Neisseria gonorrhoeae: un patógeno díscolo: conceptos microbiológicos, resistencia a antimicrobianos y su vigilancia epidemiológica en Chile / Neisseria gonorrhoeae: a wayward pathogen: microbiological concepts, antimicrobial resistance and its epidemiological surveillance in Chile

Resumen Neisseria gonorrhoeae es un diplococo gramnegativo, no móvil, esporulado, aerobio o anaerobio facultativo, catalasa y oxidasa positivas. Las infecciones de transmisión sexual causadas por este microorganismo son un problema de salud pública definido como tal desde el siglo XIX, representando una gran amenaza para la salud humana debido a la su alta prevalencia y multirresistencia a antimicrobianos. En las últimas décadas han aumentado los reportes de cepas resistentes a penicilina, fluoroquinolonas, sulfonamidas, tetraciclina, macrólidos, y más recientemente a cefalosporinas y azitromicina. Tal panorama ha generado preocupación a nivel mundial, debido al aumento de casos de gonorrea asociados a cepas multirresistentes. En Chile se desarrolló desde el 2010 hasta el 2018 el Programa de Vigilancia de N. gonorrhoeae a nivel nacional con el objeto de caracterizar esta infección en las regiones y registrar la resistencia a los antimicrobianos. Esta revisión presenta un análisis sistemático bibliográfico, actualizado, de los principales aspectos de este microorganismo, su respuesta a antimicrobianos, y entrega pautas de diagnóstico y tratamiento, a la espera de avanzar en la comprensión del mecanismo molecular y las interacciones metabólicas e inmunológicas que determinan la infección, con miras a diseñar una vacuna efectiva.

Abstract Neisseria gonorrhoeae is a nonmotile, sporulated, aerobic or facultative anaerobic gram-negative diplococcus, catalase and oxidase positive. Sexually transmitted infections caused by this microorganism were established as public health problem since the 19th century, representing a great threat to human health due to its high prevalence and multi-resistance to antimicrobials. In recent decades, reports of strains resistant to penicillin, fluoroquinolones, sulfonamides, tetracycline, macrolides, and more recently to cephalosporins and azithromycin have increased. Such a panorama has generated concern worldwide, due to the increase in cases of gonorrhea associated with multi-resistant strains. In Chile, from 2010 to 2018, the National Surveillance Program for N. gonorrhoeae was developed in order to characterize this infection in the regions and record antimicrobial resistance. This review presents an updated, systematic bibliographic analysis of the main aspects of this microorganism, its response to antimicrobials, and provides diagnostic and treatment guidelines, while waiting to advance in the understanding of the molecular mechanism and the metabolic and immunological interactions that determine infection, with a view to designing an effective vaccine.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: doença inflamatória pélvica / Brazilian Protocol for Sexually Transmitted Infections 2020: pelvic inflammatory disease / Protocolo Brasileño para Infecciones de Transmisión Sexual 2020: enfermedad inflamatoria pélvica

O tema doença inflamatória pélvica está contemplado no Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis, publicado pelo Ministério da Saúde do Brasil em 2020. A doença inflamatória pélvica é a infecção aguda do trato genital superior feminino decorrente da ascensão canalicular de microrganismos cervicovaginais endógenos e, principalmente, os de transmissão sexual. Entre os agentes etiológicos envolvidos, destacam-se Chlamydia trachomatis e Neisseria gonorrhoeae. As sequelas mais importantes são dor pélvica crônica, infertilidade e gravidez ectópica. O diagnóstico clínico apresenta-se como a abordagem prática mais importante. O tratamento com antibióticos deve ser iniciado imediatamente diante da suspeição clínica. Descrevem-se orientações para gestores e profissionais de saúde sobre testes diagnósticos, tratamento preconizado, seguimento, aconselhamento, notificação, manejo de parcerias sexuais e de populações especiais. Com a maior disponibilidade da técnica de biologia molecular no Brasil, recomenda-se o rastreio de C. trachomatis e N. gonorrhoeae como estratégia preventiva da doença.

