Antibiotic Resistance Profile of RT 027/176 Versus Other Clostridioides difficile Isolates in Silesia, Southern Poland.

Clostridioides difficile is an important health care-associated pathogen. The aim of this study was to analyze the antibiotic susceptibility of C. difficile isolates from feces of patients from 13 hospitals in Silesia, Poland. The incidence of CDI per 100.000 people in Silesia in 2018−2019 was higher than the average in Poland (39.3−38.7 vs. 30.2−29.5, respectively). The incidence doubled from 26.4 in 2020 to 55.1 in 2021. Two hundred and thirty stool samples tested positive for GDH (glutamate dehydrogenase) and toxins were cultured anaerobically for C. difficile. The isolates were characterized, typed, and tested for susceptibility to 11 antibiotics by E-test (EUCAST, 2021). The genes of toxins A/B and binary were detected by mPCR. Of 215 isolates, 166 (77.2%) were classified as RT 027 and 6 (2.8%) as related RT 176. Resistance to ciprofloxacin (96.7%), moxifloxacin (79.1%), imipenem (78.1%), penicillin (67%), and rifampicin (40.5%) was found. The ermB gene was detected in 79 (36.7%) strains. Multidrug resistance (MDR) was confirmed in 50 (23.3%) strains of RT 027 (94%). We concluded that a high prevalence of MDR among hypervirulent RT 027/176 C. difficile was found in the Silesian region of Poland, emphasizing the need to enhance regional infection control on CDI and antibiotic stewardships.

Retrospective Study of the Etiology, Laboratory Findings, and Management of Patients with Urinary Tract Infections and Urosepsis from a Urology Center in Silesia, Southern Poland Between 2017 and 2020.

BACKGROUND Recent studies have shown that up to 25% of sepsis cases originate in the urinary tract. Urosepsis can be associated with cystitis, lower urinary tract infections (UTIs), and upper UTIs and is most commonly caused by gram-negative bacteria. This retrospective study from a urology center in southern Poland, was conducted between 2017 and 2020 and aimed to investigate the causes, microbiology laboratory findings, and management in 138 patients with UTIs and urosepsis. MATERIAL AND METHODS Records of patients with UTIs with urosepsis admitted to the Urology Department of the hospital in Silesia, Poland, between 2017 and 2020 were analyzed retrospectively, and clinical and laboratory data were evaluated. RESULTS The 138 included patients were admitted to the hospital between 2017 and 2020. The median age of patients was 67 (20-94) years, and 59.9% (82/137) were men. The most common reasons for admission to the Urology Department were hydronephrosis due to dysfunction of urinary drainage in 36.5% (50/137) of patients and hydronephrosis due to urolithiasis in 22.6% (31/137) of patients. The main etiological agents responsible for the development of urosepsis were strains of Enterobacteriaceae in 85% of patients, of which 41.4% (48/116) produced extended-spectrum beta-lactamases (ESBL), accounting for 35.0% (48/137) of patients with urosepsis. In 83.3% (80/96) of patients, the pathogen cultured from the urine was identical to that cultured from the blood. CONCLUSIONS The identification of an increasing prevalence of urosepsis associated with ESBL-producing gram-negative rods from this single-center study highlights the importance of infection monitoring, rapid diagnosis, and multidisciplinary patient management.

Detection of Clostridioides difficile in hospital environment by using C diff Banana Broth™.

116 environmental samples from a 504 bed clinical hospital obtained in 2017/19 were inoculated into C diff Banana Broth™. Six C. difficile and 12 C. pefringens strains were isolated. Antibiotic-resistant Clostridium spp. dominated in hospital environment. To determine Clostridium spp. in hospital environment suitable medium like C diff Banana Broth™ should be used.

Clostridioides difficile Ribotype 027 (RT027) Outbreak Investigation Due to the Emergence of Rifampicin Resistance Using Multilocus Variable-Number Tandem Repeat Analysis (MLVA).

