Correction to: Multiresistant Neisseria gonorrhoeae: a new threat in second decade of the XXI century.

The article "Multiresistant Neisseria gonorrhoeae: a new threat in second decade of the XXI century", written by Beata Mlynarczyk­Bonikowska, Anna Majewska, Magdalena Malejczyk, Grazyna Mlynarczyk, Slawomir Majewski was originally published electronically on the publisher's internet portal (currently SpringerLink) on December 04, 2019 without open access.

Multiresistant Neisseria gonorrhoeae: a new threat in second decade of the XXI century.

Neisseria gonorrhoeae is an etiologic agent of gonorrhoea, one of the most common sexually transmitted diseases caused by bacteria. For many years, infections caused by N. gonorrhoeae were considered to be relatively easy to treat; however, resistance has emerged successively to all therapeutic agents used in treatment of the disease, e.g., penicillin, ciprofloxacin or azithromycin. Currently, the global problem is the emergence and a threat of spread of N. gonorrhoeae strains resistant to extended-spectrum cephalosporins (ESC), such as injectable ceftriaxone and oral-used cefixime. Especially, dangerous are multi-resistant strains resistant simultaneously to ESC and azithromycin. Three strains with high-level resistance to azithromycin and resistant to ESC were first time isolated in 2018. Moreover, in 2018, the first ESBL was described in N. gonorrhoeae and that makes the threat of appearing the ESBL mechanism of resistance in N. gonorrhoeae more real, even though the strain was sensitive to ceftriaxone. Molecular typing revealed that variants resistant to ESC occurred also among strains belonging to epidemic clonal complex CC1 (genogroup G1407) distinguished in NG-MAST typing system. The G1407 genogroup, in particular the ST1407 sequence type, is currently dominant in most European countries. The presence of different mechanisms of drug resistance significantly affects clinical practice and force changes in treatment regimens and introduction of new drugs.

Chronic Inflammatory Microenvironment in Epidermodysplasia Verruciformis Skin Lesions: Role of the Synergism Between HPV8 E2 and C/EBPß to Induce Pro-Inflammatory S100A8/A9 Proteins.

Persistent genus ß-HPV (human papillomavirus) infection is a major co-factor for non-melanoma skin cancer in patients suffering from the inherited skin disease epidermodysplasia verruciformis (EV). Malignant EV lesions are particularly associated with HPV type 5 or 8. There is clinical and molecular evidence that HPV8 actively suppresses epithelial immunosurveillance by interfering with the recruitment of Langerhans cells, which may favor viral persistence. Mechanisms how persistent HPV8 infection promotes the carcinogenic process are, however, less well understood. In various tumor types chronic inflammation has a central role in tumor progression. The calprotectin complex consisting of S100A8 and S100A9 proteins has recently been identified as key driver of chronic and tumor promoting inflammation in skin carcinogenesis. It induces chemotaxis of neutrophil granulocytes and modulates inflammatory as well as immune responses. In this study, we demonstrate that skin lesions of EV-patients are massively infiltrated by inflammatory cells, including CD15+ granulocytes. At the same time we observed a very strong expression of S100A8 and S100A9 proteins in lesional keratinocytes, which was mostly confined to the suprabasal layers of the epidermis. Both proteins were hardly detected in non-lesional skin. Further experiments revealed that the HPV8 oncoproteins E6 and E7 were not involved in S100A8/A9 up-regulation. They rather suppressed differentiation-induced S100A8/A9 expression. In contrast, the viral transcription factor E2 strongly enhanced PMA-mediated S100A8/A9 up-regulation in primary human keratinocytes. Similarly, a tremendous up-regulation of both S100 proteins was observed, when minute amounts of the PMA-inducible CCAAT/enhancer binding protein ß (C/EBPß), which is expressed at low levels in the suprabasal layers of the epidermis, were co-expressed together with HPV8 E2. This confirmed our previous observation that C/EBPß interacts and functionally synergizes with the HPV8 E2 protein in differentiation-dependent gene expression. Potent synergistic up-regulation of S100A8/A9 was seen at transcriptional and protein levels. S100A8/A9 containing supernatants from keratinocytes co-expressing HPV8 E2 and C/EBPß significantly induced chemotaxis of granulocytes in migration assays supporting the relevance of our finding. In conclusion, our data suggest that the HPV8 E2 protein actively contributes to the recruitment of myeloid cells into EV skin lesions, which may support chronic inflammation and progression to skin cancer.

