Influence of Incubation Time on Ortho-Toluidine Blue Mediated Antimicrobial Photodynamic Therapy Directed against Selected Candida Strains-An In Vitro Study.

(1) Background and the aim: The appropriate incubation time in the antimicrobial photodynamic therapy protocol seems to have a huge impact on the efficacy of this process. This is particularly important in relation to Candida strains, due to the size of these cells and the presence of the cell wall. The aims of this study were to determine the optimal incubation time needed for the absorption of toluidine blue by cells of C. albicans, C. glabrata, C. krusei and C. parapsilosis using direct observation by optical microscopy, and to evaluate the efficacy of TBO-mediated aPDT on planktonic cells of these strains. (2) Methods: The microscopic evaluation consisted of taking a series of images at a magnification of 600× and counting the % of stained cells. The in vitro effect of TBO-mediated aPDT combined with a diode laser (635 nm, 400mW, 12 J/cm2, CW) on the viability of yeast cells with different incubation times was evaluated. (3) Results: The presence of TBO within the cytoplasm was observed in all tested Candida strains and at all microscopic evaluation times. However, the highest percentages of cells were stained at 7 and 10 min. The highest % reduction of CFU/mL after TBO-mediated aPDT against Candida was obtained for the strain C. albicans ATCC 10,231 and it was 78.55%. (4) Conclusions: TBO-mediated aPDT against Candida was effective in reducing the number of CFU/mL at all assessed incubation times. However, the most efficient period for almost all strains was 7-10 min.

Oral cavity fungal flora among HIV-positive people.

BACKGROUND: Immunosuppressed patients, also those who are HIV-positive patients, are susceptible to oral cavity fungal infections. AIM OF STUDY: In this study, we aimed to show differences in qualitative composition of oral cavity flora between HIV-positive people and healthy controls and identify factors which affect fungal oral cavity flora. MATERIALS AND METHODS: The study group contained HIV-positive people and a control group of healthy people. All cultured species were analysed using MALDI-TOF MS. RESULTS: More HIV-positive people had two or more fungus species present than controls (p=0.008). Seven species were cultured in the study group compared to three in the control group. Smoking was associated with higher prevalence of C. albicans (p=0.03), C. glabrata (p=0.026), C. tropicalis (p=0.01). Dental prosthesis or braces was also associated with presence of more species (p=0.04).The lower level of lymphocytes CD4+ was not associated with fungus presence in oral cavity. CONCLUSIONS: HIV infection is associated with changes to oral cavity fungal flora. Given the higher number of non-albicans species among HIV-positive patients it is important to individually choose a treatment for such patients' fungal infections. Proper oral hygene and not smoking can reduce prevalence of fungi in oral cavity. Patients' immunological status did not have an impact on the frequency of Candida species isolation from the oral cavity.

Effects of Nd:YAG laser irradiation on the growth of Candida albicans and Streptococcus mutans: in vitro study.

The purpose of this study was to evaluate the effects of Nd:YAG laser with flat-top handpiece on the in vitro growth of Candida albicans and Streptococcus mutans. The incidence of C. albicans (opportunistic commensal) and S. mutans (facultatively anaerobic) infections is increasing, despite available treatments. Cultures of Streptococcus mutans and Candida albicans were irradiated using Nd:YAG laser (LightWalker, Fotona) with flat-top handpiece (Genova, LightWalker, Fotona) at the following parameters: group G1: 0.25 W, 10 Hz, 15 s, 3 J and group G2: 1 W, 10 Hz, 60s, 59 J. The results were evaluated directly and 24 h after irradiation using a quantitative culture method (estimation of colony-forming units in 1 ml of suspension, cfu/ml), and microscopic analysis with Janus green stain and compared with control group in which laser was not applied. C. albicans was reduced by 20 up to 54% for G1, and for G2 by 10 up to 60% directly after the application. The cfu/ml values for S. mutans decreased by 13% (p = 0.1771) for G1 and 89% (p < 0.0001) for G2. In both test groups 24 h after the application, the number of colony-forming units decreased by 15-46% for G1 and by 15-64% for G2. The arrested cell division, increasing the surface area and increasing the number of metabolically inactive cells, were observed in morphometric analysis. Macroscopic and microscopic analyses revealed a reduction in cell number and a significant decrease of cell metabolism after laser application for both C. albicans and S. mutans.

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.

Three-Dimensional-Printed Modular Titanium Alloy Plates for Osteosynthesis of the Jawbone.

