Candida albicans vaginitis in a murine model is reduced by polypeptide-enriched Gastrodia elata extracts.

Aim: The primary objective of this study was to evaluate the effects of polypeptide-enriched Gastrodia elata extracts (GE) on vulvovaginal candidiasis (VVC). Materials & methods: A VVC model induced by Candida albicans (C. albicans) infection was successfully developed in BALB/c mice. After treatment, the colony-forming unit (CFU) of vaginal lavage was measured by plating. The extent of the inflammatory response was assessed by hematoxylin-eosin (H&E) staining and enzyme-linked immunosorbent assay (ELISA). Results: GE had an inhibitory effect on the proliferation of C. albicans and inflammatory reaction. Meanwhile, it had a potentially beneficial effect on the growth of Lactobacillus. Conclusion: These results showed the potential application of GE as an antifungal agent in VVC treatment.

Impaired Th17 cell proliferation and decreased pro-inflammatory cytokine production in CXCR3/CXCR4 double-deficient mice of vulvovaginal candidiasis.

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.

Mannose-Binding Lectin Gene Polymorphism and Its Association with Susceptibility to Recurrent Vulvovaginal Candidiasis.

Recurrent vulvovaginal candidiasis (RVVC) is a common illness influencing childbearing women worldwide. Most women suffering from RVVC develop infection without specified risk factors. Mannose-binding lectin (MBL) is an important component of innate immune defense against Candida infection. Innate immunity gene mutations and polymorphisms have been suggested to play a role in susceptibility to RVVC. This study aimed to investigate the association between MBL 2 gene exon 1 codon 54 polymorphism and susceptibility to RVVC in childbearing women. Whole blood and serum samples were obtained from 59 RVVC cases and 59 controls. MBL serum level was measured by enzyme-linked immune-sorbent assay (ELISA). MBL2 exon 1 codon 54 polymorphism was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). It was shown that MBL serum level was nonsignificantly different between RVVC cases and controls. The risk of RVVC was 3 times higher in those carrying MBL2 exon 1 codon 54 variant allele (B). It could be concluded that the carrying of MBL2 exon 1 codon 54 variant allele (B) was shown to be a risk factor for RVVC in childbearing women.

[Eosinophilia in peripheral blood of women with recurrent vaginal candidiasis]. / Eosinofilia no sangue periférico de mulheres com candidiase vaginal recorrente.

PURPOSE: To quantify the number of defense cells and immunoglobulin E (IgE) levels in peripheral blood sampled from women with recurrent vulvovaginal candidiasis. METHODS: A cross-sectional study was conducted on 60 women, 40 with vulvovaginal candidiasis and 20 controls. The defense cells were identified using an impedance system combined with flow cytometry and total and specific IgE was measured by chemiluminescence. The Mann-Whitney test was used for nominal variables and the Spearman test was used to determine the correlation of IgE concentration and eosinophils in peripheral blood. RESULTS: The number of eosinophils in peripheral blood from patients with recurrent vulvovaginal candidiasis, 302.60 (± 253.07), was significantly higher compared to control, 175.75 (± 109.24) (p=0.037). Serum levels of total and specific IgE were similar in the groups of women with and without recurrent vulvovaginal candidiasis (p=0.361). However, there was a moderate positive correlation between eosinophils and total serum IgE in the candidiasis group (r=0.25). CONCLUSION: Women with recurrent vulvovaginal candidiasis are more likely to have eosinophils in peripheral blood.

Eosinofilia no sangue periférico de mulheres com candidiase vaginal recorrente / Eosinophilia in peripheral blood of women with recurrent vaginal candidiasis

OBJETIVO: Quantificar o número de células de defesa e os níveis de imunoglobulina E (IgE) no sangue periférico em amostra de mulheres com candidíase vaginal recorrente. MÉTODOS: Estudo de corte transversal com 60 mulheres, 40 com candidíase vulvovaginal e 20 do grupo controle (sem doença). As células de defesa foram identificadas utilizando um sistema de impedância combinada com a citometria de fluxo, os níveis de IgE total e específica foram medidos por meio de técnicas de quimiluminescência, o teste de Mann-Whitney foi utilizado para variáveis nominais e do teste de Spearman para correlações das concentrações de IgE e de eosinófilos no sangue periférico. RESULTADOS: O número de eosinófilos no sangue periférico de pacientes com candidíase vulvovaginal, 302,60 (±253,07), foi significativamente maior do que o grupo controle, 175,75 (±109,24) (p=0,037). Os níveis séricos de IgE total e específica foram semelhantes em ambos os grupos de mulheres com e sem candidíase vulvovaginal recorrente (p=0,361). Entretanto, observou-se uma correlação positiva moderada entre eosinofilia e níveis de IgE total no sangue periférico de mulheres com candidíase vaginal recorrente (r=0,25). CONCLUSÃO: Mulheres com candidíase vaginal recorrente parecem ter maior concentração de eosinófilos no sangue periférico que as assintomáticas.

