A Combination of ß-Aescin and Newly Synthesized Alkylamidobetaines as Modern Components Eradicating the Biofilms of Multidrug-Resistant Clinical Strains of Candida glabrata.

The current trend in microbiological research aimed at limiting the development of biofilms of multidrug-resistant microorganisms is increasingly towards the search for possible synergistic effects between various compounds. This work presents a combination of a naturally occurring compound, ß-aescin, newly synthesized alkylamidobetaines (AABs) with a general structure-CnTMDAB, and antifungal drugs. The research we conducted consists of several stages. The first stage concerns determining biological activity (antifungal) against selected multidrug-resistant strains of Candida glabrata (C. glabrata) with the highest ability to form biofilms. The second stage of this study determined the activity of ß-aescin combinations with antifungal compounds and alkylamidobetaines. In the next stage of this study, the ability to eradicate a biofilm on the polystyrene surface of the combination of ß-aescin with alkylamidobetaines was examined. It has been shown that the combination of ß-aescin and alkylamidobetaine can firmly remove biofilms and reduce their viability. The last stage of this research was to determine the safety regarding the cytotoxicity of both ß-aescin and alkylamidobetaines. Previous studies on the fibroblast cell line have shown that C9 alkylamidobetaine can be safely used as a component of anti-biofilm compounds. This research increases the level of knowledge about the practical possibilities of using anti-biofilm compounds in combined therapies against C. glabrata.

Methoxychalcones as potential anticancer agents for colon cancer: Is membrane perturbing potency relevant?

Chalcones are naturally produced by many plants, and constitute precursors for the synthesis of flavons and flavanons. They were shown to possess antibacterial, antifungal, anti-cancer, and anti- inflammatory properties. The goal of the study was to assess the suitability of three synthetic methoxychalcones as potential anticancer agents. In a panel of colon cancer cell lines they were demonstrated to be cytotoxic, proapoptotic, causing cell cycle arrest, and increasing intracellular level of reactive oxygen species. Anticancer activity of the compounds was not diminished in the presence of stool extract containing microbial enzymes that could change the structure of chalcones. Moreover, methoxychalcones interacted strongly with model phosphatidylcholine membranes as detected by differential scanning calorimetry. Metohoxychalcones particularly affected the properties of lipid domains in giant unilamellar liposomes formed from raft-mimicking lipid composition. This may be of importance since many molecular targets for therapy of metastatic colon cancer are raft-associated receptors (e.g., receptor tyrosine kinases). The importance of membrane perturbing potency of methoxychalcones for their biological activity was additionally corroborated by the results obtained by molecular modelling.

Palmitic Acid Modulates Microglial Cell Response to Metabolic Endotoxemia in an In Vitro Study.

Metabolic endotoxemia (ME) is characterized by a 2-3-fold increase in blood endotoxin levels and low-grade systemic inflammation without apparent infection. ME is usually accompanied by metabolic syndrome, characterized by central obesity and hyperlipidemia. According to numerous studies, ME may lead to functional brain disorders, including cognitive decline, depression, and dementia. In the current in vitro study, we aimed to determine the direct and indirect impact of endotoxin (LPS) and palmitic acid (PA), representing saturated fatty acids, on the inflammatory and oxidative stress response in the human microglial HMC3 cells unstimulated and stimulated with IFNγ. The study's results revealed that direct HMC3 cell exposition to endotoxin and PA increased inflammatory response measured as levels of IL-6 and MCP-1 released into the medium and PGE2 levels in cell lysates. Moreover, direct HMC3 cell treatment with PA and LPS induced oxidative stress, i.e., ROS and COX-2 production and lipid peroxidation. On the contrary, an indirect effect of LPS and PA on microglial cells, assessed as the impact of macrophage metabolites, was much lower regarding the inflammatory response, although still associated with oxidative stress. Interestingly, IFNγ had a protective effect on microglial cells, reducing the production of pro-inflammatory mediators and oxidative stress in HMC3 cells treated directly and indirectly with LPS and PA.

The Role of Palmitic Acid in the Co-Toxicity of Bacterial Metabolites to Endothelial Cells.

