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1.
AAPS PharmSciTech ; 21(5): 185, 2020 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-32632542

RESUMO

The present study aimed to develop, characterize and evaluate the amphotericin B-loaded nanostructured lipid carriers (AmB-NLCs) for topical treatment of cutaneous leishmaniasis (CL) and vulvovaginal candidiasis (VVC). AmB-NLCs were characterized for particle size, zeta potential, encapsulation efficiency and surface morphology. Prepared NLCs were also characterized for in vitro drug release, ex vivo skin permeation and deposition before evaluating their in vitro and in vivo efficacy. Cytotoxicity of NLCs was assessed on MRC-5 cells, whereas skin irritation potential was evaluated in vivo using rats. Significant accumulation of drug in to the skin supported the topical application potential of drug-loaded NLCs. Encapsulation of AmB in NLCs resulted in enhanced in vitro potency against promastigotes and intracellular amastigotes of L. major JISH 118 (IC50 ± SEM = 0.02 ± 0.1 µM for both) compared with free drug (IC50 ± SEM = 0.15 ± 0.2 & 0.14 ± 0.0, respectively). Similar improved potency of AmB-NLCs was also observed for other Leishmania and fungal strains compared with drug solution. Topical application of AmB-NLCs on L. major-infected BALB/c mice caused a significant reduction in parasite burden per mg of lesion (65 × 108 ± 13) compared with the control group (> 167.8 × 108 ± 11). Topical AmB-NLCs gel demonstrated superior efficacy in the vaginal C. albicans rat model for VVC as compared with plain AmB gel. Moreover, results of in vitro cytotoxicity assay and in vivo skin irritation test confirmed AmB-NLCs to be non-toxic and safe for topical use. In conclusion, NLCs may have promising potential as carrier for topical treatment of various conditions of skin and mucosa.


Assuntos
Anfotericina B/administração & dosagem , Candidíase Vulvovaginal/tratamento farmacológico , Leishmaniose Cutânea/tratamento farmacológico , Nanoestruturas/administração & dosagem , Administração Tópica , Animais , Candidíase Vulvovaginal/metabolismo , Portadores de Fármacos/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Liberação Controlada de Fármacos , Feminino , Géis/metabolismo , Humanos , Lipídeos/administração & dosagem , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Tamanho da Partícula , Ratos , Pele/metabolismo , Absorção Cutânea
2.
Sci Rep ; 9(1): 14095, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31575935

RESUMO

We characterized the vaginal ecosystem during common infections of the female genital tract, as vulvovaginal candidiasis (VVC, n = 18) and Chlamydia trachomatis infection (CT, n = 20), recruiting healthy (HC, n = 21) and bacterial vaginosis-affected (BV, n = 20) women as references of eubiosis and dysbiosis. The profiles of the vaginal microbiome and metabolome were studied in 79 reproductive-aged women, by means of next generation sequencing and proton based-nuclear magnetic resonance spectroscopy. Lactobacillus genus was profoundly depleted in all the genital infections herein considered, and species-level analysis revealed that healthy vaginal microbiome was dominated by L. crispatus. In the shift from HC to CT, VVC, and BV, L. crispatus was progressively replaced by L. iners. CT infection and VVC, as well as BV condition, were mainly characterised by anaerobe genera, e.g. Gardnerella, Prevotella, Megasphaera, Roseburia and Atopobium. The changes in the bacterial communities occurring during the genital infections resulted in significant alterations in the vaginal metabolites composition, being the decrease of lactate a common marker of all the pathological conditions. In conclusion, according to the taxonomic and metabolomics analysis, we found that each of the four conditions is characterized by a peculiar vaginal microbiome/metabolome fingerprint.


Assuntos
Candidíase Vulvovaginal/microbiologia , Infecções por Chlamydia/microbiologia , Metaboloma , Microbiota , Vagina/microbiologia , Doenças Vaginais/microbiologia , Adulto , Candidíase Vulvovaginal/metabolismo , Estudos de Casos e Controles , Infecções por Chlamydia/metabolismo , Chlamydia trachomatis , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Espectroscopia de Ressonância Magnética , Microbiota/genética , Vagina/metabolismo , Doenças Vaginais/metabolismo , Adulto Jovem
3.
Sci Transl Med ; 11(496)2019 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-31189718

RESUMO

Candida vaginitis is a frequent clinical diagnosis with up to 8% of women experiencing recurrent vulvovaginal candidiasis (RVVC) globally. RVVC is characterized by at least three episodes per year. Most patients with RVVC lack known risk factors, suggesting a role for genetic risk factors in this condition. Through integration of genomic approaches and immunological studies in two independent cohorts of patients with RVVC and healthy individuals, we identified genes and cellular processes that contribute to the pathogenesis of RVVC, including cellular morphogenesis and metabolism, and cellular adhesion. We further identified SIGLEC15, a lectin expressed by various immune cells that binds sialic acid-containing structures, as a candidate gene involved in RVVC susceptibility. Candida stimulation induced SIGLEC15 expression in human peripheral blood mononuclear cells (PBMCs) and a polymorphism in the SIGLEC15 gene that was associated with RVVC in the patient cohorts led to an altered cytokine profile after PBMC stimulation. The same polymorphism led to an increase in IL1B and NLRP3 expression after Candida stimulation in HeLa cells in vitro. Last, Siglec15 expression was induced by Candida at the vaginal surface of mice, where in vivo silencing of Siglec15 led to an increase in the fungal burden. Siglec15 silencing was additionally accompanied by an increase in polymorphonuclear leukocytes during the course of infection. Identification of these pathways and cellular processes contributes to a better understanding of RVVC and may open new therapeutic avenues.