PURPOSE: To quantify the number of defense cells and immunoglobulin E (IgE) levels in peripheral blood sampled from women with recurrent vulvovaginal candidiasis. METHODS: A cross-sectional study was conducted on 60 women, 40 with vulvovaginal candidiasis and 20 controls. The defense cells were identified using an impedance system combined with flow cytometry and total and specific IgE was measured by chemiluminescence. The Mann-Whitney test was used for nominal variables and the Spearman test was used to determine the correlation of IgE concentration and eosinophils in peripheral blood. RESULTS: The number of eosinophils in peripheral blood from patients with recurrent vulvovaginal candidiasis, 302.60 (±253.07), was significantly higher compared to control, 175.75 (±109.24) (p=0.037). Serum levels of total and specific IgE were similar in the groups of women with and without recurrent vulvovaginal candidiasis (p=0.361). However, there was a moderate positive correlation between eosinophils and total serum IgE in the candidiasis group (r=0.25). CONCLUSION: Women with recurrent vulvovaginal candidiasis are more likely to have eosinophils in peripheral blood.

Frequency of vaginal candida colonization and relationship between metabolic parameters in children with type 1 diabetes mellitus.

OBJECTIVES: Patients with diabetes mellitus (DM) are at risk for vulvovaginal candidasis. The aim of this study was to determine the species-specific prevalence rate and risk of candidiasis in patients with type 1 DM. METHODS: Children aged between 8-16 years were included in the study. Clinical and laboratory features of diabetes and evidence of genital symptoms were recorded. Vaginal swabs were taken from patients and placed on Sabouraud's dextrose agar and incubated. Following fasting overnight for 12 hours venous blood samples were taken simultaneously for analyses of blood glucose, HbA1c, and lipid profile. A simple 1-time fluconazole treatment regimen was used to treat patients with vulvo-vaginal candidiasis. RESULTS: Candida species were isolated in 30 of 76 (39%) swabs of patients with type 1 DM. Subjects who had candida colonization and candidiasis were all acute. The predominant candida species isolated from patients with type 1 DM were C.albicans 50%, C. glabrata 36.6%, C.crusei 3.3%, C.spesies 6.6%, and C.dubliniensis 3.3%. 42 patients had symptoms.The prevalence of candidiasis in symptomatic patients was 59.2%. Subjects with vulvo-vaginal candidiasis had higher mean HbA1c when compared to those who had no such infection (P = .047). CONCLUSIONS: There seems to be a significant link between hyperglycemia and vulvo-vaginal candidiasis in patients with type 1 DM. Improving glucose control may reduce the risk of candidiasis and potentially symptomatic infection among children with diabetes. Because of high rate of colonization with candida species in diabetes, patients should undergo periodic screening for genital candidiasis.

Severe vulvovaginitis as a presenting problem of type 2 diabetes in adolescent girls: a case series.

This article describes the presentation of 4 adolescent girls who sought medical attention for severe vulvovaginitis and were subsequently found to have type 2 diabetes. Symptomatic vulvovaginitis is rare in adolescent girls, and its presence should alert health care providers to test for underlying hyperglycemia. These 4 girls represent 8.5% of the females with new-onset type 2 diabetes during a 3-year period (2007-2009). The 4 cases fulfilled the current Canadian Diabetes Association screening criteria for type 2 diabetes in youth, yet none of these girls had been screened. These cases highlight the need for better awareness of screening criteria for type 2 diabetes in adolescents. Consideration should be given in clinical practice guidelines to including the presence of unusual or severe infections as a risk factor for type 2 diabetes in youth.

Mannan-binding lectin in women with a history of recurrent vulvovaginal candidiasis.

