Assessment of Microbiological and Clinical Findings of Two-Stage Revision Arthroplasty Performed Due to Prosthetic Joint Infection.

BACKGROUND: Two-stage revision arthroplasty remains the main surgical strategy for the treatment of prosthetic joint infections (PJI). Sonicate fluid culture has improved sensitivity compared to standard periprosthetic tissue culture, but its usefulness is questionable during the second stage of revision arthroplasty. MATERIAL AND METHODS: Twenty-seven patients with prosthetic joint infection were investigated. Tissue and sonicate fluid cultures were examined to detect bacteria on the removed spacer during the second stage of exchange arthroplasty. Microbiological findings were analyzed and patients were assessed within an average of 5 years follow up. RESULTS: Tissue cultures in second-stage revision arthroplasties were positive in 6/27 cases (22.2%) growing CNS in 4 cases (14.8%), Staphylococcus aureus in 1 case (3.7%), and Enterococcus faecalis in 1 case (3.7%). In 3 cases (11.1%) infection was identified as a result of a sonication procedure. Clinical failures at final follow-up were recorded in 4 (14.8%) patients, with reinfection noted in 3 of them. Arthrodesis and spacer exchange followed by suppressive antibiotic therapy were performed in 2 cases. CONCLUSIONS: 1. Tissue cultures remain the gold standard in diagnosis of PIJ although a negative culture does not rule out the presence of bacteria on spacers removed during second stage revision for PJI. 2. Sonication appears to have limited ability to detect persistent infection before reimplantation and was not associated with recurrent infection; however, it can be considered a complementary method that could make it possible to identify new microorganisms and facilitate the selection of appropriate treatment options. 3. The positive results of sonication should be interpreted as the detection of actual pathogens in the light of the clinical, microbiological and histopathological data, especially for patients with immunodeficiency.

Occurrence of urogenital mycoplasmas in men with the common genitourinary diseases.

Many serious and fatal infections with urogenital mycoplasmas in immunocompromised patients have been reported. M. genitalium is recognized as a cause of male urethritis and other common genitourinary diseases. The aim of the study was to estimate prevalence of urogenital mycoplasmas which can cause complications in men with common genitourinary diseases. Study included 85 men with genitourinary tract carcinoma (n = 35), urolithiasis (n = 36), and BPH (benign prostatic hyperplasia) (n = 14). The control group consisted of 50 healthy men. FVU (first void urine) samples were examined by PCR for the presence of urogenital mycoplasmas DNA. Occurrence of urogenital mycoplasmas was significantly more common in study group compared with control 24/85 (28.2%) and 7/50 (14%), respectively (p = 0.05). In men with urolithiasis, positive results for mycoplasmas DNA were significantly more frequent than in control: 33.3% vs. 14% (p < 0.05). In patients with urolithiasis DNA of U. urealyticum was most often found, while in the genitourinary carcinoma and BPH groups, U. parvum was more frequent. Incidence of M. fermentans was also significantly higher in the urolithiasis group vs. control (p = 0.03). A higher percentage of positive results for urogenital mycoplasma DNA in study group has been found. Further studies are required to confirm the role of urogenital mycoplasmas in the development of infectious complications among patients with urolithiasis, genitourinary carcinoma, and BPH.

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.

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.

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.

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.

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.

Occurrence of Ureaplasma parvum and Ureaplasma urealyticum in women with cervical dysplasia in Katowice, Poland.

The aim of this study was to evaluate the occurrence of genital mycoplasmas, especially Ureaplasma parvum and Ureaplasma urealyticum, in women with atypical squamous cells of undetermined significance (ASCUS), low grade squamous intraepithelial lesions (LSIL) and high grade squamous intraepithelial lesions (HSIL), compared to women with normal cytology living in Katowice, Poland. Two sterile swabs were used to obtain material from the posterior vaginal fornix of 143 women with squamous intraepithelial lesions and 39 healthy women: first for general bacteriology, second for detection of urogenital mycoplasmas using Mycoplasma IST2 kit. From each positive Mycoplasma IST2 culture DNA was isolated and PCR was performed for identification of U. parvum and U. urealyticum. Mycoplasma IST was positive in 34.1% cases. Urogenital mycoplasmas were demonstrated in women with HSIL significantly more often compared to women with LSIL, ASCUS, and with normal cytology. DNA of U. parvum was demonstrated in majority of Mycoplasma IST2-positive cases, U. urealyticum DNA-only in 9 (4.9%). Predominance of 3/14 serovars of U. parvum was demonstrated. U. urealyticum biovar 2 was present more often in women with squamous intraepithelial lesions.

Mycoplasma genitalium: a significant urogenital pathogen ?

Detection of mycoplasma infections, especially those caused by Mycoplasma genitalium (M. genitalium) is not a part of routine microbiological diagnostics, as commercial strip tests based on microculture in test wells do not include M. genitalium in their identification spectrum. Also classical methods of culture are especially demanding in the case of M. genitalium strains. PCR technique with application of specific starter oligonucleotides is currently used in the diagnostics of M. genitalium. In view of growing number of reports on the significance of this mycoplasma in etiopathogenesis of mainly urogenital disorders, an increasing number of laboratories uses molecular methods in diagnostics of M. genitalium infection. In this review we summarize epidemiological, clinical and microbiological research data of M. genitalium in the light of recent publications and our own observations.

Prevalence of urogenital mycoplasmas and ureaplasmas in women after kidney transplantation.

BACKGROUND: The prevalence of urogenital mycoplasmas and ureaplasmas in kidney transplant and hemodialyzed patients was studied. METHODS: Vaginal and cervical swabs taken from 40 women of the study group and 40 women of the control group were investigated. Identification of ureaplasmas, Mycoplasma genitalium, and human papillomavirus was performed by polymerase chain reaction. Each vaginal slide was evaluated for bacterial vaginosis. RESULTS: Urogenital mycoplasmas and ureaplasmas were significantly more common in patients from the study group (40%) compared with the control group (27.5%). Mycoplasma hominis and M. genitalium were demonstrated only in a few cases. Ureaplasma parvum was isolated predominantly, but Ureaplasma urealyticum was more common in patients from study group (10%) compared with control group (2.5%). In all U. urealyticum-positive women from the study group, human papillomavirus DNA was detected. CONCLUSION: Our observation showed the necessity of careful examination of possible atypical pathogens in diagnostic materials from hemodialyzed and kidney transplant patients.

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.