Assuntos
Candida albicans/patogenicidade , Genômica/métodos , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/microbiologia , Animais , Candidíase Vulvovaginal/genética , Candidíase Vulvovaginal/metabolismo , Citocinas/metabolismo , Feminino , Predisposição Genética para Doença/genética , Humanos , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo
4.
J Cell Physiol ; 234(8): 13894-13905, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30656691

RESUMO

Vulvovaginal candidiasis (VVC) is a common observed infection, affecting approximately 75% of women of reproductive age. Drug resistance represents a troublesome stumbling block associated with VVC therapy. Thus the aim of the present study was to provide information regarding the selection of potential drug targets for VVC. CXCR3-, CXCR4-, or CXCR/CXCR4 double-deficient mouse models of VVC were subsequently established, with changes to the load of Candida Albicans evaluated accordingly. The biological behaviors of the vaginal epithelial cells were characterized in response to the CXCR3-, CXCR4-, or CXCR3/CXCR4 double-knockout in vivo. Our initial observations revealed that in mice with VVC, CXCR3-, CXCR4-, or CXCR3 - CXCR4 double-knockout resulted in a decreased load of C. Albicans as well as reduced levels and proportion of Th17 cells. Proinflammatory cytokine production was found to be inhibited by CXCR3-, CXCR4-, or CXCR3/CXCR4 double-knockout whereby the mRNA and protein expressions CXCR3, CXCR4, IL-17, IL-6, and TNF-α exhibited decreased levels. CXCR3-, CXCR4-, or CXCR3/CXCR4 double-knockout appeared to function as positive proliferation factors, while playing a negative role in the processes of apoptosis and the cell cycle of vaginal epithelial cells. Taken together, the key findings of the study suggested that CXCR3/CXCR4 double-knockout could act to hinder the progression of VVC, highlighting its promise as a novel therapeutic target in the treatment of VVC. CXCR3 and CXCR4 genes may regulate Th17/IL-17 immune inflammatory pathways to participate in antifungal immunity.


Assuntos
Candidíase Vulvovaginal/imunologia , Candidíase Vulvovaginal/metabolismo , Citocinas/biossíntese , Mediadores da Inflamação/metabolismo , Receptores CXCR3/deficiência , Receptores CXCR4/deficiência , Células Th17/patologia , Animais , Apoptose , Candida albicans/fisiologia , Candidíase Vulvovaginal/sangue , Candidíase Vulvovaginal/microbiologia , Ciclo Celular , Proliferação de Células , Citocinas/sangue , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Células Epiteliais/patologia , Feminino , Camundongos Knockout , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores CXCR3/sangue , Receptores CXCR3/metabolismo , Receptores CXCR4/sangue , Receptores CXCR4/metabolismo , Vagina/microbiologia , Vagina/patologia
5.
J Diabetes Investig ; 10(2): 439-445, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30136398

RESUMO

AIMS/INTRODUCTION: The prevalence and risk of vaginal candidiasis before and after initiating sodium-glucose cotransporter 2 (SGLT2) inhibitors, although some clinical trials have been carried out, have not been adequately shown in real-world practice. We investigated the incidence of vaginal Candida colonization and symptomatic vaginitis, and the clinical risk factors including diabetic microvascular complications. MATERIALS AND METHODS: The participants were 114 women with type 2 diabetes who were free of vaginitis symptoms and started SGLT2 inhibitors. Vaginal candidiasis tests through self-administered swabs were carried out at baseline, 6 and 12 months. RESULTS: Before starting SGLT2 inhibitors, 17 participants (14.9%) had positive vaginal Candida colonization. Younger age and the presence of microangiopathy were significantly associated with the positive colonization in multivariate analysis. Among all participants, 23 (20.2%, 8 because of vaginitis and 15 for other reasons) discontinued SGLT2 inhibitors before reaching the 6-month test. Of 65 participants who were negative for Candida at baseline and received the 6-month test, 24 (36.9%) converted to a positive culture, and multivariate analysis showed older age as an independent risk for developing Candida colonization. There were 18 participants (15.8%) who developed symptomatic vaginitis, and they showed similar characteristics to the 24 participants. Most of those with negative cultures at 6 months showed negative results at 12 months and vice versa. CONCLUSIONS: The rates of developing positive colonization and symptomatic vaginitis after starting SGLT2 inhibitors appear to be higher in real-world practice than the rates of 31% and 5-10% in clinical trials, respectively. Risk factors of vaginal Candida colonization might be different before and after taking SGLT2 inhibitors.


