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1.
Med Microbiol Immunol ; 209(5): 641, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32382801

RESUMO

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.

2.
Med Microbiol Immunol ; 209(2): 95-108, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31802195

RESUMO

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.


Assuntos
Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Farmacorresistência Bacteriana Múltipla/genética , Gonorreia/tratamento farmacológico , Gonorreia/microbiologia , Neisseria gonorrhoeae/efeitos dos fármacos , Neisseria gonorrhoeae/genética , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Azitromicina/farmacologia , Azitromicina/uso terapêutico , Cefixima/farmacologia , Cefixima/uso terapêutico , Ceftriaxona/farmacologia , Ceftriaxona/uso terapêutico , Ciprofloxacina/farmacologia , Ciprofloxacina/uso terapêutico , Europa (Continente)/epidemiologia , Genótipo , Gonorreia/epidemiologia , Humanos , Neisseria gonorrhoeae/classificação , Neisseria gonorrhoeae/isolamento & purificação , Penicilinas/farmacologia , Penicilinas/uso terapêutico
3.
PLoS Pathog ; 13(6): e1006406, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28640877

RESUMO

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.


Assuntos
Proteína alfa Estimuladora de Ligação a CCAAT/metabolismo , Transformação Celular Viral/genética , Regulação da Expressão Gênica/fisiologia , Queratinócitos/virologia , MicroRNAs/biossíntese , Proteínas Oncogênicas Virais/metabolismo , Linhagem Celular , Imunoprecipitação da Cromatina , Ensaio de Desvio de Mobilidade Eletroforética , Epidermodisplasia Verruciforme/complicações , Epidermodisplasia Verruciforme/virologia , Humanos , Immunoblotting , Imuno-Histoquímica , Hibridização In Situ , Queratinócitos/metabolismo , Infecções por Papillomavirus/complicações , Reação em Cadeia da Polimerase em Tempo Real
4.
Postepy Dermatol Alergol ; 35(6): 346-551, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30618519

RESUMO

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.

5.
Med Dosw Mikrobiol ; 69(1): 49-54, 2017.
Artigo em Polonês | MEDLINE | ID: mdl-30351624

RESUMO

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.


Assuntos
Neoplasias do Colo/virologia , Pólipos do Colo/virologia , Infecções por Papillomavirus/complicações , Neoplasias do Colo/etiologia , Pólipos do Colo/etiologia , Papillomavirus Humano 11/isolamento & purificação , Papillomavirus Humano 6/isolamento & purificação , Humanos , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/epidemiologia , Prevalência
6.
Med Dosw Mikrobiol ; 68(1): 34-38, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28146620

RESUMO

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.


Assuntos
Proteínas de Bactérias/genética , Neisseria gonorrhoeae/isolamento & purificação , Resistência às Penicilinas/genética , Penicilinase/genética , Plasmídeos , Gonorreia/diagnóstico , Gonorreia/enzimologia , Gonorreia/genética , Humanos , Neisseria gonorrhoeae/enzimologia , Neisseria gonorrhoeae/genética , Polônia
7.
Med Dosw Mikrobiol ; 68(1): 57-62, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28146623

RESUMO

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.


Assuntos
Canal Anal/virologia , Genitália/virologia , Herpes Genital/epidemiologia , Herpesvirus Humano 1/isolamento & purificação , Herpesvirus Humano 2/isolamento & purificação , Adulto , Idoso , DNA Viral/análise , Feminino , Herpes Genital/diagnóstico , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa/virologia , Pele/virologia , Adulto Jovem
8.
Postepy Dermatol Alergol ; 33(6): 475-479, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28035227

RESUMO

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.

9.
Med Dosw Mikrobiol ; 67(1): 23-8, 2015.
Artigo em Polonês | MEDLINE | ID: mdl-26084072

RESUMO

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.


Assuntos
Neisseria gonorrhoeae/classificação , Neisseria gonorrhoeae/efeitos dos fármacos , Espectinomicina/farmacologia , Resistência Microbiana a Medicamentos , Testes de Sensibilidade Microbiana , Neisseria gonorrhoeae/isolamento & purificação , Polônia , Especificidade da Espécie
10.
Med Dosw Mikrobiol ; 67(3-4): 173-80, 2015.
Artigo em Polonês | MEDLINE | ID: mdl-27019911

RESUMO

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.


