Microbiota of female genital tract - functional overview of microbial flora from vagina to uterine tubes and placenta.

Microorganisms and eukaryotic human cells coexist in synergistic relationships in nearly every niche of the human body. The female genital tract consisting of the vagina, uterus with its cervix and endometrium, uterine tubes and ovaries - harbors its own typical microbiota, which accounts for 9 % of the total bacterial population in females. To this organ system, we also assigned the microbiome of the placenta, which has not been studied much until now. Among the spectrum of microbial species, the female genital tract is mainly dominated by Lactobacillus species, which are considered to be one of the simplest yet most important microbial communities. However, this relationship between macro- and micro-organisms seems to have a number of physiological functions, e.g., the vaginal and cervical microbiota have unique impact on reproductive health. The aim of this review was to provide current view on female genital tract microbiota and its role in reproductive health. We describe in detail the association of vaginal or tubal epithelium with microbiota or the role of microbiota in normal placental function.

Gradual cold acclimation induces cardioprotection without affecting ß-adrenergic receptor-mediated adenylyl cyclase signaling.

Novel strategies are needed that can stimulate endogenous signaling pathways to protect the heart from myocardial infarction. The present study tested the hypothesis that appropriate regimen of cold acclimation (CA) may provide a promising approach for improving myocardial resistance to ischemia/reperfusion (I/R) injury without negative side effects. We evaluated myocardial I/R injury, mitochondrial swelling, and ß-adrenergic receptor (ß-AR)-adenylyl cyclase-mediated signaling. Male Wistar rats were exposed to CA (8°C, 8 h/day for a week, followed by 4 wk at 8°C for 24 h/day), while the recovery group (CAR) was kept at 24°C for an additional 2 wk. The myocardial infarction induced by coronary occlusion for 20 min followed by 3-h reperfusion was reduced from 56% in controls to 30% and 23% after CA and CAR, respectively. In line, the rate of mitochondrial swelling at 200 µM Ca2+ was decreased in both groups. Acute administration of metoprolol decreased infarction in control group and did not affect the CA-elicited cardiprotection. Accordingly, neither ß1-AR-Gsα-adenylyl cyclase signaling, stimulated with specific ligands, nor p-PKA/PKA ratios were affected after CA or CAR. Importantly, Western blot and immunofluorescence analyses revealed ß2- and ß3-AR protein enrichment in membranes in both experimental groups. We conclude that gradual cold acclimation results in a persisting increase of myocardial resistance to I/R injury without hypertension and hypertrophy. The cardioprotective phenotype is associated with unaltered adenylyl cyclase signaling and increased mitochondrial resistance to Ca2+-overload. The potential role of upregulated ß2/ß3-AR pathways remains to be elucidated.NEW & NOTEWORTHY We present a new model of mild gradual cold acclimation increasing tolerance to myocardial ischemia/reperfusion injury without hypertension and hypertrophy. Cardioprotective phenotype is accompanied by unaltered adenylyl cyclase signaling and increased mitochondrial resistance to Ca2+-overload. The potential role of upregulated ß2/ß3-adrenoreceptor activation is considered. These findings may stimulate the development of novel preventive and therapeutic strategies against myocardial ischemia/reperfusion injury.

Genetic variations of interleukin-8, CXCR1 and CXCR2 genes and risk of acute pyelonephritis in children.

Acute pyelonephritis (APN) is the most severe form of urinary tract infection, the etiopathogenesis of which is still not well understood. Previous studies demonstrated that chemotaxis of neutrophils into the tissue and across the infected epithelial layer is a key step in rapid bacterial clearance. Variations within genes encoding the major chemokine interleukin-8 and its receptors CXCR1 and CXCR2 are therefore attractive candidates for participation in genetic predisposition to APN. The aim of our study was to evaluate the association of single nucleotide polymorphisms (SNPs) -251 T/A, +781 C/T, +1633 C/T and +2767 A/T in the IL-8 gene, +2608 G/C in the CXCR1 gene and +1208 C/T in the CXCR2 gene with susceptibility to APN in the Slovak population. PCR-SSP and PCR-RFLP were used to genotype SNPs in 147 children with APN (62 with recurrent and 85 with episodic form) and 215 healthy individuals. Statistical analysis revealed significantly increased frequency of CXCR1 +2608 C allele (P = 0.0238, OR = 2.452, 95% CI = 1.147-5.243) and GC genotype (P = 0.0201, OR = 2.627, 95% CI = 1.188-5.810) and lower frequency of CXCR2 +1208 T allele (P = 0.0408, OR = 0.645, 95% CI = 0.429-0.972) and TT+TC genotypes (P = 0.0497, OR = 0.5273, 95% CI = 0.288-0.964) in patients with recurrent APN compared with healthy controls. Furthermore, the A allele of IL-8 -251 T/A SNP was also significantly overrepresented in patients with recurrent APN when compared with those with only single episode of APN (P = 0.0439, OR = 1.627, 95% CI = 1.019-2.599). Our results indicate that the minor CXCR1 +2608 C allele is associated with significantly increased susceptibility to APN in childhood, while the CXCR2 +1208 T allele confers protection from recurrent APN. Moreover, allele A of the IL-8 -251 T/A may also increase the risk of developing recurrent attacks after the first-time APN.

