Association of human papillomavirus and bacterial vaginosis with increased risk of high-grade squamous intraepithelial cervical lesions.

OBJECTIVE: To assess the rates of co-infections between human papillomavirus (HPV) and 13 key markers of bacterial vaginosis in cervical samples by multiplex polymerase chain reaction in a population with a high rate of abnormal cytology and a positive HPV test. METHODS: The study included a total of 213 women aged 18-72 years screened using Papanicolaou smears for determining cervical abnormalities and for HPV and bacterial vaginosis by single-target and multiplex polymerase chain reaction. RESULTS: A total of 83 (39%) women were negative for intraepithelial lesion or malignancy cytology and 130 (61%) had abnormal cytology. HPV-DNA prevalence was 69.9% and bacterial vaginosis was 72.7 %. Co-infections between bacterial vaginosis with HPV-DNA and high-risk HPV were associated with an increased risk for squamous intraepithelial lesions of low-grade cytology and high-grade squamous intraepithelial lesions plus cervical cancer. The most frequent bacterial vaginosis agent was Gardnerella vaginalis (33.8%), and co-infection with HPV-DNA and high-risk HPV increased the risk for squamous intraepithelial lesions of low grade cytology and high-grade squamous intraepithelial lesions plus cervical cancer. Co-infection between Megasphaera type I and high-risk HPV increased the risk for high-grade squamous intraepithelial lesions plus cervical cancer. CONCLUSIONS: Our results reinforce the hypothesis that some bacterial vaginosis agents may play a role as co-factors in HPV-mediated cervical carcinogenesis, at least in some populations.

Propolis increases neutrophils response against Candida albicans through the increase of reactive oxygen species.

AIM: To investigate whether Brazilian green propolis improves the immune response against recurrent form isolate recurrent vulvovaginal candidiasis (RVVC) caused by Candida albicans by increasing neutrophil oxidative burst. MATERIALS & METHODS: We evaluated oxidant species production, oxygen consumption, microbicidal activity and myeloperoxidase activity in neutrophils previously treated with propolis and activated with different isolates of C. albicans (RVVC), vulvovaginal candidiasis, asymptomatic isolates and the reference strain. RESULTS: Propolis significantly increased oxidant species production, oxygen consumption, microbicidal activity and myeloperoxidase activity of neutrophils against different isolates of C. albicans including RVVC isolate that are considered resistant to the microbicidal activity of neutrophils. CONCLUSION: Brazilian green propolis may increase neutrophil burst oxidative response to RVVC leading to an efficient removal of C. albicans.

Nanoparticles of Waste Material of Propolis and Gelatin as a Novel System for Delivery of L-Ascorbic Acid.

BACKGROUND: The waste material from the preparation of propolis extracts is a potential natural compound for application in pharmaceutical and medicine nanostructured products. Ascorbic acid is an excellent antioxidant and an important cofactor of several physiological and biochemical processes. OBJECTIVE: The aim of this study was to develop and characterize nanoparticles containing L-ascorbic acid prepared with propolis byproduct. METHOD: Nanoparticle's physicochemical characteristics (surface morphology, particle size, zeta potential, and entrapment efficiency), antioxidant activity, in vitro release profile, and in vitro cytotoxicity were evaluated. RESULTS: Nanoparticles showed to be spherical, with agglomeration, mean diameter between 110.93 and 480.59 nm, zeta potential near zero and good entrapment efficiency. Antioxidant activity of L-ascorbic acid increased when nanoencapsulated and the drug release was prolonged, controlled mainly by the phenomenon of relaxation of polymer chains and dependent of propolis residue concentration. The application of technology provided a reduction in the level of cytotoxicity of L-ascorbic acid, and the nanoparticles showed a protective effect on macrophages.

Microbicidal activity of neutrophils is inhibited by isolates from recurrent vaginal candidiasis (RVVC) caused by Candida albicans through fungal thioredoxin reductase.