Antibiotic resistance and NG-MAST sequence types of Neisseria gonorrhoeae isolates in Poland compared to the world.

Gonorrhoea is one of the most common sexually transmitted infections and in 2012, the World Health Organization estimated about 78 million of new global urogenital cases among adults per year. The main concern during the latest decade has been the emergence and spread of multidrug-resistant strains of Neisseria gonorrhoeae. Resistance has emerged internationally to the extended-spectrum cephalosporins, ceftriaxone and cefixime, which are the last remaining options for empiric first-line monotherapy of gonorrhoea. In Poland, the levels of resistance to ciprofloxacin, benzylpenicillin and tetracycline are high, and the prevalence of azithromycin resistance has increased. However, no resistance to ceftriaxone has been identified. The currently spread multidrug-resistant strains frequently represent epidemic clones. The present paper reviews and describes the antimicrobial resistance and N. gonorrhoeae multiantigen sequence typing (NG-MAST) sequence types of N. gonorrhoeae strains spreading in Poland compared to the world.

Identification of C/EBPα as a novel target of the HPV8 E6 protein regulating miR-203 in human keratinocytes.

Patients suffering from Epidermodysplasia verruciformis (EV), a rare inherited skin disease, display a particular susceptibility to persistent infection with cutaneous genus beta-human papillomavirus (beta-HPV), such as HPV type 8. They have a high risk to develop non-melanoma skin cancer at sun-exposed sites. In various models evidence is emerging that cutaneous HPV E6 proteins disturb epidermal homeostasis and support carcinogenesis, however, the underlying mechanisms are not fully understood as yet. In this study we demonstrate that microRNA-203 (miR-203), a key regulator of epidermal proliferation and differentiation, is strongly down-regulated in HPV8-positive EV-lesions. We provide evidence that CCAAT/enhancer-binding protein α (C/EBPα), a differentiation-regulating transcription factor and suppressor of UV-induced skin carcinogenesis, directly binds the miR-203 gene within its hairpin region and thereby induces miR-203 transcription. Our data further demonstrate that the HPV8 E6 protein significantly suppresses this novel C/EBPα/mir-203-pathway. As a consequence, the miR-203 target ΔNp63α, a proliferation-inducing transcription factor, is up-regulated, while the differentiation factor involucrin is suppressed. HPV8 E6 specifically down-regulates C/EBPα but not C/EBPß expression at the transcriptional level. As shown in knock-down experiments, C/EBPα is regulated by the acetyltransferase p300, a well-described target of cutaneous E6 proteins. Notably, p300 bound significantly less to the C/EBPα regulatory region in HPV8 E6 expressing keratinocytes than in control cells as demonstrated by chromatin immunoprecipitation. In situ analysis confirmed congruent suprabasal expression patterns of C/EBPα and miR-203 in non-lesional skin of EV-patients. In HPV8-positive EV-lesions both factors are potently down-regulated in vivo further supporting our in vitro data. In conclusion our study has unraveled a novel p300/C/EBPα/mir-203-dependent mechanism, by which the cutaneous HPV8 E6 protein may expand p63-positive cells in the epidermis of EV-patients and disturbs fundamental keratinocyte functions. This may drive HPV-mediated pathogenesis and may potentially also pave the way for skin carcinogenesis in EV-patients.

[The prevalence of human papillomaviruses in patients with colon polyps].

