Central Carbon Metabolism in Candida albicans Biofilms Is Altered by Dimethyl Sulfoxide.

The effect of dimethyl sulfoxide (DMSO) on fungal metabolism has not been well studied. This study aimed to evaluate, by metabolomics, the impact of DMSO on the central carbon metabolism of Candida albicans. Biofilms of C. albicans SC5314 were grown on paper discs, using minimum mineral (MM) medium, in a dynamic continuous flow system. The two experimental conditions were control and 0.03% DMSO (v/v). After 72 h of incubation (37 °C), the biofilms were collected and the metabolites were extracted. The extracted metabolites were subjected to gas chromatography-mass spectrometry (GC/MS). The experiment was conducted using five replicates on three independent occasions. The GC/MS analysis identified 88 compounds. Among the 88 compounds, the levels of 27 compounds were markedly different between the two groups. The DMSO group exhibited enhanced levels of putrescine and glutathione and decreased levels of methionine and lysine. Additionally, the DMSO group exhibited alterations in 13 metabolic pathways involved in primary and secondary cellular metabolism. Among the 13 altered pathways, seven were downregulated and six were upregulated in the DMSO group. These results indicated a differential intracellular metabolic profile between the untreated and DMSO-treated biofilms. Hence, DMSO was demonstrated to affect the metabolic pathways of C. albicans. These results suggest that DMSO may influence the results of laboratory tests when it is used as a solvent. Hence, the use of DMSO as a solvent must be carefully considered in drug research, as the effect of the researched drugs may not be reliably translated into clinical practice.

Comparative analysis of dental trauma in contact and non-contact sports: A systematic review.

Dental traumas in sports are common and have physical, social, psychological, and economic impacts. The aim of this study was to determine, through a systematic review, the prevalence of dental trauma in contact and non-contact sports. This review was submitted to PROSPERO (CRD42023421206). Included studies addressed the prevalence of dental trauma in young athletes and adults above 18 years, excluding reviews, editorials, symposiums, or those evaluating athletes under 18 years. A literature search was conducted using the databases PubMed, Web of Science, Scopus, Embase, LIVIVO, SPORTDiscus, Dentistry & Oral Sciences Source (via EBSCO), and Lilacs and BBO, as well as gray literature. Bias risk was assessed using the Joanna Briggs Institute's Critical Appraisal Checklist. Data were synthesized considering study characteristics, population, sport, and outcomes. R Statistics software was used for all meta-analyses. A total of 1707 articles were identified. After applying eligibility criteria, eight were selected. Three studies, not previously observed, were later added after reading four systematic reviews on a similar topic. Fourteen contact sports and five non-contact sports were analyzed. The prevalence of dental trauma was 11.38% in contact sports and 5.24% in non-contact sports. Regardless of the type of sport, athletes face risks of dental trauma, with contact sports showing higher prevalence. The use of mouthguards is essential across all contact and non-contact sports as a preventive measure.

Cleaning and disinfection of the root canal system provided by four active supplementary irrigation methods.

This in vitro study evaluated the bacterial reduction provided by the EndoActivator (EA), Easy Clean (EC), passive ultrasonic irrigation (PUI), and XP-Endo Finisher. Eight-four mesial roots of mandibular first molars were instrumented, inoculated with Enterococcus faecalis, and divided into four groups (n. 20). Bacterial reduction in the main canals and dentinal tubules were respectively determined by MTT assays and Live/Dead BackLight technique through confocal laser scanning microscopy (CLSM) at 50, 100, and 150 µm in-depth (n. 10 per group). Statistical analyses were conducted following a significance level of 95% (P < 0.05). A significant statistical difference was just identified between XPF and EC in the main canals. In the dentinal tubules from the main root canals, at 100 and 150 µm in-depths, significant statistical differences were only observed between XPF and EC (P = 0.027) for the former and between XPF and EC (P = 0.011) and XPF and PUI (P = 0.021) for the latter. In the dentinal tubules from the isthmus, at 100 µm in-depth, statistically relevant differences did occur between XPF and EC (P = 0.038) and EC and EA (P = 0.029). At 150 µm in-depth, these differences were only significant by comparing XPF and PUI (P = 0.025) and XPF and EC (P = 0.036). Although no irrigation method could thoroughly disinfect the RCS, bacterial reduction indexes were generally better after using XPF.

Utilizing intramammary Melaleuca alternifolia as an organic internal sealant for dry-off therapy in Murrah buffaloes.

