR7. The reported group II mGluR agonist L-CCG-I was a highly potent mGluR8 agonist (EC50=0.35 microM), with significant agonist activities at both mGluR4 (EC50=3.7 microM) and mGluR7 (EC50=47 microM). The antagonist potency of the purported group III mGluR antagonist MPPG also varied among the receptors being human mGluR8 >> mGluR4 = mGluR7. The expression and second messenger coupling of human group III mGluRs expressed in the RGT cell line are useful to clearly define the subtype selectivities of mGluR ligands.

ATP-Binding Cassette Transporters↗
PubMedDate not supplied

[Hydrodynamic and biochemical adaptation to perfusion pressure profile in isolated hypothermic renal perfusion].

OBJECTIVES: To evaluate hydrodynamic, biochemical and pathologic consequences of positive forced pressure on hypothermic isolated renal perfusions in comparison to softly progressive administration of perfusion solution. MATERIAL AND METHOD: 16 hypothermic experimental pig renal perfusion o 180 min completed under constant flow conditions. A vacuum computer-controlled pump was used. A comparison was performed of the results obtained by two groups of organs (forced perfusion pressure -I.B.- and soft perfusion pressure -I.S.). RESULTS: Even though perfusion flor did not offer significant differences among both groups at the end of the procedure, I.B. group showed medium perfusion pressure and renal vascular resistance values significantly higher. Nitric oxide metabolites concentration was also greater in I.B. group. Finally, these kidneys showed more tubular cell aedema and glomerular shrinkage in comparison to I.S. group. CONCLUSIONS: Sharp and forced hypothermic perfusion of the isolated kidney don't increase flow perfusion values. Inversely, hydrodynamic and pathologic results are worst in comparison to soft and progressive organ perfusion.

Adaptation, Physiological↗
PubMed2001

[Plants and pathogenic agents, a refined and dangerous relationship: the example of fungi].

Plant-fungus interactions are highly diverse, either being beneficial to the host plant such as those leading to mycorhizal symbiosis, or very detrimental when leading to severe diseases. Since the beginning of agriculture, improvement of plant resistance to pathogens has remained a major challenge. Breeding for resistance, first conducted empirically in the past centuries, was then performed on a more theoretical basis after the statement of heredity laws by Mendel at the end of the XIXth century. As a result, most cultivated species contain various cultivars whose resistance or susceptibility to a given pathogen species depend on their interaction with various races of that pathogen. Such highly specific race-cultivar systems are particularly suited for understanding the molecular dialogue which underlies compatible (host susceptible/pathogen virulent) or incompatible (host resistant/pathogen avirulent) interactions. During the twentieth century, one of the major events that paved the way for future research was the statement by Flor [1946, 1947] of the gene-for-gene concept. Studying inheritance of the disease phenotype in the interaction between flax and Melampsora lini he showed that resistance in the host and avirulence in the pathogen are dictated by single dominant genes which correspond one to one, i.e. one resistance gene for one avirulence gene. The fact that incompatibility may depend on the presence of only one resistance (R) gene in the host and one avirulence (Avr) gene in the pathogen was fully confirmed about 40 years later. Molecular genetics and complementation experiments have allowed to isolate numerous R and Avr genes from various plant-pathogen systems, and to verify the gene-for-gene concept. These studies have enlightened the elicitor/receptor concept, formerly introduced to account for the specificity of the compatible and incompatible interactions. The present knowledge of R and Avr genes also allows to predict how such genes have evolved and how they could be used to improve disease resistance. At the beginning of the twenty first century, this remains a major challenge in view of the severe losses caused by pests and pathogens to most crops on the earth.

Fungal Proteins↗
PubMed2000

Ecological impact of antibiotic use in animals on different complex microflora: environment.

Different means of interaction between microecological systems in different animal hosts (including humans) and the environment may occur during the transfer of resistant bacteria and their resistance genes. Spread of resistance takes place in different ways with respect to clonal spread of resistance strains by the spread of wide host range plasmids and translocatable elements. Commensals in ecosystems have a special significance and a pronounced capacity for acquisition and transfer of resistance genes as with Enterococcus faecium and Escherichia coli in the gut flora or Pseudomonas spp. in aquatic environments. The route of transmission from animals to humans by meat products is well established. Other routes via water and food plants (vegetables) have been investigated less, although resistance genes transfer in aquatic environments as evidenced from sequence comparison of such genes (e.g. tetR, floR in Salmonella typhimurium DT104). Whether this is due to rare but important transfer events or whether there is a more frequent exchange in aquatic or terrestrial environments needs further elucidation.

Animal Feed↗
PubMed2002

Yeast-induced inhibition of (+)-catechin and (-)-epicatechin degradation in model solutions.

(+)-catechin and (-)-epicatechin degradation in water-alcohol solutions containing Fe2+ and tartaric acid was studied in the presence and absence of yeasts. On the basis of the results, yeast partially inhibited the degradation of both flavans, with much slower formation of browning products absorbing at 420 and 520 nm. In comparative terms, yeast was found to be more efficient toward the degradation products of (+)-catechin absorbing at the latter wavelength. Likewise, the presence of yeast decreased the yield of a group of colored compounds eluting at high retention times in HPLC and indicated these as important contributors to color darkening in white wines. This inhibitory effect may in part account for the resistance to browning observed over periods of several years in sherry wines subjected to biological aging under flor yeast.

Catechin↗