A Combination of ß-Aescin and Newly Synthesized Alkylamidobetaines as Modern Components Eradicating the Biofilms of Multidrug-Resistant Clinical Strains of Candida glabrata.

The current trend in microbiological research aimed at limiting the development of biofilms of multidrug-resistant microorganisms is increasingly towards the search for possible synergistic effects between various compounds. This work presents a combination of a naturally occurring compound, ß-aescin, newly synthesized alkylamidobetaines (AABs) with a general structure-CnTMDAB, and antifungal drugs. The research we conducted consists of several stages. The first stage concerns determining biological activity (antifungal) against selected multidrug-resistant strains of Candida glabrata (C. glabrata) with the highest ability to form biofilms. The second stage of this study determined the activity of ß-aescin combinations with antifungal compounds and alkylamidobetaines. In the next stage of this study, the ability to eradicate a biofilm on the polystyrene surface of the combination of ß-aescin with alkylamidobetaines was examined. It has been shown that the combination of ß-aescin and alkylamidobetaine can firmly remove biofilms and reduce their viability. The last stage of this research was to determine the safety regarding the cytotoxicity of both ß-aescin and alkylamidobetaines. Previous studies on the fibroblast cell line have shown that C9 alkylamidobetaine can be safely used as a component of anti-biofilm compounds. This research increases the level of knowledge about the practical possibilities of using anti-biofilm compounds in combined therapies against C. glabrata.

Microbial load of heritage dermatological moulages of the historic university department in Wroclaw, Poland.

Introduction: Many historic dermatology departments keep and preserve valuable collections of dermatological moulages. Aim: The aim of the present research was to find out whether the specimens collected in the Museum of the Department of Dermatology, Venereology, and Allergology of Wroclaw Medical University are colonized by microorganisms, and whether these organisms can pose a risk of damage to this heritage or a health risk to visitors. Material and methods: In the study 32 historic moulages and their environment (museum) were subjected to microbiological evaluation. Results: Swabs from moulages turned to be positive in 28% of cases. Micrococcus luteus was mainly isolated. The flora isolated from the air and the external surfaces of the museum display cases was much richer. Environmental bacteria and fungi were determined, as well as organisms probably associated with the hospital flora: Pseudosomonas spp., Paebacillus sp., Acinetobacter sp. Conclusions: The close proximity of clinical wards probably influences the composition of the museum environment. The surprisingly low contamination of the moulages may be due to the antiseptic properties of the bee wax from which they were made. Conservation work on the moulages as well as people visiting the museum do not pose significant health risks. However, the small number of studies devoted to this topic limits the conclusions. Further research on medical collections is needed to provide 'evidence-based care' for this heritage.

Tinea pedis-An embarrassing problem for health and beauty-A narrative review.

Fungal infections present with a broad spectrum of diseases in humans (from relatively mild superficial infections of the skin and mucous membranes to the invasive or chronic infections of internal organs, which have a high mortality rate). Globally, up to 1.6 million people die each year as a result of various types of mycoses. Currently, many scientific studies focus on the best possible understanding of the aspects of the epidemiology and pathogenesis of invasive mycoses and effective methods to combat them. However, mycoses of the skin and its appendages remain a relatively less explored area. In some communities, superficial mycoses are a frequent problem as they affect nearly 70% of the population, an example of which is the athlete's foot. It involves the nails (onychomycosis) and skin (tinea pedis). It is mainly caused by keratin-decomposing dermatophyte fungi. Less often, infections are caused by non-dermatophyte moulds (Fusarium, Aspergillus, Scopulariopsis) or yeasts. Several factors have been listed as having substantial influence on the development of dermatophytosis, including those related to climate, season, geographical region, as well as to demography, socioeconomic and cultural customs, professions or contact with animals. In this review, we summarise the current knowledge about aetiology, epidemiology, diagnostics and therapy of tinea pedis with a special focus to the role of podologic management in spreading, prevention and therapy of mycoses. The article presents up-to-date knowledge on the management of the patient from the diagnosis, treatment and skincare, to counselling on how to prevent fungal skin infections in the long term.

Physicochemical and Antifungal Properties of Clotrimazole in Combination with High-Molecular Weight Chitosan as a Multifunctional Excipient.

