Allergic vulvovaginitis: a systematic literature review.

PURPOSE: Despite the vaginal mucosa is able to respond to allergenic stimuli, vaginal allergic responses have been under investigated in clinical practice. Thus, we aimed to identify the most frequent etiological agents responsible for vulvovaginal allergies, the prevalent signs/symptoms, and the diagnostic tests applied in this clinical condition. METHODS: Literature search was performed on PubMed, Scopus, Scielo, Web of Science, and EMBASE. The study protocol was registered on PROSPERO (CRD42020167238). Studies were divided in two groups depending on allergen exposure route. Due to a significant number of studies correlating allergy to Candida infection, subgroup analysis was included. RESULTS: In direct exposure cases, Human Seminal Plasma was the most prevalent allergen, sensitizing 73% of affected women. These women presented localized swelling and burning as prevalent symptoms, affecting 42/68 and 36/68 women, respectively. Cutaneous Prick tests were applied in 58/68 women, either alone or combined with IgE measurements. Regarding cases of indirect/unidentified exposure, house dust mites was the most prevalent allergen (54%), followed by pollen (44%). Predominant symptoms were vulvar pruritus and burning, affecting 67/98 and 52/98 women. Skin prick test was the most prevalent diagnostic method used among different studies. Hypersensitivity toward Candida antigen was present in only half (163/323) of women presenting concomitant allergy and Candida infection. CONCLUSION: From the two types of allergen exposure that can cause vulvovaginal allergic responses, direct contact of the antigen with the vulva and/or vagina was the most prevalent. Still, allergens can also sensitize the vaginal mucosa secondarily to other exposure route, specifically aeroallergens.

Development of a new multiplex PCR to detect prevalent species of house dust mites in house dust.

Dermatophagoides pteronyssinus and Dermatophagoides farinae are the most common House Dust Mite (HDM) species in home environments worldwide and responsible for HDM allergy. Since the prevalence of HDM-related clinical conditions is linked to exposure to the mite itself, the detection of HDM in the human households gains importance. We aimed to develop a fast and accessible multiplex PCR to detect and distinguish two relevant HDM species in house dust. New primers were designed, and sensitivity analysis was performed. Sequencing of PCR products was also performed to confirm the method's specificity. The limit of detection of the multiplex PCR for both species was as low as 30 pg µL-1. The application of the multiplex PCR to dust samples also resulted in the identification of both species with high sensitivity. The protocol required small amount of template, reagents and a short reaction time thus presenting an alternative to classically used techniques for HDM identification.

Evaluation of overtime phenotypic variation of yeasts in chronic vulvovaginal candidosis cases.

Chronic vulvovaginal candidosis results either from reinfection or from the ability of Candida spp. to persist in the vulva and/or vagina. Persistence is usually associated with increased antifungal (mainly azoles) resistance rates, which can explain treatment failure, and/or increased expression of virulence factors by Candida spp. The aim of this study was to assess the mechanisms leading to Candida spp persistence, by studying sequential isolates from women with chronic vulvovaginal candidosis, focusing on strains genotypes, azole resistance, and ability to form biofilms along the period of clinical evaluation. The strains were identified at species level by automated analysis of biochemical profiles and molecular typing evaluated by polymorphic DNA analysis. The capacity to form biofilm was assessed with a microtiter plate assay. Fluconazole susceptibility was determined by the microdilution broth assay at both pH 7 (following the recommended guideline) and pH 4.5 (as representative of vaginal pH). We studied samples from 17 clinically recurrent cases. In 53% of the chronic cases there were two or more isolates that had a phylogenetic relationship while the remaining (47%) were caused by different species. In those cases where related strains were involved in recurrence, we verified an increase in MIC at pH 7 and also an increased capacity to form biofilms over time. Significant correlation between these two parameters was observed only in cases caused by C. glabrata, evidencing the importance of these two factors to enhance persistence in the vaginal mucosa for this particular species.

Chronic vulvovaginal candidosis (VVC) affects millions of women worldwide. We found that persistence of the same Candida strain on the vaginal mucosa does not account for the great majority of VVC cases. Moreover, modulation of biofilm formation and azole resistance overtime was investigated.

Virulence Factors as Promoters of Chronic Vulvovaginal Candidosis: A Review.