Introduction: Metabolic endotoxemia most often results from obesity and is accompanied by an increase in the permeability of the intestinal epithelial barrier, allowing co-absorption of bacterial metabolites and diet-derived fatty acids into the bloodstream. A high-fat diet (HFD) leading to obesity is a significant extrinsic factor in developing vascular atherosclerosis. In this study, we evaluated the effects of palmitic acid (PA) as a representative of long-chain saturated fatty acids (LCSFA) commonly present in HFDs, along with endotoxin (LPS; lipopolysaccharide) and uremic toxin indoxyl sulfate (IS), on human vascular endothelial cells (HUVECs). Methods: HUVECs viability was measured based on tetrazolium salt metabolism, and cell morphology was assessed with fluorescein-phalloidin staining of cells' actin cytoskeleton. The effects of simultaneous treatment of endothelial cells with PA, LPS, and IS on nitro-oxidative stress in vascular cells were evaluated quantitatively with fluorescent probes. The expression of vascular cell adhesion molecule VCAM-1, E-selectin, and occludin, an essential tight junction protein, in HUVECs treated with these metabolites was evaluated in Western blot. Results: PA, combined with LPS and IS, did not influence HUVECs viability but induced stress on actin fibers and focal adhesion complexes. Moreover, PA combined with LPS significantly enhanced reactive oxygen species (ROS) production in HUVECs but decreased nitric oxide (NO) generation. PA also considerably increased the expression of VCAM-1 and E-selectin in HUVECs treated with LPS or IS but decreased occludin expression. Conclusion: Palmitic acid enhances the toxic effect of metabolic endotoxemia on the vascular endothelium.

Human Gut Microbiota in Coronary Artery Disease: A Systematic Review and Meta-Analysis.

In recent years, the importance of the gut microbiome in human health and disease has increased. Growing evidence suggests that gut dysbiosis might be a crucial risk factor for coronary artery disease (CAD). Therefore, we conducted a systematic review and meta-analysis to determine whether or not CAD is associated with specific changes in the gut microbiome. The V3-V4 regions of the 16S rDNA from fecal samples were analyzed to compare the gut microbiome composition between CAD patients and controls. Our search yielded 1181 articles, of which 21 met inclusion criteria for systematic review and 7 for meta-analysis. The alpha-diversity, including observed OTUs, Shannon and Simpson indices, was significantly decreased in CAD, indicating the reduced richness of the gut microbiome. The most consistent results in a systematic review and meta-analysis pointed out the reduced abundance of Bacteroidetes and Lachnospiraceae in CAD patients. Moreover, Enterobacteriaceae, Lactobacillus, and Streptococcus taxa demonstrated an increased trend in CAD patients. The alterations in the gut microbiota composition are associated with qualitative and quantitative changes in bacterial metabolites, many of which have pro-atherogenic effects on endothelial cells, increasing the risk of developing and progressing CAD.

The Use of Endo-Cellulase and Endo-Xylanase for the Extraction of Apple Pectins as Factors Modifying Their Anticancer Properties and Affecting Their Synergy with the Active Form of Irinotecan.

Pectin constitutes an essential component of dietary fiber. Modified pectins from various sources possess potent anticancer and immunomodulatory activities. In this study, two pectins isolated from apple pomace by Trichoderma enzyme treatment, PX (with endo-xylanase) and PCX (with both endo-cellulase and endo-xylanase), were studied in colon cancer cell lines (HCT 116, Caco-2, and HT-29). Both pectins reduced colon cancer cell viability, induced apoptosis, and increased intracellular amounts of reactive oxygen species. Additionally, synergy between pectin and an active form of irinotecan, SN-38, in all aspects mentioned above, was discovered. This drug is a common component of cytotoxic combinations recommended as treatment for colon cancer patients. PX and PCX demonstrated significant anti-inflammatory activity in lipopolysaccharide-stimulated cells. Interaction of apple pectins with galectin-3 and Toll-like Receptor 4 (TLR4) was suggested to be responsible for their anticancer and anti-inflammatory effect. Since PCX was more active than PX in almost all experiments, the role of the enzyme used to obtain the pectin for its biological activity was discussed. It was concluded that co-operation between both enzymes was needed to obtain the molecule of the most beneficial properties. The low molecular mass of PCX together with a high proportion of rhamnogalacturonan I (RG I) regions seemed to be crucial for its superior activity.

Co-toxicity of Endotoxin and Indoxyl Sulfate, Gut-Derived Bacterial Metabolites, to Vascular Endothelial Cells in Coronary Arterial Disease Accompanied by Gut Dysbiosis.