The effects of intramammary dry cow therapy based on the administration of 5% Melaleuca alternifolia tea tree essential oil (TTO) as an internal teat sealant to Murrah cows were evaluated. A longitudinal prospective and retrospective negative control study was performed using 12 buffaloes from a total of 20 Murrah buffaloes on an organic farm, with the cow used as a control for herself. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for treatments with pure oil (TTO) and medication containing 5% TTO (O5) were determined. The buffaloes were clinically examined, and the teats were evaluated using thermography and ultrasound. Udder health was monitored during the first 100 days in milk (DIM) using milk somatic cell count (SCC) and California mastitis test (CMT). Laboratory tests against standard strains Staphylococcus aureus ATCC®25,923™, Escherichia coli ATCC®25,922™, and wild bacterial strains showed maximum MIC values of 50 µL/mL for the TTO and O5 treatments. One wild-type S. aureus strain showed no MBC. No adverse effects were observed after the intramammary application of TTO. The CMT and SCC values were similar (P > 0.05) for all observations. The medication containing 5% TTO was effective in vitro and compatible with the intramammary tissue in vivo of Murrah buffaloes. TTO was safe, not inducing inflammatory processes or other modifications of the teat detectable by thermography or ultrasound. It was able to protect buffaloes during the dry period under field conditions, demonstrating potential use as a teat sealant for organic farms.

Oral Colonization and Virulence Factors of Candida spp. in Babies With Cleft Palate.

OBJECTIVE: To examine oral colonization and virulence factors of Candida spp. in patients aged from 0 to 18 months with cleft palate (CP). MATERIALS AND METHODS: Sixty babies were allocated into 3 groups: CP, CP with orthodontic plate (CPwP), and control group (Ctrl) without CP. Information on feeding habits, hygiene, and history of candidosis was collected. The presence of Candida spp. was investigated in samples of saliva. Fungal hydrophobicity, protease, esterase, phospholipase, and hemolysin were evaluated in a semiquantitative manner. RESULTS: Positive oral isolations of Candida spp. were detected in CP (89.5%), CPwP (100%), and Ctrl (44%) groups. Candidosis was more reported in the cleft groups than in the Ctrl group (P ≤ .023). There was a higher prevalence of Candida albicans, followed by Candida krusei, Candida tropicalis, and Candida parapsilosis in all groups. There was no uniformity of expression of virulence factors, either among different species or among different groups. CONCLUSION: Candida spp. colonization occurred in all groups, being superior in CPwP group. Candidosis episodes were more reported in patients from CPwP than in other groups, although candidosis was also registered in other groups. Candida albicans was the predominant species and virulence factors did not exhibit any pattern for species or groups of patients.

Randomized clinical trial of complete denture cleaning methods in residents of care institutions

Aim: To evaluate the efficacy of ultrasonic bath and traditional brushing in the hygiene of complete dentures of dependent residents in long-term care institutions (LTCIs). Methods: A randomized cross-over clinical study was conducted in 17 maxillary complete denture wearers living in LTCIs. Cleaning protocols were brushing or ultrasonic bath, both applied with neutral liquid soap. Biofilm biomass were estimated by MTT reduction assay and specific microbial load (CFU) of Streptococcus spp., Staphylococcus spp. and Candida spp. were quantified by selective and differential culture media. Results: Ultrasound method showed higher percentage reduction of biofilm biomass and specific microbial loads of Streptococcus spp. compared to brushing (p<0.01). Reduction of microbial loads of the other microorganisms were not different between cleaning methods (p>0.05). Conclusions: The ultrasonic bath proved to be a feasible alternative method for the mechanical cleaning of complete dentures in LTCIs

Bacteriophages as tools for biofilm biocontrol in different fields.

Microbial biofilms are difficult to control due to the limited accessibility that antimicrobial drugs and chemicals have to the entrapped inner cells. The extracellular matrix, binds water, contributes to altered cell physiology within biofilms and act as a barrier for most antiproliferative molecules. Thus, new strategies need to be developed to overcome biofilm vitality. In this review, based on 223 documents, the advantages, recommendations, and limitations of using bacteriophages as 'biofilm predators' are presented. The plausibility of using phages (bacteriophages and mycoviruses) to control biofilms grown in different environments is also discussed. The topics covered here include recent historical experiences in biofilm control/eradication using phages in medicine, dentistry, veterinary, and food industries, the pros and cons of their use, and the development of microbial resistance/immunity to such viruses.

Direct detection of microorganisms in sonicated orthopedic devices after in vitro biofilm production and different processing conditions.