OBJECTIVES: To determine the serum concentration of mannan-binding lectin (MBL), a component of the innate immune system, in women with a history of recurrent vulvovaginal candidiasis (RVVC) and to correlate the result to candida-cultures, contraceptive use, if any, and to different antifungal therapies. STUDY DESIGN: Twenty-nine women with a history of RVVC were investigated. Cultures of vulvar and vaginal samples were grown on chromogenic agar. Serum levels of MBL were determined by a sandwich time-resolved immunofluorometric assay, using anti-MBL coated microtiter wells containing samples, which were washed, incubated with biotinylated anti-MBL followed by europium-labeled streptavidin and measured by time-resolved flourometry. RESULTS: The median MBL level was higher in the RVVC cases than in 30 women with no history of genital candida infection who served as a comparison group (p=0.006). It was also higher in the candida-positive than in the culture-negative RVVC (p=0.02). The median concentration of MBL was also higher in hormonal contraceptive users as compared to condom-users and those using no contraceptive at all (p=0.03). CONCLUSION: The result indicates a role of MBL in RVVC and the production may correlate to vulvar/vaginal colonization by Candida, hormonal contraceptive use, and antifungal therapies.

MBL2 genetic screening in patients with recurrent vaginal infections.

PROBLEM: Mannose-binding lectin (MBL) is an important component of the innate immunity, present at the mucosal level in vagina: a common pathogen's entry point. METHOD OF STUDY: We used a rapid genotyping method based on melting temperature assay to search for three single nucleotide polymorphisms (SNPs) located in the first exon of the MBL2 gene and we also measured MBL serum levels in patients with recurrent bacterial vaginosis (rBV) and recurrent vulvovaginal candidiasis (rVVC). RESULTS: Detected frequencies of MBL2 SNPs were comparable to the ones already reported for the Italian population and no significant differences were found between rVVC, rBV and controls. MBL serum levels did not show significant differences between the studied groups. CONCLUSION: No correlation for the screened mutations has been found neither in protecting nor in favoring the infection in rVVC and rBV patients. Our data demonstrate a lack of association between functional polymorphisms in the first exon of MBL2 gene, MBL deficiency, VVC and rBV.

Clinical aspects and luteal phase assessment in patients with recurrent vulvovaginal candidiasis.

OBJECTIVE: This study was undertaken to characterize the patients with recurrent vulvovaginal candidiasis. STUDY DESIGN: Basic data of personal history and history of recurrent vulvovaginal candidiasis, lower genital tract symptoms and signs in 50 patients were analyzed in this longitudinal follow-up study including the determination of midluteal serum progesterone and urinary pregnanediol levels during the luteal phase in 84 cycles (recurrent vulvovaginal candidiasis) and 60 cycles (healthy controls). RESULTS: All patients suffered primary idiopathic form of recurrent vulvovaginal candidiasis. Frequently, there was a striking discrepancy between severe symptoms and clinical finding, which was often negligible or normal. There was no redness and no or minimum discharge in 52% of culture documented attacks. In contrast to the healthy controls, the patients had significantly lower levels of progesterone (p<0.01) as well as those of urinary pregnanediol (p<0.05). CONCLUSION: Culture positive attacks in patients with recurrent vulvovaginal candidiasis represented rather a form of vulvovaginal discomfort than attacks of vulvovaginal candidiasis with typical inflammatory changes. Significantly lower progesterone levels in the RVVC patients as compared to the healthy controls suggest a link between an altered hormonal status and one of possible causes of RVVC in these women.

Candida albicans HWP1 gene expression and host antibody responses in colonization and disease.

In vivo expression of the developmentally regulated Candida albicans hyphal wall protein 1 (HWP1) gene was analysed in human subjects who were culture positive for C. albicans and had oral symptoms (n=40) or were asymptomatic (n=29), or had vaginal symptoms (n=40) or were asymptomatic (n=29). HWP1 mRNA was present regardless of symptoms, implicating hyphal and possibly pseudohyphal forms in mucosal carriage as well as disease. As expected, in control subjects without oral symptoms (n=10) and without vaginal symptoms (n=10) who were culture negative in oral and vaginal samples, HWP1 mRNA was not detected. However, exposure to Hwp1 in healthy culture-negative controls, as well as in oral candidiasis and asymptomatic mucosal infections, was shown by the existence of local salivary and systemic adaptive antibody responses to Hwp1. The results are consistent with a role for Hwp1 in gastrointestinal colonization as well as in mucosal symptomatic and asymptomatic infections. Overall, Hwp1 and hyphal growth forms appear to be important factors in benign and invasive interactions of C. albicans with human hosts.

Single-dose fluconazole for vulvovaginal candidiasis: impact on prothrombin time in women taking warfarin.