Assuntos
Biomarcadores/análise , Candida/isolamento & purificação , Candidíase Vulvovaginal/epidemiologia , Diabetes Mellitus Tipo 2/tratamento farmacológico , Angiopatias Diabéticas/epidemiologia , Inibidores do Transportador 2 de Sódio-Glicose/efeitos adversos , Glicemia/análise , Candidíase Vulvovaginal/metabolismo , Candidíase Vulvovaginal/microbiologia , Diabetes Mellitus Tipo 2/metabolismo , Feminino , Seguimentos , Hemoglobina A Glicada/análise , Humanos , Incidência , Japão , Pessoa de Meia-Idade , Prognóstico , Fatores de Risco
6.
PLoS One ; 13(8): e0202401, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30133508

RESUMO

OBJECTIVE: To characterize the lipid profile in vaginal discharge of women with vulvovaginal candidiasis, cytolytic vaginosis, or no vaginal infection or dysbiosis. DESIGN: Cross-sectional study. SETTING: Genital Infections Ambulatory, Department of Tocogynecology, University of Campinas, Campinas, São Paulo-Brazil. SAMPLE: Twenty-four women were included in this study: eight with vulvovaginal candidiasis, eight with cytolytic vaginosis and eight with no vaginal infections or dysbiosis (control group). METHODS: The lipid profile in vaginal discharge of the different study groups was determined by liquid chromatography-mass spectrometry and further analyzed with MetaboAnalyst 3.0 platform. MAIN OUTCOME MEASURES: Vaginal lipids concentration and its correlation with vulvovaginal candidiasis and cytolytic vaginosis. RESULTS: PCA, PLS-DA and hierarchical clustering analyses indicated 38 potential lipid biomarkers for the different groups, correlating with oxidative stress, inflammation, apoptosis and integrity of the vaginal epithelial tissue. Among these, greater concentrations were found for Glycochenodeoxycholic acid-7-sulfate, O-adipoylcarnitine, 1-eicosyl-2-heptadecanoyl-glycero-3-phosphoserine, undecanoic acid, formyl dodecanoate and lipoic acid in the vulvovaginal candidiasis group; N-(tetradecanoyl)-sphinganine, DL-PPMP, 1-oleoyl-cyclic phosphatidic, palmitic acid and 5-aminopentanoic acid in the cytolytic vaginosis group; and 1-nonadecanoyl-glycero-3-phosphate, eicosadienoic acid, 1-stearoyl-cyclic-phosphatidic acid, 1-(9Z,12Z-heptadecadienoyl)-glycero-3-phosphate, formyl 9Z-tetradecenoate and 7Z,10Z-hexadecadienoic acid in the control group. CONCLUSIONS: Lipids related to oxidative stress and apoptosis were found in higher concentrations in women with vulvovaginal candidiasis and cytolytic vaginosis, while lipids related to epithelial tissue integrity were more pronounced in the control group. Furthermore, in women with cytolytic vaginosis, we observed higher concentrations of lipids related to bacterial overgrowth.


Assuntos
Apoptose , Candidíase Vulvovaginal , Metabolismo dos Lipídeos , Estresse Oxidativo , Vagina/metabolismo , Adolescente , Adulto , Candidíase Vulvovaginal/diagnóstico , Candidíase Vulvovaginal/metabolismo , Candidíase Vulvovaginal/patologia , Cromatografia Líquida , Estudos Transversais , Citodiagnóstico , Feminino , Humanos , Espectrometria de Massas , Projetos Piloto , Vagina/microbiologia
7.
J Ethnopharmacol ; 223: 51-62, 2018 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-29775695

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Pulsatilla decoction (PD) is a classical prescription in Traditional Chinese Medicine (TCM) and has been reported to have inhibitory effects on Candida albicans proliferation. STUDY AIM: To investigate the therapeutic effects of PD in the treatment of Vulvovaginal candidiasis (VVC) and elucidate the potential mechanism. MATERIALS AND METHODS: Female BALB/c mice (N = 90) were randomized to six treatment groups, including the Control group, Model group, three PD groups and Fluconazole group which served as a positive control (20 mg/kg weekly). The three PD groups (low dose group, medium dose group and high dose group) were given a daily intragastric gavage of PD at doses of 5, 10 and 20 g/kg, respectively. Five animals from each group were euthanized on Day 4, Day 7 and Day 14 after treatment. Colony forming unit (CFU) was measured by the serial dilution method. The degree of infection was assessed by Gram staining, Periodic acid schiff (PAS) staining, Hematoxylin and eosin (H&E) staining and Scanning electron microscopy (SEM). The serum inflammation levels were determined by a Luminex assay. Gene and protein expression levels of components of the Dectin-1 signaling pathway were determined by Real-time PCR, Western-blot and immunohistochemistry, respectively. RESULTS: The administration of PD significantly decreased the fungal load from Day 7 post-infection onwards and decreased the number of visible microorganisms based on findings from Gram staining, PAS staining and SEM. H&E staining indicated that the impaired histological profiles were improved in all three PD groups. PD led to a significantly lower level of IL-23 in the serum; the levels of IL-10 and TNF-α were also decreased, although the differences were not significant. Furthermore, a substantial downregulation of Dectin-1, CARD9 and NF-κB mRNA levels and Dectin-1, Syk, CARD9 and NF-κB protein levels was observed after the administration of PD. CONCLUSION: This study suggests that PD exerts inhibitory effects on C. albicans proliferation, adhesion and inflammation and simultaneously downregulates the expression levels of important genes and proteins associated with the Dectin-1 pathway, highlighting the potential application of PD to improve the clinical management of VVC.