Pelvic Inflammatory Disease is a topic included in the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections, published by the Brazilian Ministry of Health in 2020. Pelvic inflammatory disease is an acute infection of the upper female genital tract due to canalicular spread of endogenous cervicovaginal microorganisms, in particular sexually transmitted organisms. Standing out among the etiological agents involved are Chlamydia trachomatis and Neisseria gonorrhoeae. The most important sequels are chronic pelvic pain, infertility and ectopic pregnancy. Clinical diagnosis is the most important practical approach. Antibiotic treatment should start immediately upon clinical suspicion. The article contains guidance for health service managers and health professionals on diagnostic tests, treatment, follow-up, counseling, notification, handling of sexual partnerships and special populations. In view of increased availability of the molecular biology technique in Brazil, C. trachomatis and N. gonorrhoeae screening is recommended as a disease prevention strategy.

El tema de la enfermedad inflamatoria pélvica está incluido en el Protocolo Clínico y Directrices Terapéuticas para la Atención Integral para Personas con Infecciones de Transmisión Sexual, publicado por el Ministerio de Salud de Brasil en 2020. La enfermedad inflamatoria pélvica es una infección aguda del tracto genital superior femenino resultante del ascenso canalicular de microorganismos cervicovaginales endógenos y, principalmente, los de transmisión sexual. Entre los agentes etiológicos involucrados, se destacan Chlamydia trachomatis y Neisseria gonorrhoeae. Las secuelas más importantes son: dolor pélvico crónico, infertilidad y embarazo ectópico. El diagnóstico clínico es el enfoque práctico más importante. El tratamiento con antibiótico debe iniciarse inmediatamente ante la sospecha clínica. Se describen pautas para gestores y profesionales de la salud sobre pruebas de diagnóstico, tratamiento, seguimiento, asesoramiento, notificación, manejo de parejas sexuales y poblaciones especiales. Con la mayor disponibilidad de la técnica de biología molecular, se recomienda el cribado de C. trachomatis y N. gonorrhoeae como estrategia preventiva para la enfermedad.

A rare ocular complication of neisseria gonorrhoeae.

BACKGROUND: Spontaneous formation of subconjunctival filtering blebs is rare and has been observed after penetrating ocular surgery, in association with connective tissue diseases and inflammatory conditions. METHOD: We describe here a case of a spontaneous formation of subconjunctival filtering bleb as sequelae of gonococcal conjunctivitis in an adult patient. RESULTS: Spontaneous filtering bleb is a rare complication of gonococcal conjunctivitis. In this case, the patient presented with a recent history of gonococcal conjunctivitis treated with moxifloxacin eye drops and intravenous cephalothin for a week. On the first exam, there was superior corneal thinning with surrounding conjunctival hyperemia and edema in the right eye, but after the patient lost the follow-up for 2 weeks, the corneal thinning had become a pseudopterygium. After another week, we observed a focal elevation on the superior conjunctiva that had the appearance of a bleb, although the anterior chamber remained deep. The filtering bleb was confirmed by ultrasound biomicroscopy examination (UBM). CONCLUSION: This case is relevant to the clinical practice because filtering blebs should be considered as a complication of gonococcal conjunctivitis because they pose a threat to the eye when substantial leakage occurs.

Macrophage-Neisseria gonorrhoeae Interactions: A Better Understanding of Pathogen Mechanisms of Immunomodulation.

Neisseria gonorrhoeae is a significant health problem worldwide due to multi-drug resistance issues and absence of an effective vaccine. Patients infected with N. gonorrhoeae have not shown a better immune response in successive infections. This might be explained by the fact that N. gonorrhoeae possesses several mechanisms to evade the innate and adaptative immune responses at different levels. Macrophages are a key cellular component in the innate immune response against microorganisms. The current information suggests that gonococcus can hijack the host response by mechanisms that involve the control of macrophages activity. In this mini review, we intend to condense the recent knowledge on the macrophage-N. gonorrhoeae interactions with a focus on strategies developed by gonococcus to evade or to exploit immune response to establish a successful infection. Finally, we discuss the opportunities and challenges of therapeutics for controlling immune manipulation by N. gonorrhoeae.

Neisseria gonorrhoeae Challenge Increases Matrix Metalloproteinase-8 Expression in Fallopian Tube Explants.