BACKGROUND: The gold standard for microbial detection in prosthetic joint infections is the multiple culture of the peri-prosthetic tissue. The fluid cultures after sonication can improve the recovery of the microorganisms. OBJECTIVE: The aim of this study was to evaluate the sonication technique with a plastic bag and the effect of refrigeration on microorganism detection with conventional culturing, MALDI-TOF MS and qPCR assay on an orthopedic screw model. METHODS: We produced biofilms of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans on orthopedic screws, which were stored under different conditions and temperatures before sonication. After sonication, the mass spectrometry by MALDI-TOF, qPCR and culture protocols was performed using the sonicated fluid, for detecting the microorganisms involved in the biofilm. RESULTS: The bacterial bioburden decreased by approximately one log after the refrigeration period, in the screws containing P. aeruginosa and S. aureus biofilms. All the microorganisms involved in the screw biofilms were detected with MALDI-TOF and qPCR. Significant reductions in CFU counts occurred only in groups stored in the plastic bag, indicating that changes in temperature and humidity may favor cell death. However, this variation is not important for this model as it did not affect the detection owing to the high counts obtained. CONCLUSION: Microbial identification by MALDI-TOF in sonicated fluid is feasible. With qPCR, there were no differences between the detection in the screws processed immediately or after refrigeration. It is necessary to consider whether or not the refrigeration period would affect microbial recovery in an explanted prosthesis.

Salivary proteome analysis of crack cocaine dependents.

OBJECTIVE: Salivary proteomic analysis may help to understand physiopathological changes in crack cocaine dependents. This study aimed to compare the salivary protein profile between crack cocaine dependents and non-drug users. DESIGN: Nine heavy smokers and alcohol consumers men admitted to rehab due to crack cocaine abuse and nine non-drug users age-matched men were evaluated. Unstimulated whole saliva was collected. Proteomic analysis was performed by mass spectrometer. Data were processed using ProteinLynx GlobalServer software. Results were obtained by searching the Homo sapiens database from the UniProt catalog. The search tool IBI-IMIM was used to identify proteins candidates for biomarkers. RESULTS: The mean age of crack cocaine and control groups was 36.89 ±â€¯7.78 and 35.78 ±â€¯6.68 years, respectively. 458 salivary proteins were identified in both groups; 305 proteins in the crack cocaine group. Among the 68 proteins presented in both groups, 29 were down-regulated (i.e. "Statherin" and "Transforming growth factor-beta-induced protein ig-h3" were down-regulated at least 10-fold) and 27 up-regulated (i.e. "Negative elongation factor" was up-regulated 19-fold) in the crack cocaine group compared to controls. 90 out of the 458 proteins found in the proteomic analysis were identified as candidates for biomarkers of diseases. Among these, 65 (72.22 %) were detected in the crack cocaine group. CONCLUSION: Crack cocaine dependents with chronic alcohol and tobacco use have a higher number of proteins in saliva compared to non-drug users. 22.3 % of salivary proteins present in crack cocaine dependents were present in controls; 3.9 % of them were expressed in similar quantity.

Late Oral Complications Caused by Head and Neck Radiotherapy: Clinical and Laboratory Study.

OBJECTIVES: The aim of presented cross-sectional and observational study was to determine the prevalence of late oral complications of patients with head and neck cancer who underwent radiotherapy, by clinical and laboratory analyses. MATERIAL AND METHODS: Fifty-five patients, 43 (78.2%) men and 12 (21.8%) women, mean age 60; range 38 to 87 years, who have completed radiotherapy for head and neck cancer for at least 6 months were enrolled. The presence of xerostomia, hyposalivation, oral candidiasis, and type of oral yeasts were correlated with post-radiotherapy period. A control group, age and gender matched, was used for comparisons. The Pearson's Chi-square or Fischer's exact test was used at a significance level of 5%. RESULTS: The mean post-radiotherapy period was 32 months. The oral complications found were xerostomia (45/55, [81.8%]), hyposalivation (44/55 [80%]) and oral candidiasis (15/55 [27.2%]). Xerostomia and hyposalivation was statistically higher in the study group when compared to the control group (P < 0.05). The presence of yeast occurred in 39 (70.9%) of the patients in the study group, and Candida albicans was the most prevalent etiological agent in 25 (64.1%) of those patients (P < 0.05). CONCLUSIONS: Xerostomia and hyposalivation were the more prevalent late oral complications related to radiotherapy. Oral candidiasis was also observed, although its prevalence was lower. The need for long-term dental follow-up of patients who underwent radiotherapy of the head and neck cancer is mandatory.

Prevalence and profile of users and non-users of anabolic steroids among resistance training practitioners.

BACKGROUND: To verify the prevalence and profile of users and non-users of anabolic steroid (AS) among resistance training practitioners. METHODS: An observational, cross-sectional survey was performed in 100 gyms in Curitiba city, involving 5773 individuals and self-administered questionnaires. The chi-square and z-tests of proportions were used for comparison between the groups (p < 0.05). RESULTS: 83.2% did not use, 9.1% formerly used, 3.4% currently used, and 4.3% intended used AS. The prevalence of former or current AS users was 16.9 and 6.5% among men and women, respectively. The prevalence ratios were as follows: 1) 2.6 male users for each woman; 2) 3.3 individuals aged 30-44 years and 2.8 individuals aged 18-29 years for each individual aged over 45 years. Beginners were not interested in using AS, but individuals who had trained longer had higher prevalence of AS use. CONCLUSIONS: The gym environment encouraged the use of AS owing to aesthetic appeal. Thus, suggesting the need for actions to prevent abusive use of AS considering the practitioners profile (practitioners were young, university and single).