Chitosans represent a group of multifunctional drug excipients. Here, we aimed to estimate the impact of high-molecular weight chitosan on the physicochemical properties of clotrimazole-chitosan solid mixtures (CL-CH), prepared by grinding and kneading methods. We characterised these formulas by infrared spectroscopy, differential scanning calorimetry, and powder X-ray diffractometry, and performed in vitro clotrimazole dissolution tests. Additionally, we examined the antifungal activity of clotrimazole-chitosan mixtures against clinical Candida isolates under neutral and acid conditions. The synergistic effect of clotrimazole and chitosan S combinations was observed in tests carried out at pH 4 on Candida glabrata strains. The inhibition of C. glabrata growth reached at least 90%, regardless of the drug/excipient weight ratio, and even at half of the minimal inhibitory concentrations of clotrimazole. Our results demonstrate that clotrimazole and high-molecular weight chitosan could be an effective combination in a topical antifungal formulation, as chitosan acts synergistically with clotrimazole against non-albicans candida strains.

Contribution of Malassezia spp. to the development of atopic dermatitis.

The prevalence of atopic dermatitis (AD) has been increasing. Whereas AD symptoms are obvious and easy to recognise, the etiopathogenesis remains not fully elucidated. Recently, the role of microorganisms and their impact on the immunology of AD have been discussed. In this review, we summarise a possible role of Malassezia in the development and persistence of eczema in patients with atopic eczema/dermatitis syndrome. A high proportion of AD patients present with a positive reaction to Malassezia allergens. Several possible pathogenic mechanisms enable Malassezia to trigger the development of AD. Malassezia spp. may release more allergens in a less acidic (pH <6), typical for AD, environment. The similarity between fungal thioredoxin and human proteins causes T-cell cross-reactivity. TLR-mediated mechanisms are involved in host response against Malassezia spp. An interaction between Malassezia spp. and keratinocytes alters the profile of cytokine release, and what is more, yeast cells can survive when absorbed by keratinocytes. Dendritic cells of AD patients induced by Malassezia are less susceptible to lysis mediated by NK cells which exerts a pro-inflammatory effect. Despite the evidence that Malassezia spp. contribute to the development of AD, the pathogenetic mechanisms and relationship between Malassezia and immune defense remain partly unexplained and require further research.

Detection of Polish clinical Aspergillus fumigatus isolates resistant to triazoles.

We studied the presence of triazole resistance of 121 Aspergillus fumigatus clinical isolates collected in two Polish cities, Warsaw and Wroclaw, to determine if resistance is emerging in our country. We identified five itraconazole resistant isolates (4.13%) carrying the TR34/L98H alteration in Cyp51A gene, four of which were cross-resistant to posaconazole and one to voriconazole. One isolate was intermediate susceptible to itraconazole and harbored no Cyp51A alterations. The study confirms the presence of azole resistant A. fumigatus strains in Poland at a level that is comparative to other European countries.

Synthesis and antimicrobial properties of 4-acylaminobenzenethiosulfoacid S-esters.

A series of esters of 4-acetyl, 4-trifluoroacetyl- and 4-(3-chloropropionyl)aminobenzenethiosulfoacids (twenty-four compounds) were synthesized and characterized by elemental analysis, 1H NMR and IR spectroscopy. The antibacterial activity of the novel candidates has been screened using the agar diffusion or serial dilution methods against representative Gram-positive (Staphylococcus aureus, Bacillus subtilis, Bacillus mesentericus, Mycobacterium sp., Mycobacterium luteum), Gram-negative (Aeromonas sp., Burkholderia cepacia, Alcaligenes faecalis, Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris) bacteria and fungi (Candida albicans, Candida tenuis, Candida glabrata, Verticillium dahliae, Trichophyton gypseum, Aspergillus niger, Aspergillus fumigatus, Penicillium chrysogenum). Particular potency has been discovered against all tested pathogenic bacteria and fungi by compounds 1l and 3l at nanomolar concentrations. Some appropriate effect of thiosulfoesters structure upon their antimicrobial activity was determined.

Detection of dermatophytes in human nail and skin dust produced during podiatric treatments in people without typical clinical signs of mycoses.