INTRODUCTION: Potential role of HPV infection in pathogenesis of colon polyps and cancer remains undetermined. The aim of the study was to investigate the prevalence of DNA of HPV- 6, -11, -16 and -18 in the biopsies from colon polyps. MATERIAL AND METHODS: We investigated the biopsies from 24 patients (23 from colon polyps and I from colon cancer) of Department of Gastroenterology Medical University of Warsaw using Real time PCR HPV-6/11. Real-TM (Sacace Biotechnologies) was performed on termocycler Smart Cycler Dx. RESULTS: We didn't detect oncogenic HPV16 and HPV18 in any of the investigated specimens, HPV-11 was present in 11 patients including all patients with adenoma tubule-villosum. We detect HPV6 in 5 samples from polyps and 1 from colon cancer. CONCLUSIONS: HPV 6 and HPV 11 could play a role in pathogenesis some colon polyps but the final conclusions demand further investigations. Oncogenic HPV 16 and 18 probably don't play any role in colon polyps pathogenesis.

Antiviral Medication in Sexually Transmitted Diseases. Part III: Hepatitis B, Hepatitis C.

In the previous parts of the series the antiviral agents used in genital herpes, genital HPV infection and therapeutic options in HIV infections were presented. The sexual contact is one of the major routes in the transmission of HBV and also possible modes of transmission of HCV. In this review we present the clinical indications, mechanisms of action, and side effects of presently available medication for the management of HBV and HCV infections. Currently a revolution is happening in the therapy of chronic hepatitis, especially caused by HCV. Direct-acting antivirals promise to open a new era in treating of chronic HCV infection. Efficacious, simplified and well tolerated interferon-free, and in some cases ribavirin-free regiments are available already and several other inhibitors currently are in the clinical trials.

Plasmid-mediated resistance to tetracyclines among Neisseria gonorrhoeae strains isolated in Poland between 2012 and 2013.

INTRODUCTION: One of two main mechanisms of resistance in tetracycline-resistant Neisseria gonorrhoeae (TRNG) is associated with the presence of TetM protein responsible for actively blocking of the tetracycline target site in the 30S ribosomal subunit. This mechanism is encoded by conjugative plasmids. The second mechanism is chromosomal in nature and due to mutations in specific genes. AIM: To determine the incidence and type of tetM determinants in TRNG strains isolated from patients presenting with gonorrhea infection to the Dermatology and Venereology Clinic in Warsaw in 2012-2013. MATERIAL AND METHODS: Tetracycline and doxycycline susceptibility was determined by E-Tests. The presence and type of the tetM gene were determined by polymerase chain reaction. RESULTS: Tetracycline resistance was detected in 50.8% of the evaluated strains. The TRNG strains containing the tetM plasmid constituted 13.8% of all the evaluated strains. Dutch type tetM constituted 12.3% and American type tetM 1.5% of all the evaluated strains. In the remaining TRNG strains, resistance to tetracyclines was presumably chromosome-encoded. The minimal inhibitory concentration (MIC) of tetracycline ranged from 0.25 to 32.0 mg/l, MIC50 = 2.0 mg/l, MIC90 = 32.0 mg/l. The MIC of doxycycline ranged from 0.25 to 32.0 mg/l, MIC50 = 4.0 mg/l, MIC90 = 16.0 mg/l. CONCLUSIONS: Unlike most of European countries, in 2012-2013 in Poland, the Dutch type tetM was found to be much more common than the American type. Minimal inhibitory concentration values of tetracycline and doxycycline were similar, with doxycycline exhibiting a somewhat lower effectiveness in vitro than tetracycline towards chromosome-mediated tetracycline resistant strains of N. gonorrhoeae.

Dominating types of penicillinase-plasmids in Neisseria gonorrhoeae strains isolated in 2010-2012 in Warsaw.