The redox state of glutathione in erythrocytes of individuals with Down syndrome.

BACKGROUND: The redox state of glutathione has been used as indicator for the redox environment of the cell. OBJECTIVES: To investigate relationships between the redox environments, the SOD activity, total antioxidant status and the oxidation stress markers production (MDA and lipofuscin). METHODS: Individuals with Down syndrome and age-matched healthy controls were enrolled into a study. Some parameters of oxidative stress in serum were determined: reduced glutathione, oxidized glutathione, redox potential of this couple (Eh), activity of superoxide dismutase in the red blood cells as well as malondialdehyde and lipofuscin. RESULTS: In the group of persons with DS statistically significant decrease in the GSH concentration was found, however, no differences in the GSSG concentration versus controls was observed. The redox potential values for couple GSH/GSSG are a statistically significantly increased in DS individuals compared to controls. CONCLUSION: In this study we highlighted the different ways of view at the role of GSH in metabolism of persons with DS. It is useful to look at the GSH and GSSG concentrations separately as well as at redox potential value, which influence total redox state of organism (Tab. 2, Fig. 3, Ref. 30) Full Text (Free, PDF) www.bmj.sk.

[Urinary tract infections in children caused by uropathogenic strains of Escherichia coli and the role of the innate immune response mediated by the toll-like receptor 4 and antimicrobial peptide cathelicidin in their clinical course]. / Infekcie mocových ciest u detí spôsobené uropatogénnymi kmenimi Escherichia coli a vplyv vrodených imunitných mechanizmov toll-like receptora 4 a antimikróbneho peptidu katelicidínu, na ich klinický priebeh.

Uropathogenic Escherichia coli (UPEC) are the causative organisms in more than 80% of urinary tract infections. The bacteria are introduced to the urinary tract that, except for the external part of the urethra, is free from microbial colonization, and thus can cause acute, potentially life-threatening infections with possible progression to chronic disease. The course of such infection depends not only on the agent involved but also on the activation of protective mechanisms. The recently described Toll-like receptor family and antimicrobial peptide cathelicidin appear to play an important role in the recognition and activation functions.

Hemolytic uremic syndrome caused by verotoxin-producing Escherichia coli O26. Case report.

Verotoxin-producing Escherichia coli (VTEC) are nowadays among the most important emerging group of food-borne pathogens (VTEC strains cause gastroenteritis that can be complicated by the hemorrhagic colitis or hemolytic uremic syndrome, HUS). Escherichia coli 026 producing verotoxin 2 was isolated and its identity confirmed by examination of phenotype and genotype; the strain was first described in Slovakia in association with the development of HUS in a 4-year-old girl.

Detection of virulence factors in alpha-haemolytic Escherichia coli strains isolated from various clinical materials.

In total, 201 alpha-haemolytic Escherichia coli isolates from various clinical materials (urine samples and vaginal and rectal swabs) were examined by PCR for the presence of genes for the virulence factors alpha-haemolysin (hly), cytotoxic necrotising factor type 1 (cnf1), P-fimbriae (pap), S/F1C-fimbriae (sfa/foc), aerobactin (aer) and afimbrial adhesin (afaI). Among vaginal isolates, 96% were positive for cnf1, compared with 80% of urine strains (p 0.02) and 63% of rectal strains (p 0.0001). Similarly, sfa/foc-specific DNA sequences were found in 97% of vaginal isolates compared with 75% of rectal strains (p 0.004). The afa1 and aer genes were associated more with rectal alpha-haemolytic E. coli strains than with extra-intestinal isolates. The results suggested that CNF1 and/or S/F1C-fimbriae contribute to colonisation and persistence of alpha-haemolytic E. coli strains in the vaginal environment.

A family outbreak of haemolytic uraemic syndrome and haemorrhagic colitis caused by verocytotoxigenic Escherichia coli O157 from unpasteurised cow's milk in Slovakia.

