Occurrence of Clostridium difficile ribotype 027 in hospitals of Silesia, Poland.

Clostridium difficile is an important healthcare-associated pathogen, responsible for a broad spectrum of diarrheal diseases. The aim of this prospective study was to determine the occurrence of C. difficile infection (CDI), to characterize cultured C. difficile strains and to investigate the association of fecal lactoferrin with CDI. Between January 2013 and June 2014, 148 stool samples were obtained from adult diarrheal patients (C. difficile as a suspected pathogen) hospitalized in different healthcare facilities of 15 Silesian hospitals. Out of 134 isolated C. difficile strains, 108 were ribotyped: 82.4% belonged to Type 027, 2.8% to Type 176, 2.8% to Type 014, 1.9% to Type 010 and 0.9% to Types 001, 018, 020 and 046 each. In total, 6.5% non-typable strains were identified. All Type 027 isolates contained both toxin genes tcdA & tcdB, and binary toxin genes (cdtA &cdtB). Susceptibility testing revealed that all Type 027 isolates were sensitive to metronidazole and vancomycin and resistant to moxifloxacin, ciprofloxacin, imipenem and erythromycin. Of 89 Type 027 strains, 16 had a ermB (688 bp) gene coinciding with high levels of erythromycin resistance (MIC >256 µg/mL). Of 16 ermB positive strains, 14 demonstrated also high level of resistance to clindamycin (>256 µg/mL). A significant difference (p = 0.004) in lactoferrin level was found between C. difficile toxin-positive (n = 123; median 185.9 µg/mL; IQR 238.8) and toxin-negative (n = 25; median 22.4 µg/mL; IQR 141.7) fecal samples. Stool samples from n = 89 patients with CDI caused by Type 027 demonstrated significantly higher (p = 0.03) lactoferrin level (median 173.0 µg/mL; IQR 237.3) than from patients with CDI caused by other ribotypes and non-typable C. difficile strains (median 189.4 µg/mL; IQR 190.8).

Prevalence of Urogenital Mycoplasmas Among Men with NGU in Upper Silesia, Poland. Preliminary Study.

The prevalence of urogenital mycoplasmas in men with NGU in Upper Silesia (Poland) was studied. Mycoplasmas were detected in 36.7% men (Ureaplasma parvum and Mycoplasma genitalium were found in 30% and 16.7% respectively). Urealyticum urealyticum was not detected. We suggest including M. genitalium in the diagnostic scheme for nongonococcal urethritis (NGU).

Urogenital mycoplasmas and human papilloma virus in hemodialysed women.

Bacterial infections, especially endogenous, are the frequent complications among hemodialyzed and renal transplant patients. In this study we assumed the prevalence of urogenital mycoplasmas and HPV among hemodialysed women. We examined 32 hemodialysed women aged 20-48 (mean 35.6 ± 8.23) and 100 healthy controls of the same ages. Two swabs were collected for detection of mycoplasmas and HPV. Culture of Ureaplasma spp. and M. hominis was performed using Mycoplasma IST2 (bioMérieux, France), Identificaton of U. parvum and U. urealyticum was performed by Kong. Primers described by Jensen were used for M. genitalium. For detection of high-risk HPV types Amplicor HPV (Roche Molecular System, CA) was used. Prevalence of urogenital mycoplasmas in the hemodialysed women (53.1%) was significantly higher (P = 0.0059), compared with controls (25%). In both groups, U. parvum was the most frequently isolated. Cooccurrence of urogenital mycoplasmas was shown in 75% of the HPV-positive hemodialysed women and in 30.4% of HPV-positive controls (P = 0.0461). Cooccurrence of urogenital mycoplasmas with HPV was significantly higher in hemodialysed women. The need to take into account these microorganisms in routine diagnostic, especially for hemodialysed patients, was demonstrated. Further studies to demonstrate the role of this cooccurrence in etiopathogenesis of infection in hemodialysed patients are required.

Fecal lactoferrin and Clostridium spp. in stools of autistic children.