Assuntos
Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis , Técnica Direta de Fluorescência para Anticorpo , Reação em Cadeia da Polimerase em Tempo Real , Canal Anal/microbiologia , Colo do Útero/microbiologia , Chlamydia trachomatis/isolamento & purificação , Feminino , Humanos , Masculino , Faringe/microbiologia , Uretra/microbiologia
11.
Med Dosw Mikrobiol ; 67(2): 89-95, 2015.
Artigo em Polonês | MEDLINE | ID: mdl-26591660

RESUMO

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.


Assuntos
Infecções por Chlamydia/complicações , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/isolamento & purificação , Coinfecção/diagnóstico , Coinfecção/epidemiologia , Gonorreia/complicações , Gonorreia/microbiologia , Adulto , Colo do Útero/microbiologia , Coinfecção/microbiologia , Feminino , Humanos , Masculino , Neisseria gonorrhoeae/isolamento & purificação , Faringe/microbiologia , Prevalência , Uretra/microbiologia
12.
Med Dosw Mikrobiol ; 67(2): 125-32, 2015.
Artigo em Polonês | MEDLINE | ID: mdl-26591664

RESUMO

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.


Assuntos
DNA Viral/isolamento & purificação , Herpesvirus Humano 1/genética , Herpesvirus Humano 1/isolamento & purificação , Herpesvirus Humano 2/genética , Herpesvirus Humano 2/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Líquido Cefalorraquidiano/virologia , DNA Polimerase Dirigida por DNA/genética , Humanos , Soro/virologia , Pele/virologia , Urina/virologia
13.
Med Dosw Mikrobiol ; 67(1): 29-38, 2015.
Artigo em Polonês | MEDLINE | ID: mdl-26084073

RESUMO

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.


Assuntos
Gonorreia/diagnóstico , Gonorreia/microbiologia , Neisseria gonorrhoeae/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Técnicas de Tipagem Bacteriana , Colo do Útero/microbiologia , Feminino , Humanos , Masculino , Faringe/microbiologia , Sensibilidade e Especificidade , Uretra/microbiologia
14.
PLoS Pathog ; 8(7): e1002833, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22911498

RESUMO

Infection with genus beta human papillomaviruses (HPV) is implicated in the development of non-melanoma skin cancer. This was first evidenced for HPV5 and 8 in patients with epidermodysplasia verruciformis (EV), a genetic skin disease. So far, it has been unknown how these viruses overcome cutaneous immune control allowing their persistence in lesional epidermis of these patients. Here we demonstrate that Langerhans cells, essential for skin immunosurveillance, are strongly reduced in HPV8-positive lesional epidermis from EV patients. Interestingly, the same lesions were largely devoid of the important Langerhans cells chemoattractant protein CCL20. Applying bioinformatic tools, chromatin immunoprecipitation assays and functional studies we identified the differentiation-associated transcription factor CCAAT/enhancer binding protein ß (C/EBPß) as a critical regulator of CCL20 gene expression in normal human keratinocytes. The physiological relevance of this finding is supported by our in vivo studies showing that the expression patterns of CCL20 and nuclear C/EBPß converge spatially in the most differentiated layers of human epidermis. Our analyses further identified C/EBPß as a novel target of the HPV8 E7 oncoprotein, which co-localizes with C/EBPß in the nucleus, co-precipitates with it and interferes with its binding to the CCL20 promoter in vivo. As a consequence, the HPV8 E7 but not E6 oncoprotein suppressed C/EBPß-inducible and constitutive CCL20 gene expression as well as Langerhans cell migration. In conclusion, our study unraveled a novel molecular mechanism central to cutaneous host defense. Interference of the HPV8 E7 oncoprotein with this regulatory pathway allows the virus to disrupt the immune barrier, a major prerequisite for its epithelial persistence and procarcinogenic activity.


