Mechanism of n-butanol alcohol extract of Baitouweng Decoction in treatment of vulvovaginal candidiasis based on negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis / 中国中药杂志

This study aimed to explore the mechanism of n-butanol alcohol extract of Baitouweng Decoction(BAEB) in the treatment of vulvovaginal candidiasis(VVC) in mice based on the negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis. In the experiment, female C57BL/6 mice were divided randomly into the following six groups: a blank control group, a VVC model group, high-, medium-, and low-dose BAEB groups(80, 40, and 20 mg·kg~(-1)), and a fluconazole group(20 mg·kg~(-1)). The VVC model was induced in mice except for those in the blank control group by the estrogen dependence method. After modeling, no treatment was carried out in the blank control group. The mice in the high-, medium-, and low-dose BAEB groups were treated with BAEB at 80, 40, and 20 mg·kg~(-1), respectively, and those in the fluconazole group were treated with fluconazole at 20 mg·kg~(-1). The mice in the VVC model group received the same volume of normal saline. The general state and body weight of mice in each group were observed every day, and the morphological changes of Candida albicans in the vaginal lavage of mice were examined by Gram staining. The fungal load in the vaginal lavage of mice was detected by microdilution assay. After the mice were killed, the degree of neutrophil infiltration in the vaginal lavage was detected by Papanicolaou staining. The content of inflammatory cytokines interleukin(IL)-1β, IL-18, and lactate dehydrogenase(LDH) in the vaginal lavage was tested by enzyme-linked immunosorbent assay(ELISA), and vaginal histopathology was analyzed by hematoxylin-eosin(HE) staining. The expression and distribution of NLRP3, PKCδ, pNLRC4, and IL-1Ra in vaginal tissues were measured by immunohistochemistry(IHC), and the expression and distribution of pNLRC4 and IL-1Ra in vaginal tissues were detected by immunofluorescence(IF). The protein expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by Western blot(WB), and the mRNA expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by qRT-PCR. The results showed that compared with the blank control group, the VVC model group showed redness, edema, and white secretions in the vagina. Compared with the VVC model group, the BAEB groups showed improved general state of VVC mice. As revealed by Gram staining, Papanicolaou staining, microdilution assay, and HE staining, compared with the blank control group, the VVC model group showed a large number of hyphae, neutrophils infiltration, and increased fungal load in the vaginal lavage, destroyed vaginal mucosa, and infiltration of a large number of inflammatory cells. BAEB could reduce the transformation of C. albicans from yeast to hyphae. High-dose BAEB could significantly reduce neutrophil infiltration and fungal load. Low-and medium-dose BAEB could reduce the da-mage to the vaginal tissue, while high-dose BAEB could restore the damaged vaginal tissues to normal levels. ELISA results showed that the content of inflammatory cytokines IL-1β, IL-18, and LDH in the VVC model group significantly increased compared with that in the blank control group, and the content of IL-1β, IL-18 and LDH in the medium-and high-dose BAEB groups was significantly reduced compared with that in the VVC model group. WB and qRT-PCR results showed that compared with the blank control group, the VVC model group showed reduced protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues of mice and increased protein and mRNA expression of NLRP3. Compared with the VVC model group, the medium-and high-dose BAEB groups showed up-regulated protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues and inhibited protein and mRNA expression of NLRP3 in vaginal tissues. This study indicated that the therapeutic effect of BAEB on VVC mice was presumably related to the negative regulation of NLRP3 inflammasome by promoting PKCδ/NLRC4/IL-1Ra axis.

Aspectos da abordagem terapêutica sobre candidíase vulvovaginal / Aspects of the therapeutic approach to vulvovaginal candidiasis / Aspectos del abordaje terapéutico de la candidiasis vulvovaginal

A candidíase vulvovaginal, é uma infecção da vulva e vagina causada por vários tipos de Candida spp. Essa patologia afeta 75% de todas as mulheres pelo menos uma vez durante a vida, ocorrendo com mais frequência durante a idade fértil. A transmissão dessa infeção fúngica ocorre por meio de contato com mucosas e secreções em pele de portadores ou doentes, contato sexual, água contaminada e transmissão vertical. Alguns outros sintomas característicos mais vistos em casos de CVV, são lesões brancas, cremosas e planas, sendo mais intensos no período pré-menstrual, quando a acidez vaginal aumenta. numerosos antifúngicos estão disponíveis no mercado, os quais são encontrados para administração oral na forma de comprimidos ou, para uso tópico, na forma de cremes, loções, comprimidos vaginais, supositórios e tampões revestidos. O objetivo geral do trabalho foi analisar através da revisão de literatura, tratamentos convencionais e alternativos para abordagem terapêutica da Candidíase Vulvovaginal contextuando a mesma, utilizando definições, dados epidemiológicos e sua sintomatologia frente à sociedade. O presente trabalho é uma revisão integrativa, que teve a coleta de dados realizada de março de 2021 a outubro de 2021 nas bases de dados Lilacs, Scielo, Google acadêmico, A busca resultou em 902 artigos, dos quais 14 atenderam ao critério de inclusão. A busca por tratamentos frente a candidíase vulvovaginal tem se mostrado ampla de acordo com os artigos selecionadas. Concluímos que a patologia candidíase vulvovaginal, vem apresentando resistência em algumas abordagens terapêuticas, assim como algumas mulheres não aderem há algum tipo de tratamento, devido à falta de conhecimento sobre a patologia.

Vulvovaginal candidiasis is an infection of the vulva and vagina caused by various types of Candida spp. This condition affects 75% of all women at least once in their lifetime, occurring more frequently during their childbearing years. The transmission of this fungal infection occurs through contact with mucous membranes and secretions on the skin of patients or patients, sexual contact, contaminated water and vertical transmission. Some other characteristic symptoms more seen in cases of VVC are white, creamy and flat lesions, being more intense in the premenstrual period, when the vaginal acidity increases. numerous antifungals are available on the market which are available for oral administration in tablet form or, for topical use, in the form of creams, lotions, vaginal tablets, suppositories and coated tampons. The general objective of the work was to analyze, through a literature review, conventional and alternative treatments for the therapeutic approach of Vulvovaginal Candidiasis in its context, using definitions, epidemiological data and its symptoms in society. The present work is an integrative review, which had data collection carried out from March 2021 to October 2021 in the Lilacs, Scielo, Google academic databases. The search resulted in 902 articles, of which 14 met the inclusion criteria. The search for treatments against vulvovaginal candidiasis has been shown to be wide according to the selected articles. We conclude that the vulvovaginal candidiasis pathology has been showing resistance in some therapeutic approaches, as well as some women do not adhere to any type of treatment, due to lack of knowledge about the pathology.

