Antifungal characteristics of warp-knitted cotton fabrics treated with various metal complexes

We investigated the efficacy of oral fluconazole, alone or in combination with oral flucytosine (5FC), in treating Candida endophthalmitis using a rabbit model. Albino rabbits were infected with an intravitreal inoculation of 1,000 CFU of susceptible Candida albicans and randomized 5 days later to receive treatment with oral fluconazole alone (80 mg/kg of body weight per day), a combination of fluconazole and 5FC (100 mg/kg/12 h), or no treatment. The treatment effect was assessed at 2 and 4 weeks after therapy by funduscopy, quantitative vitreous culture, and histopathology. Intravitreal levels of fluconazole, 2 to 24 h after the first dose, were measured to be > 10 times the MIC of the drug for C. albicans. Among rabbits treated with fluconazole for 2 weeks, 67% had a > 90% reduction in fungal load (P < 0.05) and 33% were sterile. After 4 weeks, all had a > 99% reduction in fungal load (P < 0.05) and 75% were sterile (P = 0.01). This treatment effect was unchanged 4 weeks after discontinuation of fluconazole. Among rabbits treated with fluconazole and 5FC for 2 weeks, 67% died during therapy. Among the surviving rabbits, 75% had a > 90% reduction in fungal load (P < 0.05) and 25% were sterile. We conclude that oral fluconazole may be useful for treatment of Candida endophthalmitis. Addition of 5FC was associated with high toxicity and minimal additional antifungal effect in our rabbit model.

Background: Candida albicans is increasingly recognized in peri-implantitis due to its capacity to form resilient biofilms on implant surfaces, limiting treatment success. Antimicrobial photodynamic therapy (aPDT) may offer a non-invasive adjunct by leveraging photosensitizer activation to produce reactive oxygen species that disrupt microbial cells. This in vitro study assessed the antifungal efficacy of QroxB2, a dual-photosensitizer containing riboflavin and curcumin, activated by 450 nm blue light against C. albicans biofilms on titanium implants. Methods: C. albicans biofilms were formed on 63 titanium implants and randomly assigned to nine groups (n = 7): untreated control (GC), chlorhexidine (CHX), riboflavin (RIB), curcumin (CUR), QroxB2 (QBX), laser only (L), and three photodynamic therapy groups combining laser irradiation with each photosensitizer (L + RIB, L + CUR, L + QBX). Treatments were followed by colony-forming unit (CFU) enumeration. Results: The L + QBX group showed the strongest antifungal effect, achieving a 94% reduction in fungal load, with median CFU counts decreasing from 49,000 in the untreated control to 2800 CFU/mL. CHX eradicated all viable cells (0 CFU/mL). Among photosensitizer-only groups, QBX produced a moderate reduction (median 21,800 CFU/mL), whereas laser irradiation alone (L) exhibited no meaningful antifungal activity, with median counts comparable to the untreated control (49,000 CFU/mL). Conclusions: QroxB2-mediated aPDT achieved a significant reduction in Candida albicans colony-forming units on implant surfaces. While not as potent as chlorhexidine, this light-activated, biocompatible approach may serve as a complementary tool in managing peri-implant fungal infections. Clinical validation is warranted.

Effect of amphotericin B lipid formulation on immune response in aspergillosis

Drug repositioning has been an important ally in the search for new antifungal drugs. Statins are drugs that act to prevent sterol synthesis in both humans and fungi and for this reason they are promissory candidates to be repositioned to treat mycoses. In this study we evaluated the antifungal activity of atorvastatin by in vitro tests to determine the minimum inhibitory concentration against azole resistant Candida albicans and its mechanisms of action. Moreover, the efficacy of both atorvastatin-loaded oral and vaginal emulgels (0.75%, 1.5% and 3% w/w) was evaluated by means of in vivo experimental models of oral and vulvovaginal candidiasis, respectively. The results showed that atorvastatin minimal inhibitory concentration against C. albicans was 31.25μg/ml. In oral candidiasis experiments, the group treated with oral emulgel containing 3.0% atorvastatin showcased total reduction in fungal load after nine days of treatment. Intravaginal delivery atorvastatin emulgel showed considerable effectiveness at the concentration of 3% (65% of fungal burden reduction) after nine days of treatment. From these findings, it is possible to assert that atorvastatin may be promising for drug repositioning towards the treatment of these opportunistic mycoses.