INTRODUCTION: The vast majority of the species of the genus Candida spp. is commensal in humans; however, some are opportunistic pathogens that can cause infection, called candidosis. Among the different types of candidosis, we highlight the vulvovaginal (VVC) which can occur in two main clinical variants: chronic (cVVC) and episodic or sporadic. The incidence of cVVC has been worrying the scientific community, promoting the research on genotypic and phenotypic causes of its occurrence. We summarize important findings on factors that favor chronic vulvovaginal candidosis with respect to molecular epidemiology and the expression of various virulence factors, while clarifying the terminology involving these infections. AIM AND METHODOLOGY: The aim of this review was to gather research that linked virulence factors to VVC and its persistence and recurrence, using two databases (Pubmed and Google Scholar). Predisposing factors in women for the occurrence of cVVC and some studies that refer new preventive and alternative therapies were also included, where appropriate. RESULTS AND DISCUSSION: Several studies have been shedding light on the increasing number of persistence and recurrences of VVC. The expression of virulence factors has been related to both chronic forms of VVC and antifungal resistance. Other studies report mutations occurring in the genome of Candida spp. during the infection phase which may be important indications for new therapies. The introduction of preventive therapies and new therapies has revealed great importance and is also highlighted here.

Recurrent vulvovaginal Candida spp isolates phenotypically express less virulence traits.

OBJECTIVE: Vulvovaginal candidosis (VVC) is a condition that impacts the quality of life of women worldwide. At least 5-8% of all VVC cases re-occur. Recurrent vulvovaginal candidosis (RVVC) can be defined as the occurrence of a VVC episode at least four times per year. The reasons for recurrence to occur are poorly understood. This work aims to identify key phenotypic traits associated with RVVC Candida spp. isolates that might be used to plan strategies to control RVVC. METHODS: The capacity to form biofilms (with the microtitration plate assay), to develop germinative tube in the presence of fetal bovine serum and to produce phospholipase (in the egg-yolk plate assay) was assessed for a collection of Candida spp. isolates obtained from 17 women diagnosed with RVVC and 16 women with non-recurrent VVC (VVC). The differences obtained regarding the proportion of isolates expressing each virulence factor was assessed by statistical analysis (χ2). RESULTS: We found that C. albicans isolates had a higher ability to form germinative tubes than RVVC isolates (29% vs 4%, p < 0.05). In addition, the ability of Candida spp. isolates to form biofilm (63% vs 51%) and to produce phospholipase (13% vs 11%) was also higher, though not statistically different (p > 0.05). CONCLUSIONS: We conclude that biofilm formation and phenotypic-switching associated with germinative tube production are particularly important C. albicans virulence factors for acute, sporadic VVC cases.

Semen supports growth of Candida albicans: A putative risk factor for recurrence of vulvovaginal infections?

AIM: Vulvovaginal candidosis (VVC) is the second most common vaginal infection (20-25%), and about 90% of all VVC cases are caused by Candida albicans. Unprotected sexual intercourse has been implicated as one of the risk factors that lead to an outbreak of VVC. To further investigate the relevance of this particular risk factor, in this study, we aim to evaluate the effect of human semen in the promotion of the growth of C. albicans. METHODS: The disposable amount of 41 samples of semen obtained from infertility patients were included in this study, with informed consent. The spermogram and physical characteristics of the samples were performed at the Unit; this information was provided with the anonymity of the samples. Samples were inoculated with a calibrated suspension of C. albicans ATCC 10231 in culture media. After the incubation time, C. albicans CFU/mL was determined. RESULTS: We found that semen allowed the growth of C. albicans (4.30 ± 1.00 CFU/mL), but not as much as the culture medium (9.45 ± 1.90 CFU/mL). Interestingly, we found that the increase in viscosity impaired significantly C. albicans growth. In addition, in what respects to the rate of multiplication of C. albicans in semen, we observed two different trends. However, we found no relation between these and the physical characteristics of the semen samples in which these behaviors were differently observed. CONCLUSION: Semen has the ability to sustain C. albicans growth, but further studies are needed to elucidate its role in VVC.

In vitro evaluation of potential benefits of a silica-rich thermal water (Monfortinho Thermal Water) in hyperkeratotic skin conditions.