INTRODUCTION: The reason of Neisseria gonorrhoeae resistance to penicillin is often production of TEM beta-lactamases encoded by plasmids. The most common types of the plasmid are Africa, Asia and Toronto/Rio. Another reason of resistance can be mutations in bacterial chromosome. The aim of the study was to investigate the types of plasmids occurring in in Neisseria gonorrhoeae strains isolated in 2010-2012 in Warsaw. MATERIAL AND METHODS: From 218 isolated in 2010, 2011 and at the beginning of 2012 from patients of Medical University in Warsaw we selected 12 strains producing beta- lactamase (penicillinase producing N. gonorrhoeae, PPNG). d B-tests to investigate bacterial sensitivity to penicillin and cefiriaxon. The types of plasmids were determined with PCR. RESULTS: The Beta-lactamases were encoded by Toronto/Rio (41,7%), Asia (33,3%) and Africa (25,0%) plasmids. All the strains were resistant to penicillin (MIC 2-8 mg/L) and sensitive to ceftriaxon (MIC 0,004-0,032 mg/L). CONCLUSIONS: All of the investigate PPNG strains were penicillin resistant and ceftriaxon sensitive. The dominating type of the penicillinase plasmid was Toronto/Rio.

The occurrence of herpes simplex viruses 1 and 2 in skin and mucosal lesions in patients with suspicion of genital herpes.

INTRODUCTION: Infection with herpes simplex viruses 1 and 2 (HSV 1 and 2 or Human herpesvirus HHV) are one of the most common infections in human. Real time PCR is a sensitive and specific method for diagnostics of HHV infections. The aim of the study was to investigate the occurrence of HHV 1 and HHV 2 DNA in patient with clinical symptoms suggesting HHV infection. METHODS: We used real time PCR to investigate swabs from genital and perianal lesions from 74 patients of the Department of Dermatology and Venereology Medical University Warsaw and of gynecological outpatient clinics in Warsaw 40 women and 34 men. RESULTS: The results were positive for HHV 2 in 25 cases (34%), for HHV 1 in 19 cases (26%) and for both viruses in 20 cases (27%). 10 samples were negative for both viruses. CONCLUSIONS: The results confirm that the main cause of symptomatic genital herpes is HHV 2, however the percentage of HHV 1 and specially of mixed HHV 1/HHV 2 infections was unexpectedly high.

Susceptibility to ceftriaxone and occurrence of penicillinase plasmids in Neisseria gonorrhoeae strains isolated in Poland in 2012-2013.

Recent years have seen rising concerns over increasing antibiotic resistance of the gonorrhea-causing bacterium, Neisseria gonorrhoeae. This is especially true for third-generation cephalosporins, which are currently recommended for the treatment of such infections. Therefore, susceptibility to these antibiotics should be monitored internationally to the greatest extent possible. The susceptibility of N. gonorrhoeae strains to ceftriaxone and penicillin, as well as production of beta-lactamase by the Cefinase test was determined. Moreover, the presence and type of penicillinase plasmids were determined by PCR. All strains were susceptible to ceftriaxone, the minimal inhibitory concentration (MIC) values ranged from 0.002 to 0.125 mg/L; MIC50 was =0.016 mg/L and MIC90 was =0.064 mg/L. As much as 7.7 % of the strains demonstrated ceftriaxone MIC of 0.125 mg/L. For penicillin, the MICs ranged from 0.064 to 32 mg/L; MIC50 was =0.5 mg/L and MIC90 was =4 mg/L. It was shown that only 1.5 % of the strains were sensitive to penicillin according to The European Committee on Antimicrobial Susceptibility Testing (EUCAST). Among the penicillin-resistant strains, six (30.0 %) produced penicillinase. The MICs of penicillin were substantially higher for penicillinase-producing than for penicillin-resistant, penicillinase-negative strains. MICs of ceftriaxone for penicillinase-producing strains were low (0.002-0.016 mg/L). Three of the penicillinase-producing strains possessed plasmids of African type (50 %) and three Toronto/Rio type (50 %). An increase of the proportion of beta-lactamase-positive strains in the last years as well as emergence of strains with elevated MIC of ceftriaxone indicate a need to constantly monitor N. gonorrhoeae strains for their susceptibility to beta-lactam antibiotics, as well as for their ability to produce beta-lactamases.