Assuntos
Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candidíase Vulvovaginal/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Lectinas Tipo C/metabolismo , Animais , Candida albicans/efeitos dos fármacos , Candidíase Vulvovaginal/metabolismo , Candidíase Vulvovaginal/microbiologia , Candidíase Vulvovaginal/patologia , Proliferação de Células/efeitos dos fármacos , Citocinas/sangue , Feminino , Camundongos Endogâmicos BALB C , Fitoterapia , Transdução de Sinais , Vagina/efeitos dos fármacos , Vagina/microbiologia , Vagina/patologia
8.
J Biomed Nanotechnol ; 14(1): 215-226, 2018 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-29463379

RESUMO

The female genital system infections have high-incidence among various gynecological diseases. The role of endogenous peptides in female reproductive tract secretion has been rarely investigated regarding gynecological infections. To identify gynecological infections associated endogenous peptides, a comparative peptidomic profiling of vaginal secretion from sexually mature and perimenopause women was conducted in this study using nanoflow liquid chromatography-tandem mass spectrometer. A total of 3096 peptide fragments originating from 1364 precursor proteins were identified, 102 of which were discrepantly expressed, which included 60 over-expressed peptides and 42 under-expressed peptides in the sexually mature group compared with the perimenopause group. 6 differentially expressed bioactive peptides were identified to be related to immunodeficiency virus (HIV) infections, human papillomavirus (HPV) infections, bacterial vaginosis (BV) and vulvovaginal candidiasis (VVC). MUC4, MUC5A, MUC5B, MUC6, MUC17 and MUC19 peptides originating from mucin family were identified to enhance vaginal mucous immunity against infections. Moreover, with remarkable development of biomedical nanotechnology, the identified potential anti-infection peptides in our study will be useful in diagnosis and treatment of gynecological infections, especially when combined with nanoparticles. In conclusion, our findings will fill the gap on peptidomics for female genital system infections and pave the way for future studies about reproductive tract diseases management.


Assuntos
Anti-Infecciosos , Genitália Feminina/metabolismo , Nanotecnologia , Peptídeos , Adulto , Candidíase Vulvovaginal/metabolismo , Candidíase Vulvovaginal/terapia , Feminino , Doenças dos Genitais Femininos/metabolismo , Doenças dos Genitais Femininos/terapia , Humanos , Incidência
9.
AAPS PharmSciTech ; 19(3): 1297-1307, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29340981

RESUMO

This paper focuses on the development and evaluation of mucoadhesive vaginal gel of fluconazole using nanolipid carriers to enhance tissue deposition in treating vulvovaginal candidiasis. Treatment of vulvovaginal candidiasis includes antimycotic agents prescribed for 1 to 7 days or longer, in relapse either orally or topically. The delivery of fluconazole as nanolipid carriers in vaginal gel can be proposed as suitable alternative to the existing conventional formulations to improve the patient acceptability, compliance and localized drug action. The nanolipid carriers of fluconazole were prepared by phase inversion temperature technique and incorporated into Carbopol 974P as gelling polymer. GRAS excipients selected and optimized were Precirol ATO 5, oleic acid and Kolliphor RH 40 to produce nanolipid dispersions. Stable nanolipid dispersions were developed using sodium dodecyl sulfate as the charge inducer. The optimized nanolipid dispersion of fluconazole had particle size, polydispersity index and zeta potential value of 158.33 ± 2.55 nm, 0.278 ± 0.003 and - 27.33 ± 0.40 mV, respectively and the average entrapment of fluconazole in the lipid carriers was found to be 67.24 ± 0.87%. The optimized vaginal gel had satisfactory mucoadhesive strength and rheological properties to facilitate vaginal application. The fluconazole release from the gel was sustained showing 30.69 ± 1.02% drug deposition in the porcine vaginal mucosa at the end of 8 h with improved antifungal activity against Candida albicans during well diffusion studies. The optimized gel was non-irritant to the vaginal mucosa of female Wistar rats with no signs of erythema or edema.


Assuntos
Antifúngicos/administração & dosagem , Candida albicans/efeitos dos fármacos , Candidíase Vulvovaginal/tratamento farmacológico , Fluconazol/administração & dosagem , Nanopartículas/administração & dosagem , Animais , Antifúngicos/metabolismo , Candida albicans/metabolismo , Candidíase Vulvovaginal/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Feminino , Fluconazol/metabolismo , Géis , Humanos , Lipídeos , Membrana Mucosa/efeitos dos fármacos , Membrana Mucosa/metabolismo , Nanopartículas/metabolismo , Tamanho da Partícula , Ratos , Ratos Wistar , Suínos
10.
Infect Immun ; 86(2)2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-29109176

RESUMO

Unlike other forms of candidiasis, vulvovaginal candidiasis, caused primarily by the fungal pathogen Candida albicans, is a disease of immunocompetent and otherwise healthy women. Despite its prevalence, the fungal factors responsible for initiating symptomatic infection remain poorly understood. One of the hallmarks of vaginal candidiasis is the robust recruitment of neutrophils to the site of infection, which seemingly do not clear the fungus, but rather exacerbate disease symptomatology. Candidalysin, a newly discovered peptide toxin secreted by C. albicans hyphae during invasion, drives epithelial damage, immune activation, and phagocyte attraction. Therefore, we hypothesized that Candidalysin is crucial for vulvovaginal candidiasis immunopathology. Anti-Candida immune responses are anatomical-site specific, as effective gastrointestinal, oral, and vaginal immunities are uniquely compartmentalized. Thus, we aimed to identify the immunopathologic role of Candidalysin and downstream signaling events at the vaginal mucosa. Microarray analysis of C. albicans-infected human vaginal epithelium in vitro revealed signaling pathways involved in epithelial damage responses, barrier repair, and leukocyte activation. Moreover, treatment of A431 vaginal epithelial cells with Candidalysin induced dose-dependent proinflammatory cytokine responses (including interleukin 1α [IL-1α], IL-1ß, and IL-8), damage, and activation of c-Fos and mitogen-activated protein kinase (MAPK) signaling, consistent with fungal challenge. Mice intravaginally challenged with C. albicans strains deficient in Candidalysin exhibited no differences in colonization compared to isogenic controls. However, significant decreases in neutrophil recruitment, damage, and proinflammatory cytokine expression were observed with these strains. Our findings demonstrate that Candidalysin is a key hypha-associated virulence determinant responsible for the immunopathogenesis of C. albicans vaginitis.