Thermal therapy has gained popularity over the years, and Portugal is one of the richest European countries in mineral therapeutic waters. The interest in the use of these natural mineral waters (NMW) for dermatologic purposes is continuously growing but there is a lack of scientific studies supporting its health benefits. The study aims to investigate the effect of a silica-rich NMW in skin cell homeostasis using two representative cell lines of the epidermis and dermis, keratinocytes and fibroblasts, respectively, in addition to a macrophage cell line. Mouse skin fibroblasts, macrophages and human keratinocytes were exposed to culture medium prepared with NMW. Cell metabolism (MTT or resazurin assays) and cell proliferation (trypan blue exclusion dye assay) were investigated. Migration (scratch-wound assay) and senescence (ß-galactosidase activity assay) of fibroblasts were also studied. Exposure to NMW compromised the cell metabolic state of all the cell lines tested. This impairment was more pronounced in skin keratinocytes (60% reduction) relatively to skin fibroblasts (45% reduction) or macrophages (25% reduction). Proliferation of macrophages was reduced threefold upon exposure to thermal water, compared to controls. No differences were observed in migration between fibroblasts exposed to NMW and controls, while a potentiation of senescence of these cells was observed. Our results shed light in the bioactive effects of a silica-rich NMW supporting its therapeutic use. A reduction in both cell metabolism and proliferation of keratinocytes and macrophages supports the empirical clinical benefits of this NMW in hyperkeratotic conditions, such as psoriasis and atopic dermatitis.

The vaginal sheet: an innovative form of vaginal film for the treatment of vaginal infections.

Objective: To develop and characterize a new form of vaginal film.Significance: This formulation is intended to overcome some known limitations of traditional dosage forms. It has an absorptive intention to control symptoms and to improve the treatment of vaginal infections characterized by excessive fluid. The vaginal sheet is a thick drug delivery system easy to manipulate, nontoxic and composed by biocompatible macromolecules and polymers, such as gelatin and chitosan.Methods: The sheets were prepared by formulating gelatin or chitosan based gels isolated or in combination, in association with a plasticizer. Gels were subsequently lyophilized. Different proportions of polymer:plasticizer were tested. Lactose was used as a surrogate to study powder incorporation in the formulation. All formulations were analyzed regarding their organoleptic characteristics, texture (hardness and resilience), in vitro absorption efficiency of vaginal fluid simulant - VFS (pH 4 and 5), pH and acid-buffering capacity.Results: Different properties were obtained by varying polymer and plasticizer proportions. Combinations including gelatin with propylene glycol showed the best organoleptic characteristics. The best proportions were 4:3 and 4:5. Up to 10% of powder was successfully incorporated in the formulation. Hardness and resilience of formulations were largely dependent on the concentration of plasticizer. Absorption of vaginal fluid was found to be highly efficient, especially at pH 5. Buffering capacity, upon dilution in normal saline and VFS, was generally higher for VFS pH 4.Conclusions: The vaginal sheet is a promising solid drug delivery system able to further incorporate drugs to treat vaginal clinical conditions characterized by excessive fluid.

Chemical signature and antimicrobial activity of Central Portuguese Natural Mineral Waters against selected skin pathogens.

The common therapeutic indications of Portuguese Natural Mineral Waters (NMWs) are primarily for respiratory, rheumatic and musculoskeletal systems. However, these NMWs have been increasingly sought for dermatologic purposes. Opposing to what is observed in the major European Thermal Centres, there are few scientific evidences supporting the use of Portuguese NMWs for clinical applications. The aim of this study was to characterize the antimicrobial profile of individual NMWs from the central region of Portugal and correlate the results with their physicochemical characterization. An extensive multivariate analysis (principal component analysis) was also performed to further investigate this possible correlation. Six collection strains representing skin microbiota, namely Staphylococcus aureus, Escherichia coli, Corynebacterium amycolatum, Candida albicans, Staphylococcus epidermidis and Cutibacterium acnes, were analysed, and their antimicrobial profile was determined using Clinical and Laboratory Standards Institute M07-A10, M45-A2, M11-A6 and M27-A3 microdilution methods. Different NMWs presented different antimicrobial profiles against the strains used; the physicochemical composition of NMWs seemed to be correlated with the different susceptibility profiles. Cutibacterium acnes showed a particularly high susceptibility to all NMWs belonging sulphurous/bicarbonated/sodic ionic profile, exhibiting microbial reductions up to 65%. However, due to the complex physicochemical composition of each water an overall conclusion regarding the effect of a specific ion on the growth of different microorganisms is yet to be known.

An update on the role of Atopobium vaginae in bacterial vaginosis: what to consider when choosing a treatment? A mini review.