Gut dysbiosis, alongside a high-fat diet and cigarette smoking, is considered one of the factors promoting coronary arterial disease (CAD) development. The present study aimed to research whether gut dysbiosis can increase bacterial metabolites concentration in the blood of CAD patients and what impact these metabolites can exert on endothelial cells. The gut microbiomes of 15 age-matched CAD patients and healthy controls were analyzed by 16S rRNA sequencing analysis. The in vitro impact of LPS and indoxyl sulfate at concentrations present in patients' sera on endothelial cells was investigated. 16S rRNA sequencing analysis revealed gut dysbiosis in CAD patients, further confirmed by elevated LPS and indoxyl sulfate levels in patients' sera. CAD was associated with depletion of Bacteroidetes and Alistipes. LPS and indoxyl sulfate demonstrated co-toxicity to endothelial cells inducing reactive oxygen species, E-selectin, and monocyte chemoattractant protein-1 (MCP-1) production. Moreover, both of these metabolites promoted thrombogenicity of endothelial cells confirmed by monocyte adherence. The co-toxicity of LPS and indoxyl sulfate was associated with harmful effects on endothelial cells, strongly suggesting that gut dysbiosis-associated increased intestinal permeability can initiate or promote endothelial inflammation and atherosclerosis progression.

Antiadherent and antibacterial properties of TiO2-coated and TiO2:Ag-coated stainless steel orthodontic wires against S. mutans bacteria.

PURPOSE: Conventional orthodontic treatment with stainless steel orthodontic wires may be detrimental to oral health, as it contributes to demineralized lesions and increases adhesion and bacterial biofilm formation, which contributes to cavity development. An alternative that has been investigated to reduce the side effects of orthodontic treatment is the use of coating materials with antimicrobial nanoparticles. This study aims to evaluate the antiadherent and antibacterial properties of TiO2-coated and TiO2:Ag-coated stainless steel orthodontic wires against S. mutans bacteria. METHODS: In the sol-gel method, TiO2:Ag thin films were deposited on stainless steel orthodontic wires. Coated archwires were analyzed for their antibacterial and antiadherent properties. The evaluation of Streptococcus mutans adhesion to the orthodontic wires' surface was conducted according to the type of coating used, biofilm formation assay, and measurement of the pH of the bacterial community. RESULTS: In the microbiological test, the TiO2:Ag coatings revealed a statistically significant difference in terms of microbial adhesion and biofilm formation by Streptococcus mutans. The TiO2:Ag coating on stainless steel wire increased pH levels in the saliva environment. CONCLUSIONS: It can be concluded that antimicrobial orthodontic wires coated with silver TiO2 nanoparticles using the sol-gel thin film are a promising choice for improving orthodontic treatment.

Newly Obtained Apple Pectin as an Adjunct to Irinotecan Therapy of Colorectal Cancer Reducing E. coli Adherence and ß-Glucuronidase Activity.

Colorectal cancer (CRC) is the second cause of cancer death worldwide. The composition and enzymatic activity of colonic microbiota can significantly affect the effectiveness of CRC chemotherapy. Irinotecan is a drug widely used to treat colon cancer. However, the transformation of a drug-glucuronide (SN-38G) back to its active form (SN-38) by bacterial ß-glucuronidase (GUS) constitutes the primary reason for the observed intestinal toxicity of irinotecan. It was demonstrated that novel enzymatically extracted apple pectin (PC) might be a promising candidate for an adjunct to irinotecan therapy. PC itself reduced the viability of HCT 116 and Caco-2 colorectal cancer cells, induced apoptosis, and increased intracellular reactive oxygen species production. Moreover, PC enhanced the cytotoxic and proapoptotic effect of irinotecan (at concentrations below its IC50), i.e., synergistic effect was recorded. Additionally, PC exhibited potent anti-inflammatory properties and prevented adhesion of prototype adherent-invasive E. coli (AIEC) LF82 strain and laboratory K-12C600 strain to colon cancer cells. PC was also identified to be an effective inhibitor of bacterial GUS activity. Altogether, novel apple pectin was identified as a promising candidate for a supplement to irinotecan therapy that might alleviate its side-effects via inhibition of bacterial GUS and thus increasing its therapeutic efficacy.

Production, Mechanical Properties and Biomedical Characterization of ZrTi-Based Bulk Metallic Glasses in Comparison with 316L Stainless Steel and Ti6Al4V Alloy.