La candidiasis vulvovaginal es una infección de la vulva y la vagina cau- sada por diversos tipos de Candida spp. Esta afección afecta al 75% de las mujeres al menos una vez en la vida, siendo más frecuente durante la edad fértil. La transmisión de esta infección fúngica se produce por contacto con mucosas y secreciones de la piel de pacientes o enfermos, contacto sexual, agua contaminada y transmisión vertical. Otros síntomas característicos más observados en los casos de CVV son las lesiones blancas, cremosas y planas, siendo más intensas en el período premenstrual, cuando aumenta la acidez vaginal. Existen en el mercado numerosos antifúngicos disponibles para adminis- tración oral en forma de comprimidos o, para uso tópico, en forma de cremas, lociones, comprimidos vaginales, supositorios y tampones recubiertos. El objetivo general del tra- bajo fue analizar, a través de una revisión bibliográfica, los tratamientos convencionales y alternativos para el abordaje terapéutico de la Candidiasis Vulvovaginal en su contexto, utilizando definiciones, datos epidemiológicos y su sintomatología en la sociedad. El pre- sente trabajo es una revisión integradora, que tuvo recolección de datos realizada de marzo de 2021 a octubre de 2021 en las bases de datos académicas Lilacs, Scielo, Google. La búsqueda resultó en 902 artículos, de los cuales 14 cumplieron los criterios de inclu- sión. La búsqueda de tratamientos contra la candidiasis vulvovaginal se ha mostrado am- plia según los artículos seleccionados. Concluimos que la patología de la candidiasis vul- vovaginal viene mostrando resistencia en algunos abordajes terapéuticos, así como algu- nas mujeres no se adhieren a ningún tipo de tratamiento, debido al desconocimiento de la patología.

Perfil de resistencia antifúngica en el tratamiento de candidiasis vaginal: Un diagnóstico de agentes etiológicos / Antifungal resistance profile in the treatment of vaginal candidiasis: A diagnosis of etiologic agents

Introducción: En las micosis, la resistencia a los antifúngicos aumenta debido a los diagnósticos y tratamientos incorrectos. Por tanto, se ha sugerido la vigilancia y el seguimiento de las cepas resistentes para garantizar una terapia adecuada. Objetivo: Determinar las especies de Candida y el perfil de resistencia a fluconazol y voriconazol, que presentan los aislados obtenidos de muestras almacenadas durante los meses diciembre 2017 y marzo 2018 de pacientes provenientes del Laboratorio del Hospital Regional "Miguel Ángel Mariscal Llerena" del departamento de Ayacucho, Perú. Material y Métodos: Estudio descriptivo transversal, en el cual se aislaron 110 cepas de Candida sp almacenadas por un período de cuatro meses, y procesadas por métodos estandarizados y aprobados por el Ministerio de Salud (MINSA) y el Instituto Nacional de Salud (INS) del Perú relacionados con el diagnóstico de agentes etiológicos de micosis humanas y la sensibilidad antifúngica con los métodos estandarizados de Clinical Laboratory Standard Institute (CLSI). Resultados: Se observó C. albicans en 86,4 % de los aislados seguida por C. glabrata con 9,1 %, C. parapsilosis 2,7 % y 0,9 % de C. tropicalis y C. krusei. El 10,5 % de C. albicans fue resistente al fluconazol y voriconazol con CMI ≥ 128 μg/mL y ≥ 16 μg/mL respectivamente, mientras que 20 % de C. glabrata mostró sensibilidad dosis dependiente y 10 % de resistencia al fluconazol. Conclusiones: Existe una gran variedad de especies de Candida, siendo la C. albicans la más común, seguida por C. glabrata, con un mayor porcentaje de aislamiento en comparación con otras especies. Se puede observar que estas especies poseen grados de vulnerabilidad considerables a la aplicación de fármacos como el fluconazol y voriconazo(AU)

Introduction: In mycoses, resistance to antifungals increases due to incorrect diagnosis and treatment. Therefore, surveillance and monitoring of resistant strains has been suggested to ensure adequate therapy. Objective: To determine the Candida species and the resistance profile to fluconazole and voriconazole presented by the isolates obtained from samples stored during the months of December 2017 and March 2018 from patients coming from the Laboratory of the Regional Hospital "Miguel Ángel Mariscal Llerena" in the department of Ayacucho, Peru. Material and Methods: Cross-sectional descriptive study in which 110 strains of Candida sp were isolated and stored for a period of four (04) months, and processed by standardized methods approved by the Ministry of Health (MINSA) and the National Institute of Health (INS) of Peru related to the diagnosis of etiological agents of human mycosis and antifungal sensitivity with the standardized methods of the Clinical Laboratory Standard Institute (CLSI). Results: C. albicans was observed in 86,4 % of isolates, followed by C. glabrata (9,1 %), C. parapsilosis (2,7 %), and 0,9 % of C. tropicalis and C. krusei; 10,5 % of C. albicans was resistant to fluconazole and voriconazole with MIC ≥ 128 μg/mL and ≥ 16 μg/mL respectively, while 20 % of C. glabrata showed dose dependent sensitivity and 10 % resistance to fluconazole. Conclusions: There is a wide variety of Candida species, with C. albicans being the most common, followed by C. glabrata, with a higher percentage of isolation compared to other species. It can be seen that these species have considerable degrees of vulnerability to the application of drugs such as fluconazole and voriconazole(AU)

Antifungal activity of linalool against fluconazole-resistant clinical strains of vulvovaginal Candida albicans and its predictive mechanism of action