BACKGROUNDS: The titanium-aluminum-vanadium alloy (Ti-6Al-4V) is frequently used in implantology due to its biocompatibility. The use of 3D printing enables the mechanical modification of implant structures and the adaptation of their shape to the specific needs of individual patients. METHODS: The titanium alloy plates were designed using the 3D CAD method and printed using a 3D SLM printer. Qualitative tests were performed on the material surface using a microcomputed tomography scanner. The cytotoxicity of the modular titanium plates was investigated using the MTT assay on the L929 cell line and in direct contact with Balb/3T3 cells. Cell adhesion to the material surface was evaluated with hFOB1.19 human osteoblasts. Microbial biofilm formation was investigated on strains of Lactobacillus rhamnosus, Staphylococcus epidermidis, Streptococcus mutans and Candida albicans using the TTC test and scanning electron microscopy (SEM). RESULTS: The surface analysis showed the hydrophobic nature of the implant. The study showed that the titanium plates had no cytotoxic properties. In addition, the material surface showed favorable properties for osteoblast adhesion. Among the microorganisms tested, the strains of S. mutans and S. epidermidis showed the highest adhesion capacity to the plate surface, while the fungus C. albicans showed the lowest adhesion capacity. CONCLUSIONS: The manufactured modular plates have properties that are advantageous for the implantation and reduction in selected forms of microbial biofilm. Three-dimensional-printed modular titanium plates were investigated in this study and revealed the potential clinical application of this type of materials, regarding lack of cytotoxicity, high adhesion properties for osteoblasts and reduction in biofilm formation. The 3D CAD method allows us to personalise the shape of implants for individual patients.

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.

Effect of Camel Peptide on the Biofilm of Staphylococcus epidermidis and Staphylococcus haemolyticus Formed on Orthopedic Implants.

The increasing bacterial drug resistance and the associated challenges in the treatment of infections warrant the search for alternative therapeutic methods. Hope is placed in antimicrobial peptides, which have a broad spectrum of action and are effective against strains which are resistant to conventional antibiotics. Antimicrobial peptides are also tested for their efficacy in the treatment of infections associated with the formation of biofilm. The aim of the present study was to examine the effect of Camel peptide on S. epidermidis and S. haemolyticus adhesion to and formation of biofilm on steel cortical bone screws and also on the process of reducing mature biofilm in orthopedic implants. The tests were performed on steel implants for osteosynthesis. The MIC value and MBEC values of the peptide were determined using the microdilution method in microtiter plates. The effect of the peptide on adhesion and biofilm formation, as well as on the activity on the preformed biofilm, was evaluated using quantitative methods and confocal microscopy. The presented research results indicate that the peptide exhibits very good antimicrobial properties against the analyzed strains. Concentrations above MIC reduced biofilm in the range of 90-99%.

Evaluation of the microbial, cytotoxic and physico-chemical properties of the stainless steel crowns used in pediatric dentistry.

Preformed stainless steel crowns are used in pediatric dentistry to obtain full crown restoration of primary molar teeth. They are consider the best restoration in terms of durability and effectiveness. The purpose of this study is to evaluate microbial, cytological and physio-chemical properties to determine whereas stainless steel crown are biocompatible, safe for surrounding tissue and helpful in avoiding micro-organisms influence on the tooth tissue. Based on the results, it was determined that stainless steel crowns used in pediatric dentistry represent no cytotoxic risk to the surrounding tissues, have a low probability of developing hypersensitivity to the coronal material and also that their biological properties make them suitable to use in pediatric dentistry for the reconstruction of damaged primary molar tissue.

Identification of Fungi Isolated from Oral Cavity of Patients with HIV Using MALDI-TOF MS.

BACKGROUND: A growing incidence of invasive fungal infections, especially among immunocompromised patients, has given increased significance to microbiological diagnostics of yeast-like fungi. More accurate and faster fungi identification methods that can compete with classical methods are being searched for. In this paper, classical microbiological methods are compared to MALDI-TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry). METHODS: The diagnostic material was collected from buccal mucosa from 98 adults, including 69 with HIV. Only positive cultures were included in the study. RESULTS: Matching results were obtained in 45 samples, and there were nonmatching results in 35 samples, with the majority of these in the study group, constituting 50% of identifications within this group. A particularly common mistake resulting from the use of classical methods is the false identification of C. dubliniensis as C. albicans. Additionally, C. tropicalis proves to be difficult to identify. CONCLUSIONS: Our results and literature data suggest that MALDI-TOF MS should be considered an effective alternative to classical methods in terms of fungi identification, especially among HIV-positive patients, due to the different morphology of fungal colonies.

Does Systemic Methotrexate Therapy Induce Azole Resistance among Endogenous Candida Strains?