Microstructure, mechanical properties, corrosion resistance, and biocompatibility were studied for rapidly cooled 3 mm rods of Zr40Ti15Cu10Ni10Be25, Zr50Ti5Cu10Ni10Be25, and Zr40Ti15Cu10Ni5Si5Be25 (at.%) alloys, as well as for the reference 316L stainless steel and Ti-based Ti6Al4V alloy. Microstructure investigations confirm that Zr-based bulk metallic samples exhibit a glassy structure with minor fractions of crystalline phases. The nanoindentation tests carried out for all investigated composite materials allowed us to determine the mechanical parameters of individual phases observed in the samples. The instrumental hardness and elastic to total deformation energy ratio for every single phase observed in the manufactured Zr-based materials are higher than for the reference materials (316L stainless steel and Ti6Al4V alloy). A scratch tester used to determine the wear behavior of manufactured samples and reference materials revealed the effect of microstructure on mechanical parameters such as residual depth, friction force, and coefficient of friction. Electrochemical investigations in simulated body fluid performed up to 120 h show better or comparable corrosion resistance of Zr-based bulk metallic glasses in comparison with 316L stainless steel and Ti6Al4V alloy. The fibroblasts viability studies confirm the good biocompatibility of the produced materials. All obtained results show that fabricated biocompatible Zr-based materials are promising candidates for biomedical implants that require enhanced mechanical properties.

Cornelian Cherry Iridoid-Polyphenolic Extract Improves Mucosal Epithelial Barrier Integrity in Rat Experimental Colitis and Exerts Antimicrobial and Antiadhesive Activities In Vitro.

BACKGROUND AND AIMS: Inflammatory bowel disease pharmacotherapy, despite substantial progress, is still not satisfactory for both patients and clinicians. In view of the chronic and relapsing disease course and not always effective treatment with adverse effects, attempts to search for new, more efficient, and safer substances are essential and reasonable. This study was designed to elucidate the impact of cornelian cherry iridoid-polyphenolic extract (CE) and loganic acid (LA) on adherent-invasive E. coli growth and adhesion in vitro and to assess the effect of pretreatment with CE or LA on the course of intestinal inflammation in rat experimental colitis compared with sulfasalazine. METHODS: Antibacterial and antiadhesive activities of CE and LA were assessed using microdilution, Int407 cell adherence, and yeast agglutination assays. The colitis model was induced by 2,4,6-trinitrobenzenesulfonic acid. Studied substances were administered intragastrically for 16 days prior to colitis induction. Body weight loss; colon index; histological injuries; IL-23, IL-17, TNF-α, and chemerin levels; and STAT3, Muc2, and TFF3 mRNA expression were evaluated. RESULTS: Only CE exerted antimicrobial and antiadhesive activities in vitro and alleviated colonic symptoms. CE coadministrated with sulfasalazine was more effective than single compounds in reversing increased concentrations of TNF-α, IL-17, and chemerin and decreased Muc2 mRNA expression. CONCLUSIONS: CE exerted a protective effect against experimental colitis via impaired mucosal epithelial barrier restoration and intestinal inflammatory response attenuation and given concomitantly with sulfasalazine counteracted colitis in a more effective way than sulfasalazine alone, which indicates their synergistic interaction. The beneficial effect of CE may also be due to its bacteriostatic and antiadhesive activities.

The prevalence of mucosa-associated diffusely adherent Escherichia coli in children with inflammatory bowel disease.

BACKGROUND: The relationship of diffusely adherent Escherichia coli (DAEC) with pediatric inflammatory bowel disease (IBD) has not been previously studied. Diffusely adherent E. coli are a common cause of long-lasting childhood diarrhea and we postulated that they may induce inflammation of the intestinal mucosa, contributing to the development of IBD in susceptible children. OBJECTIVES: The aim of the study was to investigate the relationship between DAEC and pediatric IBD, including Crohn's disease (CD) and ulcerative colitis (UC). Diffusely adherent E. coli isolates were also assessed regarding their pathogenicity. MATERIAL AND METHODS: Diffusely adherent E. coli were screened among 130 E. coli strains isolated from intestinal biopsy specimens from 26 children with IBD using polymerase chain reaction (PCR) with primers specific to the pathotype and adherence assays to HEp-2 cells. Diffusely adherent E. coli were further analyzed for their ability to adhere to and invade polarized Caco-2 cells. The immunomodulatory effect of DAEC on the secretion of tumor necrosis factor α (TNF-α) by human monocyte-derived macrophages (MDM) was assessed using an immunoenzymatic assay. RESULTS: Diffusely adherent E. coli were recovered from 18 (69.2%) of the 26 intestinal biopsy specimens from both CD and UC patients. Most DAEC isolates carried AfaE3 adhesin, adhered to and were internalized by Caco-2 cells, and induced secretion of elevated levels of TNF-α. CONCLUSIONS: The study demonstrated the internalization of DAEC by intestinal epithelial cells and their ability to induce secretion of increased level of TNF-α in a Caco-2/macrophage compartmentalized culture. This indicated that the pathovar should be considered a pathobiont inducing inflammation of the intestinal mucosa in pediatric patients with IBD.