INTRODUCTION: Bacterial vaginosis (BV) is the most common vaginal disorder in reproductive-age women. The condition is characterised by the replacement of a healthy, lactobacilli-dominated vaginal microbiota by anaerobic and facultative anaerobic bacteria. BV increases the risk of acquisition of STIs and is associated with pregnancy complications. Although the composition of the bacteria in BV varies between individuals, there are some species such as Gardnerella, Atopobium, Mycoplasma, Snethia, Megasphera, Dialister, etc., that are found most frequently. MATERIAL AND METHODS: Literature research to the importance of Atopobium vaginae in BV and treatment options. RESULTS: Atopobium (A.) vaginae is an important component of the complex abnormal vaginal flora in BV; even though A. vaginae, like Gardnerella vaginalis, has also been detected in the normal flora, it is much more common in BV patients. A. vaginae has been shown to play an important role in the pathophysiology of BV and is thought to be at least a partial cause of the known negative sequelae. The presence of A. vaginae in the BV-associated biofilms and its resistance to some antimicrobial substances has been described - this seems to have a major impact on treatment outcome. CONCLUSION: Current scientific data demonstrate that dequalinium chloride (Fluomycin®) is one of the valid therapeutic options for BV treatment, since it displays a broad antimicrobial spectrum against relevant vaginal pathogens, especially against G. vaginalis and A. vaginae, without having safety concerns.

Development and validation of a new one step Multiplex-PCR assay for the detection of ten Lactobacillus species.

Lactobacillus sp. are well-known colonizers of human mucosa and frequently used as probiotics. Accurate species identification is crucial both for fundamental studies and biotechnology applications; however, it has been thus far challenging. The aim of this work was to develop a one-step multiplex-PCR assay for detection of ten Lactobacillus species (L. jensenii, L. fermentum, L. acidophilus, L. crispatus, L. reuteri, L. iners, L. casei, L. gasseri, L. plantarum, L. rhamnosus) directly in complex bacterial genomic DNA. A multiplex-PCR assay was optimized based on Box-Behnken experimental design, which showed to be efficient for optimization of all crucial reaction components. Nineteen Lactobacillus strains, including six collection strains and thirteen human isolates were used in order to verify the specificity and sensitivity of the assay. In addition, a set of PCR adjuvants was introduced to remove non-specific amplifications and enhance reaction yield. Among them, Triton™ X-100, Tween® 20, BSA, and dithiothreitol showed beneficial effects when compared with other adjuvants. The application of the developed method to samples that resulted from the mixing of DNA from the ten strains, resulted in amplicons of the expected sizes (from about 100 to 1000 bp). The detection limit was 1.25 ng/µl for all species with the exception of L. gasseri (0.31 ng/µl). In order to confirm the method applicability on human samples, ten vaginal fluids were enrolled in this study showing that the method can be successfully used on these biological materials. The proposed multiplex-PCR assay was shown to be selective, sensitive and efficient for detection of ten Lactobacillus species directly in human vaginal samples. This method provides a cost-effective and accessible methodology applicable to the detection of Lactobacillus species to different environments. At the same time, this approach represents a considerable improvement over other PCR-based approaches for identification of these species.

Anti-Candida activity of antidepressants sertraline and fluoxetine: effect upon pre-formed biofilms.

As an opportunistic fungal pathogen Candida spp. has the ability to form biofilms. The most prescribed drugs for Candida infections, azoles, have shown to be less effective when biofilms are present. In addition, increasing treatment costs and the fact that most prescribed antifungal drugs have only fungistatic activity justify the search for new treatment strategies. One promising approach is third generation antidepressants, selective serotonin re-uptake inhibitors (SSRIs), because of their proven antifungal activity against several Candida spp. Thus, the aim of this work was to determine the ability of two commonly used SSRIs, fluoxetine and sertraline, to impair both biofilm metabolic viability and biofilm biomass. The in vitro effect of fluoxetine and sertraline was individually tested against biofilm metabolic viability and biofilm biomass using the MTT assay and the Crystal Violet assay, respectively. For both drugs, a dose-dependent reduction on both biofilm metabolism and biomass was present. At high concentrations, fluoxetine was able to reduce biofilm metabolism by 96% (C. krusei) and biofilm biomass by 82% (C. glabrata), when compared to the control. At similar conditions, sertraline achieved a reduction of 88% on biofilm biomass (C. glabrata) and 90% on biofilm metabolism (C. parapsilosis). Moreover, fluoxetine showed interesting anti-biofilm activity at previously reported planktonic MIC values and even at sub-MIC values. These results reinforce the potential interest of SSRIs as anti-biofilm agents to be study to counteract resistance phenomena on candidosis.