BACKGROUND: Research confirms that Candida spp. incubated with methotrexate develop multi-drug resistance to azoles, but it is not clear whether this phenomenon occurs in vivo in patients treated with cytostatics. The aim of the study was to assess whether systemic methotrexate therapy induces resistance to azoles among endogenous Candida strains in patients with rheumatological diseases. METHODS: The test group consisted of 52 rheumatological patients on methotrexate therapy, who have never been exposed to fluconazole. The control group was composed of 49 individuals who have never been exposed to either methotrexate or fluconazole. Oral swab and clinical information were obtained from each participant. The acquired material was cultured, then each strain was isolated and identified (MALDI TOF). Subsequently, minimal inhibitory concentration (MIC) for fluconazole was determined. RESULTS: MIC values ranged from <0.125 to 64 µg/mL with the most common result <0.125 µg/mL. Samples obtained from 4 patients of the test group and 2 patients of the control group contained strains resistant to fluconazole. CONCLUSIONS: Despite slightly higher incidence of fluconazole-resistant strains among patients on systemic methotrexate therapy, we found no solid evidence to support the hypothesis that methotrexate induces resistance to azoles among endogenous Candida strains in patients with rheumatological diseases.

rDNA-based genotyping of clinical isolates of Candida albicans.

The study presents an analysis of the restriction pattern ofrDNA fragments of 95 C. albicans isolates previously classified on the basis of the presence of the intron in rDNA into genotypes A (62 isolates), B (28), and C (5). Most isolates (61) with genotype A were classified as "subtype a" and one as "subtype d" (Karahan and Akar; 2005). No differences were observed in the restriction patterns of the tested genotype B isolates. Similarly, most genotype C strains (4/5) showed the same restriction pattern. The results indicate low subtyping variations of the analyzed isolates, which is in contrast to published data obtained from a Turkish collection of yeasts.

Assessment of cytotoxic and antimicrobial activity of selected gingival haemostatic agents - in vitro study.

PURPOSE: The present research aimed to determine whether and how the aluminium chloride - based materials affect the cell line of the bacterial line and fungi. METHODS: Cytotoxicity of haemostatic astringents: Alustat (liquid), Alustat (gel), Alustat (foam), Alustin, Hemostat, Racestyptine and Traxodent containing AlCl3 was conducted on L929 cell line with the use of MTT and SRB assays. The antimicrobial activity (CFU and MIC) against C. albicans, S. mutans, L. rhamnosus was determined. RESULTS: In the MTT results, cell viability for all agents were very low. In SRB, the lowest cytotoxicity was demonstrated for Hemostat and Alustat (foam), Traxodent and Racestyptine. Total reduction of the CFU of S. mutans was observed. Alustat (gel) and Alustat (liquid) completely inhibited the growth of C. albicans, S. mutans and L. rhamnosus. CONCLUSIONS: The viability of L929 cells obtained in the SRB assay is more reliable than that obtained in the MTT assay, in the case of gingival haemostatic agents.

The Influence of Ozonated Olive Oil-Loaded and Copper-Doped Nanohydroxyapatites on Planktonic Forms of Microorganisms.

The research has been carried out with a focus on the assessment of the antimicrobial efficacy of pure nanohydroxyapatite, Cu2+-doped nanohydroxyapatite, ozonated olive oil-loaded nanohydroxyapatite, and Cu2+-doped nanohydroxyapatite, respectively. Their potential antimicrobial activity was investigated against Streptococcus mutans, Lactobacillus rhamnosus, and Candida albicans. Among all tested materials, the highest efficacy was observed in terms of ozonated olive oil. The studies were performed using an Ultraviolet-Visible spectrophotometry (UV-Vis), electron microscopy, and statistical methods, by determining the value of Colony-Forming Units (CFU/mL) and Minimal Inhibitory Concentration (MIC).

Study of Surface Structure Changes for Selected Ceramics Used in the CAD/CAM System on the Degree of Microbial Colonization, In Vitro Tests.

In the article has been presented an analysis of susceptibility of selected dental materials, made in the CAD/CAM technology. The morphology and structural properties of selected dental materials and their composites were determined by using XRPD (X-ray powder diffraction) techniques, as well as the IR (infrared) spectroscopy. Moreover, an adhesion as well as development of biofilm by oral microorganisms has been studied. It has been shown that a degree of the biofilm development on the tested dental materials depended on microorganism genus and species. Streptococcus mutans has demonstrated the best adhesion to the tested materials in comparison with Candida albicans and Lactobacillus rhamnosus. However, the sintered materials such as IPS e.max® and the polished IPS e.max® have showed the best "anti-adhesive properties" in relation to S. mutans and L. rhamnosus that have not formed the biofilm on the polished IPS e.max® sample. Furthermore, S. mutans have not formed the biofilm on both surfaces. On the contrary to S. mutans and L. rhamnosus, C. albicans has demonstrated the adhesive properties in relation to the above-mentioned surfaces. Moreover, in contrast to S. mutans and C. albicans, L. rhamnosus has not formed the biofilm on the polished IPS Empress material.