This report describes a family outbreak of verocytotoxigenic Escherichia coli O157 (VTEC) infection, involving nine persons from one extended family, which occurred in eastern Slovakia. Three children suffered from haemolytic uraemic syndrome, two children had bloody diarrhoea, and four adults were asymptomatic carriers. Fourteen sorbitol-non-fermenting E. coli O157 isolates harbouring the vtx2, eae and ehxA genes were obtained. Verocytotoxin 2 activity was demonstrated in all 14 isolates. After epidemiological surveillance, the source of infection was identified as unpasteurised cow's milk.

Influence of 2,5-dihydroxybenzylidene aminoguanidine on lipid oxidative damage and on antioxidant levels in model diabetes mellitus.

2,5-Dihydroxybenzylidene aminoguanidine (BAG) is a structural analogue of the antidiabetic compound aminoguanidine, and is an example of a substance protecting diabetic rats from lipoprotein oxidation arising in oxidative stress conditions characteristic of diabetes mellitus. We found that administration of BAG to diabetic rats decreases their susceptibility to lipoprotein oxidation, decreases formation of conjugated dienes and 4-hydroxy-2-nonenal, and increases antioxidant potential of plasma. On the other hand, our results show that BAG has a negative influence on lipoprotein oxidation in control rats. Increased formation of 4-hydroxy-2-nonenal and conjugated dienes and a decrease in plasma antioxidant potential was observed when BAG was administered to control rats. It is therefore necessary to search for other structural modifications of this substance that would combine higher antidiabetic activity with less toxicity in healthy individuals.

Influence of beta-resorcylidene aminoguanidine on selected metabolic parameters and antioxidant status of rats with diabetes mellitus.

We studied the effects of administration of beta-resorcylidene aminoguanidine (RAG) to Wistar strain rats with experimental diabetes mellitus (DM) induced by streptozotocin. The effects studied included antioxidant levels in plasma and the liver, oxidative damage of lipids represented by the formation of substances reacting with thiobarbituric acid (TBARP) and selected biochemical indicators. The administration of RAG did not significantly affect antioxidant status of diabetic rats or hemoglobin glycation and plasma concentration of fructosamine. In diabetic rats, application of RAG decreased formation of TBARP in plasma but not in the liver. Moderate steatosis of liver and increased plasma levels of triacylglycerols in diabetic rats were significantly improved by application of RAG.

Detection of Shiga toxins, intimin and enterohemolysin in Escherichia coli strains isolated from children in eastern Slovakia.

Fifty Escherichia coli strains isolated from stool samples of 51 healthy children, 143 strains isolated from stool samples of 327 children with diarrhea and 24 strains isolated from stool samples of 21 children with suspected hemolytic uremic syndrome were examined for the presence of Shiga toxin-producing E. coli virulence factors (shiga toxin 1 and 2, intimin and enterohemolysin) and their genes. Vero-cell assay and latex agglutination were used for detection of Shiga toxin 1 and 2, TSB agar with washed erythrocytes was used for detection of enterohemolysin; genes encoding shiga toxin 1 and 2, intimin and enterohemolysin were detected using multiplex PCR. The presence of E. coli strains harboring genes encoding shiga toxin 1 and 2 (12 strains), intimin (34 strains) and enterohemolysin (12 strains) was demonstrated.

Occurrence and genetic association of selected virulence factors in clinical Escherichia coli isolates.

Occurrence of cnf1+ E. coli pathogenic strains among extraintestinal E. coli isolates was evaluated to explain an impact of cytotoxic necrotizing factor type 1 (CNF1) in human infections. A total of 120 E. coli isolates were characterized for presence of virulence factors cnf1- and pap--specific sequences by PCR, and the production of alpha-hemolysin using blood agar-plate test. Different association patterns among the detected virulence factors were obtained by comparison of various groups of clinical E. coli isolates. These differences probably reflect a potential impact of CNF1 in the colonization of vaginal environment.

Effects of aminoguanidine Schiff's base on biomarkers of the oxidative stress, 4-hydroxy-2-nonenal and conjugated dienes, in the model diabetes mellitus.

Diabetes mellitus evoked by streptozotocine in rats is associated with the oxidative stress. We examined the effect of Schiff's base 2,5-dihydroxybenzaldehyde with a well-known antidiabetic drug aminoguanidine, 2,5-dihydroxybenzilideneaminoguanidine (BAG) on the production of markers of oxidative stress such as 4-hydroxy-2-nonenal (4HNE) and conjugated dienes in diabetic rats. BAG administration did not affect glucose level in diabetic rats but significantly decreased the production of 4HNE and conjugated dienes. On the other hand, BAG caused the elevation of conjugated dienes and an insignificant increase of 4HNE levels in the control animals.