Assuntos
Candida albicans/patogenicidade , Células Epiteliais/microbiologia , Proteínas Fúngicas/metabolismo , Membrana Mucosa/microbiologia , Animais , Candidíase Vulvovaginal/imunologia , Candidíase Vulvovaginal/metabolismo , Citocinas/metabolismo , Células Epiteliais/metabolismo , Feminino , Proteínas Fúngicas/farmacologia , Humanos , Camundongos , Membrana Mucosa/patologia , Infiltração de Neutrófilos/imunologia , Transdução de Sinais , Vagina/imunologia , Vagina/metabolismo , Vagina/microbiologia , Fatores de Virulência
11.
Sci Rep ; 7(1): 17877, 2017 12 19.
Artigo em Inglês | MEDLINE | ID: mdl-29259175

RESUMO

The expression of host inflammatory and Candida albicans putative virulence factors was studied in women with vulvovaginal candidiasis (VVC; twenty) or colonized by the fungus but asymptomatic (carriers; fifteen) or non-colonized asymptomatic (ten subjects). Overexpression of genes encoding NLRP3 and caspase-1 inflammasome components sharply differentiated VVC patients from asymptomatic colonized or non-colonized women. Inflammasome expression was coupled with neutrophils recruitment in the vagina of VVC women and IL-1ß and IL-8 production. Both cytokines were present, though to a lower concentration, also in the vaginal fluid of colonized and non-colonized women. Secretory aspartyl proteinases (SAPs) and hyphae associated genes HWP1 and ECE1 were upregulated in VVC but with some differences among infected women. The most overexpressed SAP gene was SAP2, that correlated with neutrophils accumulation. Our data provide clinical evidence that the intracytoplasmic activation of NLRP3 inflammasome complex plays a critical, pathogenesis-relevant role in human VVC.


Assuntos
Candida albicans/metabolismo , Candidíase Vulvovaginal/metabolismo , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Adulto , Candidíase Vulvovaginal/microbiologia , Citocinas/metabolismo , Feminino , Proteínas Fúngicas/metabolismo , Humanos , Hifas/metabolismo , Interleucina-1beta/metabolismo , Pessoa de Meia-Idade , Infiltração de Neutrófilos/fisiologia , Neutrófilos/metabolismo , Vagina/microbiologia , Fatores de Virulência/metabolismo , Adulto Jovem
12.
Acta Obstet Gynecol Scand ; 96(3): 295-301, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28052314

RESUMO

INTRODUCTION: Recurrent vulvovaginal candidiasis is defined as having three to four episodes per year and causes substantial suffering. Little is known about the mechanisms leading to relapses in otherwise healthy women. Nitric oxide is part of the nonspecific host defense and is increased during inflammation. Nitric oxide levels were measured and the expression of inducible nitric oxide synthase was analyzed in the vagina during an acute episode of recurrent vulvovaginal candidiasis and after treatment with fluconazole. MATERIAL AND METHODS: Twenty-eight women with symptoms of recurrent vulvovaginal candidiasis were enrolled together with 31 healthy controls. Nitric oxide was measured with an air-filled 25-mL silicon catheter balloon incubated in the vagina for five minutes and then analyzed by chemiluminescence technique. Vaginal biopsies were analyzed for the expression of inducible nitric oxide synthase. Symptoms and clinical findings were surveyed using a scoring system. The measurements and biopsies were repeated in patients after six weeks of fluconazole treatment. RESULTS: Nitric oxide levels were increased during acute infection (median 352 ppb) compared with controls (median 6 ppb), p < 0.0001. The levels decreased after treatment (median 18 ppb) but were still higher than in controls. Increased expression of inducible nitric oxide synthase was observed in the epithelial basal layer in patients before and after treatment compared with controls. Before treatment, there were positive correlations between nitric oxide and symptom (rs  = 0.644) and examination scores (rs  = 0.677), p < 0.001. CONCLUSIONS: Nitric oxide is significantly elevated in patients with recurrent vulvovaginal candidiasis during acute episodes of infection and decreases after antifungal treatment. The results illustrate the pronounced inflammatory response in recurrent vulvovaginal candidiasis correlating to symptoms of pain and discomfort.