Stools from autistic and healthy children were studied for fecal lactoferrin, Clostridium difficile toxins, Clostridium perfringens enterotoxin and cultured for Clostridium spp. Elevated level of FLA was demonstrated in 24.4% stools, all from boys (31.25%). No toxins were detected. Clostridium spp. was isolated with similar frequency from all samples. C. perfringens were isolated significantly often from the autistic stools, intermediate sensitive strains to penicillin 19%, to clindamycin 11.3%, and to metronidazole 7.5% were detected. Further studies on fecal microflora and inflammatory mediators, with larger groups of patients, are required in order to explain their role in neurological deficits.

[Intestinal microflora of autistic children]. / Mikroflora jelitowa dzieci autystycznych.

Autistic behavior is often accompanied by numerous disturbing symptoms on the part of gastrointestinal system, such as abdominal pain, constipation or diarrhea. These problems are often connected with deregulation of physiological microflora in intestine. The aim of this study was to determine differences in intestinal microflora of autistic and healthy children. Strains of Clostridium spp. and enterococci were isolated more frequently from stool samples of autistic children and rarely lactobacilli. Quantitative differences were observed maliny among staphylococci, Candida spp. and Clostridium perfringens. Monitoring and stabilization of intestinal microflora and knowledge about role of particular strains in etiology of autistic disorders can increase the chances for appropriate therapy.

[HBD-1 and hBD-2 are expressed in cervico-vaginal lavage in female genital tract due to microbial infections]. / Ekspresja hBD-1 i hBD-2 w popluczynach pochwowo-szyjkowych z dróg rodnych kobiet w stanach zapalnych wywolanych drobnoustrojami.

OBJECTIVES: The aim of this study was to evaluate and compare concentration of selected human beta-defensins (hBD-1, hBD-2) in cervico-vaginal lavage (CVL), obtained from women with candidiasis, chlamydiasis and other bacterial infections. MATERIAL AND METHODS: beta-defensins were detected quantitatively by RT-PCR (7000 Taqman, Applied Biosystems) in cervico-vaginal lavage collected from 120 (79 women in the study group and 41 controls) non-pregnant women, aged 18-40 (mean age 28.5 +/- 6.29). The study group patients were divided into three subgroups on the basis of clinical and microbiological diagnosis: women with candidiasis (n=13); with chlamydiasis (n=13), and with other bacterial infections (n=12). RESULTS: The highest count of hBD-1 RNA copies was found in women with bacterial infections and candidiasis (335.84 and 320.10 respectively), and hBD-2--with chlamydiasis. The difference between RNA copies of hBD-1/microg in candidiasis, chlamydiasis and bacterial pathogens was statistically significant; for hBD-2 only in case of chlamydiasis. CONCLUSIONS: Chlamydia trachomatis infection activates the production of hBD-2. Candida albicans, Chlamydia trachomatis, and bacterial pathogens induced variable increases of hBD-1 concentration.

From mild cognitive impairment to Alzheimer's disease - influence of homocysteine, vitamin B12 and folate on cognition over time: results from one-year follow-up.

BACKGROUND AND PURPOSE: People with mild cognitive impairment (MCI) have higher risk of developing dementia than the general population. Currently known risk factors for dementia include older age, low education level, gait disorders, hippocampal atrophy, and apolipoprotein E allele. Vascular risk factors may modify the neurodegenerative process. The aim of this study was therefore to assess the influence of vascular (genetic and environmental) risk factors on progression to dementia in an MCI group during a one-year period. MATERIAL AND METHODS: Fifty-five MCI patients (30 men and 25 women) and 44 controls (25 men and 19 women) matched for age, gender and education were studied. Mild cognitive impairment was diagnosed according to Petersen criteria (Mayo Clinic Group). Neuropsychological evaluation was made. Assessed vascular risk factors included hypertension, cardiovascular disease, diabetes, cigarette smoking, hyperlipidaemia, hyperhomocysteinaemia with vitamin B12 and folate deficiency. Genetic risk factors (APOE polymorphism, C677T and A1298C MTHFR polymorphisms) were also assessed. RESULTS: Vascular risk factors were found significantly more often in the MCI group (p = 0.041), including APOE4 allele (p = 0.018), hyperhomocysteinaemia (p = 0.012) and folate deficiency (p = 0.023). Discriminant function analysis showed that only age and hypertension are potential factors which may have an influence on progression to dementia in the MCI group within one year of prospective observation. CONCLUSION: Vascular risk factors are associated with cognitive impairment but do not have a significant influence on progression to dementia in the MCI group.