OBJECTIVE: The aim of this study was Clostridioides difficile outbreak investigation due to the emergence of rifampicin resistant ribotype 027 (RT 027) fecal isolates from patients of Polish tertiary care hospital between X. 2017 and II. 2018 using multilocus variable tandem repeat analysis (MLVA). MATERIALS AND METHODS: Twenty-nine C. difficile fecal isolates from patients of tertiary care hospital in Southern Poland were ribotyped and analyzed by MLVA. Multiplex PCR (mPCR) for genes encoding GDH (gluD), toxins A (tcdA)/ B (tcdB), 16S rDNA and binary toxin genes (ctdA and ctdB) was performed. The antibiotic susceptibility profile was determined by E-test. RESULTS: The A, B and binary toxins encoding genes were detected in all 29 C. difficile strains which were sensitive to metronidazole, vancomycin and were resistant to erythromycin, clindamycin, and moxifloxacin; resistance to imipenem demonstrated 97%, to rifampicin - 45% isolates. C. difficile strains could be grouped by MLVA into 5 distinct clusters, and the largest cluster II contains 16 strains. The comparison of rifampicin GM MIC of cluster II (n=16 strains) with all others (n=13) showed that strains from clusters I, III, IV and V possessed significantly (p <0.005) higher GM MIC and were more resistant to rifampicin. CONCLUSION: MLVA analysis proved transmission and recognized outbreak due to multidrug-resistant RT 027 C. difficile among patients of tertiary care hospital in Southern Poland. The reason for this is probably the widespread occurrence of spores in the hospital environment, which includes, among others, neglect of hygienic procedures and epidemic supervision. High resistance to imipenem (97%) and to rifampicin (45%) among C. difficile RT 027 Silesian isolates is threatening and requires further studies to elucidate this phenomenon.

The Bioinformatic and In Vitro Studies of Clostridioides Difficile Aminopeptidase M24 Revealed the Immunoreactive KKGIK Peptide.

Clostridioides difficile (CD) is a Gram-positive pathogen responsible for CD-associated disease (CDAD), which is characterized by symptoms ranging from mild diarrhea to pseudomembranous colitis. This work is an attempt to respond to the need of novel methods for CD infection (CDI) prevention, since the number of CDI cases is still rising. A bioinformatics approach was applied to design twenty-one peptides consisting of in silico predicted linear B-cell and T-cell epitopes of aminopeptidase M24 from CD. These peptides were mapped for epitopes exploiting PEPSCAN procedure and using sera obtained from CD infected patients, umbilical cord blood, and healthy volunteers. Two new CD epitopes, 131KKGIK135 and 184KGTSTHVIT192, were identified and characterized. Immunoreactivity of the synthetic biotinylated 131KKGIK135 epitope was significantly higher compared to 184KGTSTHVIT192 epitope in Enzyme-Linked Immunosorbent Assay (ELISA) with umbilical cord blood and CDI patients' sera. Hereafter, the conjugate of bovine serum albumin and epitope 131KKGIK135 was evaluated in vitro on lung epithelial cell line. In vitro, a significant induction of IL-6 by conjugate was observed, thereby we postulate that this new 131KKGIK135 epitope possesses immunostimulating properties suggesting possibility of its use in a vaccine against Clostridioides difficile.

Epitopes identified in GAPDH from Clostridium difficile recognized as common antigens with potential autoimmunizing properties.

Clostridium difficile (CD) infections are a growing threat due to the strain resistance to antibiotic treatment and the emergence of hypervirulent strains. One solution to this problem is the search for new vaccine antigens, preferably surface-localized that will be recognized by antibodies at an early stage of colonization. The purpose of the study was to assess the usefulness of novel immunoreactive surface proteins (epitopes) as potential vaccine antigens. Such approach might be tough to pursue since pathogens have acquired strategies to subvert adaptive immune response to produce humoral response against non-essential proteins for their survival. In this study CD surface proteins were isolated, immunoreactive proteins identified and mapped to select potential epitopes. The results of the study exclude the use of CD glyceraldehyde 3-phosphate dehydrogenase as a vaccine antigen, especially as a whole protein. Sequences P9 (201AAGNIVPNTTGAAKAI218) and P10 (224KGKLDGAAQRVPVVTG241) recognized by patients sera are conserved and widespread among CD strains. They show cross-reactivity with sera of people suffering from other bacterial infections and are recognized by sera of autoimmune disease patients. Our study documents that special care in analyzing the sequence of new epitope should be taken to avoid side effects prior to consider it as a vaccine antigen.