Background:Neisseria gonorrhoeae (Ngo) is the etiological agent of gonorrhea, a sexually transmitted infection that initially infects the female lower genital tract. In untreated women, the bacteria can ascend to the upper genital reproductive tract and infect the fallopian tube (FTs), which is associated with salpingitis and can lead to impaired FT function and infertility. The extracellular matrix (ECM) plays an important role in cell migration and differentiation in the female genital tract, and some pathogens modify the ECM to establish successful infections. The ECM is regulated by matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), their endogenous inhibitors; MMP deregulation causes pathological conditions in a variety of tissues. Results: The aim of this work was to analyze the expression and localization of MMP-3, MMP-8, MMP-9, and TIMP-1 in FT explants during Ngo infection using real-time PCR, immunohistochemistry, zymography and ELISA. No significant variations in MMP-3, MMP-9, and TIMP-1 transcript levels were observed. In contrast, a significant increase (p < 0.05) was observed for MMP-8 expression and was accompanied by stromal immunoreactivity in infected explants. ELISA results supported these findings and showed that MMP-8 release increased upon gonococcal infection. Conclusions: Our results indicate that gonococcal infection induces increased MMP-8 expression, which might contribute to FT damage during infection.

ATP Induces IL-1ß Secretion in Neisseria gonorrhoeae-Infected Human Macrophages by a Mechanism Not Related to the NLRP3/ASC/Caspase-1 Axis.

Neisseria gonorrhoeae (Ngo) has developed multiple immune evasion mechanisms involving the innate and adaptive immune responses. Recent findings have reported that Ngo reduces the IL-1ß secretion of infected human monocyte-derived macrophages (MDM). Here, we investigate the role of adenosine triphosphate (ATP) in production and release of IL-1ß in Ngo-infected MDM. We found that the exposure of Ngo-infected MDM to ATP increases IL-1ß levels about ten times compared with unexposed Ngo-infected MDM (P < 0.01). However, we did not observe any changes in inflammasome transcriptional activation of speck-like protein containing a caspase recruitment domain (CARD) (ASC, P > 0.05) and caspase-1 (CASP1, P > 0.05). In addition, ATP was not able to modify caspase-1 activity in Ngo-infected MDM but was able to increase pyroptosis (P > 0.01). Notably ATP treatment defined an increase of positive staining for IL-1ß with a distinctive intracellular pattern of distribution. Collectively, these data demonstrate that ATP induces IL-1ß secretion by a mechanism not related to the NLRP3/ASC/caspase-1 axis and likely is acting at the level of vesicle trafficking or pore formation.

Estado actual de la gonorrea / Current state of gonorrhea

La gonorrea es una infección de transmisión sexual conocida desde la antigüedad que se ha perpetuado en el tiempo. Su agente etiológico, Neisseria gonorrhoeae, fue adquiriendo resistencia a la mayoría de los antimicrobianos utilizados para su tratamiento empírico. Las resistencias emergentes en los últimos años son a fluorquinolonas, macrólidos y cefalosporinas orales y parenterales. Hay aislamientos multirresistentes que plantean un desafío para su tratamiento. En países donde estos hallazgos ocurrieron precedentemente, la experiencia de tratamientos con antibióticos no convencionales, por ejemplo aminoglucósidos, es limitada y también emergieron cepas resistentes. Una de las estrategias utilizadas frente a la sensibilidad disminuida a ceftriaxona es aumentar la dosis o utilizar tratamientos combinados. En el marco actual de la multirresistencia de este microorganismo es importante promover la prevención de la infección, realizar vigilancia epidemiológica y buscar nuevas estrategias para su tratamiento.

Gonorrhea is a sexually transmitted infection known since antiquity that has been perpetuated over time. Its etiologic agent, Neisseria gonorrhoeae, was becoming resistant to most antimicrobials agents used for empiric therapy. Emerging resistance in recent years are fluoroquinolones, macrolides, oral and parenteral cephalosporins. There are multiresistant isolates that represent a challenge for its treatment. In countries where these findings occurred previously, the experience with unconventional treatments such as aminoglycosides is limited and resistant strains have emerged. One of the strategies used against the reduced susceptibility to ceftriaxone is to increase the dose or use combined treatments. In the current context of multidrug resistance of this organism it is important to promote the prevention of infection, epidemiological surveillance and look for new strategies for treatment.

History of gonorrhea and prostate cancer in a population-based case-control study in Mexico.