Vulvovaginal candidiasis (VVC) is characterized by an infection of the vulva and vagina, mainly caused by Candida albicans, a commensal microorganism that inhabits the vaginal, digestive, and respiratory mucosae. Vulvovaginal candidiasis affects approximately 75% of women, and 5% develop the recurrent form (RVVC). The aim of the present study was to evaluate whether neutrophils microbicidal response is triggered when activated with RVVC isolates caused by C. albicans. Our results showed that RVVC isolates induced neutrophil migration but significantly decrease the microbicidal activity of neutrophils, compared with VVC and ASS isolates. The microbicidal activity of neutrophils is highly dependent on the production of reactive oxygen species/reactive nitrogen species (ROS/RNS). However, this isolate induced detoxification of ROS/RNS produced by neutrophils, reflected by the high level of thiol groups and by the oxygen consumption. Therefore, RVVC isolates induced biochemical changes in the inflammatory response triggered by neutrophils, and these effects were mainly related to the detoxification of ROS/RNS through the thioredoxin reductase (TR), a key antioxidant enzyme in fungi. This might be one of the resistance mechanisms triggered by RVVC caused by C. albicans.

Prevalence of clue cells suggestive for Gardnerella vaginalis in population-based cervical screening in the public versus private health care in Maringá, Paraná, Brazil.

OBJECTIVES: To determine the prevalence of clue cells suggestive for Gardnerella vaginalis in population-based cervical screening in the public (SUS) versus private (RP) health care utilization in Maringá, Paraná, Brazil. METHODS: A retrospective study on the results of 133,966 cytology examinations of patients from the Brazilian Public Health System-SUS (better socio-economic status-SES) and from the Private Network-RP (low SES) done by the Souza Anatomy Laboratory in Maringá, Brazil, from 2004 to 2007. RESULTS: In patients using the SUS, the prevalence of clue cells was 6.75% (n = 3,573), higher (p < 0.05) than for RP users (3.53%, n = 2,813). The clue cells predominated in age range of the 20-49 years from the SUS and RP (p > 0.05). In both the populations, the prevalence and age range did not vary among the 4 years (p > 0.05) and the prevalence declines in age above 45 years (p < 0.05). CONCLUSIONS: The prevalence of clue cells was low and stable in both populations in the 4 years, indicating equilibrium in terms of numbers of bacterial vaginosis (BV) in this localization. The results indicate that SES may influence the epidemiology of BV and that the prevalence declines expressively in older women.

Susceptibilidad de levaduras vaginales a los antifúngicos más utilizados en Maringá, Paraná, Brasil / Susceptibility to vaginal yeast in most used antifungal in Maringá, Paraná, Brazil

Se determinó la susceptibilidad antifúngica in vitro de 78 cepas de levaduras aisladas de mujeres de la ciudad de Maringá/Paraná, Brasil, con candidiasis vulvovaginal (CVV), atendidas en el Laboratorio de Enseñanza e Investigación en Análisis Clínicos (LEPAC) de la Universidad Estatal de Maringá, desde el 1 de enero 2005 al 31 de diciembre 2006. Su sensibilidad in vitro fue investigada por el método de microdilución frente a ketoconazol (KETO), fluconazol (FLU), itraconazol (ITRA), nistatina (NIS) y anfotericina B (AMB). Para KETO, 41,5% de las cepas de C. albicans y 96% de Candida no-albicans presentaron resistencia (100% de C. glabrata) y para FLU solamente el 3,8% de los aislamientos de C. albicans y el 8,0% de C. glabrata fueron resistentes. Sólo 1,9% de las cepas de C. albicans y 20% de las de C. no-albicans fueron resistentes a ITRA y el 5,7% de las C. albicans y el 8% de las C. no-albicans (sólo C. glabrata) fueron resistentes a AMB. No hubo aislamientos resistentes a NIST, pero sí una elevada frecuencia de sensibilidad dosis dependiente "in vitro". Estos datos avalan la creciente necesidad de la realización de pruebas de identificación y susceptibilidad in vitro a los antifúngicos para establecer el correcto tratamiento de la CVV.