Candida albicans is the most frequently isolated opportunistic pathogen in the female genital tract, with 92.3% of cases in Brazil associated with vulvovaginal candidiasis (VVC). Linalool is a monoterpene compound from plants of the genera Cinnamomum, Coriandrum, Lavandula, and Citrus that has demonstrated a fungicidal effect on strains of Candida spp., but its mechanism of action is still unknown. For this purpose, broth microdilution techniques were applied, as well as molecular docking in a predictive manner for this mechanism. The main results of this study indicated that the C. albicans strains analyzed were resistant to fluconazole and sensitive to linalool at a dose of 256 µg/mL. Furthermore, the increase in the minimum inhibitory concentration (MIC) of linalool in the presence of sorbitol and ergosterol indicated that this molecule possibly affects the cell wall and plasma membrane integrity of C. albicans. Molecular docking of linalool with proteins that are key in the biosynthesis and maintenance of the cell wall and the fungal plasma membrane integrity demonstrated the possibility of linalool interacting with three important enzymes: 1,3-β-glucan synthase, lanosterol 14α-demethylase, and Δ 14-sterol reductase. In silico analysis showed that this monoterpene has theoretical but significant oral bioavailability, low toxic potential, and high similarity to pharmaceuticals. Therefore, the findings of this study indicated that linalool probably causes damage to the cell wall and plasma membrane of C. albicans, possibly by interaction with important enzymes involved in the biosynthesis of these fungal structures, in addition to presenting low in silico toxic potential.

Distribution and antifungal susceptibility of Candida species isolated from clinical samples in southern Brazil

Abstract Invasive infections caused by Candida species have been strongly associated with poor prognosis and high resistance rates to some antifungals. This study aimed to identify Candida species isolated from different anatomical sites and to describe their susceptibility profile to antifungals. Ninety-four clinical isolates of Candida were obtained from a Medical Laboratory of Santa Catarina/Brazil. Species identification was performed by MALDI-TOF MS. Susceptibility assays were performed as described by Clinical Laboratory Standard Institute (CLSI) microboth method. Among the analyzed samples, C. albicans was the pathogen most incident (59.9%) followed by C. parapsilosis complex (14.9%), C. glabrata complex (8.5%), and C. tropicalis (6.3%). 37 Candida strains were isolated from vaginal content (39.3%), 21 from the nail (22.4%), 8 from tracheal aspirates (8.5%), and 7 from urine (7.4%). Together, the Candida isolates presented decreased susceptibility to azole drugs, mainly to fluconazole and itraconazole. Amphotericin B showed sensibility in 95.7% of samples analyzed. Previous knowledge about etiology and antifungal susceptibility becomes indispensable to conduct an efficient treatment.

Relationship between sleep quality and risk of complicated vulvovaginal candidiasis in women / 中华健康管理学杂志

Objective:To investigate the correlation between sleep quality and risk of female complicated vulvovaginal candidiasis (VVC).Methods:From January 2021 to June 2021, patients in the gynecological clinic of Xiangya Hospital of Central South University were continuously enrolled as the research objects using a cross-sectional survey. A self-made questionnaire was used to collect the age, marital status, education level, family monthly income, place of residence in the past two years, maternity history, number of births, intrauterine device, number of abortions, frequency of sex life, use of contraceptives within two months, use of antibacterial drugs within two weeks. Generalized Anxiety Disorder Scale, Patient Health Questionnaire, Health Questionnaire Somatic Symptom Group Scale, Pittsburgh Sleep Quality Index were used to collect patients′ anxiety, depression, somatization symptoms, and sleep quality conditions. The total scores of sleep quality and the scores of each dimension were used as observation indicators. Three logistic regression analysis models were constructed to explore the relationship between sleep quality and complicated VVC groups.Results:Patients in the complex VVC group were significantly higher in age, married, middle school education, rural area of residence in the last two years, birth history, number of births ≥3, sexual frequency≥1/week, and no antibiotic use within two weeks compared to those in the control group (all P<0.05). Without adjusting for confounding factors, women with poor subjective sleep quality had a 6.73-fold increased risk ( OR=7.73, 95% CI: 3.22-18.55) of complex VVC compared with those with good subjective sleep quality. After adjusting for confounding factors, the risk was further increased to 9.08 fold ( OR=10.08, 95% CI: 3.47-29.33)(all P<0.05). Compared with women without sleep disorders, women with mild sleep disorders had a 97% increased risk of complex VVC ( OR=1.97, 95% CI: 1.15-3.37). After adjusting for confounders, the risk remained 97% higher ( OR=1.97, 95% CI: 1.10-3.55)(all P<0.05). Conclusion:Poor subjective sleep quality and mild sleep disorder may be associated with the risk of complex VVC.

Butyl alcohol extract of Baitouweng Decoction alleviates vulvovaginal candidiasis in mice by downregulating NLRP3 inflammasome and related signal pathways / 中国中药杂志

This study aims to explore the effect of butyl alcohol extract of Baitouweng Decoction(BAEB) on vulvovaginal candidiasis(VVC) in mice and to clarify the mechanism from Toll-like receptors(TLRs)/MyD88 and Dectin-1/Syk signal pathways and NLRP3 inflammasome. To be specific, female KM mice were randomized into control group(i.g., normal saline), model group, fluco-nazole group(i.g., 20 mg·kg~(-1)), and low-dose, medium-dose, and high-dose BAEB groups(i.g., 20, 40, and 80 mg·kg~(-1), respectively). VVC was induced in mice except the control group. After the modeling, administration began and lasted 7 days. The ge-neral conditions and body weight of mice were recorded every day. On the 1 st, 3 rd, 7 th, and 14 th after vaginal infection by Candida albicans, the fungal load in the vaginal lavage fluid of the mice was measured with the plate method, and the morphology of C. albicans in vaginal lavage fluid was observed based on Gram staining. After the mice were killed, vaginal tissues were subjected to hematoxylin-eosin(HE) staining and periodic acid-Schiff(PAS) staining for vaginal histopathological analysis. The content of cytokines in vaginal lavage fluid, such as interleukin(IL)-1β, IL-18, tumor necrosis factor-α(TNF-α), IL-6, and S100 a8, was determined by enzyme-linked immunosorbent assay(ELISA), and content of reactive oxygen species(ROS) in vaginal tissues by tissue ROS detection kit. The protein expression of NLRP3, ASC, caspase-1, Dectin-1, Syk, MyD88, TLR2, TLR4, and nuclear factor-κB(NF-κB) in vaginal tissues was detected by Western blot, and the levels and distribution of NLRP3, Dectin-1, Syk, MyD88, TLR2, and TLR4 in vaginal tissues were determined with the immunohistochemical method. The results show that BAEB can improve the general conditions of VVC mice, reduce the fungal load and C. albicans hyphae in vaginal secretion, decrease ROS content in vaginal tissues and content of cytokines in vaginal lavage fluid, and down-regulate the expression of NLRP3, ASC, caspase-1, Dectin-1, Syk, MyD88, TLR2, TLR4, and NF-κB in vaginal tissues. The above results indicate that BAEB exerts therapeutic effect on VVC mice by down-regulating the key proteins in the TLRs/MyD88 and Dectin-1/Syk signal pathways and NLRP3 inflammasome.