Toxin profile of fecal Clostridium perfringens strains isolated from children with autism spectrum disorders.

Infectious factors are taken into consideration in pathophysiology of autism spectrum disorders (ASD). ASD patients often suffer from gastrointestinal disorders. The intestinal microbiota of autistic patients significantly differs from that in healthy individuals. The aim of the study was to compare the profile of toxins produced by C. perfringens strains isolated from feces of children with ASD, with healthy individuals and obese subjects. This study included 111 strains of C. perfringens: 49 isolates from 29 children with ASD, 30 - from 17 healthy individuals and 32 - from 24 young obese subjects. Alpha, beta, beta2, epsilon, iota and enterotoxin genes were detected using appropriate PCRs. The alpha toxin gene (cpa) was present in all 111 examined strains (100%). The beta2 gene (cpb2) was detected in 45/49 strains (91.8%) isolated from children with ASD, 17/30 (56.7%) isolates from healthy subjects, and 12 of 32 (37.5%) isolates from obese subjects. C. perfringens strains with cpb2 gene were detected in 27/29 ASD patients (93.1%), 10/17 healthy subjects (58.8%) and 11/24 (45.8%) obese subjects. Beta2 toxin encoding cpb2 gene was significantly more common in strains isolated from ASD patients, with no significant difference between control subjects regardless of diet. Further research to explain observed phenomena and pathomechanism of beta2 toxin is required.

Identification of Asymptomatic Prosthetic Joint Infection: Microbiologic and Operative Treatment Outcomes.

BACKGROUND: This prospective study investigated the role of mechanical and biological factors in aseptic implant loosening or presumed silent prosthetic joint infection (PJI). METHODS: Thirty-seven patients were investigated. Microbiologic and molecular methods were used to detect bacteria on the surface of the failed implants removed during revision arthroplasty. Histopathologic analysis was performed. The influence of body mass index (BMI) and various co-morbidities on implant failure also was determined. RESULTS: The results of sonicated fluid cultures were positive for bacteria in 29.7% and the results of intra-operative tissue and joint liquid cultures in 18.9% and 16.2%, respectively. Molecular detection with 16S rRNA sequencing revealed a large variety of bacteria. The most frequent organism was coagulase-negative Staphylococcus (CNS). The outcomes of histopathologic tests of peri-prosthetic tissue showed evidence of the infection type in all culture-positive joints and in 41.4% of the cases with negative culture results. Overweight status or obesity was present in 82.8% of the culture-negative patients. CONCLUSIONS: The results of this study proved the presence of micro-organisms on the surface of implants in both aseptic and presumed PJI cases. Inclusion of the sonication procedure in the diagnostic algorithm increased the ability to identify the pathogen. The results of our study suggest the co-existing roles of BMI and the time to implant loosening as well as biological agents in causing prosthesis loosening.

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).

Comparison of cultures and 16S rRNA sequencing for identification of bacteria in two-stage revision arthroplasties: preliminary report.

BACKGROUND: The use of a prefabricated spacer in two-stage revision arthroplasty remains one of the few surgery strategies for infected-joint arthroplasty treatment, despite the many unidentified microorganisms in the infected joint replacements reported in some recent studies. The aim of this prospective survey was to investigate if the sonication followed by polymerase chain reaction (PCR) can improve bacterial identification on the surfaces of prefabricated spacers and if the systemic laboratory mediators of infection and positive microbiological results can take a role of predictive factors of infection and clinical failures in 2-years follow-up. METHODS: Thirteen patients with prosthetic joint infection were investigated. Bacterial culture and deoxyribonucleic acid (DNA) sequencing were used to detect bacteria on the surface of prefabricated spacers removed during the second stage of revision arthroplasty. The results of pre- and intraoperative culture and DNA sequencing were compared. Minimum follow-up was 2 years. RESULTS: The result of tissue cultures in second-stage revision arthroplasties revealed positive results in 15 % of patients with Coagulase-negative Staphylococci (CNS) growth. Bacterial DNA was found in over 90 % of patients with negative synovial fluid culture. Positive PCR results revealed potential pathogenic bacteria and species of human and environmental microflora with low virulence. Clinical failures at final follow-up were recorded in 2 (16.6 %) patients. CONCLUSION: The lack of clinical signs of infection, negative culture of preoperative joint aspirate, and intraoperative specimens do not exclude the presence of bacteria on the surfaces of spacers. The positive results of sonication and molecular tests should be interpreted as real pathogenicity factors in the light of the clinical and laboratory data, especially for patients with immunodeficiency. We confirmed our previous results that sonication followed by PCR and sequencing improved bacterial identification.