OBJECTIVE: To estimate the effect of a single oral 150-mg dose of fluconazole on the prothrombin time of women on long-term warfarin therapy. METHODS: Women on warfarin therapy for 6 months or more with no change in dose within 4 weeks of the study, and a prothrombin time (PT) with an International Normalized Ratio (INR) between 2 and 3 were invited to participate. Two consecutive baseline PTs were obtained (days -1 and 0), and women were given 150 mg of fluconazole. Prothrombin times were measured on days 2, 5, and 8 of the study. The change in PT was calculated from the difference between the baseline PT on day 0 and the PT during the study period. To detect a 10% difference in a PT (approximately 2.1 seconds) of a patient with an INR of 2.0, at P < .05 and a power of 90%, 5 subjects are required. RESULTS: Six women participated. The mean (+/- standard deviation) PT for day 0 was 27.7 +/- 4.1 seconds or INR 2.6 +/- 0.4. The PT increased 11% at day 2, 34% at day 5, and 2% at day 8; these differences were not statistically significant. However, one half of the women had either a clinically relevant increase of the INR greater than 4, or bleeding that required their dosage of warfarin to be decreased. CONCLUSION: A single 150-mg oral dose of fluconazole may increase the PT to a clinically relevant level in a woman on chronic warfarin therapy. Clinicians should monitor the PT carefully after a single dose of fluconazole. LEVEL OF EVIDENCE: II-2.

The serum levels of calcium, magnesium, iron and zinc in patients with recurrent vulvovaginal candidosis during attack, remission and in healthy controls.

The real cause of recurrent vulvovaginal candidosis (RVVC) is concealed and the etiopathogenesis of this disease remains to be determined. In a cohort study, concentrations of metals in 44 patients with RVVC and 30 healthy age-matched women were measured and compared. The concentrations of serum calcium (Ca), magnesium (Mg) and iron (Fe) were measured photometrically, the zinc (Zn) levels were determined using flame atomic absorption spectrometry. For statistical analysis were used the Student's t-tests (paired analysis for attack vs. remission; non-paired analysis for patient vs. control). Although all measured metals were within normal ranges the patients with RVVC had in contrast to the healthy controls significantly lower levels of serum Ca, Mg and Zn and insignificantly higher levels of Fe. These relative changes may contribute to the development of attacks in patients with RVVC.

Glucose tolerance in pregnant women with vaginal candidiasis.

BACKGROUND: The use of traditional historic risk factors to identify gestational diabetes mellitus (GDM) will miss half of women with gestational diabetes mellitus. Our aim was to evaluate whether impaired glucose tolerance is a risk factor for vaginal candidiasis in pregnant women. PATIENTS AND METHODS: In a cross-sectional study, we compared the prevalence of impaired glucose tolerance in 64 pregnant women with vaginal candidiasis (positive microscopy) and 59 Candida-negative control subjects. Subjects underwent standardized 75-gram oral glucose tolerance testing between the 24th and 28th weeks of their pregnancies. Patients were included only if they had no known diabetes mellitus or historic risk factors for gestational diabetes mellitus, and had not been receiving antibiotic or steroid therapy. We compared glucose levels at fasting, 30 minutes, 60 minutes and 120 minutes, and perinatal and neonatal outcomes in the two groups. RESULTS: There were no statistical differences between cases and controls in demographic characteristics. Glucose concentrations were higher in pregnant women with vaginal candidiasis than in control subjects at fasting (89 vs. 84 mg/dL, P=0.021), 30 minutes (139 vs. 126 mg/dL, P=0.050), and 60 minutes (124 vs. 106 mg/dL, P= 0.018) after intake of 75 gram of glucose. The two groups did not differ in glucose level at 120 minutes after glucose administration. Gestational diabetes prevalence was 3.1% and 3.4% in the study and control group, respectively (P=0.274). CONCLUSION: The tolerance to glucose in pregnant women with vaginal candidiasis seems discretely impaired.

[Genital infections in girls with type 1 diabetes]. / Genitalni infektsii pri momicheta s tip 1 diabet.