Espécies de Candida causadoras de vulvovaginites e resistência aos antifúngicos utilizados no tratamento / Candida species cause vulvovaginitis and resistance to antifungals used for treatment

Neste estudo foram identificadas espécies de Candida em isolados de secreção vaginal, avaliados os perfis de suscetibilidade in vitro a antifúngicos e correlacionados com os antifúngicos prescritos para pacientes em um serviço de atenção primária. A identificação das espécies pela Reação em Cadeia da Polimerase mostrou que 36,5% dos isolados foram caracterizados como espécie não-C.albicans. Nos testes de sensibilidade a maioria dos isolados foi suscetível a cetoconazol, fluconazol e itraconazol, contudo cerca de 40% e 50% apresentaram resistência ou sensibilidade dose-dependente a miconazol e nistatina, respectivamente. A análise dos fármacos prescritos para as pacientes revelou que 34,2% dos isolados foram considerados resistentes aos agentes utilizados no tratamento. Diversas espécies de Candida podem causar vulvovaginite com variados perfis de suscetibilidade aos antifúngicos comumente utilizados no tratamento. A identificação das espécies de Candida é relevante para o gerenciamento epidemiológico das infecções, além de ser útil, assim como os testes de suscetibilidade, na escolha do tratamento farmacológico mais eficaz para a paciente.

In this study were identified Candida species from vaginal secretion isolates, evaluated their in vitro antifungal susceptibilities, and correlated these features with antifungal agents prescribed for patients assisted in a primary care service. Species identification by Polymerase Chain Reaction showed that 36.5% of isolates were characterized as non-C. albicans species. In antifungal susceptibility tests most isolates were susceptible to ketoconazole, fluconazole, and itraconazole, although between 40% and 50% of isolates show resistance or dose-dependent susceptibility to miconazole and nystatin, respectively. Analysis of drugs prescribed to patients revealed that 34.2% of the isolates were considered resistant to agents used in treatment. Several Candida species can cause vulvovaginitis and exhibit different susceptibility profiles to antifungal drugs used in treatment. The identification of Candida species is relevant and useful to the epidemiological management of infections. The antifungal susceptibility test may also be useful for choosing most effective drug treatment for each patient.

Laboratorial aspects of cytolytic vaginosis and vulvovaginal candidiasis as a key for accurate diagnosis: a pilot study / Aspectos laboratoriais da vaginose citolítica e candidíase vulvovaginal como uma chave para o diagnóstico preciso: Um estudo piloto

Abstract Objective To identify clinical, microscopic, and biochemical characteristics that differentiate cytolytic vaginosis (CV) from vulvovaginal candidiasis (VVC). Methods The present cross-sectional study analyzed the vaginal contents of 24 non-pregnant women aged 18 to 42 years who were attended at the Genital Infections Clinic at Centro de Atenção Integral à Saúde da Mulher da Universidade Estadual de Campinas (CAISM-UNICAMP). They were diagnosed either with (CV = 8, VVC = 8) or without vulvovaginitis or vaginal dysbiosis (controls). The socio-demographic, clinical, and gynecological data were obtained from a detailed patient interview. Samples of the vaginal contents were collected for analysis of vaginal pH, gram stain, and specific fungal culture. The Kruskal-Wallis and Fisher exact tests were used to compare the differences between the groups. Odds ratios were used to compare the categorical variables. The significance level was considered at p < 0.05. Results Both women with CV and VVC had a lumpy vaginal discharge (p = 0,002) and vaginal hyperemia (p = 0.001), compared with controls. The inflammatory process was more intense in the VVC group (p = 0.001). In the CV group, there was statistical significance for the lactobacillus amount (p = 0.006), vaginal epithelium lysis (p = 0.001), and vaginal pH (p = 0.0002). Conclusion Cytolytic vaginosis and VVC diagnoses rarely differ on clinical characteristics but have different laboratorial findings. The present study highlights the importance of conducting an accurate investigation through laboratory tests rather than clinical criteria to avoid misdiagnosis.

Resumo Objetivo Identificar características clínicas, microscópicas e bioquímicas que diferenciam a vaginose citolítica (VC) da candidíase vulvovaginal (CVV). Métodos O presente estudo de corte transversal analisou o conteúdo vaginal de 24 mulheres não grávidas, com idades entre 18 e 42 anos, atendidas no ambulatório de Infecções Genitais do Centro de Atenção Integral à Saúde da Mulher da Universidade Estadual de Campinas (CAISM-UNICAMP). Elas foram diagnosticadas com (CV = 8, CVV = 8) ou sem vulvovaginite ou disbiose vaginal (controles = 8). Os dados sociodemográficos, clínicos e ginecológicos foram obtidos em uma entrevista detalhada do paciente. Amostras do conteúdo vaginal foram coletadas para análise do pH vaginal, coloração de Gram e cultura específica de fungos. Os testes exatos de Kruskal-Wallis e Fisher foram utilizados para comparar as diferenças entre os grupos. A razão de chances foi utilizada para comparar as variáveis categóricas. O nível de significância considerado foi de p < 0,05. Resultados As mulheres com VC e CVV apresentaram corrimento vaginal irregular (p = 0,002) e hiperemia vaginal (p = 0,001), em comparação aos controles. O processo inflamatório foi mais intenso no grupo CVV (p = 0,001). No grupo VC, houve significância estatística para a quantidade de lactobacilos (p = 0,006), lise do epitélio vaginal (p = 0,001) e pH vaginal (p = 0,0002). Conclusão Os diagnósticos de VC e CVV raramente diferem nas características clínicas, mas apresentam achados laboratoriais diferentes. O presente estudo destaca a importância de conduzir uma investigação precisa por meio de testes laboratoriais, em vez de critérios apenas clínicos, a fim de evitar erros de diagnóstico.