[The Neisseria gonorhoeae and Chlamydia trachomatis coinfection in patients of Department of Dermatology and Venereology Medical University of Warsaw]. / Wsólistnienie zakazen Neisseria gonorhoeae i Chlamydia trachomatis u pacjentów zglaszajacych sie do Kliniki Dermatologii i Wenerologii Warszawskiego Uniwersytetu Medycznego.

INTRODUCTION: The aim of the study was to investigate the coexistence of N. gonorrhoeae and C. trachomatis infections in patients of Department of Dermatology and Venereology in Warsaw. METHODS: We investigated the urethral, cervical, anal and pharyngeal speci mens from 140 symptomatic and asymptomatic patients of Department of Dermatology and Venereology in Warsaw using the Real-Time PCR method. Real Time PCR DUPLICα® RealTime Neisseria gonorrhoeae and DUPLICα® RealTime Chlamydia trachomatis 2nd Generation Detection Kits (Euroclone®)was performed on termocycler Smart Cycler® Dx. For DNA isolation the Bact Extra Pure Kit (Euroclone®) was used. RESULTS: 22 samples were positive for C. trachomatis and 28 were positive for N. gonorrhoeae. Both infections coexisted in nine patients (6.4%). In our investigations, in opposition to results from other centers, gonorrhoea was more prevalent than C. trachomatis infection. The chlamydial infection coexisted in 32.1% with gonorrhea and gonorrhea coexisted with chlamydial infection in 40.9%. CONCLUSIONS: The coinfection with Neisseria gonorrhoeae and Chlamydia trachomatis occurs very often. According IUSTI and CDC recommendations patients with one kind of sexually transmitted disease (STI) diagnosed should be tested for the others. It is especially true in the case of gonorrhea and Chlamydia trachomatis infection. In IUSTI and CDC recommendations treatment for Chlamydia trachomatis is indicated in patients with gonorrhea.

[Novel multiplex real-time PCR assay for detection and differentiation of herpes simplex virus type 1 and 2 DNA]. / Wykorzystanie nowej techniki multiplex real-time PCR do wykrywania i róznicowania DNA wirusów opryszczki typu 1 i 2.

INTRODUCTION: Herpes simplex viruses type 1 and 2 are the cause of world spread multiple infections with different course and severity. The aim of this work was to design and to optimize multiplex real-time PCR assay for simultaneous detection of HSV-1 and HSV-2. The second aim of the project was to check if the designed method is laboratory useful analyzing different clinical specimens. MATERIALS AND METHODS: In this experiment primers and probes were designed to specific viral sequences: for HSV-1 to the gene of viral DNA polymerase; for HSV-2 to the UL5 sequence. For performing qPCR assay TaqMan chemistry was used. Reference strains HSV-I McIntyre and HSV-2 MS were used as a positive control. To test laboratory utility of the designed method 58 different clinical specimens were analyzed. RESULTS: Developed multiplex real-time PCR gave positive result only in the samples containing genetic material of HSV-1/2. Of the 58 clinical samples tested, 27 proved to be positive for HSV-1 and 17 for HSV-2. The 7 samples showed the presence of both types of DNA herpes simplex virus, and 7 others were found for both HSV-1 and HSV-2 negative. CONCLUSIONS: Obtained results show that the designed method is highly specific and can possibly be used to simultaneously detect and differentiate HHV-1/2. Both high specificity and very short time of analysis have great importance in diagnosing immunocompromised patients, which ought to be diagnosed quickly and effectively in order to provide appropriate treatment.

[Susceptibility of Neisseria gonorrhoeae strains isolated in Poland in 2012-2013 to spectinomycin]. / Wrazliwosc na spektynomycyne szczepów Neisseria gonorrheae izolowanych w Polsce w latach 2012-2013.