Pedicures are the most common cosmetic foot treatment. Many pedicurists and podiatrists suffer from respiratory infections and diseases such as asthma, sinusitis, chronic cough and bronchitis. Skin and nail dust may play an important role in the development of occupational diseases and the transmission of mycosis to other clients. To examine the presence of dermatophytes in nail and skin dust produced during podiatric treatments of people without typical symptoms of mycosis and to assess the epidemiological hazards of tinea pedis for podiatrists as well as other clients. Seventy-seven samples underwent direct microscopy and culture. The results of direct microscopy were positive in 28/77 samples (36.36%) and doubtful in 3/77 (3.9%). Fungi were cultured from 36/77 samples (46.75%), including 8/77 (10.3%) positive for dermatophytes (Trichophyton rubrum-6 isolates and Trichophyton mentagrophytes-2). Material collected during podiatric treatments is potentially infected by pathogenic fungi; thus, there is a need to protect both workers who perform such treatments, as well as other clients, to prevent the transmission of pathogens in the Salon environment. Exposure to this occupational hazard may increase not only the risk of respiratory infections but also increase asthmatic or allergic reactions to Trichophyton.

New monomeric and dimeric uridinyl derivatives as inhibitors of chitin synthase.

This study described the synthesis and in vitro evaluation of eight new derivatives of uridine as antifungal agents and inhibitors of chitin synthase. Dimeric uridinyl derivatives synthesized by us did not exhibit significant activity. One of the studied monomeric derivative, 5'-(N-succinyl)-5'-amino-5'-deoxyuridine methyl ester (compound 7) showed activities against several fungal strains (MIC range 0.06-1.00 mg/mL) and inhibited chitin synthase from Saccharomyces cerevisiae (IC50=0.8mM). Moreover compound 7 exhibited synergistic interaction with caspofungin against Candida albicans (FIC index=0.28).

Activity of fluconazole and its Cu(II) complex towards Candida species.

Candida species, although they are present as commensal organisms in the digestive tract of healthy individuals, can produce a broad spectrum of serious illnesses in compromised hosts. Fluconazole, a water-soluble triazole with bioavailability greater than 90 %, has been extensively used to treat a wide range of Candida infections. However, a growing resistance of microorganisms in the treatment leads to the discovery of new drugs or modifications of existing ones. The aim of the present study was to investigate whether coordination of Cu(II) ions to fluconazole affects its antifungal activity. The in vitro susceptibility tests and antifungal studies were performed with two Candida spp.: Candidaglabrata and Candida albicans. Overall, 34 strains of the former and 16 strains of the latter were treated with fluconazole, its Cu(II) complex and free Cu(II) ions. The obtained MIC values in 16 cases of the C. glabrata and in 5 cases of the C. albicans were lower for the complex in comparison to the drug. This implies that the complex is more effective against particular strains than the parent drug. The most significant improvement in the complex drug efficacy was observed for fluconazole-resistant species.

Antifungal Susceptibility of Saccharomyces cerevisiae Isolated from Clinical Specimens.

(1) Background: Despite being considered a non-pathogenic yeast, recently, a growing occurrence of Saccharomyces cerevisiae infections has been noted. There is little knowledge about the drug susceptibility of this species. Therefore, the objective of this research was to expand it and determine the drug susceptibility profile of a local collection of clinical isolates of this species. (2) Methods: This study contained 55 clinical isolates identified as Saccharomyces cerevisiae using the MALDI-TOF method. The susceptibility of Saccharomyces cerevisiae was tested to 10 antifungals (amphotericin B, flucytosine, fluconazole, voriconazole, posaconazole, micafungin, anidulafungin, caspofungin, and itraconazole) using MICRONAUT-AT tests and manogepix, a new drug, using the microdilution method according to EUCAST. (3) Results: Overall, most strains were classified as sensitive to amphotericin B and flucytosine (MIC ranges of ≤0.03-1 and ≤0.06-0.125, respectively) and also to echinocandins. However, five isolates expressed high MIC values for all of the tested azoles, indicating cross-resistance. The MIC range for manogepix was 0.001-0.125 mg/L, with an MIC50 of 0.03 mg/L and an MIC90 of 0.06 mg/L. (4) Conclusions: The occurrence of resistance to azoles may be a concerning problem and therefore should be investigated further. However, the new antifungal manogepix appears to be an interesting new therapeutic option for treating such infections.

Assessing the Interpretation of Molecular Test Results in the Diagnosis of Bloodstream Infections.