UNLABELLED: We evaluated the association between a history of sexually transmitted diseases (STDs) and the risk for prostate cancer (PC) among Mexican males. METHODS: PC incident cases (n=402) that were identified at six public hospitals in Mexico City were matched by age (±5 years) with 805 population controls with no history of PC. By face-to-face interview, we obtained information about sexual history, previous STDs, sociodemographic characteristics, and familial history of PC. An unconditional logistic regression model was used to estimate the risk for PC. RESULTS: A total of 16.6% of men reported having had at least one previous STD, and the most frequently reported STD was gonorrhea (10.5%). After adjusting by PC familial history, the history of STD was associated with a two-fold greater risk of PC: odds ratio (OR)=2.67; 95% confidence interval (95% CI=1.91-3.73). When each STD was evaluated separately, only gonorrhea was associated with a significant increase in PC risk (OR=3.04; 95% CI=1.99-4.64). These associations were similar when we stratified by low-risk PC (Gleason <7) and high-risk PC (Gleason ≥7). CONCLUSION: These results confirm that STDs, and particularly gonorrhea, may play an etiological role in PC among Mexican males, which is consistent with a previous report from a multiethnic cohort.

Prevalence of bla TEM-220 gene in Penicillinase-producing Neisseria gonorrhoeae strains carrying Toronto/Rio plasmid in Argentina, 2002 - 2011.

BACKGROUND: Penicillinase-producing Neisseria gonorroheae (PPNG) was first isolated in 1976. PPNG strains carrying bla TEM-1 and bla TEM-135 gene have been described in different countries. Recently, a novel bla TEM-220 allele was detected in PPNG isolates carrying Toronto/Rio plasmid. The prevalence and characteristics of TEM-220 strains worldwide are unknown, and therefore, it needs to be studied. The purpose of this study was to detect bla TEM-220 gene in PPNG strains possessing Toronto/Rio plasmid over a period of ten years in Argentina, and to evaluate the proportion of isolates producing non-TEM-220 containing the T539C substitution in the bla TEM allele. METHODS: One hundred and fifty one PPNG isolates carrying Toronto/Rio plasmid were studied between 2002 and 2011. A mismatch amplification mutation assay (MAMA) PCR was used to identify the T539C substitution in the bla TEM allele and a MAMA-PCR protocol was developed to detect the G547A substitution in the bla TEM-220. The reference agar dilution method of the Clinical and Laboratory Standard Institute (CLSI) was used for susceptibility testing to five ß-lactams antibiotics, ciprofloxacin, tetracycline and azithromycin. In all TEM-220-producing isolates, the whole bla TEM gene was sequenced and the isolates were typed using N. gonorroheae multiantigen sequence typing (NG-MAST). RESULTS: MAMA PCR successfully identified the G547A substitution in the bla TEM-220 allele. The proportion of isolates that possessed the bla TEM-220 allele was 2.6 %, and 93.2 % MAMA TEM-220 PCR-negative isolates showed the T539C substitution in the bla TEM gene. No differences in the susceptibility to five beta-lactam antibiotics tested were observed in PPNG isolates TEM-220-producing and PPNG isolates carrying the T539C substitution in the bla TEM gene. All TEM-220 isolates were indistinguishable by NG-MAST. CONCLUSION: This is the first study which shows the prevalence of bla TEM-220 in N. gonorrhoeae isolates carrying Toronto/Rio plasmid in Argentina. Although the bla TEM-220 allele does not appear to be associated with an extended spectrum beta-lactamase (ESBL) phenotype of resistance, a single nucleotide polymorphism added to the bla TEM-220 or bla TEM containing the T539C substitution could lead to the emergence of ESBL. Thus, it is imperative to investigate in surveillance programs, not only the plasmid type in PPNG isolates and the bla TEM allele associated, but phenotypical characteristics and geographical distribution of isolates.

Rho GTPases as pathogen targets: Focus on curable sexually transmitted infections.