Effect of amyloid curli fibrils and curli CsgA monomers from Escherichia coli on in vitro model of intestinal epithelial barrier stimulated with cytokines.

Amyloid curli fibrils produced by Escherichia coli are well-known virulence factor influencing E. coli adhesion and biofilm formation. However, the impact of curli on intestinal epithelial barrier stimulated with proinflammatory cytokines is unknown. In the study, we examined the effect of curli produced by nonpathogenic E. coli K-12 and wild-type E. coli EC32 strains, and purified CsgA proteins on differentiated Caco-2 cell monolayers stimulated with a mixture of IL-1ß, TNF-α, and INFγ cytokines as a model of 'inflamed intestinal epithelial barrier' in vitro. The results of the study indicated that curliated E. coli adhered better to polarized Caco-2 cells than their curli-deficient mutants and the adherence was further augmented by stimulation of epithelial cells with proinflammatory cytokines. Interestingly, curli reduced internalization but enhanced intracellular survival of the wild-type E. coli strain EC32 within intestinal epithelial cells. Curli-expressing E. coli, as well as purified CsgA proteins, attenuated IL-8 secretion by unstimulated Caco-2 cells, although the effect was barely observed on cytokine-stimulated cells. The findings of the study revealed that curli fibrils are an important virulence factor enabling curliated E. coli to effectively colonize intestinal epithelium especially in individuals with inflammatory intestinal disorders.

A twelve-year retrospective analysis of prevalence and antimicrobial susceptibility patterns of Ureaplasma spp. and Mycoplasma hominis in the province of Lower Silesia in Poland.

OBJECTIVE: Genital mycoplasmas are opportunistic pathogens that have been associated with urogenital infections in humans. Only a few groups of antimicrobials are available for treatment of urogenital tract infections caused by genital mycoplasmas. However, emerging resistance of mycoplasmas to antimicrobial agents has been reported worldwide. The aim of the study was a retrospective analysis of the prevalence and antimicrobial susceptibility patterns of M. hominis and Ureaplasma spp. in patients with urogenital tract infections during a twelve-year period between 2003 and 2015. STUDY DESIGN: Mycoplasma IST2 test was used for the detection, enumeration, identification and antimicrobial susceptibility testing of genital mycoplasmas in 1182 samples from 778 women and 404 men with genitourinary tract infection. Indicative enumeration in the test determines whether the mycoplasma count in the sample is equal or higher than the threshold set at 104 colony forming units. RESULTS: A total of 152 (12.8%) samples were found to be positive for genital mycoplasmas. M. hominis was detected only in three samples and Ureaplasma spp. in 141 samples. Both, M. hominis and Ureaplasma spp. were detected in the remaining eight samples. In the analyzed period between 2003 and 2015, a gradually increasing resistance of ureaplasmas to ciprofloxacin and clarithromycin and decreasing resistance to ofloxacin, erythromycin and tetracycline were observed. Pristinamycin, josamycin and doxycycline were most active against Ureaplasma spp. In contrast, fluoroquinolones had the lowest efficacy against Ureaplasma spp. and as many as 116 (82.3%) and 77 (54.6%) of Ureaplasma spp. isolates were resistant to ciprofloxacin and ofloxacin, respectively. M. hominis isolates were uniformly resistant to azithromycin, clarithromycin and erythromycin but susceptible to josamycin, ofloxacin, doxycycline and pristinamycin. One-third of these isolates were resistant to ciprofloxacin and tetracycline. CONCLUSION: In the study Ureaplasma spp. and M. hominis were detected with relatively low frequency in comparison with other studies however, most of these isolates were resistant to ciprofloxacin indicating the need for better management of ciprofloxacin prescription. Important limitations of Mycoplasma IST2 assay concerning antimicrobial susceptibility testing and divergences between breakpoints in the test and EUCAST guidelines point the need to introduce new methodologies to improve evaluation of resistant strains at our region.