In vitro antifungal susceptibility was determined in the 78 yeasts isolated from patients with vulvovaginal candidiasis (VVC) from the city of Maringá/Parana/Brazil, assisted in the Laboratory of Teaching and Research in Clinical Analysis of the State University of Maringá, from 01 January 2005 to December 31, 2006. Its sensibility in vitro was tested according to microdilution method in front of ketoconazol (KETO), fluconazole (FLU), itraconazole (ITRA), nistatin (NIS) and amphotericin B (AMB). For KET, 41.5% of the C. albicans and 96.0% of the C. non-albicans showed resistance (100.0% of C. glabrata) and for FLU, only 3.8% of the isolates of C. albicans and 8.0% of C. glabrata showed resistance. Only 1.9% of the C. albicans and 20% of the C. no-albicans were resistant. For AMB, 5.7% of the C. albicans and 8% of the C. no-albicans (only C. glabrata), were resistant. There were no isolations resistant from NIST, however, there was a high frequency of dose-dependent sensibility (SDD) in vitro. These data makes it possible to confirm the growing necessity of the performance of identification tests and in vitro antifungal susceptibility to antifungals to establish the correct treatment of CVV.

Oncotic colpocytology stained with Harris-Shorr in the observation of vaginal microorganisms.

The purpose of this work was to evaluate the efficacy of oncotic colpocytology stained with Harris-Shorr in the identification of the cervicovaginal microflora and infectious agents. Results of microbiologic evaluation carried out in colpocytology exams, bacterioscopy (Gram), and direct exams of 2,017 women aged from 13 to 80 years were compared. Of these, 83.1% agreed between cytology and Gram, 3.6% partially agreed, and 12.8% disagreed. The predominant microflora was of lactobacilli (63.3%), followed by mixed flora (32.1%). The results of sensitivity to lactobacilli were 96.1% and to mixed flora 88.0%; the specificity values were 91.2 and 92.0%, respectively. Colpocytology detected all the instances of trichomoniasis observed at direct exam (0.6%). The most frequent infectious agents were of candidiasis (14.8%, sensitivity 80.3%) and bacterial vaginosis (11.9%, sensitivity 68.1%). Thus, Harris-Shorr stained cytology was shown to be an excellent diagnostic method for T. vaginalis, lactobacilli, mixed flora, and candidiasis.

A simplified technique for evaluating the adherence of yeasts to human vaginal epithelial cells.

Thousands of women all over the world annually suffer of vulvovaginal candidiasis (VVC), an infection caused by yeasts, which mainly affect the mucosa of the vulva and vagina. The adherence of the yeasts to the mucosae is an essential step for colonization and predisposes the start of the infectious process. In this study, a technique capable of evaluating the adhesion of yeasts to human vaginal epithelial cells (HVEC) was employed. Twenty-five vaginal yeast isolates (10 Candida albicans, nine C. glabrata, two C. parapsilosis, one C. tropicalis, two Saccharomyces cerevisiae, and one Trichosporon sp.) were evaluated. A suspension of each yeast was co-incubated with HVEC obtained from a healthy donor in the ovulatory phase. After 1 hr, smears were made, stained with crystal violet and Papanicolaou, and the number of yeasts that adhered to 600 HVEC was evaluated. The adhesion of C. albicans was significantly greater than that of the other species and occurred mainly in the intermediate HVEC, rather than the superficial. In addition, the proposed technique, easy to execute and of low cost showed to be reproducible and enables the determination of the adherence capacity of different isolates, whose adhesion was confirmed by scanning electron microscopy.

Correlation of Candida species and symptoms among patients with vulvovaginal candidiasis in Maringá, Paraná, Brazil.

Vulvovaginal candidiasis (VVC) is an infection caused by abnormal yeast growth in the mucosa of the female genital tract which is commonly diagnosed in gynecology. The aim of this study was to correlate the frequency of yeasts and their respective species in asymptomatic women with different clinical manifestation of VVC; evaluate possible relationships between number of fungus colonies and symptoms in this pathology. All patients who visited the laboratory within a period of five months, for routine examinations of vaginal secretion, independent of the presence or absence of symptoms of VVC were included in this study. Of these, women with immunodeficiency or with an infection of the genital tract by another agent were excluded. Candida albicans was the most frequently yeast isolated (60%). Among non-C. albicans yeasts, 61.5% were isolated of the asymptomatic women, 38.7% from patients with VVC and 11.1% of those from patients with RVVC. C. albicans was associated with symptoms of VVC and while, the presence of non-C. albicans yeasts with asymptomatic women. However, there was no association between the number of fungal colonies and symptoms.