INTRODUCTION: A rapid growth of the antibiotic resistance among Neisseria gonorrhoeae strains was recently observed in many countries. The common resistance or decreased susceptibility to penicillin, tetracyclines, ciprofloxacin, azithromycin as well as emergence of the first strains resistant to ceftriakson and cefixim is a cause of an anxiety worldwide. Spectinomycin may constitute an alternative therapy of gonorrhoea except of pharyngeal infection. METHODS: The susceptibility to spectinomycin of 65 N. gonorrhoeae strains isolated in the second half of 2012 and the first half of 2013 in Dermatology and Venereology Clinic in Warsaw was investigated. The E-Tests (bioMerieux) were performed and the results (sensitive or resistant) were interpreted according to EUCAST and CLSI recommendations. RESULTS: The MIC (Minimal inhibitory concentration) values of spectinomycin for the investigated strains ranged from 4,0 mg/L to 32 mg/L, MIC50=16,0 mg/L and MIC=90=16,0 mg/L. It was shown that 100% of the strains was sensitive to spectinomycin according to EUCAST as well CLSI tables. CONCLUSIONS: High susceptibility of the investigated strains to spectinomycin suggests that the drug can be used in the therapy of the resistant gonorrhoea in monotherapy or combined with other drugs.

[The Comparison of Real-Time PCR and bacterial culture in laboratory diagnostics of gonorrhoea in patients of Department of Dermatology and Venereology Medical University of Warsaw]. / Porównanie metody Real-Time PCR i hodowli bakteryjnej w laboratoryjnej diagnostyce rzezaczki u pacjentów Kliniki Dermatologii i Wenerologii Warszawskiego Uniwersytetu Medycznego.

INTRODUCTION: Gonorrhoea is one of the most common sexually transmitted diseases in the world. Fast and effective laboratory diagnostics of the infection is very important. The aim of the study was to compare Real-Time PCR and bacterial culture in diagnostics of Neisseria gonorrhoeae infections in patients of Department of Dermatology and Venereology Medical University of Warsaw. METHODS: The urethral, cervical, anal and pharyngeal specimens from 93 patients of Department of Dermatology and Venereology Medical University of Warsaw were examined by two methods. The bacterial culture was performed on chocolate agar plates with antibiotics in the 5% CO2 atmosphere at 37°C. N. gonorrhoeae was identified by colony-morphology, Gram stain and oxidase reaction, followed by carbohydrate utilization test. The DNA Bact Extra Pure Kit (Euroclone®) to isolate DNA was used. Real-Time PCR DUPLICα® Real-Time Neisseria gonorrhoeae 2nd Generation Detection Kit (Euroclone®) was performed using Smart Cycler® Dx. RESULTS: In 85.9% of results of Real-Time PCR and culture were identical. The remaining 14.1% of samples were culture negative and PCR positive. The results ofthe Real-Time PCR and culture were more often concordant in the case of samples obtained from men (96.8%) than from woman (85%). In patients after treatment with antibiotics the concordance of the results obtained with these two methods was 81%. CONCLUSIONS: In laboratory diagnostics of N. gonorrhoeae infections, Real-Time PCR was more sensitive than the culture, but in some cases it was less specific.

Antiviral medication in sexually transmitted diseases. Part II: HIV.

This is a second part of a review under a main title Antiviral medication in sexually transmitted diseases. In the part we published in Mini Rev Med Chem. 2013,13(13):1837-45, we have described mechanisms of action and mechanism of resistance to antiviral agents used in genital herpes and genital HPV infection. The Part II review focuses on therapeutic options in HIV infection. In 1987, 6 years after the recognition of AIDS, the FDA approved the first drug against HIV--zidovudine. Since then a lot of antiretroviral drugs are available. The most effective treatment for HIV is highly active antiretroviral therapy--a combination of several antiretroviral medicines that cause a reduction of HIV blood concentration and often results in substantial recovery of impaired immunologic function. At present, there are over 20 drugs licensed and used for the treatment of HIV/AIDS, and these drugs are divided into one of six classes. Investigational agents include GS-7340, the prodrug of tenofovir and BMS-663068--the first in a novel class of drugs that blocks the binding of the HIV gp120 to the CD4 receptor.