Background/Objectives: Oral candidiasis is frequently encountered in pediatric populations, particularly in infants and toddlers, where the development of immunity and inconsistent oral hygiene contribute to disease susceptibility. While topical antifungal agents remain the standard of care, treatment challenges persist, especially regarding adherence and recurrence. Aloe vera, recognized for its antimicrobial, anti-inflammatory, and mucosal healing properties, may offer therapeutic benefits when used in conjunction with standard regimens. This study aimed to evaluate the adjunctive effect of topical Aloe vera gel, when added to standard antifungal therapy, on reducing fungal load and improving treatment adherence in children with moderate oral candidiasis. Methods: A prospective controlled study was conducted among 54 children diagnosed with moderate oral candidiasis. Participants were randomly assigned to receive either standard topical nystatin or nystatin in conjunction with Aloe vera gel over a 7 day treatment duration. Fungal load was assessed using colony-forming units (CFU) counts from oral swabs collected at baseline and day 7, analyzed via ANCOVA. Additional parameters included treatment adherence, compared using an independent t-test, and clinical recurrence at a 14-day post-treatment follow-up, assessed through logistic regression. Results: Baseline characteristics were similar across groups. By day 7, children in the Aloe vera group exhibited a greater reduction in fungal load compared to those receiving standard therapy alone. Adherence was significantly higher in the aloe group (92.73% vs. 89.21%; p < 0.0001). Regression analysis identified both baseline fungal burden and adherence as factors associated with an increased risk of recurrence. Conclusions: The addition of Aloe vera gel to standard therapy may support a more effective fungal clearance and improved treatment adherence in children with moderate oral candidiasis, suggesting its potential as a complementary treatment option. Given the single-center design, short follow-up, and underpowered recurrence analysis, these findings should be considered preliminary, pending confirmation in larger studies with symptom-anchored endpoints.

The kinetics of various parameters of fungal load and cytokines were investigated, in order to acquire insight into the pathogenesis of invasive pulmonary aspergillosis (IPA) during antifungal treatment with amphotericin B. Neutropenic rats with left-sided IPA received either treatment with amphotericin B or remained untreated. At 0, 4, 8, 16, 24, 48, 72 and 120 h after fungal inoculation, the rats were dissected. The size of the macroscopic pulmonary lesions, the number of cfu and amounts of chitin were determined in the infected left lung. Galactomannan concentrations were measured both in the left lung and serum. The cytokines tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, interferon (IFN)-gamma, IL-4, IL-10, and the chemokines macrophage inflammatory protein (MIP)-2 and monocyte chemoattractant protein (MCP)-1 were determined quantitatively by ELISA in the infected left lung, uninfected right lung and serum. Amphotericin B treatment of IPA resulted in changed aspect of pulmonary lesions and significantly reduced levels of left lung chitin (72 and 120 h), left lung galactomannan (72 and 120 h) and serum galactomannan (120 h), but not left lung cfu, compared with untreated infected rats. In addition, amphotericin B treatment resulted in a significant decrease in levels of left lung IL-6 (at 72 and 120 h), MIP-2 (at 120 h) and MCP-1 (at 120 h). No local or systemic increases in TNF-alpha, IL-1beta or IFN-gamma were observed during infection. It is concluded that treatment with amphotericin B results in decreased fungal load in the infected lung. This reduction in fungal load probably results in a decreased local inflammatory response, as measured by decreased levels of IL-6, MIP-2 and MCP-1 in the infected lung.

In this study, the author used endoscopic sinus photography to study the effects of reduction of fungi in the nose, and in environmental air, on the sinus mucosa of 639 patients diagnosed with chronic rhinosinusitis. Sinus mucosal photographs were taken before and after reduction of fungal load in the nose and air, to determine if there was an optimum environmental air fungal load associated with sinus mucosal recovery to normal appearance. Systemic symptoms associated with fungal exposure, which resolved when fungus was removed from the patient and the environmental air and reappeared with recurrent environmental fungal exposure, are also discussed and are termed systemic fungal symptoms. Interventions consisted of nasal fungal load reduction with normal saline nasal irrigations and antimicrobial nasal sprays, and environmental air fungal load reduction with high-efficiency particulate air (HEPA) filtration in combination with ionizers or evaporation of a solution of botanical extract. Main outcome measures were obtained with environmental air 1-hr gravity-plate fungal colony counts, laser air particle counts, and endoscopic sinus photography. Blood levels of immunoglobulins IgG and IgE for 7 common molds were also determined. After intervention, 94% of patients who used antimicrobial nasal sprays and who reduced their environmental fungal air count to 0-4 colonies per 1-hr agar gravity-plate exposure (n = 365) exhibited normal sinus mucosa by endoscopic exam. Environmental air fungal counts that exceeded 4 colonies resulted in sinus mucosal abnormalities ranging from edema, to pus and/or nasal polyps at higher counts. Neutralization of allergy, and/or surgery, were used as appropriate following implementation of environmental measures. On the basis of these observations, as well as detailed clinical experience and a review of the current literature, the author hypothesizes that the pathogenesis of chronic rhinosinusitis, allergic fungal sinusitis, and systemic fungal symptoms is a genetic defect at the variable beta chain helper T-cell receptor (TCR Vbeta) site which requires the presence of an antigen (fungus). Chronic sinusitis patients who have recurring exposure to environmental air that contains fungal concentrations in excess of 4 colonies per 1-hr agar plate exposure appear to have an increased risk of persistent chronic sinusitis and/or systemic symptoms, regardless of the medical treatment provided.