Intestinal epithelial barrier: The target for pathogenic Escherichia coli.

Diarrheagenic Escherichia coli strains are included in 9 pathotypes (pathovars) that present different virulence factors responsible for the patomechanism of infections they cause. As all other intestinal pathogens, E. coli exerts a significant effect on intestinal epithelium. To initiate the infection, these microorganisms have evolved countless strategies to subvert the epithelial barrier and efficiently colonize the intestinal epithelium. The barrier function of the intestinal epithelium is achieved by the presence of a tight junction protein network surrounding individual cells around their circumference that links neighboring cells and seals the intracellular space. Pathogenic E. coli strains may impair intestinal epithelial barrier in 3 different pathways: (i) through a direct effect of their virulence factors on tight junctions proteins, (ii) by disrupting host cell actin cytoskeleton that indirectly damages epithelial barrier, and (iii) via stimulation of the secretion of proinflammatory cytokines that directly disrupt epithelial tight junctions or trigger neutrophils migration through intestinal epithelium, thus disrupting the intestinal barrier. Most pathogenic E. coli generates all these 3 pathways concomitantly upon interaction with intestinal epithelium.

Relevance of serology for Mycoplasma pneumoniae infection among children with persistent cough.

BACKGROUND: Mycoplasma pneumoniae is an important cause of upper and lower respiratory tract infections. Cough and tracheobronchitis are the commonest features of M. pneumoniae infection but diagnosis based on clinical symptoms that may be due to other respiratory pathogens is impossible. Thus laboratory testing for M. pneumoniae is particularly important. Correct and rapid diagnosis of M. pneumoniae infections is of prime importance to introduce appropriate antibiotic treatment. OBJECTIVES: Evaluation of the incidence of IgM and IgG antibodies specific to M. pneumoniae among children with pneumonia and/or chronic cough. MATERIAL AND METHODS: Serum samples from 148 children with a history of chronic cough (lasting at least one month), recurrent respiratory tract infections, allergic rhinitis, and/or inflammatory changes on X-chest ray. First, all sera were screened for specific anti-M. pneumoniae antibodies using agglutination test following the detection of specific IgM and IgG anti-M. pneumoniae antibodies using immunoenzymatic assays. RESULTS: Out of the 148 serum samples, 57 (38.5%) gave positive screening results. However, the presence of M. pneumoniae-specific IgM and/or IgG antibodies was confirmed by immunoenzymatic assays in only 30 (52.6%) of these 57 positive samples. These results indicated that in as many as 27 (47.4%) out of the 57 serum samples screened, false-positive results occurred. CONCLUSIONS: Evaluation of acute- and convalescent-phase sera is necessary to make possible accurate interpretation of the serological testing results.

Colonization of the lower urogenital tract with Ureaplasma parvum can cause asymptomatic infection of the upper reproductive system in women: a preliminary study.

PURPOSE: Genital ureaplasmas are considered opportunistic pathogens of human genitourinary tract involved in adverse pregnancy sequelae and infertility. While association of Ureaplasma urealyticum with urogenital tract infections is well established, the role of Ureaplasma parvum in these infections is still insufficient. In the study, we compared how often cervicovaginal colonization with U. parvum is associated with the presence of these microorganisms in the upper genitourinary tract of fertile and infertile women. METHODS: We used PCR assay to determine the prevalence of U. parvum and U. urealyticum in pairs of specimens, i.e., vaginal swabs and Douglas' pouch fluid samples from consecutive 40 women with no symptoms of genital tract infection. RESULTS: In total, 19 (47.5 %) of the 40 samples were positive for ureaplasmas. U. parvum was simultaneously detected in pairs of samples in five (55.5 %) of the nine (47.4 %) women positive in PCR assay. As many as 5 (18.5 %) of the 27 infertile women and 1 (7.7 %) of the 13 fertile women showed infection of the upper genital tract with U. parvum. CONCLUSION: The results of the study demonstrated that colonization of the lower genital tract with U. parvum can produce asymptomatic infection of the upper reproductive system in women. These findings also imply that U. parvum may be present in the upper genital tract at the time of conception and might be involved in adverse pregnancy outcomes.