Pathogens have evolved highly specialized mechanisms to infect hosts. Several microorganisms modulate the eukaryotic cell surface to facilitate their engulfment. Once internalized, they hijack the molecular machinery of the infected cell for their own benefit. At different stages of phagocytosis, particularly during invasion, certain pathogens manipulate pathways governed by small GTPases. In this review, we focus on the role of Rho proteins on curable, sexually transmitted infections caused by Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis and Treponema pallidum. Despite the high, worldwide frequencies of these sexually-transmitted diseases, very little is known about the strategies developed by these microorganisms to usurp key eukaryotic proteins that control intracellular signaling and actin dynamics. Improved knowledge of these molecular mechanisms will contribute to the elucidation of how these clinically important pathogens manipulate intracellular processes and parasitize their hosts.

Colonização nasal em profissionais de enfermagem de unidades especializadas em HIV/aids / Colonización nasal en unidades de enfermeras especializadas en el VIH/SIDA / Nasal colonization in nursing professionals from units specialized in HIV/AIDS

RESUMO Objetivo: investigar a presença de micro-organismos nas narinas dos profissionais de enfermagem de um hospital de ensino brasileiro. Método: estudo transversal, em duas unidades de internação especializadas em HIV/aids. Foram coletadas amostras de secreção nasal de profissionais de enfermagem no período de um mês. As amostras foram processadas no laboratório de microbiologia da instituição e a análise dos dados resultantes por meio do software Statistical Package for the Social Sciences (SPSS) versão 19.0. Os aspectos éticos foram contemplados. Resultados: dos 73 profissionais de enfermagem do serviço, foram coletadas amostras de secreção nasal de 61 (80,2%). Foram isolados seis tipos de micro-organismos em 22 (41,0%) culturas positivas. Destaca-se que o Staphylococcus aureus representou 22,9%, sendo quatro resistentes à oxacilina (MRSA). Conclusão: o Staphylococcus aureus foi o micro-organismo de maior prevalência nos indivíduos deste estudo. .

RESUMEN Objetivo: investigar la presencia de microorganismos en las fosas nasales del personal de enfermería de un hospital universitario brasileño. Método: estudio transversal en dos unidades de hospitalización especializados en VIH/SIDA. Muestras de secreción nasal de enfermeras fueron recolectados durante un mes. Las muestras fueron procesadas en el laboratorio de microbiología de la institución y se analizaron con el paquete estadístico para el software de Ciencias Sociales (SPSS) versión 19.0. Los aspectos éticos fueron cubiertos. Resultados: 73 de los profesionales de enfermería, se recogieron muestras de las secreciones nasales de 61 (80,2%). Se aislaron seis tipos de microorganismos en 22 (41,0%) cultivos positivos. Es de destacar que el Staphylococcus aureus representó el 22,9%, cuatro oxacilina-resistente (MRSA). Conclusión: Staphylococcus aureus fue la prevalencia más microorganismo en los individuos de este estudio. .

ABSTRACT Objective: to investigate the presence of microorganisms in the nostrils of the nursing professionals of a Brazilian teaching hospital. Method: cross-sectional study in two inpatient units specialized in HIV/AIDS. Nasal secretion samples of nursing professionals were collected in one month. The samples were processed at the microbiology laboratory of the institution and analyzed using the Statistical Package for the Social Sciences (SPSS) software, version 19.0. Ethical aspects were abided. Results: from the 73 members of the nursing staff, samples of nasal secretions were collected from 61 (80.2%). Six types of microorganisms were isolated in 22 (41.0%) positive cultures. It is noteworthy that Staphylococcus aureus accounted for 22.9%, four of them oxacillin-resistant (MRSA). Conclusion: Staphylococcus aureus microorganism accounted for the largest prevalence in individuals of this study. .

Neisseria gonorrhoeae induces a tolerogenic phenotype in macrophages to modulate host immunity.

Neisseria gonorrhoeae is the etiological agent of gonorrhoea, which is a sexually transmitted disease widespread throughout the world. N. gonorrhoeae does not improve immune response in patients with reinfection, suggesting that gonococcus displays several mechanisms to evade immune response and survive in the host. N. gonorrhoeae is able to suppress the protective immune response at different levels, such as B and T lymphocytes and dendritic cells. In this study, we determined whether N. gonorrhoeae directly conditions the phenotype of RAW 264.7 murine macrophage cell line and its response. We established that gonococcus was effectively phagocytosed by the RAW 264.7 cells and upregulates production of immunoregulatory cytokines (IL-10 and TGF- ß 1) but not the production of proinflammatory cytokine TNF- α , indicating that gonococcus induces a shift towards anti-inflammatory cytokine production. Moreover, N. gonorrhoeae did not induce significant upregulation of costimulatory CD86 and MHC class II molecules. We also showed that N. gonorrhoeae infected macrophage cell line fails to elicit proliferative CD4+ response. This implies that macrophage that can phagocytose gonococcus do not display proper antigen-presenting functions. These results indicate that N. gonorrhoeae induces a tolerogenic phenotype in antigen-presenting cells, which seems to be one of the mechanisms to induce evasion of immune response.