Aspergillus fumigatus causes life-threatening lung infections in immunocompromised patients. Mouse models are extensively used in research to assess the in vivo efficacies of antifungals. In recent years, there has been an increasing interest in the use of noninvasive imaging techniques to evaluate experimental infections. However, single imaging modalities have limitations concerning the type of information they can provide. In this study, magnetic resonance imaging and bioluminescence imaging were combined to obtain longitudinal information on the extent of developing lesions and fungal load in a leukopenic mouse model of invasive pulmonary aspergillosis (IPA). This multimodal imaging approach was used to assess changes occurring within lungs of infected mice receiving voriconazole treatment starting at different time points after infection. The results showed that IPA development depends on the inoculum size used to infect animals and that disease can be successfully prevented or treated by initiating intervention during early stages of infection. Furthermore, we demonstrated that a reduction in fungal load is not necessarily associated with the disappearance of lesions on anatomical lung images, especially when antifungal treatment coincides with immune recovery. In conclusion, multimodal imaging allows an investigation of different aspects of disease progression or recovery by providing complementary information on dynamic processes, which are highly useful for assessing the efficacy of (novel) therapeutic compounds in a time- and labor-efficient manner.

Synthesis and antifungal activity of 1,2,3-triazole containing fluconazole analogues

We evaluated the pre-clinical efficacy of simvastatin (SVT)-loaded emulgels in an in vivo model of azole-resistant Candida albicans (ATCC 10231)-induced vaginal candidiasis and conducted in vitro biopharmaceutical characterization of the most effective formulation. The efficacy of SVT-loaded emulgels (15, 30, and 60mg/g) was assessed in immunosuppressed Wistar rats with induced candidiasis and compared to commercially available vaginal creams containing clotrimazole (10mg/g) or nystatin (25,000 IU/g). Products were administered once daily for seven days (n = 5 per group). A high-resolution and sensitive HPLC-PDA method was developed and validated to quantify SVT, determine its solubility in the release medium, and evaluate its in vitro release profile and kinetics. The 60mg/g SVT-loaded emulgel achieved a 100% reduction in fungal load after 7 days (p < 0.05). SVT solubility in the release medium was 881 ± 36µg/mL, and its content in the emulgel was 114.95 ± 4.46% m/m. The controlled release kinetics followed Makoid-Banakar's model, indicating an average release rate of 0.036%/h. Therefore, the 60mg/g SVT-loaded emulgel shows potential as a therapeutic strategy for treating resistant vulvovaginal candidiasis, warranting further clinical studies.

Efficacy of intrathecal administration of liposomal amphotericin B combined with voriconazole in a murine model of cryptococcal meningitis

Cryptococcusalbidus and Cryptococcuslaurentii are uncommon species of this genus that in recent decades have increasingly caused opportunistic infections in humans, mainly in immunocompromised patients; the best therapy for such infection being unknown. Using a murine model of systemic infection by these fungi, we have evaluated the efficacy of amphotericin B (AMB) at 0.8mg/kg, administered intravenously, fluconazole (FLC) or voriconazole (VRC), both administered orally, at 25mg/kg and the combination of AMB plus VRC against three C.albidus and two C.laurentii strains. All the treatments significantly reduced the fungal burden in all the organs studied. The combination showed a synergistic effect in the reduction in fungal load, working better than both monotherapies. The histopathological study confirmed the efficacy of the treatments.