Fatores associados à presença de Candida spp. em amostras de fluido vaginal de mulheres residentes em comunidades quilombolas / Factors associated with the presence of Candida spp. in vaginal fluid samples from women living in quilombola settlements

RESUMO: Objetivo: Investigar a frequência e fatores associados à detecção de Candida spp. em fluido vaginal de mulheres residentes em uma comunidade quilombola. Materiais e Método: Um estudo transversal foi conduzido com 177 mulheres residentes em uma comunidade quilombola no Nordeste do Brasil. Foram coletados dados sociodemográficos, hábitos de vida, saúde geral e saúde reprodutiva. O material coletado na região do colo do útero foi submetido a exame citopatológico. Amostras de fluido vaginal foram coletadas para a análise microbiológica. Os testes Qui-quadrado, exato de Fisher e modelos de Re-gressão Logística foram utilizados na análise estatística. Resultados: Foi detectado Candida spp. em 28,9% das amostras. A espécie mais frequente nos casos positivos foi a Candida albicans (49%), seguida da Candida krusei (39,2%). Após o ajustamento, observou-se que mulheres com idade igual ou maior que 50 anos apresentaram maior chance da detecção de Candida spp. no fluido vaginal (OR ajustada = 3,46; IC95% = 1,68-7,12), enquanto as mulheres com queixa de corrimento vaginal apresentaram uma chance menor de detecção de Candida spp. (OR ajustada = 0,29; IC95% = 0,11-0,78). Conclusão: Os achados sugerem uma elevada detecção de Candida spp. no fluido vaginal entre mulheres residentes em comunidades quilombolas, o que sinaliza a necessidade do planejamento de medidas para prevenção e rastreamento de caso de candidíase vulvovaginal neste grupo populacional. Além disso, a presença de Candida spp. foi mais elevada em mulheres a partir da quinta década de vida, menor nas mulheres com queixas de corrimento vaginal. (AU)

ABSTRACT: Objective: Investigate the frequency and factors associated with the detection of Candida spp. in vaginal fluid from women living in a quilombola settlement. Materials and Method: A cross-sectional study was conducted with 177 women living in a quilombola settlement in Northeast, Brazil. Socio-demographic data, living habits, general health, and reproductive health were collected. The material collected in the region of the cervix was submitted to cytopathological examination. Vaginal fluid samples were collected for microbiological analysis. Chi-square test, exact Fisher's test, and logistic regression models were used for the statistical analysis. Results: Candida spp. was detected in 28.9% of the samples. The most frequent species in positive cases were Candida albicans (49%), followed by Candida krusei (39.2%). After adjustment, it was observed that women aged over 50 years had a higher chance of detecting Candida spp. in vaginal fluid (OR adjusted = 3.46; 95% CI = 1.68-7.12), while women complaining of vaginal discharge had a lower chance of detecting Candida spp. (OR adjust-ed = 0.29; 95% 95% CI= 0.11-0.78). Conclusion: The findings suggest a high detection of Candida spp. in vaginal fluid among women living in quilombola settlements, which indicates the need for planning measures for prevention and track the cases of vulvovaginal candidiasis in this population group. Besides, the presence of Candida spp. was higher in women from the fifth decade of life, lower in women with complaints of vaginal discharge. (AU)

Identification of various Candida species isolated from patients with vulvovaginal candidiasis attending a tertiary care hospital in North India

Background: Vulvovaginal Candidiasis (VVC) is a condition diagnosed in a large proportion of women presenting with complaints of abnormal vaginal discharge. The risk of VVC is high in women with diabetes mellitus, pregnancy, using oral contraceptives or broad-spectrum antibiotics and those having Human Immunodeficiency Virus infection.Objective of the present study was to identify the various species of Candida isolated from patients with Vulvovaginal Candidiasis.Methods: A cross-sectional study was done from January to June 2018. A total of 69 clinically suspected VVC cases were included in the study whose high vaginal swabs were collected and subjected to direct microscopy and cultured on Sabouraud Dextrose Agar (SDA). Creamy white pigmented colonies on SDA were confirmed as Candida by doing Gram抯 staining. Speciation of positive cultures was confirmed by conventional methods like Corn Meal agar culture, Germ tube test (GTT), Candida CHROM Agar Medium (CAM), sugar assimilation and fermentation test.Results: Out of 69 clinically suspected VVC cases included in the study, only 14 cultures were positive for Candida species. All the 14 isolates were found to be Non-albicans Candida (NAC). Most common species isolated were Candida glabrata 7(50%), followed by Candida tropicalis 3(21.4%), Candida lusitaniae 3(21.4%), and Candida parapsilosis 1 (7.1%).Conclusions: Candida albicans was known to be the most common causative agent of VVC as it was isolated in 90% cases. However, this study showed that all isolates from VVC were Non-albicans Candida. Therefore, species identification should be done in all microbiology laboratories for accurate diagnosis of VVC.