Abordagem atual da doença inflamatória pélvica / Current approaches to pelvic inflammatory disease

A doença inflamatória pélvica (DIP) é um processo inflamatório de natureza infecciosa que pode atingir estruturas e órgãos do trato genital superior. Devido à sua importância epidemiológica e de suas graves complicações, este artigo atualiza e propõe uma abordagem sistemática da DIP. Os principais agentes etiológicos são a Neisseria gonorrhoeae,Chlamydia trachomatis e outros agentes etiológicos de uretrites, cervicites, vulvovaginites e vaginoses, em geral, polimicrobiana, o que é a base de sua terapêutica. A mulher deve ser investigada para DIP quando apresenta, especialmente, desconforto abdominal, dor lombar, dispareunia e nódoas ou manchas ao exame ginecológico, previamente a procedimentos transcervicais. A classificação clínico-laparoscópica deDIP pode ser dividida em: a) estágio I (endometrite/salpingite sem peritonite); estágio II (salpingite aguda com peritonite); estágio III (salpingite aguda com oclusão tubária ou abscesso tubo-ovariano); estágio IV (abscesso tubo-ovariano roto). A definição do estágio orienta a conduta e o tratamento, pois em formas leves (estágio I) o tratamento e seguimento podem ser feitos ambulatorialmente, enquanto para os casos moderadosou graves a internação hospitalar está indicada para início do tratamento por via endovenosa e monitorização da resposta ao tratamento. O tratamento suportivo, retirada de dispositivo intrauterino (DIU), abstinência sexual e repouso também são indicados, além de orientações sobre as implicações da doença e abordagem do parceiro.

Pelvic inflammatory disease (PID) is an inflammatory process of infectious nature that can affect structures and organs of the upper genital tract. Considering this disease's epidemiological relevance and severe complications, this article provides an update and proposes a systematic approach to PID. The main etiological agents are Neisseria gonorrhoeae, Chlamydia trachomatis and other etiological agents of urethritis, cervicitis, vulvovaginitis and vaginoses. These are generally of polymicrobial origin, which determines the treatment basis for pelvic inflammatory diseases.Women must be checked for PID when experiencing abdominal discomfort, backache, dyspareunia, or presenting with stains during gynecological examination and prior to transcervical procedures. The clinical and laparoscopic classification of PID can be divided into: a) stage I (endometritis/salpingitis without peritonitis), stage II (acute salpingitis with peritonitis), stage III (acute salpingitis with tubal occlusion or tube-ovarian abscess), and stage IV (tube-ovarian abscess rupture). Defining the stage guides procedures and treatment, given that in mild forms (stage I) the treatment and follow-up can be performed in the ambulatory environment while moderate to severe cases require hospitalization so that intravenous treatment and treatment outcome monitoring can be started. Supportive treatment, removal of intrauterine device (IUD), sexual abstinence and rest are also indicated, as well as counseling on the implications of the disease and partner approach.

Molecular basis of high-level ciprofloxacin resistance in Neisseria gonorrhoeae strains from Shandong Province, China