A retrospective study at the 4th Military Clinical Hospital in Wroclaw, Poland, assessed PCR testing alongside blood cultures to guide antimicrobial therapy decisions in hospitalized patients, to determine how much time the results of the molecular tests preceded conventional methods. Among 118 patients, Staphylococcus aureus (37%) and Escherichia coli (21%) were the most common bloodstream infection agents. Blood cultures utilized the BacT/ALERT 3D system, and molecular diagnostics were conducted using the FilmArray platform with the BIOFIRE BCID2 panel. Methicillin susceptibility was observed in 66% of S. aureus strains, while 26% of Gram-negative bacilli exhibited an ESBL phenotype. Therapeutic decisions based on molecular test results were often incorrect for S. aureus infections, particularly MSSA (64.5%), but generally accurate for Gram-negative bacilli. The median times from positive blood culture to BCID2 and pathogen identification/susceptibility were 10 h and 52 h, respectively. Molecular diagnostics facilitated faster initiation of appropriate antibiotic therapy, highlighting the need to educate medical staff on proper interpretation. Consulting within an antimicrobial stewardship program (ASP) could enhance the benefits of implementing molecular methods in bloodstream infection diagnostics.

Evaluation of the in vitro susceptibility of clinical isolates of NDM-producing Klebsiella pneumoniae to new antibiotics included in a treatment regimen for infections.

Background: Due to the growing resistance to routinely used antibiotics, the search for new antibiotics or their combinations with effective inhibitors against multidrug-resistant microorganisms is ongoing. In our study, we assessed the in vitro drug susceptibility of Klebsiella pneumoniae strains producing New Delhi metallo-ß-lactamases (NDM) to antibiotics included in the Infectious Diseases Society of America (IDSA) and European Society of Clinical Microbiology and Infectious Diseases (ESCMID) recommendations. Methods: A total of 60 strains of NDM-producing K. pneumoniae were obtained from different patients hospitalized at the 4th Military Hospital in Wroclaw between 2019 and 2022 and subjected to drug susceptibility to selected antibiotics, including the effects of drug combinations. Results: Among the tested antibiotics, the highest sensitivity (100%) was observed for cefiderocol, eravacycline (interpreted according to the European Committee on Antimicrobial Susceptibility Testing [EUCAST]), and tigecycline. Sensitivity to intravenous fosfomycin varied depending on the method used. Using the "strip stacking" method, determining cumulative sensitivity to ceftazidime/avibactam and aztreonam demonstrated 100% in vitro sensitivity to this combination among the tested strains. Conclusion: The in vitro susceptibility assessment demonstrated that, the best therapeutic option for treating infections caused by carbapenemase-producing strains seems to be a combination of ceftazidime/avibactam with aztreonam. Due to the safety of using both drugs, cost effectiveness, and the broadest indications for use among the tested antibiotics, this therapy should be the first-line treatment for carbapenemase-producing Enterobacterales infections. Nevertheless, a comprehensive evaluation of the efficacy of treating infections caused by NDM-producing K. pneumoniae strains should include not only in vitro susceptibility assessment but also an analysis of clinical cases.

Pluronic F-127 Enhances the Antifungal Activity of Fluconazole against Resistant Candida Strains.

Candida strains as the most frequent causes of infections, along with their increased drug resistance, pose significant clinical and financial challenges to the healthcare system. Some polymeric excipients were reported to interfere with the multidrug resistance mechanism. Bearing in mind that there are a limited number of marketed products with fluconazole (FLU) for the topical route of administration, Pluronic F-127 (PLX)/FLU formulations were investigated in this work. The aims of this study were to investigate (i) whether PLX-based formulations can increase the susceptibility of resistant Candida strains to FLU, (ii) whether there is a correlation between block polymer concentration and the antifungal efficacy of the FLU-loaded PLX formulations, and (iii) what the potential mode of action of PLX assisting FLU is. The yeast growth inhibition upon incubation with PLX formulations loaded with FLU was statistically significant. The highest efficacy of the azole agent was observed in the presence of 5.0 and 10.0% w/v of PLX. The upregulation of the CDR1/CDR2 genes was detected in the investigated Candida strains, indicating that the efflux of the drug from the fungal cell was the main mechanism of the resistance.

Candidaemia in polish hospitals - a multicentre survey.