Assessment of selected B cells populations in the workers of X-ray departments.

OBJECTIVES: Workers of X-ray departments are occupationally exposed to long-term low levels of ionizing radiation (LLIR), which may affect their humoral immunity. The aim of the study was to assess the influence of LLIR on the number and proportion of B cells (CD19+), B1 cells (CD5+CD19+) and memory B cells (CD27+CD19+) in peripheral blood of such workers. MATERIALS AND METHODS: In the study group of 47 X-ray departments workers and the control group consisting of 38 persons, the number and percentage of CD19+, CD5+CD19+, CD27+CD19+ cells as well as CD5+CD19+/CD19+ and CD27+CD19+/CD19+ cell ratios were assessed using flow cytometry. Additionally, the study group was divided into 2 groups by the length of employment below and over 15 years and analysis adjusted for age and smoking habit was performed. RESULTS: The total number of CD19+ cells showed significant increase in the group of workers in comparison with the persons from the control group, whereas the percentage of CD5+CD19+ cells as well as CD27+CD19+/CD19+ and CD5+CD19+/CD19+ cell ratios were lower. Percentage, number of CD5+CD19+ cells and CD5+CD19+/CD19+ cell ratio were significantly lower in the workers with length of employment longer than 15 years in comparison with those employed below 15 years. Moreover, we found positive associations between the number of CD19+ cells and employment as well as smoking habit, whereas the number of CD5+CD19+ cells was positively associated with cigarette smoking alone. Percentage of CD5+CD19+ cells as well as CD5+CD19+/CD19+ and CD27+CD19+/CD19+ cell ratios were negatively correlated with employment. CONCLUSIONS: The study suggests association between the suppressive influence of low level ionizing radiation on circulating in peripheral blood, especially of B1 cells as well as of memory B cells, in workers of X-ray units, which is adverse in relation to microbiological threat.

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.

Prevalence of cervical high-risk human papillomavirus infections in kidney graft recipients.

BACKGROUND: Female kidney graft recipients are regarded as a group at risk of cervical cancer development. The objective of this study was to assess the prevalence of cervical high-risk human papilloma virus (HR-HPV) infection and cervical intraepithelial neoplasia (CIN) in female kidney graft recipients in comparison to healthy controls. MATERIAL AND METHODS: We assessed the prevalence of HR-HPV and CIN in 60 female kidney graft recipients of reproductive age in outpatient care of the First Department of Obstetrics and Gynecology, Medical University of Warsaw. The control group consisted of 60 healthy women. Cervical swabs were analyzed with use of Amplicor HPV Roche Molecular Systems test, detecting DNA of 13 types of high-risk HPV: 6, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 68. RESULTS: HR-HPV was detected in 11/60 kidney recipients and 15/60 healthy women (p=0.37). There was no correlation between HR-HPV presence and immunosuppressive regimen, underlying disease, graft function, or time interval from transplantation. In both groups, higher prevalence of HR-HPV was observed in females with ≥2 lifetime sexual partners. Abnormal Pap test results in 2 kidney recipients (2 cases of HSIL) and 2 healthy women (LSIL and HSIL) required colposcopy-guided cervical biopsy, which in all cases revealed CIN2+. CONCLUSIONS: Female kidney graft recipients seem to be equally exposed to cervical infection with HPV of high oncogenic potential as the healthy population.

Gut microbiota, microinflammation, metabolic profile, and zonulin concentration in obese and normal weight subjects.