In the study, the ciprofloxacin resistance rate was 100%. High-level ciprofloxacin resistance rate was 63.55%. Sixteen different mutation patterns involved in the formation of ciprofloxacin resistance were identified. The most prevalent were patterns P7 (25.2%), P8 (15.0%), P9 (11.2%), P1 (10.3%), and P5 (10.3%). All of the 107 NG isolates analyzed for mutations in the study have demonstrated a change of Ser-91 → Phe in the gyrA gene, and all except one have demonstrated a change in position 95 of the amino acid sequence. All of the 68 high-level QRNG isolates had double mutations in gyrA gene combined with a single or two mutations in parC gene. It is most important that a new mutation site of Ile-97 → Met in gyrA and a new mutation of Leu-106 → Ile in parC were found in the study, both leading to high-level ciprofloxacin resistance (MIC values, 8 µg/mL, 32 µg/mL, respectively). Therefore, we confim that gyrA mutations are necessary for the fluoroquinolone resistance phenotype and parC mutations are correlated intimately with high-level fluoroquinolone resistance. In China fluoroquinolone resistance in Neisseria gonorrhoeae strains is very serious and the new mutation sites in the fluoroquinolone resistance-determining regions emerge more and more quickly. Hence, in China fluoroquinolones, which are used to treat gonorrhoea presently, should be substituted by a new antibiotics.(AU)

Neisseria gonorrhoeae pilus attenuates cytokine response of human fallopian tube explants.

BACKGROUND: A role for pilus during attachment of Neisseria gonorrhoeae to epithelia of the female reproductive tract is currently assumed. However, Pil⁻ gonococci have been observed during infection of the reproductive tract, which prompted us to examine the effect of pili on the dynamics of infection and the inflammatory responses of mucosal explants of the human fallopian tube. METHODS: Mucosal explants were infected in vitro with Opa negative Pil⁻ and Pil⁺N. gonorrhoeae strains. RESULTS: Piliation enhanced gonococcal adherence to the epithelium within 3 h of infection (P < 0.05) but thereafter did not offer advantage to gonococci to colonize the epithelial cell surface (P > 0.05). No differences were found between the strains in numbers of gonococci inside epithelial cells. Pil⁻ bacteria induced higher levels (P < 0.05) of IL-1ß, TNF-α, GM-CSF, MCP-1, and MIP-1ß than Pil⁺ bacteria. There were no differences between both strains in LOS pattern, and Pil expression did not change after coincubation with mucosal strips. CONCLUSIONS: Results show that gonococcal invasion of the human fallopian tube can occur independently of pilus or Opa expression, and suggest that pilus, by inhibition of several key elements of the initial inflammatory response, facilitates sustained infection of this organ.

Neisseria gonorrhoeae induced disruption of cell junction complexes in epithelial cells of the human genital tract.

Pathogenic microorganisms, such as Neisseria gonorrhoeae, have developed mechanisms to alter epithelial barriers in order to reach subepithelial tissues for host colonization. The aim of this study was to examine the effects of gonococci on cell junction complexes of genital epithelial cells of women. Polarized Ishikawa cells, a cell line derived from endometrial epithelium, were used for experimental infection. Infected cells displayed a spindle-like shape with an irregular distribution, indicating potential alteration of cell-cell contacts. Accordingly, analysis by confocal microscopy and cellular fractionation revealed that gonococci induced redistribution of the adherens junction proteins E-cadherin and its adapter protein ß-catenin from the membrane to a cytoplasmic pool, with no significant differences in protein levels. In contrast, gonococcal infection did not induce modification of either expression or distribution of the tight junction proteins Occludin and ZO-1. Similar results were observed for Fallopian tube epithelia. Interestingly, infected Ishikawa cells also showed an altered pattern of actin cytoskeleton, observed in the form of stress fibers across the cytoplasm, which in turn matched a strong alteration on the expression of fibronectin, an adhesive glycoprotein component of extracellular matrix. Interestingly, using western blotting, activation of the ERK pathway was detected after gonococcal infection while p38 pathway was not activated. All effects were pili and Opa independent. Altogether, results indicated that gonococcus, as a mechanism of pathogenesis, induced disruption of junction complexes with early detaching of E-cadherin and ß-catenin from the adherens junction complex, followed by a redistribution and reorganization of actin cytoskeleton and fibronectin within the extracellular matrix.

The incidence and correlates of symptomatic and asymptomatic Chlamydia trachomatis and Neisseria gonorrhoeae infections in selected populations in five countries.