The aim of the study was to investigate the relation between genital infections (frequency, etiology, clinical presentation) among girls with type 1 diabetes and some factors as age of the girls, duration and metabolic control of diabetes. Forty-three girls with type 1 diabetes and twelve healthy girls for controls were involved in the study. Methods used are: genital tract inspection, direct microscopy and cultures from genital discharge and urine. Glycaemic control in the diabetic girls was assessed by measuring total glycosylated haemoglobin. The results show vulvovaginal candidiasis in 27 (62.8%) in diabetic girls versus 2 (16.6%) in controls (p < 0.01). There was clear prevalence of non-albicans Candida species. Concomitant infection with bacterial pathogens were proved in only 6 (13.9%) of diabetic girls and 1 (8.5%) of the controls. The clinical presentation of genital candidiasis was vulvovaginitis and most of the affected diabetic girls (88.9%) were in puberty period. There was no relation between duration and metabolic control of diabetes and genital infections.

Partial protection against experimental vaginal candidiasis after mucosal vaccination with heat-killed Candida albicans and the mucosal adjuvant LT(R192G).

The effectiveness of a mucosal vaccine composed of heat-killed Candida albicans (HK-CA) or C. albicans culture filtrate (CaCF) in conjunction with the mucosal adjuvant LT(R192G) against vulvovaginal candidiasis was examined in an estrogen-dependent murine model. Mice vaccinated intranasally with HK-CA + LT(R192G) exhibited a significant but short-lived protection accompanied by a vigorous delayed-type hypersensitivity response as well as high titers of circulating C. albicans-specific antibodies. Surprisingly, the levels of antigen-specific antibodies in the vaginal secretions of protected mice were negligible and no correlates of vaginal-associated Type 1 or Type 2 cytokines were observed. Vaginal priming with C. albicans before vaccination did not alter the protective outcome. Immunization with CaCF + LT(R192G) induced a discrete level of protection when administered intrarectally but not intranasally. These results suggest that mucosal vaccination can afford partial protection against vulvovaginal candidiasis, but the precise immune mechanisms responsible for protection are complex and as yet, not well understood.

Prevalence and risk factors for vaginal Candida colonization in women with type 1 and type 2 diabetes.

BACKGROUND: Diabetes mellitus increases the rate of vaginal colonization and infection with Candida species METHODS: We surveyed women with diabetes receiving care at either an urban or suburban diabetes clinic to examine the relationship between vaginal Candida colonization, diabetes type and duration, and HbA1c level. 101 participants completed the self-administered questionnaire and self-collected a vaginal swab for Candida culture. Candida colonization was similar by age and race. RESULTS: Type 1 diabetics were three times as likely as type 2 diabetics to be colonized with any Candida species (OR = 3.4; 95% CI: 1.03, 11.41; p = 0.04); even after adjusting for abnormal HbA1c, which had an independent effect (OR = 1.4; 95% CI: 1.04, 1.76; p = 0.02). Recent antibiotic use (OR = 4.5; 95% CI: 1.18, 16.79; p = 0.03), lifetime history of chlamydia (OR = 5.8; 95% CI: 1.09, 30.54; p = 0.04), and performing oral sex during the past 2 weeks (OR = 4.9; 95% CI:0.84, 28.27; p = 0.08) were also associated with Candida carriage after adjusting for diabetic type and abnormal HbA1c. C. albicans was isolated from the majority of colonized type 1 participants (56%), while C. glabrata was the most common isolate among colonized type 2 participants (54%). CONCLUSIONS: Improving glucose control and possibly modifying sexual behavior may reduce risk of Candida colonization, and potentially symptomatic infection, among women with diabetes.

Cellular immunity in recurrent vulvovaginal candidiasis.

Impaired T cell function has been reported to predispose women to recurrent vulvovaginal candidiasis, but conflicting results have been noted in the literature. Most clinical episodes occur in the late luteal phase, suggesting hormonal influence on host resistance. The present study assesses the cellular immune responses of 28 women with recurrent vaginal candidiasis (patients) and 25 control women (controls), noting results in relation to whether the women were in the follicular or luteal phase of the menstrual cycle at the time of sampling. Candida-stimulated peripheral blood lymphocyte proliferation was significantly reduced in patients compared with controls. Interferon-gamma (IFN-gamma) production in response to both Candida and purified protein derivative (PPD) stimulation was significantly lower in patients compared with controls. Skin test responses were comparable in both groups. A significant reduction in Candida-stimulated IFN-gamma production was seen in patients but not controls in the follicular phase compared with those in the luteal phase. There was also a trend towards lower proliferation in response to Candida in patients but not controls in the follicular phase compared with patients in the luteal phase. These results suggest that there is a partial T cell dysregulation in recurrent vaginal candidiasis which may be exacerbated by the hormonal balance present during the follicular phase, correlating with the risk of clinical infection.