[Influence of Lactobacillus casei on biofilm formation by enteroaggregative Escherichia coli strains isolated from irritable bowel syndrome (IBS)]. / Wplyw Lactobacillus casei na tworzenie biofilmu przez enteroagregacyjne szczepy Escherichia coli izolowane od osób z zespolem jelita nadwrazliwego.

INTRODUCTION: Many pathogenic bacterial species have the ability to biofilm formation. In our study we determined the influence of Lactobacillus casei on biofilm formation by enteroaggregative Escherichia coli (EAEC) strains obtained from irritable bowel syndrome patients. METHODS: The ability of EAEC isolates to biofilm formation was assessed in the presence of various concentrations of the probiotic L. casei strain in an a semi- quantitative microtitre plate assays under culture conditions, similar to those prevailing in the human intestine. RESULTS: Depending on the concentrations L. casei inhibited biofilm formation of the majority (> 80%) of the EAEC strains. Concentration of 4.5 x 10(7) cfu/ml of L. casei was the most effective inhibitory dose, although a few strains (approximately 18%) formed the biofilm regardless of the presence and concentration of the probiotic L. casei strain. CONCLUSION: The inhibitory effect of L. casei on biofilm formation at most of studied EAEC strains suggest that L. casei may reduce the risk of developing persistent intestinal infections in humans.

Association of untypeable enteropathogenic Escherichia coli (EPEC) strains with persistent diarrhea in children from the region of lower Silesia in Poland.

Enteropathogenic Escherichia coli strains (EPEC) carrying the eae gene encoding intimin are divided into typical strains producing bundle forming pili, encoded by the bfpA gene, and atypical strains lacking the gene. In the study typical and atypical EPEC that did not agglutinated with EPEC polyvalent antisera but carrying virulence factors characteristic to other pathogenic E. coli i.e. diffusely adhering and enteroaggregative E. coli were isolated from 24 (43.6%) of 55 children > 10 years old with persistent diarrhea. These results indicated that non-typeable typical and atypical EPEC can contribute to chronic intestinal infections in teenagers.

ESBL-producing Escherichia coli isolated from children with acute diarrhea - antimicrobial susceptibility, adherence patterns and phylogenetic background.

BACKGROUND: Escherichia coli remains the principal bacterial pathogen in childhood diarrhea and constitutes an important public health problem, especially in developing countries. Diarrheagenic E. coli strains often display resistance to beta-lactams due to the production of extended-spectrum beta-lactamases (ESBLs). OBJECTIVES: A total of thirty ESBL-producing E. coli strains colonizing the gastrointestinal tracts of children with acute diarrhea were studied in order to determine their antimicrobial susceptibility, adherence patterns to the HEp-2 cell line and phylogenetic background. MATERIAL AND METHODS: ESBL production was detected by the double disk synergy test (DDST). The minimal inhibitory concentrations (MICs) of antibacterial drugs were determined by an agar dilution technique on Mueller-Hinton agar. The presence of bla(TEM), bla(SHV) and bla(CTX-M) determinants in the strains studied was ascertained by polymerase chain reaction (PCR). RESULTS: The strains displayed the resistance pattern typical of ESBL producers. The majority of them (23 out of 30) were found to produce CTX-M-type ESBLs conferring a high level of resistance to oxyimino-beta-lactams, especially to cefotaxime and ceftriaxone. In many cases, the strains exhibited resistance to non-beta-lactam antimicrobials, such as gentamicin, amikacin, co-trimoxazole and tetracycline. On the other hand, these strains were uniformly susceptible to carbapenems, to oxyimino-beta-lactams combined with clavulanic acid and to tigecycline. The E. coli strains were distributed among the four main phylogenetic groups: A, B1, B2 and D. The in vitro adhesion assay revealed that all but two of the strains adhered to the HEp-2 epithelial cell line. Aggregative and diffuse adherence patterns were found to be the most prevalent. CONCLUSIONS: CTX-M-type enzymes were the most prevalent ESBLs among the strains studied. As many as 40% of the diarrheagenic E. coli isolates were found to belong to phylogenetic group D, which usually comprises E. coli strains associated with extra intestinal infections. The effectiveness of tigecycline against ESBL-producing E. coli strains was similar to that of imipenem and meropenem.