[The Comparison of Real-Time PCR and direct immunofluorescence in laboratory diagnostics of chlamydiosis in patients of Department of Dermatology and Venereology Medical University of Warsaw]. / Porównanie metody Real-Time PCR z metoda bezposredniej immunofluorescencji w laboratoryjnej diagnostyce chlamydiozy u pacjentów Kliniki Dermatologii i Wenerologii Warszawskiego Uniwersytetu Medycznego.

INTRODUCTION: Chlamydial infection is one of the most common bacterial sexually transmitted diseases. The aim of the study was to compare Real Time PCR and Direct Immunofluorescence (DIF) in laboratory diagnostics of Chlamydia trachomatis infection in patients of Department of Dermatology and Venereology in Warsaw. METHODS: We investigated the urethral, cervical, anal and pharyngeal specimens from 152 patients of Department of Dermatology and Venereology in Warsaw. We used DNA Bact Extra Pure Kit (Euroclone®) to isolate DNA. Real Time PCR DUPLICα® 2nd Generation Detection Real Time Advanced Dual Easy Chlamydia trachomatis Kit (Euroclone®) was performed on termocycler Smart Cycler® Dx. For direct immunofluorescence we used MicroTrack Chlamydia trachomatis Direct Specimen Test (Trinity Biotech). RESULTS: In 90% of cases of Real Time PCR and DIF were consistent. 9% of samples were DIF negative and PCR positive and 1% were PCR negative and DIF positive. The results of PCR and DIF were identical more often in woman (98%)than in men (84%). In patients after antibiotics treatment the results of the two tests were consistent in 88%. CONCLUSIONS: In laboratory diagnostics of C. trachomatis infections the results of the Real Time PCR and DIF were highly consistent although Real Time PCR. was more sensitive than DIF.

[Resistance to ciprofloxacin of Neisseria gonorrhoeae strains isolated in Poland in 2012-2013]. / Opornosc na ciprofloksacyne szczepów Neisseria gonorrhoeae izolowanych w Polsce w latach 2012-2013.

INTRODUCTION: Ciprofloxacin is commonly used in Poland specially for the treatment of urinary tract infections including urethritis. Patients are often treated without pathogen identification and antimicrobial resistance tests. Neisseria gonorrhoeae infection is one of the most common causes of urethritis in Poland. The resistance of bacteria to a wide range of antibiotics including ciprofloxacine makes the therapy of gonorrhoea more difficult. The mechanism of ciprofloxacine action depends on inactivation of bacterial topoisomerase II (gyrase) and topoisomerase IV. A resistance to ciprofloxacine occurring in Neisseria gonorrhoeae is mainly due to mutations in bacterial gyrA (encoding topoisomerase II) and/or parC (encoding topoisomerase IV ) genes. High level resistance is an effect of combination of three or four mutations. Another, less important mechanism of ciprofloxacin resistance, that can coexist with mutations in gyrA and parC genes related to the overproduction of membrane pumps proteins. MATERIAL AND METHODS: 65 Neisseria gonorrhoeae strains isolated from patients of Department of Dermatology and Wenereology in Warsaw in the second half of 2012 and first of 2013 was investigated. The strains were cultured on chocolate agar plates in a 5% CO2 atmosphere at 37 degrees C and identified by colony morphology, Gram stain and oxidase reaction, followed by carbohydrate utilization test. Ciprofloxacin susceptibility was determined by E-Tests (bioMerieux). Bacteria were incubated at 35 degrees C in 5% CO2 for 24 h on chocolate agar plates. Tests were performed according to producers recommendations. The results (sensitive or resistant) were interpreted according to EUCAST recommendations. RESULTS: The MIC (Minimal inhibitory concentration) of Ciprofloxacin in investigated strains ranged from 0,002 to > 32 mg/L, MIC50 = 8 mg/L, MIC90 = > 32 mg/L. It was shown that only 38.5% of the strains were sensitive to ciprofloxacin according to EUCAST criteria from 2013 year. CONCLUSIONS: Due to the high percentage of ciprofloxacin resistant Neisseria gonorrhoeae strains (more than 61%) the antibiotic should not be used for the treatment of gonorrhoea in Poland.