Factores de riesgo asociados a infección vaginal en mujeres embarazadas / Risk factors associated to vaginal infections in pregnant women / Fatores de risco associados à infecção vaginal em gestantes

RESUMEN La infección vaginal constituye una de las complicaciones médicas más frecuentes asociadas al embarazo. Se realizó un estudio epidemiológico, observacional, analítico, longitudinal de casos y controles con mujeres embarazadas atendidas en el Policlínico Jimmy Hirzel, Bayamo, Granma, período junio 2016 - diciembre 2017, con el objetivo de identificar factores de riesgo hipotéticamente relacionados con la aparición de infección vaginal. Los datos se obtuvieron a partir de una encuesta aplicada a las pacientes y su historia clínica, previo consentimiento informado. Se aplicó modelo de riesgo proporcional de Cox para análisis univariado y multivariado, y el Chi cuadrado, con una confiabilidad del 95% y una probabilidad menor de 0.05. La historia previa de infección de transmisión sexual (OR 2,25), más de tres gestaciones (OR 3,20), los abortos previos (9,88) y el no empleo de preservativos en las relaciones sexuales (OR 5, 35) se constituyeron en los antecedentes gineco-obstétricos relacionados con la aparición de infección vaginal en mujeres embarazadas. El inicio precoz de la vida sexual (OR 2, 25) se constituyó en un factor demográfico y de conducta sexual relacionado con la aparición de infección vaginal en mujeres embarazadas. Haber recibido algún tipo de tratamiento vaginal previo al estudio (OR 1,26), constituyó un factor de riesgo relacionado con los procedimientos externos, aunque no de forma significativa. La presencia de infección de vías urinarias y de diabetes mellitus se constituyeron en factores de riesgo relacionados con la aparición de infección vaginal en mujeres embarazadas.

ABSTRACT Vaginal infection is one of the most frequent medical complications associated with pregnancy. An epidemiological, observational, analytical, longitudinal study of cases and controls was carried out with pregnant women attended at the Jimmy Hirzel Polyclinic, Bayamo, Granma, June 2016 - December 2017, with the objective of identifying risk factors hypothetically related to the appearance of vaginal infection. The data was obtained from a survey applied to patients and their clinical history, with prior informed consent. Cox proportional risk model was applied for univariate and multivariate analysis, and Chi square, with a 95% reliability and a probability lower than 0.05. The previous history of sexually transmitted infection (OR 2.25), more than three pregnancies (OR 3.20), previous abortions (9.88) and the non-use of condoms in sexual relations (OR 5, 35) were constituted in the gynecological-obstetric history related to the appearance of vaginal infection in pregnant women. The early onset of sexual life (OR 2, 25) became a demographic and sexual behavior factor related to the appearance of vaginal infection in pregnant women. Having received some type of vaginal treatment prior to the study (OR 1.26), constituted a risk factor related to external procedures, although not in a significant way. The presence of urinary tract infection and diabetes mellitus constituted risk factors related to the appearance of vaginal infection in pregnant women.

RESUMO A infecção vaginal é uma das complicações médicas mais frequentes associadas à gravidez. Dezembro de 2017, com o objetivo de identificar fatores de risco hipoteticamente relacionados ao aparecimento de - uma epidemiológica,,, estudos de caso-controle observacionais analíticos longitudinais com mulheres grávidas que frequentam Policlínica Jimmy Hirzel, Bayamo, Granma, período de junho de estudo 2016 foi realizado infecção vaginal. Os dados foram obtidos a partir de uma pesquisa aplicada aos pacientes e sua história clínica, com consentimento prévio informado. O modelo de risco proporcional de Cox foi aplicado para análise univariada e multivariada, e o qui quadrado, com 95% de confiabilidade e probabilidade menor que 0,05. história prévia de doença sexualmente transmissível (OR 2,25), mais de três gestações (OR 3,20), abortos anteriores (9,88) e não-uso de preservativos durante as relações sexuais (OR 5, 35) foram constituídos na história gineco-obstétrica relacionada ao aparecimento de infecção vaginal em gestantes. O início precoce da vida sexual (OR 2, 25) tornou-se um fator de comportamento demográfico e sexual relacionado ao aparecimento de infecção vaginal em gestantes. Ter recebido algum tipo de tratamento vaginal antes do estudo (OR 1,26), constituiu um fator de risco relacionado aos procedimentos externos, embora não de forma significativa. A presença de infecção do trato urinário e diabetes mellitus constituiu fatores de risco relacionados ao aparecimento de infecção vaginal em gestantes.

Prevalence of vulvovaginal candidiasis in females in the reproductive age group

Background: Vulvovaginal candidiasis (VVC) is a common infection among reproductive age group females. The objective of present study is to determine the prevalence of vulvovaginal candidiasis, its distribution and association of risk factors among reproductive age group females, attending the outpatient department of obstetrics and gynaecology of our Prime Medical Centre, Sharjah attached with Prime Hospital, Dubai, United Arab Emirates (UAE).Methods: It was cross-sectional descriptive study over a period of six months. Patients who came to our outpatient department with complains of vaginal discharge and itching in reproductive age group were included in this study. Patients characteristics i.e. age, parity, risk factors like diabetes, pregnancy, use of oral contraceptive pills (OCPills) and intrauterine contraceptive device (IUCD) were noted. High vaginal swabs (HVS) were collected and sent for culture. Candida positive cases were noted, and results were analyzed.Results: A total of 224 high vaginal swabs were collected. Prevalence of vulvovaginal candidiasis was found to be 31.6%. It was found more in 26-30 years age group and multiparous women. Previous history of candidiasis and diabetes were the commonest risk factors. Frequency of C. albicans was more (76.05%) than non-albicans candida (23.94%).Conclusions: Present study concluded that vulvovaginal candidiasis is more prevalent in reproductive age group females, therefore a routine high vaginal swab culture must be performed in every woman presenting with vaginal discharge and itching for correct diagnosis. Women should be educated on clinical symptoms.

Preparation and Evaluation of Amphotericin B Vaginal Nanosuspension Foam / 中国药学杂志

OBJECTIVE: To prepare amphotericin B vaginal nanosuspension foam and study the physicochemical properties and in vivo pharmacodynamics. METHODS: Using the single factor method, the concentration of egg lecithin and the technical parameters of foaming agent in formulation were gradually optimized. Then, the physicochemical properties, including drug particle size, appearance of foam and formed capability, were studied. The therapeutic effect on vaginal inflammatory infection was investigated with the mouse model of Candida albicans vaginitis. RESULTS: The optimal formulation of amphotericin B nanosuspension foam included 5 mg•mL-1 amphotericin B, 60 mg•mL-1 egg lecithin and 30 mg•mL-1 APG0810. The drug particle size, range of micro-foam diameter and foam half-life were about 240 nm, 200-400 μm and 43.5 min, respectively. It was found that amphotericin B nanosuspension vaginal foam had significant therapeutic effect on mouse vaginal inflammation infection. CONCLUSION: The amphotericin B nanosuspension foam has good physicochemical properties and excellent pharmacological activity in vivo, which shows potential use for the therapy of vulvovaginal candidiasis.