Application of Coffee Silverskin in cosmetic formulations: physical/antioxidant stability studies and cytotoxicity effects.

CONTEXT: Currently, there is an increasing interest of cosmetic industry on natural extracts. The inclusion of antioxidants in topical formulations can contribute to minimize oxidative stress in the skin, which has been associated with aging. Also, questions of sustainability are leading to the study of new cosmetic ingredients obtained from food by-products. Coffee Silverskin (CS) is a food by-product with established antioxidant activity that has not yet been incorporated into a topical formulation. OBJECTIVE: The aim of this study was to evaluate the physical and microbiological stabilities and antioxidant activity of a hand cream formulation containing 2.5% (w/w) of CS extract upon production and after 6 months of shelf-life and in vitro safety/cytotoxicity on skin cell lines after production. MATERIALS AND METHODS: The in vitro cytotoxicity was evaluated with MTS and LDH assays, at different concentrations, in HaCaT and HFF-1 cells. Formulations were stored at 25 °C/65% RH and 40 °C/75% RH. Physical, microbiological, and antioxidant stabilities were evaluated by centrifugation, viscosity, total colony count, DPPH and total phenolic content (TPC). RESULTS: The hand cream containing 2.5% (w/w) of CS extract showed stable physical characteristics independently of the storage conditions. The DPPH activity and TPC of the CS formulation were significantly higher compared with those of the base formulation. However, during storage, the antioxidant activity decreases slightly. Microbiological quality was also confirmed. No cytotoxic effects were observed. CONCLUSION: It is possible to suggest that this formulation is stable under extreme conditions and safe for topical use.

Women's experiences, preferences and perceptions regarding vaginal products: Results from a cross-sectional web-based survey in Portugal.

OBJECTIVE: We aimed to identify Portuguese women's experiences, preferences and perceptions regarding vaginal products. METHODS: A descriptive cross-sectional study was conducted (February-May 2013) among Portuguese women (aged 18 to 65 years) using an online questionnaire. Descriptive and chi-squared statistics were applied. RESULTS: Among 2529 women, 85.4% had used vaginal products, mostly to manage vulvovaginal infections (75.3%). Gels, creams and ointments (semi-solids) were the most frequently used (82%), followed by vaginal suppositories (56.5%) and tablets/capsules (41.8%), while vaginal rings were used by 10% of women. Semi-solids were preferred as an intravaginal medication both by women who had previously used them and by women who had never used an intravaginal product, while preference for vaginal rings was higher only among women who had previously used them. Even though 87.1% of all women considered vaginal drug delivery to be advantageous, the majority preferred to use oral products. Leakage (84.8%) and insertion difficulties (58.4%) were the main problems reported for vaginal products. CONCLUSIONS: Overall, semi-solids were the most used and preferred vaginal products, while vaginal rings were highly acceptable for women who had previously used them. Although they considered the vaginal route to be more efficient and safe, many women felt it to be less appealing than the oral route, particularly due to comfort issues.

Coffee silverskin: a possible valuable cosmetic ingredient.

CONTEXT: Currently, there is a great tendency in cosmetic area to use natural extracts. Coffee silverskin (CS) is the most abundant solid by-product generated during roasting of coffee processing. OBJECTIVES: To evaluate different CS extracts as promising cosmetic ingredients, regarding antioxidant, antimicrobial, and cytotoxic properties. MATERIALS AND METHODS: Aqueous, hydroalcoholic and ethanolic CS extracts were obtained by an environmentally friendly procedure considering costs and pollution. Extracts were characterized for total phenolic and flavonoid contents (TPC and TFC, respectively), antioxidant activity by 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), antimicrobial activity expressed as minimal inhibitory concentration (MIC) and cytotoxicity using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) and lactate dehydrogenase (LDH) assays in two skin cell lines (fibroblasts and keratinocytes). RESULTS: The TPC of extracts was 18.33-35.25 mg of gallic acid equivalents per g of material on a dry basis (mg GAE/g db). The TFC of extracts was 1.08-2.47 µg cathechin equivalents per g dry material (µg CE/g db). The antioxidant activity was high, with values ranging between 95.95 and 216.40 µmol Fe(2+)/g for aqueous and alcoholic samples, respectively. Preliminary assays for antimicrobial potential showed that extracts display antibacterial activity. The MIC varied from 31.3 to 250 µg/mL for Gram-positive, and from 31.3 to 1000 µg/mL for Gram-negative. Extracts did not affect in vitro cell viability, with values near 100% in all concentrations tested. CONCLUSION: RESULTS seem show that CS is a safe source of natural antioxidants with antifungal and antibacterial activity and no cytotoxicity, with potential usefulness for cosmetic applications.