Assuntos
Candidíase Vulvovaginal/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico/metabolismo , Adolescente , Adulto , Antifúngicos/administração & dosagem , Antifúngicos/uso terapêutico , Candidíase Vulvovaginal/tratamento farmacológico , Estudos de Casos e Controles , Feminino , Fluconazol/administração & dosagem , Fluconazol/uso terapêutico , Humanos , Recidiva , Inquéritos e Questionários , Adulto Jovem
13.
Clin Exp Obstet Gynecol ; 44(1): 7-10, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29714856

RESUMO

Purpose of the investigation: Vulvovaginal candidosis (VVC) is a common vaginal infection affecting almost 75% of all women once per lifetime. Vaginal associated immunity is important in the protection against VVC. The purpose of this study was to evaluate a potential role of IL-23, IFN-α, and IFN-ß in the local immune response against VVC. MATERIALS AND METHODS: The study included 202 non-pregnant women; 71 patients with clinical symptoms of VVC and 131 asymptomatic patients served as control. IL-23, IFN-α, and IFN-ß were measured in the vaginal fluid by ELISA. Microbiological cultures were used for Candida detection. RESULTS: C. albicans was detected in 67.6% of patients, C. glabrata in 2 1.1% of patients, and 5.6% were infected with C. krusei or coinfected with C. albicans and C. krusei. Levels of IL-23 (p < 0.001) and IFN-ß (p < 0.017) were significantly lower in the VVC group. IFN-α was elevated in the VVC group compared to the asymptomatic patients (p < 0.001). CONCLUSION: IL-23 and IEFN-ß seem to play a protective role against VVC. Decreased levels in VVC patients suggest a compromised local immune response at the time of occurrence of symptoms. In contrast, IFN-α seems to be released once the infection has occurred. These cytokines may be prospective targets in the treatment and prevention of primary and recurrent vaginal infections with Candida species.


Assuntos
Candidíase Vulvovaginal/metabolismo , Muco do Colo Uterino/metabolismo , Interferon-alfa/metabolismo , Interferon beta/metabolismo , Interleucina-23/metabolismo , Candida albicans/isolamento & purificação , Candida glabrata/isolamento & purificação , Estudos de Casos e Controles , Feminino , Humanos
14.
J Mycol Med ; 26(3): 255-60, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27544321

RESUMO

OBJECTIVE OF THE STUDY: This study aims to characterize phospholipase and proteinase activities of Candida isolates from 82 vulvovaginal candidiasis (VVC) and to study the relationship of these activities with vulvovaginitis. METHODS: Totally 82 Candida isolates from vagina samples of VVC patients were randomly collected over the period between September and December 2014 from hospitalized patients at the general hospitals of Lorestan province, Iran. Isolates were previously identified by conventional mycological methods. The phospholipase and proteinase activities were evaluated by Egg yolk agar, Tween 80 opacity medium and agar plate methods. RESULTS: The most common Candida species was identified Candida albicans (n=34, 41.5%), followed by Candida famata (n=13, 15.8%), Candida tropicalis (n=11, 13.4%), and Candida parapsilosis (n=9, 11%). The most phospholipase activity was observed in Candida colliculosa (40%), followed by C. famata (38.5%), and Candida krusei (33.3%). The findings revealed that the correlation between phospholipase production by Candida spp. and the presence of VVC was not found to be statistically significant (P=0.91). All Candida spp. exhibited considerable proteinase activity; so that 100% of C. colliculosa, C. parapsilosis, Candida kefyr, and Candida intermedia isolates produced high proteinase activity with Pz 4+ scores. There was a significant correlation between proteinase production by Candida spp. and the presence of VVC (P=0.009). CONCLUSION: The obtained findings revealed that Candida spp. isolates may produce both virulence factors, phospholipase and proteinase. Although the phospholipase production was only observed in <40% of the isolates; however there was a significant association between proteinase production by Candida spp. and VVC.


Assuntos
Candida/enzimologia , Candida/isolamento & purificação , Candidíase Vulvovaginal/microbiologia , Peptídeo Hidrolases/metabolismo , Fosfolipases/metabolismo , Candida albicans/enzimologia , Candida albicans/isolamento & purificação , Candidíase Vulvovaginal/metabolismo , Feminino , Humanos , Irã (Geográfico)
15.
Zhonghua Fu Chan Ke Za Zhi ; 51(7): 530-4, 2016 Jul 25.
Artigo em Chinês | MEDLINE | ID: mdl-27465873

RESUMO

OBJECTIVE: To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. METHODS: (1)Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis(VVC)murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group(n=15)and control group(n=15). Suspension(30 µl, 100 µg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline(PBS)was given to the control group.(2)Tweenty-four hours after treatment, the fungal burden and colony-forming unit(CFU)of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. RESULTS: (1)VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group.(2)Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×10(4) CFU/ml]than that in the control group [(8.5±2.1)×10(4) CFU/ml, P=0.017]. IFN-γ level of the test group was increased [(257±11)vs(197±4)pg/ml, P=0.000], while the level of IL-10 was reduced [(287 ± 15)vs(379 ± 17)pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group(0.892±0.008 vs 0.496±0.013, P=0.000). CONCLUSION: Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γ and IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.


Assuntos
Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Candidíase Vulvovaginal/tratamento farmacológico , Catelicidinas/genética , Interleucina-10/genética , Animais , Anti-Infecciosos/farmacologia , Candidíase Vulvovaginal/metabolismo , Catelicidinas/metabolismo , Catelicidinas/farmacologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interferon gama , Interleucina-10/metabolismo , Camundongos , Células Procarióticas , Distribuição Aleatória , Proteínas Recombinantes
16.
Eur J Clin Microbiol Infect Dis ; 35(5): 797-801, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26951262

RESUMO

Vaginal infections with Candida spp. frequently occur in women of childbearing age. A small proportion of these women experience recurrent vulvovaginal candidosis (RVVC), which is characterized by at least three episodes of infection in one year. In addition to known risk factors such as antibiotics, diabetes, or pregnancy, host genetic variation and inflammatory pathways such as the IL-1/Th17 axis have been reported to play a substantial role in the pathogenesis of RVVC. In this study, we assessed a variable number tandem repeat (VNTR) polymorphism in the NLRP3 gene that encodes a component of the inflammasome, processing the proinflammatory cytokines IL-1ß and IL-18. A total of 270 RVVC patients and 583 healthy controls were analyzed, and increased diseases susceptibility was associated with the presence of the 12/9 genotype. Furthermore, functional studies demonstrate that IL-1ß production at the vaginal surface is higher in RVVC patients bearing the 12/9 genotype compared to controls, whereas IL-1Ra levels were decreased and IL-18 levels remained unchanged. These findings suggest that IL-1ß-mediated hyperinflammation conveyed by the NLRP3 gene plays a causal role in the pathogenesis of RVVC and may identify this pathway as a potential therapeutic target in the disease.