Human neutrophil peptides in vaginitis/cervicitis of different etiology.

Development of female genito-urinary infections depends on many factors, such as immune system activity, virulence of microorganism and production of factors inhibiting the growth of microorganisms. Taking into account the possibility of relapses or severe complications, it is very important to appropriately diagnose and treat infections. Because of recently observed increase of microbial resistance to antibiotics, researchers are looking for alternatives. In our study we evaluated and compared the concentration of human neutrophil peptides (HNP 1-3) in cervico-vaginal lavages (CVL), obtained from women with vaginitis/cervicitis. Swabs from the posterior vaginal fornix and from the endocervical canal as well as CVL samples were obtained from 32 patients with vaginitis/cervicitis and 29 healthy women (control group). Supernatants of CVL were used for determination of concentration of HNP by ELISA. The difference between concentrations of HNP 1-3 in studied and control groups was statistically significant (p = 0.018). The maximal concentration was determined in patients with mixed infections (28.41 ng/ml), and Group B Streptococci, GBS, (28.06 ng/ml), the minimal concentrations in cases of C. trachomatis (mean concentrations did not differ from those in the control group: 16.93 ng/ml and 16.39 ng/ml, respectively). Maximal correlation was determined for control-studied group with isolation of GBS (r = 0.79), and very high negative correlation for group of GBS - C. trachomatis (r = -0.98).

[Cervical dysplasia and microflora of sexual tract]. / Zmiany dysplastyczne nablonka szyjki macicy a mikroflora dróg rodnych.

The aim of this study was to evaluate microflora of the vaginal part of the cervix uteri (including high-oncogenic HPV types and C. trachomatis) in women with ASCUS, LSIL comparing with women with normal cytology. The results demonstrated that infections with high-oncogenic HPV types and C. trachomatis don't reduce significantly Lactobacillus spp.

[The influence of AMPs from HaCaT keratinocytes on some bacteria strains derived from skin changes]. / Ocena dzialania peptydów antydrobnoustrojowych z hodowli keratynocytów HaCaT na wybrane szczepy bakterii pochodzace ze zmian skórnych.

The aim of this study was to evaluate antimicrobial activity of protein extracts from HaCaT cell line against bacterial strains, isolated from clinical materials, obtained from patients with clinical symptoms of acne (Propionibacterium acnes) and gas gangrene (Clostridium perfringens and Sterptococcus pyogenes). Reference strain of Staphylococcus aureus ATCC 25923 also was used. Protein extracts from cultured HaCaT cells were obtained by 3-fold freezing/defreezing cells in dry ice following by centrifugation and incubated with appropriate bacterial suspension (0.5 McFarland scale) during 6 and 24 hours. We observed time-depending and strain-depending activity of HaCaT--protein extract. Interestingly, high activity was demonstrated against strains of S. pyogenes and C. perfringens. Because of increasing bacterial resistance to antibiotics further studies in the field of antimicrobial peptides are required.

[Antimicrobial peptides]. / Peptydy antydrobnoustrojowe.

For many years, attention has been focused on adaptive immunity as the main antimicrobial defense system. The discovery of antimicrobial peptides turned this point of view. In general, these peptides act on microorganisms directly by disrupting the structure of the microbial cytoplasmatic membranes and indirectly acting on immune system. This review provides an overview of some natural antimicrobial peptides and probability of their clinical usage.

Co-occurrence of urogenital mycoplasmas and group B streptococci with chlamydial cervicitis.

The aim of our study was to evaluate whether in women with chlamydial cervicitis urogenital mycoplasmas and group B streptococci (GBS) colonization is found more often than among women with non-chlamydial cervicitis. This study included 351 (mean age 31.7 +/- 6.82) not pregnant, menstruating, sexually active women. We confirmed a high frequency (49.3%) of C. trachomatis infection among women with cervicitis. Cervical ectopia was confirmed in 26.5% of examined women, in half of them ectopia was associated with chlamydial infection. We did not notice differences in frequency of colonization by urogenital mycoplasmas and GBS among women with chlamydial and non-chlamydial cervicitis.