Anti-Candida activity of fluoxetine alone and combined with fluconazole: a synergistic action against fluconazole-resistant strains.

The purpose of this work was to determine the antimicrobial activity of fluoxetine, alone and combined with fluconazole, against 29 Candida strains isolated from patients with vulvovaginal candidiasis. MIC and minimum lethal concentration values ranged from 9.8 to 625 µg/ml for all strains tested. The combination of fluconazole with fluoxetine resulted in synergistic activity against six Candida strains, with fractional inhibitory index (FIX) values between 0.15 and 0.31. An indifferent effect was found for the remaining strains, with FIX values between 0.63 and 1.

Humulus lupulus aqueous extract and hydrolate as a potential ingredient for cosmetics: chemical characterization and in vitro antimicrobial, cytotoxicity, antioxidant and anti-inflammatory assessment.

Humulus lupulus extracts have in their composition different molecules, such as polyphenols, α-acids, ß-acids, and hydrocarbons, which contribute to the plant's medicinal properties. These molecules are associated with antimicrobial, antioxidant and anti-inflammatory activities. OBJECTIVE: This work focuses on the evaluation of H. lupulus biological activities, with the aim of evaluating its potential for inclusion in cosmetic formulations. METHODS: Two distinct aqueous extracts and two hydrolates obtained via hydrodistillation were evaluated. These include the flower parts (FE, FH) and the mix of aboveground parts (ME, MH). The chemical profiles for both aqueous extracts and hydrolates were identified by high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS). Antimicrobial, antioxidant, cytotoxicity, and anti-inflammatory activity were tested in vitro using standard methods. RESULTS: Rutin was the major compound found in FE (40.041 µg mg-1 of extract) and ME (2.909 µg mg-1 of extract), while humulenol II was the most abundant compound in hydrolates (FH: 20.83%; MH: 46.80%). Furthermore, FE was able to inhibit the growth of Staphylococcus aureus and Staphylococcus epidermis with MIC values of 50% and 25% (v/v), respectively. FH showed the same effect in Staphylococcus aureus (50% v/v). FH evidenced poor antioxidant potential in DPPH scavenging test and demonstrated significant antioxidant and anti-inflammatory effects by reducing (***p < 0.001) intracellular reactive oxygen species (ROS), NO (nitric oxide) levels (***p < 0.001) and cyclooxygenase-2 (COX-2) protein expression (***p < 0.001) in lipopolysaccharide (LPS)-stimulated macrophages. Nevertheless, it is important to note that FH exhibited cytotoxicity at high concentrations in 3T3 fibroblasts and RAW 264.7 macrophages. CONCLUSION: The studied H. lupulus aqueous extracts and hydrolates revealed that FH stands out as the most promising bioactive source for cosmetic formulations. However, future research addressing antimicrobial activity is necessary to confirm its potential incorporation into dermatological and cosmetic formulations.

The Role of Yeasts in Human Health: A Review.

The microbiome consists mostly of bacteria, but new evidence and developments in sequencing methods have shown that fungi play an important role in human health and in the stability of the microbiota. Scientific knowledge about the role of commensal fungi in intestinal, oral, vaginal and cutaneous communities has been increasing; however, more studies are still needed to better understand their action in these niches. To date, fungal research focuses primarily on opportunistic diseases caused by fungal species, leaving unclear the possible role of fungi as an integral part of the microbiota. Although they are much less abundant than bacteria, fungi such as species belonging to the genus Candida, Malassezia, Rhodotorula and Cryptococcus are some of the yeasts that have been in the focus of the scientific community because they inhabit various niches. In this review, we have summarized the current information about the yeasts that inhabit the human body, including some of the diseases that they can cause when the microbiota becomes unstable.

Thymus mastichina (L.) L. and Cistus ladanifer L. for skin application: chemical characterization and in vitro bioactivity assessment.