Assuntos
Candidíase Vulvovaginal/genética , Candidíase Vulvovaginal/microbiologia , Estudos de Associação Genética , Predisposição Genética para Doença , Repetições Minissatélites , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Alelos , Candidíase Vulvovaginal/metabolismo , Estudos de Casos e Controles , Citocinas/metabolismo , Feminino , Genótipo , Humanos , Íntrons
17.
Peptides ; 71: 211-21, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26238597

RESUMO

Vulvovaginal candidiasis (VVC) is a frequent gynecological condition caused by Candida albicans and a few non-albicans Candida spp. It has a significant impact on the quality of life of the affected women also due to a considerable incidence of recurrent infections that are difficult to treat. The formation of fungal biofilm may contribute to the problematic management of recurrent VVC due to the intrinsic resistance of sessile cells to the currently available antifungals. Thus, alternative approaches for the prevention and control of biofilm-related infections are urgently needed. In this regard, the cationic antimicrobial peptides (AMPs) of the innate immunity are potential candidates for the development of novel antimicrobials as many of them display activity against biofilm formed by various microbial species. In the present study, we investigated the in vitro antifungal activities of the cathelicidin peptides LL-37 and BMAP-28 against pathogenic Candida spp. also including C. albicans, isolated from vaginal infections, and against C. albicans SC5314 as a reference strain. The antimicrobial activity was evaluated against planktonic and biofilm-grown Candida cells by using microdilution susceptibility and XTT [2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide] reduction assays and, in the case of established biofilms, also by CFU enumeration and fluorescence microscopy. BMAP-28 was effective against planktonically grown yeasts in standard medium (MIC range, 2-32µM), and against isolates of C. albicans and Candida krusei in synthetic vaginal simulated fluid (MIC range 8-32µM, depending on the pH of the medium). Established 48-h old biofilms formed by C. albicans SC5314 and C. albicans and C. krusei isolates were 70-90% inhibited within 24h incubation with 16µM BMAP-28. As shown by propidium dye uptake and CFU enumeration, BMAP-28 at 32µM killed sessile C. albicans SC5314 by membrane permeabilization with a faster killing kinetics compared to 32µM miconazole (80-85% reduced biofilm viability in 90min vs 48h). In addition, BMAP-28 at 16µM prevented Candida biofilm formation on polystyrene and medical grade silicone surfaces by causing a >90% reduction in the viability of planktonic cells in 30min. LL-37 was overall less effective than BMAP-28 against planktonic Candida spp. (MIC range 4-≥64µM), and was ineffective against established Candida biofilms. However, LL-37 at 64µM prevented Candida biofilm development by inhibiting cell adhesion to polystyrene and silicone surfaces. Finally, Candida adhesion was strongly inhibited when silicone was pre-coated with a layer of BMAP-28 or LL-37, encouraging further studies for the development of peptide-based antimicrobial coatings.


Assuntos
Antifúngicos , Peptídeos Catiônicos Antimicrobianos , Biofilmes/efeitos dos fármacos , Candida albicans/fisiologia , Candidíase Vulvovaginal/tratamento farmacológico , Antifúngicos/química , Antifúngicos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Candida albicans/isolamento & purificação , Candidíase Vulvovaginal/metabolismo , Feminino , Humanos
18.
Reprod Sci ; 22(8): 964-8, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25670719

RESUMO

OBJECTIVE: Neutrophil gelatinase-associated lipocalin (NGAL) is a component of innate immunity that prevents iron uptake by microorganisms. We evaluated whether NGAL was present in vaginal fluid and whether concentrations were altered in women with bacterial vaginosis (BV) or vulvovaginal candidiasis (VVC). METHODS: Vaginal secretions from 52 women with VVC, 43 with BV, and 77 healthy controls were assayed by enzyme-linked immunosorbent assay for NGAL and for concentrations of L-lactic acid. RESULTS: The median concentration of NGAL in vaginal fluid was significantly higher in control women (561 pg/mL) than in women with BV (402 pg/mL; P = .0116) and lower in women with VVC (741 pg/mL; P = .0017). Median lactic acid levels were similar in controls (0.11 mmol/L) and women with VVC (0.13 mmol/L) and were lower in women with BV (0.02 mmol/L; P < .0001). The NGAL and lactic acid concentrations were highly correlated (P < .0001). CONCLUSION: A decrease in Lactobacilli and/or lactic acid plus the absence of leukocytes results in lower vaginal NGAL levels that might facilitate the growth of bacteria associated with BV.