BACKGROUND: Asymptomatic Chlamydia trachomatis (chlamydia) and Neisseria gonorrhoeae (gonorrhea) infections pose diagnostic and control problems in developing countries. METHODS: Participants in China, India, Peru, Russia, and Zimbabwe were screened for C. trachomatis and N. gonorrhoeae infections and symptoms. RESULTS: A total of 18,014 participants were evaluated at baseline, 15,054 at 12 months, and 14,243 at 24 months. The incidence of chlamydia in men was 2.0 per 100 person years both from baseline to 12 months and from 12 to 24 months, and in women, 4.6 from baseline to 12 months and 3.6 from 12 to 24 months; a range of 31.2% to 100% reported no symptoms across the 5 countries. The incidence of gonorrhea in men was 0.3 per 100 person years both from baseline to 12 months and from 12 to 24 months, and in women, 1.4 from baseline to 12 months and 1.1 from 12 to 24 months; a range of 66.7% to 100% reported no symptoms. Being female, aged 18 to 24 years, and having more than 1 partner were associated with both the infections. In addition, being divorced, separated, or widowed was associated with gonorrhea. Being male, having 6+ years of education, and reporting only 1 partner were associated with having no symptoms among those infected with chlamydia. No variables correlated with asymptomatic gonorrhea among those infected. CONCLUSION: A high prevalence and incidence of asymptomatic sexually transmitted infections was identified among men and women in a wide variety of settings. More effective programs are needed to identify and treat chlamydia and gonorrhea infections, especially among women, young adults, those with multiple partners, those repeatedly infected, and particularly those at risk without symptoms. The risk of transmission from persons with no symptoms requires further study.

2nd international workshop on Neisseria vaccines (Neisseria vaccines 2009).

A 5-day workshop on Neisseria vaccines was held in Varadero, Cuba, 17-22 May 2009, and hosted by the Latin American Association for Immunology and the Cuban Society for Immunology, which attracted more than 100 scientists from different parts of the world. The meeting summarizes current knowledge regarding Neisseria species vaccine development, including plain polysaccharide vaccines, conjugate polysaccharides and protein-based vaccines. Main discussion topics were the discovery of new potential antigens with protective capacity, current and new vaccines, pathogenesis and immune response, clinical trials and alternative correlates of protection and manufacture, control and regulation, gonococcus vaccines, and adjuvant as the main component of Neisseria vaccines. Seven keynote addresses, 32 oral presentations, 49 posters and an International Vaccine Course were presented. The meeting was concluded with a 'Hot Topics' session discussing future challenges. This article highlights the most important issues presented.

Infection of human fallopian tube epithelial cells with Neisseria gonorrhoeae protects cells from tumor necrosis factor alpha-induced apoptosis.

Following infection with Neisseria gonorrhoeae, bacteria may ascend into the Fallopian tubes (FT) and induce salpingitis, a major cause of infertility. In the FT, interactions between mucosal epithelial cells and gonococci are pivotal events in the pathogen's infection cycle and the inflammatory response. In the current study, primary FT epithelial cells were infected in vitro with different multiplicities of infection (MOI) of Pil+ Opa+ gonococci. Bacteria showed a dose-dependent association with cells and induced the secretion of tumor necrosis factor alpha (TNF-alpha). A significant finding was that gonococcal infection (MOI = 1) induced apoptosis in approximately 30% of cells, whereas increasing numbers of bacteria (MOI = 10 to 100) did not induce apoptosis. Apoptosis was observed in only 11% of cells with associated bacteria, whereas >84% of cells with no adherent bacteria were apoptotic. TNF-alpha was a key contributor to apoptosis, since (i) culture supernatants from cells infected with gonococci (MOI = 1) induced apoptosis in naïve cultures, suggesting that a soluble factor was responsible; (ii) gonococcal infection-induced apoptosis was inhibited with anti-TNF-alpha antibodies; and (iii) the addition of exogenous TNF-alpha induced apoptosis, which was inhibited by the presence of increasing numbers of bacteria (MOI = 10 to 100). These data suggest that TNF-alpha-mediated apoptosis of FT epithelial cells is likely a primary host defense mechanism to prevent pathogen colonization. However, epithelial cell-associated gonococci have evolved a mechanism to protect the cells from undergoing TNF-alpha-mediated apoptosis, and this modulation of the host innate response may contribute to establishment of infection. Understanding the antiapoptotic mechanisms used by Neisseria gonorrhoeae will inform the pathogenesis of salpingitis and could suggest new intervention strategies for prevention and treatment of the disease.