Initial prevalence of anal human papilloma virus infection in liver transplant recipients.

Although liver transplant recipients are at increased risk of human papilloma virus (HPV)-related anal cancer, limited data are available regarding the initial prevalence of anal HPV infection in this population. Anal swabs collected from 50 liver transplant recipients within the first three postoperative weeks were subjected to real-time polymerase chain reaction for detection of the four HPV genotypes: 6, 11, 16, and 18. Predictors of any, low-risk, and high-risk anal HPV infection were evaluated. Overall, the prevalence of any anal HPV infection was 18.0%, with the corresponding rates for high- and low-risk HPV genotypes being 8.0% and 10.0%, respectively. Infection with any type of anal HPV was higher in patients with hepatitis B virus (HBV) infection (P = 0.027), ≥3 sexual partners (P = 0.031), and alcoholic liver disease (P = 0.063). HBV infection was the only factor significantly associated with high-risk HPV infection (P = 0.038). Male sex (P = 0.050), age ≥52 years (P = 0.016), ≥30 sexual partners (P = 0.003), age at first intercourse ≤18 years (P = 0.045), and time since first intercourse ≥38 years (P = 0.012) were identified as predictors of low-risk HPV infection. These results indicate that HPV vaccination of liver transplant candidates and screening for anal HPV infection in high-risk groups should be considered.

Antimicrobial susceptibility/resistance and genetic characteristics of Neisseria gonorrhoeae isolates from Poland, 2010-2012.

BACKGROUND: In Poland, gonorrhoea has been a mandatorily reported infection since 1948, however, the reported incidences are likely underestimated. No antimicrobial resistance (AMR) data for Neisseria gonorrhoeae has been internationally reported in nearly four decades, and data concerning genetic characteristics of N. gonorrhoeae are totally lacking. The aims of this study were to investigate the AMR to previously and currently recommended gonorrhoea treatment options, the main genetic resistance determinant (penA) for extended-spectrum cephalosporins (ESCs), and genotypic distribution of N. gonorrhoeae isolates in Poland in 2010-2012. METHODS: N. gonorrhoeae isolates cultured in 2010 (n = 28), 2011 (n = 92) and 2012 (n = 108) in Warsaw and Bialystok, Poland, were examined using antimicrobial susceptibility testing (Etest), pyrosequencing of penA and N. gonorrhoeae multi-antigen sequence typing (NG-MAST). RESULTS: The proportions of N. gonorrhoeae isolates showing resistance were as follows: ciprofloxacin 61%, tetracycline 43%, penicillin G 22%, and azithromycin 8.8%. No isolates resistant to ceftriaxone, cefixime or spectinomycin were found. However, the proportion of isolates with an ESC MIC = 0.125 mg/L, i.e. at the resistance breakpoint, increased significantly from none in 2010 to 9.3% and 19% in 2012 for ceftriaxone and cefixime, respectively. Furthermore, 3.1% of the isolates showed multidrug resistance, i.e., resistance to ciprofloxacin, penicillin G, azithromycin, and decreased susceptibility to cefixime (MIC = 0.125 mg/L). Seventy-six isolates (33%) possessed a penA mosaic allele and 14 isolates (6.1%) contained an A501V/T alteration in penicillin-binding protein 2. NG-MAST ST1407 (n = 58, 25% of isolates) was the most prevalent ST, which significantly increased from 2010 (n = 0) to 2012 (n = 46; 43%). CONCLUSIONS: In Poland, the diversified gonococcal population displayed a high resistance to most antimicrobials internationally previously recommended for gonorrhoea treatment and decreasing susceptibility to the currently recommended ESCs. The decreasing susceptibility to ESCs was mostly due to the introduction of the internationally spread multidrug-resistant NG-MAST ST1407 in 2011. It is essential to promptly revise the gonorrhoea treatment guidelines, improve the gonorrhoea laboratory diagnostics, and implement quality assured surveillance of gonococcal AMR (ideally also treatment failures) in Poland.