Introduction: Recurring vulvovaginal candidiasis (RVVC) is an infectious disease of the lower genitourinary tract, that occurs at least 4 times per year. Drug treatment can last months and favor the appearance of adverse effects and increase the chances of recurrence. Blue Light Diode (LED) is an electromagnetic spectrum light, with antimicrobial functions. Case Report: A case report was made of a patient with RVVC whose treatment consisted of three sessions of 60-minutes each, of application of blue LED in intervals of 15 days. Evaluation was carried out before, at the end of third application and three months after the last session by means of fungal culture, cervical cytology; analysis of patient´s clinical condition; and measurement of vaginal pH. After the third session, there was a reduction in fungal load and vaginal pH; absence of symptoms (pruritus, burning and dyspareunia) and vulvovaginal edema, but there were no alterations in the cytology and microflora, which remained with inflammatory markers. Three months after the end of the treatment, there was no Candida in vaginal secretion, neither signs nor symptoms of candidiasis, and vaginal pH was normal. As for cytology and microflora, cellular alterations associated with cytolysis and presence of lactobacilli was observed. Conclusion: The blue LED 401 nm may be a promising alternative to treat RVVC by eliminating signs and symptoms in women.

Isogenic mouse strains have previously been characterized as susceptible or resistant to Paracoccidioides brasiliensis infection; the former presented anergy in delayed-type hypersensitivity reactions (DTH) and progressive disease with high numbers of colony-forming units (CFU), while the later presented preserved DTH responses and control of the infectious process. Here, we studied whether susceptible mice infected with P. brasiliensis and treated with the antifungal drug trimethoprim-sulfamethoxazole (SXT) had their behavior pattern altered to the one observed in infected resistant mice. Therapy with either 30 or 150 mg SXT day(-1 )kg(-1), instituted 24 h after infection, elicited more adequate DTH responses than those of non-treated mice, and also diminished the number of viable fungi in the spleen and lungs, but not in epiploo and liver, indicating a partial control of the infectious process. This phenomenon was confirmed by histopathological analyses, in which the spleen was found to be the organ in which differences between the treated and non-treated groups were most remarkable. In control non-treated mice, the spleen parenchyma showed multiple granulomatous foci presenting giant cells, plasmocytes and many yeasts of P. brasiliensis with well-preserved morphology and abundant budding, whereas SXT-treated mice, independently of the dosage used, had no granulomas within the parenchyma and only few capsular lesions, mainly composed of pseudoxantomatous macrophages. Treatment with 150 mg day(-1 )kg(-1) (the dose considered to evoke best responses in CFU assays when therapy was instituted 24 h after infection), initiated at different times after infection, did not led to sustained DTH reactions, but provided an effective control of the disease when therapy began until the 15(th) day post infection, as showed by CFU assays. We conclude that reversal from the susceptible to the resistant pattern in experimental paracoccidioidomycosis can occur, but only when therapy with an adequate SXT dosage is instituted at a very initial phase of the infection. These protocols may constitute a model for further investigations concerning responses during antifungal therapy.

The fungal unfolded protein response consists of a two-component relay in which the ER-bound sensor, IreA, splices and activates the mRNA of the transcription factor, HacA. Previously, we demonstrated that hacA is essential for Aspergillus fumigatus virulence in a murine model of fungal keratitis (FK), suggesting the pathway could serve as a therapeutic target. Here we investigate the antifungal properties of known inhibitors of the mammalian Ire1 protein both in vitro and in a treatment model of FK. The antifungal activity of Ire1 inhibitors was tested against conidia of several A. fumigatus isolates by a microbroth dilution assay and against fungal biofilm by XTT reduction. The influence of 4μ8C on hacA mRNA splicing in A. fumigatus was assessed through gel electrophoresis and qRT-PCR of UPR regulatory genes. The toxicity and antifungal profile of 4μ8C in the cornea was assessed by applying drops to uninfected or A. fumigatus-infected corneas 3 times daily starting 4 hours post-inoculation. Corneas were evaluated daily through slit-lamp imaging and optical coherence tomography, or at endpoint through histology or fungal burden quantification via colony forming units. Among six Ire1 inhibitors screened, the endonuclease inhibitor 4μ8C displayed the strongest antifungal profile with an apparent fungicidal action. The compound both blocked conidial germination and hyphal metabolism of A. fumigatus Af293 in the same concentration range that blocked hacA splicing and UPR gene induction (60-120 μM). Topical treatment of sham-inoculated corneas with 0.5 and 2.5 mM 4μ8C did not impact corneal clarity, but did transiently inhibit epithelialization of corneal ulcers. Relative to vehicle-treated Af293-infected corneas, treatment with 0.5 and 2.5 mM drug resulted in a 50% and >90% reduction in fungal load, respectively, the latter of which corresponded to an absence of clinical signs of infection or corneal pathology. The in vitro data suggest that 4μ8C displays antifungal activity against A. fumigatus through the specific inhibition of IreA. Topical application of the compound to the murine cornea can furthermore block the establishment of infection, suggesting this class of drugs can be developed as novel antifungals that improve visual outcomes in FK patients.