Assuntos
Betapapillomavirus/patogenicidade , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Quimiocina CCL20/genética , Quimiocina CCL20/metabolismo , Queratinócitos/metabolismo , Células de Langerhans/fisiologia , Infecções por Papillomavirus/imunologia , Neoplasias Cutâneas/virologia , Betapapillomavirus/imunologia , Betapapillomavirus/metabolismo , Sítios de Ligação , Proteína beta Intensificadora de Ligação a CCAAT/genética , Linhagem Celular Tumoral , Movimento Celular , Células Cultivadas , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Elementos Facilitadores Genéticos , Epiderme/metabolismo , Epiderme/virologia , Epidermodisplasia Verruciforme/virologia , Humanos , Queratinócitos/imunologia , Proteínas Oncogênicas Virais/metabolismo , Regiões Promotoras Genéticas
15.
BMC Infect Dis ; 14: 65, 2014 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-24502606

RESUMO

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.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Gonorreia/microbiologia , Neisseria gonorrhoeae/efeitos dos fármacos , Neisseria gonorrhoeae/genética , Adolescente , Adulto , Idoso , Proteínas de Bactérias/genética , Ciprofloxacina/uso terapêutico , Feminino , Gonorreia/epidemiologia , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Neisseria gonorrhoeae/classificação , Neisseria gonorrhoeae/isolamento & purificação , Polônia/epidemiologia , Prevalência , Adulto Jovem
16.
Transpl Int ; 27(8): 816-23, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24750339

RESUMO

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.


Assuntos
Doenças do Ânus/epidemiologia , Transplante de Fígado/efeitos adversos , Infecções por Papillomavirus/epidemiologia , Adulto , Fatores Etários , Idoso , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Papillomaviridae/classificação , Prevalência , Comportamento Sexual
17.
Med Dosw Mikrobiol ; 66(2): 99-104, 2014.
Artigo em Polonês | MEDLINE | ID: mdl-25369656

RESUMO

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.


Assuntos
Ciprofloxacina/farmacologia , Resistência Microbiana a Medicamentos , Neisseria gonorrhoeae/classificação , Neisseria gonorrhoeae/efeitos dos fármacos , Antibacterianos/farmacologia , Contraindicações , Gonorreia/tratamento farmacológico , Humanos , Testes de Sensibilidade Microbiana , Neisseria gonorrhoeae/isolamento & purificação , Polônia , Especificidade da Espécie
18.
Med Dosw Mikrobiol ; 66(3-4): 209-14, 2014.
Artigo em Polonês | MEDLINE | ID: mdl-25804074

RESUMO

INTRODUCTION: Azithromycin is one of the most commonly used macrolide antibiotics. As other macrolides it inhibits bacterial proteins synthesis by binding with V domene of bacterial 23S rRNA. Resistance to azithromycin can be related to: 1. Mutations in gene encoding 23S rRNA. Significant effect on azithromycin MIC (minimal inhibitory concentration) is not observed when the mutation occurs only in 1 allele. In case of mutations occurring in 4 alleles, more common mutation C2611T is associated with MIC 2-16 mg/L and the second mutation A2059T results in high level resistance to azithromycin MIC > 256 mg/l 2. Over- production of membrane pumps proteins MtrCDE and MacAB, that remove antibiotics from bacterial cells. The mechanism is not able to cause azithromycin resistance itself but coexisting with other mechanisms of resistance can additionally increase MIC. 3. Synthesis of 23S rRNA methylases ErmB, ErmF, ErmC, ErmA. These enzymes cause demethylation of adenine (A2058) in V domain of 23S rRNA. The mechanism was common in the past, but it has been replaced by mutations in in V domain of 23S rRNA. Nowadays 23S rRNA methylases are very rare in N. gonorhoeae. METHODS: Sixty five Neisseria gonorrhoeae strains isolated from patients of the Department of Dermatology and Venereology in Warsaw in the second half of 2012 and first of 2013 were investigated. The strains were cultured on chocolate agar plates in a 5% CO2 atmosphere at 37 °C and identified by colony morphology, Gram staining and oxidase reaction, followed by carbohydrate utilization test. Azithromycin susceptibility was determined by E-Tests (bioMerieux). Bacteria were incubated at 37°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 of azithromycin in investigated strains ranged from 0,064 to 4 mg/L, MIC50 = 0.5 mg/L, MIC90 = 2 mg/L. It was shown that only 38.5% of the strains were sensitive to azithromycin according to EUCAST criteria from 2014 year and 89.3% of the strains were sensitive to azithromycin according to CDC criteria from 2014 year. CONCLUSIONS: The percentage of azithromycin resistant Neisseria gonorrhoeae strains is increasing in Poland and the antibiotic should not be used in monotherapy as gonorrhoea patients. It should only be used in combination with ceftriaxone or cefixime.