Study on the antifungal and mucosal repair effects of chitosan gynecological soft gel on vulvovaginal candidiasis in rats / 中国实用妇科与产科杂志

OBJECTIVE: To establish the rat model of vulvovaginal candidiasis(VVC)and evaluate the antifungal and mucosal repair effects of water-soluble chitosan gynecological soft gel(Funingkang)on VVC.METHODS: From September 2018 to December 2018,animal experiments were conducted in Beijing Obstetrics and Gynecology Hospital,Capital Medical University.Thirty female Sprague Dawley(SD)rats were randomized into five treatment groups:control,infection model,Funingkang,nystatin and estrogen groups.The drug was administered continuously for 7 days according to the grouping.Vaginal secretions were collected after treatment and the clearance rate of pathogens was calculated.Vaginal specimens were assessed by hematoxylin-eosin(HE)staining,immunohistochemistry and transmission electron microscopy.RESULTS: The negative conversion rate of pathogen in Funingkang group was 100%(6/6)one day and four days after drug withdrawal.After treatment with Funingkang,the mean thickness of vaginal mucosa was significantly increased compared with that of infected rats[(114.15±12.65)μm vs.(74.47± 28.90)μm,P=0.001],and the morphology of vaginal epithelium returned to normal.In addition,the expression of interleukin(IL)-4 and interferon(IFN)-γ kept unchanged,but IL-17(P=0.035)and non B-IgG(P=0.003)were up-regulated after treatment with Funingkang.CONCLUSION: Funingkang can effectively eliminate the pathogen of Candida albicans,repair vaginal mucosa,and regulate the innate immune response.

Establishment and application of loop-mediated isothermal amplification for Candida albicans / 临床检验杂志

Objective@#To establish a simple and rapid loop-mediated isothermal amplification (LAMP) to detect Candida albicans (CA) and explore its clinical value.@*Methods@#The Primer Explorer 5.0 software was used to design 4 primers for amplification of CA by LAMP. The system and conditions of LAMP reaction were optimized to evaluate its specificity and the minimum limit in the detection. The vaginal swabs were collected from 123 vulvovaginal candidiasis (VVC) patients and 42 healthy individuals. Fungal culture, LAMP test, PCR test and 1.79 mol/L KOH microscopy were conducted in parallel. Fungal culture was used as the reference method for VVC diagnosis. The positive rates between two groups were compared by Pearson χ² test. The consistency of the results from LAMP, PCR, microscopy and culture were analyzed by Kappa test and P＜0.05 was considered as statistically significant difference. ROC curve analysis was used to evaluate LAMP and PCR test for diagnosis of VVC.@*Results@#The optimum reaction temperature of LAMP was 61 ℃ with high specificity. No cross reaction with other strains was found. The minimum detection limit was 10³ copies/ul. The positive rates of LAMP, PCR and microscopy between VVC and healthy group showed statistically significant difference（LAMP: χ²=68.576；PCR: χ²=64.918；microscopy: χ²=50.076，P＜0.01）. LAMP detection and PCR showed good consistency (κ=0.744, 0.720), but microscopy examination showed poor consistency (κ=0.533). LAMP showed diagnostic sensitivity of 87.62%, specificity of 88.33%, positive predictive value of 92.93% and negative predictive value of 80.30%. The area under the curves of LAMP and PCR were 0.873 and 0.888, respectively. No difference in efficacies between LAMP and PCR was found (Z=0.849, P=0.395 6), but the lowest detection time of LAMP was shorter than 1 hour.@*Conclusion@#A rapid, reliable, sensitive and specific LAMP technique for detecting CA was established. The comprehensive screening performance should be superior to the routine method in laboratories, so LAMP could be used for the supplementary diagnosis and monitoring therapeutic effects in CA infection.

In vitro anti-Candida activity of Quercus infectoria gall extract-based vaginal cream and its local tissue effects in vivo

Aims@#Aqueous extract of Quercus infectoria (QI) galls has been reported to possess anti-fungal and anti-inflammatory activities. Hence, this study aimed to determine in vitro antimicrobial activity of formulated QI gall extract-based vaginal cream against Candida albicans and to evaluate the possible side effects on the cervicovaginal epithelium of healthy rats. @*Methodology and results@#Three different cream formulations containing 10%, 20%, and 30% of QI gall extract respectively were tested for their antimicrobial activity against C. albicans (ATCC 10231) by using disc diffusion test. Microbroth serial dilution method was performed in determining the minimum inhibitory concentration (MIC) and fungicidal concentration (MFC). The 30% formulated extract cream (FEC) was applied topically on the cervicovaginal surface of healthy Sprague Dawley (SD) rats and examined for local tissue effects histologically. The mean scores of inhibition zone diameter were compared by one-way ANOVA and post-hoc test using PRISM software. All extract cream formulations displayed a relatively good anti-Candida activity. The MIC values exhibited by 10%, 20%, and 30% FEC against C. albicans were 1.094 mg/mL, 0.547 mg/mL, and 0.068 mg/mL, respectively. The 10% and 20% FECs showed a significant difference (P=0.0254) in the mean of inhibition zone diameter. The lowest MFC value (0.068 mg/mL) was shown by 30% FEC. There were no abnormal changes seen at the vagina and cervical mucosa after 2 weeks application of 30% FEC. @*Conclusion, significance and impact of study@#QI gall extract formulated in the cream base has an anti-Candida activity in vitro and the present finding suggests that this herbal cream formulation is potentially useful in preventing vaginal candidiasis without causing any unwanted local side effects.