Significant changes in the frequency of candidaemia and the distribution of causative species have been noted worldwide in the last two decades. In this study, we present the results of the first multicentre survey of fungaemia in Polish hospitals. A total of 302 candidaemia episodes in 294 patients were identified in 20 hospitals during a 2-year period. The highest number of infections was found in intensive care (30.8%) and surgical (29.5%) units, followed by haematological (15.9%), 'others' (19.2%) and neonatological (4.6%) units. Candida albicans was isolated from 50.96% of episodes; its prevalence was higher in intensive care unit and neonatology (61.22% and 73.33%, respectively), and significantly lower in haematology (22%; P < 0.001). The frequency of C. krusei and C. tropicalis was significantly higher (24% and 18%) in haematology (P < 0.02); whereas, the distribution of C. glabrata (14.1%) and C. parapsilosis (13.1%) did not possess statistically significant differences between compared departments. Obtained data indicates that species distribution of Candida blood isolates in Polish hospitals reflects worldwide trends, particularly a decrease in the prevalence of infections due to C. albicans.

Synthesis and preliminary biological evaluations of 5'-substituted derivatives of uridine as glycosyltransferase inhibitors.

New derivatives of uridine which contain a b-ketoenol motif were synthesized, characterized and biologically tested. Synthesized compounds 1-4 showed no activity against bovine milk ß-1,4-galactosyltransferase I at concentrations up to 2.0 mM and were not active against Candida albicans and Aspergilus fumigatus up to the maximum tested concentration of 1,000 µg/mL.

Quantitative and Qualitative Airborne Mycobiota Surveillance in High-Risk Hospital Environment.

(1) Background: The primary aim of the presented study was to assess the prevalence of fungi in the indoor air of selected hospital wards, and the additional goal was to evaluate the susceptibility of cultured isolates of Aspergillus fumigatus to triazoles. (2) Methods: Three hematology departments and a hospital for lung diseases were surveyed in 2015 and/or 2019. Air samples were taken with a MicroBio MB1 air sampler on Sabouraud agar. The susceptibility of Aspergillus fumigatus isolates to voriconazole, posaconazole and itraconazole was tested with a microdilution method, according to EUCAST. (3) Results: The amount of fungi cultured from rooms equipped with sterile air circulation, as well as flow devices for air disinfection, was significantly lower compared to that from unprotected rooms. The areas most contaminated with fungi were corridors and bathrooms. The dominant species were Cladosporium and Penicillium. A. fumigatus was rare in hematological departments (6/61, 9.8% examinations performed in 2014 and 2/40, 5% in 2019), whereas in the hospital for lung diseases an outbreak of A. fumigatus spores with up to 300 CFU/m3 was noted in March 2015. No triazole-resistant A. fumigatus isolate was detected. (4) Conclusions: Regular microbiological testing of the hospital environment can contribute to the detection of spore outbreaks, and thus enable the implementation of corrective procedures (e.g., additional disinfection, changing of HEPA filters).

Impact of the COVID-19 Pandemic on the Epidemiology of Bloodstream Infections in Hospitalized Patients-Experience from a 4th Military Clinical Hospital in Poland.

Bloodstream infections (BSIs) are associated with high mortality and inappropriate or delayed antimicrobial therapy. The purpose of this study was to investigate the impact of the COVID-19 pandemic on the epidemiology of BSIs in hospitalized patients. The research aimed to compare the incidence of BSIs and blood culture results in patients hospitalized before and during the COVID-19 pandemic. METHODS: Retrospective and prospective data were collected from blood cultures obtained from 4289 patients hospitalized between June 2018 and July 2022. Two groups of patients were distinguished: those with BSIs admitted during the pre-COVID-19 period and those admitted during the COVID-19 surge. Demographic and clinical data, blood cytology, and biochemistry results were analyzed, and the usefulness of PCT was assessed in patients with COVID-19. RESULTS: The study showed a significant increase in the incidence of BSIs during the pandemic compared to the pre-COVID-19 period. Positive blood cultures were obtained in 20% of patients hospitalized during the pandemic (vs. 16% in the pre-COVID-19 period). The incidence of BSIs increased from 1.13 to 2.05 cases per 1000 patient days during COVID-19, and blood culture contamination was more frequently observed. The mortality rate was higher for patients hospitalized during the COVID-19 pandemic. An increased frequency of MDRO isolation was observed in the COVID-19 period. CONCLUSIONS: The incidence of BSIs increased and the mortality rate was higher in the COVID-19 period compared to the pre-COVID-19 period. The study showed limited usefulness of procalcitonin in patients with COVID-19, likely due to the administered immunosuppressive therapy.