Assuntos
Proteínas da Fase Aguda/análise , Líquidos Corporais/química , Candidíase Vulvovaginal/metabolismo , Lipocalinas/análise , Proteínas Proto-Oncogênicas/análise , Vagina/metabolismo , Vaginose Bacteriana/metabolismo , Adulto , Biomarcadores/análise , Líquidos Corporais/metabolismo , Líquidos Corporais/microbiologia , Candidíase Vulvovaginal/diagnóstico , Candidíase Vulvovaginal/imunologia , Candidíase Vulvovaginal/microbiologia , Estudos de Casos e Controles , Regulação para Baixo , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunidade Inata , Ácido Láctico/análise , Lipocalina-2 , Vagina/imunologia , Vagina/microbiologia , Vaginose Bacteriana/diagnóstico , Vaginose Bacteriana/imunologia , Vaginose Bacteriana/microbiologia , Adulto Jovem
19.
BJOG ; 122(12): 1580-5, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25196575

RESUMO

OBJECTIVE: Do metabolites in vaginal samples vary between women with different vaginal disorders. DESIGN: Cross-sectional study. SETTING: Campinas, Brazil. SAMPLE: Seventy-seven women (39.9%) with no vaginal disorder, 52 women (26.9%) with vulvovaginal candidiasis (VVC), 43 women (22.3%) with bacterial vaginosis (BV), and 21 women (10.9%) with cytolytic vaginosis (CTV). METHOD: Concentrations of D- and L-lactic acid, extracellular matrix metalloproteinase inducer (EMMPRIN), and matrix metalloproteinase-8 (MMP-8), and the influence of Candida albicans on EMMPRIN production by cultured vaginal epithelial cells, were determined by enzyme-linked immunosorbent assay (ELISA). Associations were determined by the Mann-Whitney U-test and by Spearman's rank correlation test. MAIN OUTCOME MEASURES: Metabolite levels and their correlation with diagnoses. RESULTS: Vaginal concentrations of D- and L-lactic acid were reduced from control levels in BV (P < 0.0001); L-lactic acid levels were elevated in CTV (P = 0.0116). EMMPRIN and MMP-8 concentrations were elevated in VVC (P < 0.0001). EMMPRIN and L-lactic acid concentrations (P ≤ 0.008), but not EMMPRIN and D-lactic acid, were correlated in all groups. EMMPRIN also increased in proportion with the ratio of L- to D-lactic acid in controls and in women with BV (P ≤ 0.009). Concentrations of EMMPRIN and MMP-8 were correlated in controls and women with VVC (P ≤ 0.0002). Candida albicans induced EMMPRIN release from vaginal epithelial cells. CONCLUSIONS: Vaginal secretions from women with BV are deficient in D- and L-lactic acid, women with VVC have elevated EMMPRIN and MMP-8 levels, and women with CTV have elevated L-lactic acid levels. These deviations may contribute to the clinical signs, symptoms, and sequelae that are characteristic of these disorders.


Assuntos
Basigina/metabolismo , Candidíase Vulvovaginal/metabolismo , Ácido Láctico/metabolismo , Metaloproteinase 8 da Matriz/metabolismo , Vagina/microbiologia , Vaginose Bacteriana/metabolismo , Adulto , Líquidos Corporais/metabolismo , Brasil , Candidíase Vulvovaginal/microbiologia , Estudos Transversais , Ensaio de Imunoadsorção Enzimática , Células Epiteliais , Feminino , Humanos , Vaginose Bacteriana/microbiologia
20.
Acta Pharm ; 63(3): 359-72, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24152896

RESUMO

The aim of the present investigation was to prepare and evaluate novel bioadhesive vaginal tablets containing clotrimazole loaded microspheres in order to provide long-term therapeutic activity at the site of infection. Tablets were prepared by incorporating drug loaded microspheres and using bioadhesive polymers hydroxypropylmethylcellulose, sodium carboxymethylcellulose and Carbopol. Microspheres were prepared by the spray drying technique using Eudragit RS-100 and Eudragit RL-100. Microspheres were characterized by SEM, DSC, FTIR, particle size analysis and evaluated for percentage yield, drug loading, encapsulation efficiency and in vitro drug release. To achieve bioadhesion to the mucosal tissue, optimized microspheres were incorporated into bioadhesive tablets and were evaluated for in vitro drug release, in vitro and in vivo mucoadhesion. FTIR and DSC studies showed that no chemical interaction occurred between the drug and polymers. The sphericity factor indicated that the prepared microspheres were spherical. Formulation Mt6 indicated a controlled in vitro drug release and good bioadhesive strength. The in vivo images confirmed the bioadhesion and retention property of tablets up to 24 h. The results indicated that this drug delivery system can be explored for controlled intravaginal drug release.


Assuntos
Antifúngicos/química , Candidíase Vulvovaginal/tratamento farmacológico , Clotrimazol/química , Portadores de Fármacos/química , Microesferas , Adesivos Teciduais/química , Resinas Acrílicas/administração & dosagem , Resinas Acrílicas/química , Resinas Acrílicas/metabolismo , Administração Intravaginal , Animais , Antifúngicos/administração & dosagem , Antifúngicos/metabolismo , Candidíase Vulvovaginal/metabolismo , Clotrimazol/administração & dosagem , Clotrimazol/metabolismo , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/metabolismo , Sistemas de Liberação de Medicamentos/métodos , Feminino , Coelhos , Ovinos , Adesivos Teciduais/administração & dosagem , Adesivos Teciduais/metabolismo , Resultado do Tratamento , Cremes, Espumas e Géis Vaginais
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