Changing virulence factors among vaginal non-albicans Candida species

Background: Vulvovaginal candidiasis (VVC) is caused by overgrowth of Candida species in the female lower genital tract and most commonly caused by Candida albicans. The production of various virulence factors may attribute to their pathogenicity. Hence, this study was aimed to determine the production of various virulence factors of Candida spp. causing VVC. Materials and Methods: A total of 51 Candida spp. were isolated prospectively from 50 patients among 211 clinically suspected cases of VVC. The haemolytic activity, biofilm production, proteinase activity, phospholipase activity and esterase activity were detected by standard methods. Statistical analysis was performed using OpenEpi version 3.01. Results: Haemolytic activity was observed in 42 Candida isolates (82.4%), biofilm activity in 21 Candida isolates (41.2%), proteinase and esterase activity in 19 Candida isolates (37.3%) each and phospholipase activity in 15 Candida isolates (29.4%). Phospholipase activity was observed in all of the C. albicans strains, whereas all strains of Candida krusei were able to produce biofilm. All strains of Candida parapsilosis and 87% strains of Candida glabrata were haemolytic. Five of the eight C. glabrata strains were found to produce strong proteinase (Prz score ?0.63). About 30.4% strains of C. glabrata and 20% strains of C. krusei were found to be positive for esterase activity. This is one of the few studies which revealed esterase activity among C. glabrata and C. krusei strains. Conclusions: This study highlighted that there is a change in the virulence factors among the non-albicans Candida species, especially C. glabrata strains which were haemolytic and produce strong proteinase activity and esterase activity. It may be one of the explanation of the most common causative agent of VVC in our study. Multicentric studies from this area might be required to get a more generalised conclusion.

Meningoencefalitis criptocóccica refractaria en una mujer joven

RESUMEN Mujer de 23 años presenta hace 2 meses cefalea que se acompaña de visión borrosa, disartria y pérdida de la conciencia. Por líquido cefalorraquídeo se confirma meningitis a criptococo. A pesar del tratamiento con anfotericina B y fluconazol presenta empeoramiento general. Durante su evolución presentó paracoccidioidomicosis pulmonar y candidiasis vaginal. A pesar de la terapia dirigida, la paciente fallece con la tardía confirmación de una inmunodeficiencia variable común.

ABSTRACT A 23-year-old woman has been presenting headache for two months that is accompanied by blurred vision, dysarthria and loss of consciousness. Cerebrospinal fluid confirms meningitis to Cryptococcus. In spite of the treatment with amphotericin B and fluconazole she presents a general deterioration. During her evolution she presented pulmonary paracoccidiomycosis and vaginal candidiasis. Despite specific therapy, the patient dies with the late confirmation of a common variable immunodeficiency.

Recurrent vaginal discharge: a myth or a fact? / Corrimento vaginal recorrente: mito ou fato?

Introduction: Vaginal discharge is a frequent gynecological complaint, and may represent a disease or not. A vaginal discharge is considered recurrent when it occurs four or more episodes per year. Among the aetiologies, physiological and infectious conditions are mentioned, being the infectious ones, particularly those caused by Candida spp. fungus, the most related to the symptom. Despite the diagnostic and therapeutic resources available, empirical clinical treatments and self-treatments are very frequent and related to ineffective therapeutic results, leading this population to question what the differences regarding women with no symptoms are. Objective: To identify sociodemographic, behavioral and microbiological differences between women with recurrent vaginal discharge and asymptomatic women. Methods: Cross-sectional study involving 126 women with recurrent discharge complaints (study group) and 155 (control group), totaling 281 evaluated women. The group included women in the menacme, sexually active, and those who fit in the criteria of recurrent vaginal discharge, without definite previous diagnosis, compared with asymptomatic women, who attended an annual routine examination. Pregnant, diabetic and immunosuppressed women were excluded. The study was based on the principle of the null hypothesis, when there are no differences between the two studied groups. Results: The average age was 29.95 years, predominantly single and without children. There was no significant difference in the analysis of relationship time with the current partner, numbers of partners throughout life, gender and contraceptive method. There was predominance of normal vaginal flora (type 1) in both groups, with average prevalence of 44.9%. The alkaline vaginal pH was predominant in the study group. Conclusion: The null hypothesis was confirmed. Biological, behavioral and sociodemographic differences in the studied populations were not identified. In women with recurrent discharge group, there were no infectious etiologic factors, suggesting that clinical diagnoses are not sufficient for the most efficient management of these situations, indicating laboratory evaluation for these cases in order to improve diagnostic accuracy

Introdução: O corrimento vaginal é queixa ginecológica frequente, podendo ou não representar doença. Conceitua-se como corrimento vaginal recorrente aquele que ocorre em quatro ou mais episódios ao ano. Entre as etiologias, citam-se condições fisiológicas e infecciosas, sendo as infecciosas, particularmente as causadas por fungo Candida spp., as mais relacionadas ao sintoma. Apesar dos recursos diagnósticos e terapêuticos disponíveis, tratamentos clínicos empíricos e autotratamentos são muito frequentes e associados a resultados terapêuticos pouco efetivos, levando essa população a questionamentos sobre quais diferenças elas teriam em relação a mulheres sem sintomas. Objetivo: Identificar diferenças sociodemográficas, comportamentais e microbiológicas entre mulheres com corrimento vaginal recorrente e mulheres assintomáticas. Métodos: Estudo transversal envolvendo 126 mulheres com queixa de corrimento recorrente (grupo de estudo) mais 155 controles, totalizando 281 mulheres avaliadas. Foram incluídas no grupo de estudo mulheres no menacme, sexualmente ativas e enquadradas nos critérios de corrimento vaginal recorrente, sem diagnóstico prévio definido, comparadas a mulheres assintomáticas, que compareciam a exame de rotina anual. Foram excluídas as gestantes, diabéticas e imunossuprimidas. Partiu-se de princípio da hipótese nula, em que não há diferenças entre os dois grupos estudados. Resultados: A média de idade foi de 29,95 anos, predominando solteiras e sem filhos. Não houve diferença significativa quando analisados: tempo de relacionamento com o atual parceiro, número de parceiros ao longo da vida, sexarca e método anticoncepcional. Houve predomínio da flora vaginal normal (tipo 1) em ambos os grupos, com prevalência média de 44,9%. O pH vaginal alcalino foi predominante no grupo de estudo. Conclusão: Confirmou-se a hipótese nula, não se identificando diferenças biológicas, comportamentais e sociodemográficas nas populações estudadas. Não se observaram, no grupo de mulheres com corrimento recorrente, fatores etiológicos infecciosos, sugerindo que diagnósticos clínicos não são suficientes para o manejo mais eficiente dessas situações, indicando-se avaliação laboratorial para esses casos com o objetivo de melhorar a acurácia diagnóstica.