Assuntos
Azitromicina/farmacologia , Farmacorresistência Bacteriana , Neisseria gonorrhoeae/efeitos dos fármacos , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Humanos , Testes de Sensibilidade Microbiana , Mutação , Neisseria gonorrhoeae/classificação , Neisseria gonorrhoeae/genética , Polônia , RNA Bacteriano/genética , RNA Ribossômico 23S/genética , Especificidade da Espécie
19.
Skin Res Technol ; 19(1): e200-8, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22671986

RESUMO

BACKGROUND/PURPOSE: The knowledge how surfactants and hydrolyzed proteins influence the elastic properties of living epidermal keratinocytes is sparse. We demonstrate that the stiffness of cells measured by atomic force microscope (AFM) can be correlated with viability test. METHODS AND MATERIALS: The effects of sodium lauryl sulphate (SLS) and hydrolyzed collagen (HK) of molecular weight 9 kDa were examined with respect to human keratinocytes viability and elasticity. MTT assay was applied to determine the survival fraction of keratinocytes treated with SLS and HK solutions of various molar ratios. The AFM measurements of the keratinocytes stiffness were carried out immediately after the exposure of cells to the SLS and HK, respectively. RESULTS: The increase of the SLS concentration resulted in the decrease of cells proliferation and this effect was inhibited by addition of HK. The strongest inhibition was observed for the SLS:HK molar ratio equals to 2:1. AFM study shows decrease in the cell stiffness for cells treated with SLS. Fluorescence microscopy reveals remodeling of actin filaments of SLS-treated cells. SLS:HK mixture treatment results in mechanical stiffness close to untreated cells. CONCLUSION: These results provide possible correlations between mechanical properties and viability of keratinocytes when the chemical stress occurs.


Assuntos
Colágeno/toxicidade , Elasticidade/efeitos dos fármacos , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Modelos Biológicos , Tensoativos/toxicidade , Citoesqueleto de Actina/efeitos dos fármacos , Citoesqueleto de Actina/fisiologia , Antígenos de Superfície/efeitos dos fármacos , Antígenos de Superfície/fisiologia , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Membrana Celular/fisiologia , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/fisiologia , Elasticidade/fisiologia , Humanos , Hidrólise , Queratinócitos/fisiologia , Microscopia de Força Atômica , Dodecilsulfato de Sódio/toxicidade
20.
Med Dosw Mikrobiol ; 63(2): 115-20, 2011.
Artigo em Polonês | MEDLINE | ID: mdl-22184905

RESUMO

In recent years the resistance of Neisseria gonorrhoeae to antibiotics is increasing in many countries. The aim of the study was to investigate penicillinase production by Neisseria gonorrhoeae strains isolated from the patients of Clinic of Dermatology and Wenerology WUM in a period between 2006 - 2009. We cultured the bacteria on Roiron medium and we used the iodometric test or BBL Cefinase discs to detect penicillinase. The enzyme was produced by 1,1% of 183, 0,9% of 111, 1,1% of 94 and 0% of 91 of investigated strains, respectively in 2006, 2007, 2008 and 2009 year - on average by 0,8%. This is the lowest result in Europe and one of the lowest in the world.


Assuntos
Neisseria gonorrhoeae/enzimologia , Penicilinase/biossíntese , Farmacorresistência Bacteriana/fisiologia , Feminino , Humanos , Masculino , Neisseria gonorrhoeae/classificação , Neisseria gonorrhoeae/isolamento & purificação , Especificidade da Espécie
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