Influence of age on activities of antioxidant enzymes and lipid peroxidation products in erythrocytes and neutrophils of Down syndrome patients.

Thirty-seven individuals with Down syndrome (DS) were divided into four age categories: (i) 1 to < 6 years, (ii) 6 to < 13 years, (iii) 13 to < 20 years, and (iv) over 20 years. Activities of antioxidant enzymes found in individual age categories were different, but the differences between age groups were not statistically significant. We confirmed significantly higher activities of Cu/Zn superoxide dismutase (SOD) and glutathione peroxidase (GPx) in blood cells of people with DS as compared to 35 controls, which consisted, for the first time, of siblings of children with DS. No significant differences were found in activities of catalase and glutathione reductase in DS vs. controls. A significant difference was observed in serum concentration of malondialdehyde (MDA) in DS vs. controls (8.39 +/- 0.34 micromol/l vs. 7.34 +/- 0.27 micromol/l; p = .021) and concentration of MDA in erythrocytes of individuals with DS between the third and fourth age group (p = .05). In DS persons, an elevated ratio of SOD to catalase plus GPx with respect to the controls in all age categories was found, suggesting oxidative imbalance, potentially contributing to accelerated aging observed in these persons.

[Effect of losartan on antioxidant status in rats with diabetes mellitus]. / Vplyv losartanu na antioxidacný stav potkanov s diabetes mellitus.

The aim of this study was to monitor the effect of the administration of antihypertensive drug losartan on: (1) the antioxidant status of rats with experimental insulin-dependent diabetes mellitus; (2) oxidative damage which is represented by the production of compounds which can react with thiobarbituric acid (TBARP), and (3) some metabolic parameters. Losartan administration did not significantly influence the concentration of glucose, cholesterol, triacylglycerol, and uric acid in the plasma of control and diabetic animals. In the liver tissue, the concentration of triacylglycerols decreased after losartan administration, but the concentration of cholesterol did not change. The present authors have found that losartan administration increased the levels of water solubile antioxidants in the plasma of diabetic rats, which can result in a decrease of the TBARP levels in the plasma of diabetic rats.

Antioxidant systems in polymorphonuclear leucocytes of Type 2 diabetes mellitus.

AIMS: To examine the effect of Type 2 diabetes mellitus (DM) on enzymes of importance for oxygen-dependent killing of microorganisms by leucocytes. METHODS: Twenty patients with Type 2 DM and 20 nondiabetic controls provided blood samples for analysis. RESULTS: The superoxide dismutase (SOD) activity was lower by 41% in polymorphonuclear leucocytes (PMNL) from patients with Type 2 DM than in controls (3.42+/-0.32 U/mg of protein vs. 5.79+/-0.71 U/mg of protein, P<0.005). Glutathione peroxidase (GSHPx) and glutathione reductase (GR) activities of Type 2 DM patients were 73.04% and 81.12% of control values (0.84+/-0.07 nkat/mg of protein vs. 1.15+/-0.10 nkat/mg of protein, P<0.003, and 2.02+/-0.12 nkat/mg of protein vs. 2.49+/-0.16 nkat/mg of protein, P < 0.023, respectively). The catalase activity showed no significant difference. A significant increase (141.37% of control) in the concentration of thiobarbituric acid reactive products was observed (9.91+/-0.78 miromol/l vs. 7.01+/-0.47 micromol/l, P<0.003). A positive correlation between thiobarbituric acid reactive products and glucose, glycated haemoglobin and fructosamine in the serum of diabetic patients was observed. CONCLUSION: These findings may explain some of the mechanisms underlying the increased susceptibility to certain infection in patients with Type 2 DM.

Free radicals derived from oxygen, and medicine.

Toxic free radicals may be produced by many reactions, which are necessary for the maintenance of normal metabolism, and the production of energy in cells. The origin, reactivity with other molecules and removal of free radicals, is in the foreground of interest since their effect is mostly toxic and result in a whole series of pathological states of cells, organs and whole organisms. Production of these radicals increases in oxidative stress and in the presence of ions of metals (chiefly iron), leads to the creation of more reactive metabolites. The generally accepted view is that the main biological actor in damaged tissues is the hydroxyl radical (OH), which is created in the iron catalyzed Haber-Weiss reaction. The balance between the increased creation of free radicals in various pathological states, or unfavourable conditions in the environment, and natural antioxidants of a low-molecular (vitamin C, E, glutathione etc.) or enzyme character (superoxide dismutase, glutathione peroxidase, glutathione reductase, catalase, etc.), plays the chief role in damage which is the cause of many diseases and ageing. (Fig. 3, Tab. 5, Ref. 62.).