The Effect of High-Pressure Homogenization Conditions on the Physicochemical Properties and Stability of Designed Fluconazole-Loaded Ocular Nanoemulsions.

The growing interest in high-energy emulsification is a result of its scalability, which is important from an industrial perspective and allows for a more reproducible and efficient production of pharmaceutical formulations. The aim of this study was to evaluate the effect of composition, mainly a fixed surfactant/cosurfactant (Smix) ratio, their concentration, and the parameters of high-pressure homogenization (HPH) processing on the quality and stability of ophthalmic fluconazole-loaded nanoemulsions. After a physicochemical analysis of nanoemulsions containing 20% w/w of oil, as optimal conditions for the HPH process, three cycles at a pressure of 1000 bar were established, obtaining formulations with an average droplet diameter size in the range of 80.63-129.68 nm and PDI values below 0.25. While it was expected that an increasing cosurfactant concentration decreased the droplet size, in the case of formulations containing Tween 20 and 10% w/w of cosurfactants, "over-processing" was observed, identified by the droplet size and polydispersity index increase. Consecutively, the selected formulations were evaluated for in vitro drug release in Franz's cell, antifungal activity, and 30-day stability using NMR spectroscopy. An antifungal activity test showed no significant difference in the antifungal activity between optimal fluconazole-loaded nanoemulsions and a 0.3% aqueous drug solution, but previously, research showed that prepared formulations were characterized by a higher viscosity and satisfactory prolonged release compared to a control. In a 30-day stability study, it was observed that higher HLB values of the used surfactants decreased the stability of the formulations in the following order: Kolliphor EL, Tween 80, Tween 20. The NMR spectra confirmed that Kolliphor EL-based formulations ensured the higher stability of the nanoemulsion composition in comparison to Tween 80 and a better stabilizing effect of propylene glycol as a cosurfactant in comparison to PEG 200. Therefore, the optimization of HPH technology should be focused on the selection of Smix and the Smix:oil ratio in order to prepare stable formulations of high quality.

Susceptibility to Clotrimazole of Candida spp. Isolated from the Genitourinary System-A Single Center Study.

The aim of this study was to determine the susceptibility to clotrimazole of 125 isolates of Candida spp. originated from the genitourinary system of hospitalized patients as well as outpatients, tested in the mycological laboratory of Wroclaw Medical University in the years 1999-2018. The minimal inhibitory concentrations of clotrimazole and fluconazole were determined with the use of the microdilution method according to EUCAST, and the MFC was determined by subsequent subculture on Sabouraud agar. For the tested population of Candida yeasts, the MIC values of clotrimazole ranged from 0.008 to 8 mg/L, and MIC90 was 1 mg/L, whereas MIC50 was 0.008 mg/L. The minimal fungicidal concentration ranged between 1 and >8 mg/L. The great majority of the isolates (88%; 110/125) displayed MIC < 1 mg/L and were classified as WT (wild-type), whereas MIC ≥ 1 mg/L was determined for 2/61 (3.2%) isolates of C. albicans, 9/38 (23.6%) of C. glabrata, 1/2 of C. tropicalis, and 3/3 of C. guilliermondii. Six isolates (four of C. glabrata and two of C. albicans), defined as non-WT for clotrimazole, were classified as resistant to fluconazole, according to CBP from EUCAST. The isolates with elevated MIC to clotrimazole originated mostly from patients of the pediatric hematology unit, and their proportion in this population amounted to 17.8% (13 out of 73 isolates). In contrast, among strains from ambulatory patients, the highest observed MIC value was 1 mg/L (1 out of 37 isolates; 2.7%). The data obtained correlate well with those of most published studies on the in vitro susceptibility of Candida spp. to clotrimazole, which is usually very high. However, the existence of reports regarding the growing prevalence of resistant isolates has also to be noted. These results support the need for routinely checking the susceptibility of Candida clinical isolates to this imidazole derivative.