The association between gut microbiota and circulating zonulin level, a marker of intestinal permeability, has not been studied yet. The aim of the study is the assessment of plasma zonulin, haptoglobin and proinflammatory cytokines (TNF- α and IL-6) levels in relation to composition of gut microbiota in obese and normal weight subjects. Circulating inflammation markers, such as TNF- α , sTNFR1, sTNFR2, IL-6, zonulin, and haptoglobin levels were measured and semiquantitative analysis of gut microbiota composition was carried out in 50 obese and 30 normal weight subjects without concomitant diseases. Higher circulating zonulin, TNF- α , sTNFR1, sTNFR2, and IL-6 levels were found in the obese subjects. Plasma zonulin level correlated positively with age (r = 0.43, P < 0.001), body mass (r = 0.30, P < 0.01), BMI (r = 0.33, P < 0.01), fat mass and fat percentage (r = 0.31, P < 0.01 and r = 0.23, P < 0.05, resp.). Positive correlations between bacterial colony count and sTNFR1 (r = 0.33, P < 0.01) and plasma zonulin (r = 0.26, P < 0.05) but not haptoglobin levels were found. Additionally, plasma zonulin level was proportional to daily energy intake (r = 0.27, P < 0.05) and serum glucose concentration (r = 0.18, P < 0.05) and inversely proportional to diet protein percentage (r = -0.23, P < 0.05). Gut microbiota-related systemic microinflammation in the obese is reflected by circulating zonulin level, a potential marker of interstitial permeability.

Identification of silent prosthetic joint infection: preliminary report of a prospective controlled study.

PURPOSE: We will test the hypothesis that ultrasound supported by polymerase chain reaction (PCR) could improve bacterial identification in non-infected prosthetic joint loosening. The aim was to detect bacterial species in non-infected prosthetic joint loosening using ultrasound and 16S rRNA gene sequencing. METHODS: A total of 16 patients (11 women and five men) aged 46-80 years (mean age 65.7) with diagnosed knee or hip implant loosening (mean implant survival of 102.1 months) were investigated. Bacterial culture and DNA sequencing were used to detect bacteria on the surface of failed implants removed during revision arthroplasty. The results of pre- and intraoperative culture and DNA sequencing were compared. Histopathological analysis was also performed. RESULTS: The number of positive cultures rises with a higher level of C-reactive protein (CRP). The results of the cultures from synovial fluid obtained through joint aspiration were consistent with sonicates from components of prostheses in 12 cases (75%). Bacterial DNA was found in 90% of patients with negative synovial fluid culture. PCR revealed two or more bacterial species, often of the same genus: Ralstonia pickettii, Pseudomonas spp., Brevibacterium spp., Lactobacillus spp., Propionibacterium spp. and Staphylococcus spp.These are micro-organisms present in the environment or on the human body and often associated with compromised immunity. CONCLUSIONS: The ultrasound procedure followed by PCR and sequencing improve bacterial identification in silent prosthetic joint infection. The lack of clinical signs of infection and negative preoperative and intraoperative cultures do not exclude the presence of micro-organisms on the implants.

Prevalence of high-risk human papillomavirus cervical infection in female kidney graft recipients: an observational study.

BACKGROUND: Immunosuppressive therapy protects the transplanted organ but predisposes the recipient to chronic infections and malignancies. Transplant patients are at risk of cervical intraepithelial neoplasia (CIN) and cervical cancer resulting from an impaired immune response in the case of primary infection or of reactivation of a latent infection with human papillomavirus of high oncogenic potential (HR-HPV). METHODS: The aim of this study was to assess the prevalence of HR-HPV cervical infections and CIN in 60 female kidney graft recipients of reproductive age in comparison to that in healthy controls. Cervical swabs were analyzed for the presence of HR-HPV DNA. HR-HPV-positive women remained under strict observation and were re-examined after 24 months for the presence of transforming HR-HPV infection by testing for HR-HPV E6/E7 mRNA. All the HR-HPV-positive patients were scheduled for further diagnostic tests including exfoliative cytology, colposcopy and cervical biopsy. RESULTS: The prevalence of HR-HPV did not differ significantly between the study group and the healthy controls (18% vs 25%, p = 0.37). There was no correlation between HR-HPV presence and the immunosuppresive regimen, underlying disease, graft function or time interval from transplantation. A higher prevalence of HR-HPV was observed in females who had had ≥ 2 sexual partners in the past. Among HR-HPV-positive patients, two cases of CIN2+ were diagnosed in each group. In the course of follow-up, transforming HR-HPV infections were detected in two kidney recipients and in one healthy female. Histologic examination confirmed another two cases of CIN2+ developing in the cervical canal. CONCLUSIONS: Female kidney graft recipients of reproductive age are as exposed to HR-HPV infection as are healthy individuals. Tests detecting the presence of HR-HPV E6/E7 mRNA offer a novel diagnostic opportunity in those patients, especially in those cases where lesions have developed in the cervical canal.