[Antibiotic susceptibility analysis of Enterococcus spp. isolated from urine]. / Analiza lekowrazliwosci Enterococcus spp. izolowanych z moczu.

Recently increase of enterococcal infections has been observed. These bacteria, mainly Enterococcus faecalis and Enterococcus faecium are members of the normal flora of gastrointestinal tract but also are typical opportunistic pathogens. Enterococci are characterized by natural resistance to numerous antibiotics (among them cephalosporins), and also by easy acquired resistance to antibiotics. Infections caused by multiresistant strains are difficult in treatment, chronic, recurrent and sometimes fatal are described. Enterococcal infections are caused often by E. faecalis, rarely by E. faecium. In the last years other species of enterococci have been isolated from different clinical materials (E. casseliflavus, E. avium, E. durans, E. gallinarum). The aim of this study was to analyze the antibiotics susceptibility of Enterococcus spp. isolated from urine of children with urinary tract infection (UTI), hospitalized in the Upper Silesian Health Center of Child and Mother or treated ambulatory. Susceptibility of the 130 strains of E. faecalis and 41 E. faecium to: penicillin, tetracycline, high level of aminoglycoside (gentamycin 120 microg and streptomycin 300 microg), glycopeptides (vancomycin and teicoplanin), ciprofloxacin and nitrofurantoin was estimated. All isolated strains of E. faecalis (100%) were sensitive to glycopeptides (vancomycin and teicoplanin) and to nitrofurantoin, 96% strains were sensitive to penicillin, 43% to ciprofloxacin and 28% to tetracycline. All strains of E. faecium were sensitive to glycopeptides (vancomycin and teicoplanin), 32% strains were sensitive to penicillin, 19% to tetracycline 14% to ciprofloxacin and 50% to nitrofurantoin. Twenty two strains of E. faecalis (17%) and twelve strains of E. faecium (29%) were demonstrated high level resistance to aminoglycosides (HLAR). Among HLAR strains of enterococci, were observed strains of high level resistance only to streptomycin, high level resistance only to gentamycin and high level resistance to both aminoglycosides. The HLAR strains of E. faecalis and E. faecium were also resistant to other antibiotics: penicillin, tetracycline and ciprofloxacin. The most frequent cause of child urinary tract infection was E. faecalis, with high sensitivity to penicillins, glycopeptides and nitrofurantoin. The increased role of E. faecium and other strains of enterococci also has been shown. These species demonstrated sensitivity to glycopeptides, but resistance to other antibiotics, routinely used for treatment of UTI as well as to nitrofurantoin.

[Urinary tract infections in children--etiologic agents and susceptibility to antibiotics]. / Czynniki etiologiczne zakazen ukladu moczowego u dzieci i ich wrazliwosc na antybiotyki.

Urinary tract infections (UTIs) are common clinical problem in pediatric hospitals. The aim of this study was to compare the incidence of UTIs during two different time periods (from February 1999 to March 2000, and from April 2000 to September 2001) in pediatric patients treated in ambulatory or hospitalized in new children hospital. The frequency of occurrence of ESBL--producing Gram-negative rods increased during second period study. The isolated ESBL--producing strains were resistant also to aminoglicosides and trimethoprim/sulfamethoxazol Gram-negative rods were predominant microorganisms. Isolated organisms from urine samples included Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Proteus penneri, Proteus vulgaris, Morganella morganii, Citrobacter freundi, Serratia spp., Enterobacter cloacae, Pseudomonas aeruginosa, Staphylococcus aureus, CNS, Enterococcus spp. Streptococcus spp, and Candida spp. Enterococcus spp. were isolated from urine samples in 20% of cases. Only 9.4% of isolated Enterococcus spp. were high-level aminoglicoside resistant (HLAR) strains. No vancomycin-resistant Enterococcus spp. were isolated. Among coagulase-negative Staphylococci (CNS) methicillin resistant strains (MRCNS) were isolated in 43.3%.