ETHNOPHARMACOLOGICAL RELEVANCE: Thymus mastichina (L.) L. (TM) and Cistus ladanifer L. (CL) are two Portuguese autochthonous species with traditional skin application in folk medicine. TM is majorly known for its antiseptic and wound healing properties, as an external anti-inflammatory agent and for its application in folk cosmetics and hygiene products. Its use in acne vulgaris has also been reported. CL is traditionally used in remedies for wounds, ulcers and other skin ailments such as psoriasis and eczema. Its application has been found useful due to its anti-inflammatory, astringent, wound healing and antiseptic properties. AIM OF THE STUDY: With this work, we aimed to investigate relevant bioactivities related with the traditional application of TM and CL essential oils (EOs) and hydrolates (by-products of EO production) in skin ailments. Specifically their in vitro antioxidant, anti-inflammatory, cytotoxic, wound healing and antimicrobial properties were evaluated. The chemical composition of both EOs and respective hydrolates was also characterized. MATERIALS AND METHODS: Chemical characterization of EOs and hydrolates was performed by GC-FID and GC-MS. Cellular biocompatibility was evaluated using the MTT assay in macrophages (RAW 264.7) and fibroblasts (L929) cell lines. Anti-inflammatory activity was investigated by studying nitric oxide (NO) production by macrophages with Griess reagent. Wound healing potential was evaluated with the scratch-wound assay. The antioxidant potential was studied by the DPPH scavenging method. Antimicrobial activity was evaluated by broth microdilution assay against relevant microbial strains and skin pathogens, namely Staphylococcus aureus, Staphylococcus epidermidis, Cutibacterium acnes, Pseudomonas aeruginosa, Escherichia coli, Candida albicans and Aspergillus brasiliensis. RESULTS: The major compounds present in TM and CL EOs were 1,8-cineole and α-pinene, respectively. 1,8-cineole and E-pinocarveol were the major compounds in the correspondent hydrolates. CL EO presented the highest anti-inflammatory potential [EC50 = 0.002% (v/v)], still with significant cytotoxicity [IC50 = 0.012% (v/v)]. TM preparations presented anti-inflammatory potential, also presenting higher biocompatibility. The same profile was present on fibroblasts regarding biocompatibility of the tested preparations. CL EO and hydrolate increased fibroblasts' migration by 155.7% and 148.4%, respectively. TM hydrolate presented a milder activity than CL hydrolate, but wound healing potential was still present, increasing cell migration by 125.1%. All preparations presented poor antioxidant capacity. CL EO presented higher antimicrobial activity, with MICs ranging from 0.06% (v/v) to 2% (v/v), against different microorganisms. CONCLUSIONS: Anti-inflammatory and skin repairing potential were present for CL preparations. TM hydrolate presented an interesting biocompatible profile on both cell lines, also presenting anti-inflammatory potential. Furthermore, EOs from both species presented antimicrobial activity against a panel of different microorganisms. These in vitro bioactivities support some of their traditional skin applications, specifically regarding their antiseptic, wound healing and anti-inflammatory uses.

Uncovering the Yeast Diversity in the Female Genital Tract: An Exploration of Spatial Distribution and Antifungal Resistance.

Candida albicans is the leading cause of vulvovaginal yeast infections; however, other species are becoming relevant in this niche. The spatial distribution of these fungi in the female genital tract remains poorly understood. In this study, swab samples were collected from 33 patients, first from the anterior vulva and then from the upper third and right lateral wall of the vagina: 16 were with symptoms of vulvovaginal candidiasis and 17 were without characteristic symptoms; furthermore, the genus and species of each isolate were identified. In vitro susceptibility testing for fluconazole and clotrimazole was performed for all isolates. Candida albicans was the most common species (63.6%), followed by Rhodotorula spp. (51.5%), and then Candida parapsilosis (15.2%). Rhodotorula spp. and C. parapsilosis were more commonly associated with colonization, and C. albicans with infection. Rhodotorula spp. isolates presented a low susceptibility to fluconazole, with the MIC ranging from 32 to >64 µg/mL. Differences in susceptibility to fluconazole and clotrimazole between the pairs of vaginal and vulvar isolates were found for Candida albicans, Rhodotorula spp., and Nakaseomyces glabratus. The results suggest that different niches may impact the susceptibility profiles of the isolates, as well as their different clinical behaviors.