Is the plasma anti-Müllerian hormone (AMH) level associated with body weight and metabolic, and hormonal disturbances in women with and without polycystic ovary syndrome?

OBJECTIVE: The aim of the study was to analyze interrelation between AMH levels and body weight, metabolic, and hormonal status in normal and overweight weight women with and without polycystic ovary syndrome (PCOS). STUDY DESIGN: Eighty-seven women (54 normal weight and 33 overweight) diagnosed with PCOS and 50 apparently healthy women - Non-PCOS (28 normal weight and 22 overweight) were enrolled. The body weight and height were measured and BMI was calculated. In addition to serum glucose, lipids, androgens, FSH, LH, SHBG and insulin, AMH were assessed in fasting state and free androgens index (FAI) was calculated. The insulin resistance was assessed based on the homeostasis model of assessment-insulin resistance (HOMA-IR). RESULTS: Plasma AMH levels were similar in normal weight and overweight PCOS groups (9.6±3.5 vs. 11.2±4.5ng/mL, respectively), and as expected markedly higher than in both Non-PCOS groups (2.5±0.8 and 2.3±0.7ng/mL, respectively). There were no correlations between BMI and AMH levels in all study groups. A significant positive correlation between HOMA-IR, free testosterone concentrations or FAI and AMH levels were found (R=0.31, p<0.001; R=0.91, p<0.001 and R=0.62, p<0.001, respectively). Moreover, there was positive correlation between total or LDL cholesterol and AMH levels (R=0.22, p<0.05 and R=0.31, p<0.05, respectively) and a negative one between HDL cholesterol and AMH levels (R=-0.17, p<0.05) in all study subjects. CONCLUSIONS: The plasma AMH level is associated with insulin resistance but not with BMI per se. Increased circulating AMH level seems to reflect the disturbances of gonadotrophins release in PCOS. It seems that AMH level may be used not only as new surrogate marker of ovarian hyperandrogenism in PCOS but also as a potential new cardiovascular risk factor.

Identification of pregnant women at risk of Streptococcus group B colonisation.

OBJECTIVES: The aims of the study were: 1) to evaluate the prevalence of recto-vaginal group B streptococci (GBS) colonisation using Todd Hewitt Broth - recommended by the Centers for Disease Control (CDC) - and Granada medium; 2) to establish the sensitivity and specificity of Granada medium for the detection of GBS colonisation; 3) to evaluate each vaginal Gram stained swab for bacterial vaginosis (BV) using Nugent criteria and for determining the amount of polymorphonuclear (PMN) leucocytes. METHODS: Eighty pregnant women between 35 and 40 gestation weeks hospitalised in the Department of Gynecology and Obstetrics, Medical University of Silesia, Poland, were included in the study. Two specimens were collected from each patient: one from the posterior vaginal fornix (Gram stain) and one from both vagina and anus to detect GBS colonisation. Each vaginal Gram stained swab was evaluated for BV using Nugent criteria as well as for PMN leucocyte count. To detect GBS colonisation, the liquid Todd Hewitt Broth, subsequently subcultured to blood agar and direct inoculation onto Granada medium, were used. Isolated GBS were identified by morphological features and by serological (Slidex Strepto-Kit, bioMerieux) and biochemical (rapid ID 32 Strep, bioMerieux) testing. RESULTS: GBS colonisation was observed in 22 (27.8%) patients in both used media. Only in one case were GBS detected in Todd Hewitt Broth and not detected in Granada medium. The sensitivity and specificity of Granada medium were established as: 95.65% and 100%, respectively, compared with Todd Hewitt Broth recommended by CDC. Nugent criteria demonstrated 6.25% of cases of BV; in one case both BV and GBS colonisation were detected. CONCLUSIONS: The selective Granada medium may be used concurrently with liquid Todd Hewitt Broth as a screening tool for prenatal group B streptococcal colonisation in pregnant women.

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.