Correction: Anabolic steroids among resistance training practitioners.

[This corrects the article DOI: 10.1371/journal.pone.0223384.].

Anabolic steroids among resistance training practitioners.

OBJECTIVE: To compare the use of anabolic steroids (AS), the motivation to use them, their side effects, the source of information and the form in which AS were obtained, the medical follow-up, and the periodic examinations in resistance training practitioners who are either current or former users of AS. METHODS: A prevalence survey was performed in the gyms of the city of Curitiba, including 719 current and former AS users who self-administered a questionnaire. The chi-square and z of proportions (p <0.05) statistical tests were conducted. RESULTS: Esthetics was the main motivation associated with AS intake, leading to satisfactory results. The information about the form in which to use AS was provided by doctors and AS were either purchased at the pharmacy with a prescription or illegally. Current users reported a higher number of cycles and doses, a longer duration of use, as well as larger economical investments into AS. This shows a higher consumption of such drugs, regardless of the medical follow-up and post-cycle therapy. CONCLUSION: Given that a change in the usage pattern was observed when increasing the AS consumption, this should be considered in the elaboration of public policies to inhibit such a trend.

Salivary proteome characterization of alcohol and tobacco dependents.

BACKGROUND: Alcohol and substances found in tobacco may alter salivary flow and amount of saliva proteins. This study aimed to compare salivary proteins between alcohol dependent smokers and controls. METHODS: This is a case-control study with men older than 18 years of age, matched by age. The alcohol-dependent group was composed by heavy smokers and alcohol consumers. Unstimulated whole saliva was collected from all subjects. Analysis of digested peptides was performed in mass spectrometer. Data were processed using ProteinLynx GlobalServer software. Results were obtained by searching theHomo sapiens database from the UniProt catalog. The search tool IBI-IMIM was used to identify candidate proteins for biomarkers. RESULTS: Alcohol-dependent and control groups were composed of nine participants each, with mean age of 36.89 ±â€¯2.57 and 35.78 ±â€¯1.64 years, respectively. 404 salivary proteins were found in both groups; 282 in the alcohol-dependent. Among the 96 proteins presented in both groups, 32 were up-regulated in the alcohol dependents (i.e. "Hemoglobin subunit beta" and "Forkhead box protein P2" were up-regulated at least 10-fold), 23 were down-regulated (i.e. "Statherin" and "RNA-binding protein 25" were down-regulated at least 10-fold), and 41 presented similar expression in both groups. 71 proteins were candidates for biomarkers of disorders 58 presented in alcohol dependents' saliva. The most common disorders were neoplasms, genetic, cardiovascular, metabolic and glandular diseases. CONCLUSIONS: Salivary protein profile undergoes strong changes in alcohol and tobacco dependents. 34% of salivary proteins present in alcohol and tobacco dependents were present in controls; 14.5% of them were expressed in similar quantity.

Sialometric and Sialochemical Changes in Morbidly Obese Patients before and after Bariatric Surgery.

AIM: To evaluate the sialometric and sialochemical changes in morbidly obese patients before and after bariatric surgery. MATERIALS AND METHODS: A total of 74 participants were divided into three groups paired by sex and age: (a) Experimental 1 (E1)-morbid obesity (n = 40)-according to the Fobi-Capella technique, corresponding to the individuals with a body mass index (BMI) of greater than 40 kg/m2 prior to bariatric surgery; (b) Experimental 2 (E2)-the same individuals after surgery; and (c) control (C) (n = 34)-individuals with a BMI of nearly 23 kg/m2. The measure of salivary flow was carried out by collecting stimulated saliva. The pH was evaluated using the pocket pH meter, while the salivary buffer capacity (SBC) was determined by the titration method. Analyses of the salivary concentration of total proteins (TPs), amylase activity, urea, calcium (Ca++), and glucose were evaluated using the calorimetric method. RESULTS: Group E1, as compared with group C, presented the highest pH levels (p = 0.03), amylase activity (p = 0.00), and calcium (p = 0.00). The opposite was observed for glucose (p = 0.00), TP (p = 0.04), and urea (p = 0.04). Group E2, as compared with group C, revealed higher levels of amylase (p = 0.00) and calcium (p = 0.00). The opposite was observed for SBC (p = 0.01), PT (p = 0.00), and glucose (p = 0.00). Group E1, as compared with group E2, presented higher values of SBC (p = 0.00) and urea (p = 0.00). The lowest values were found for calcium and urea (p = 0.03). CONCLUSION: Both weight gain and bariatric surgery are risk factors for the oral condition, causing change in some important salivary components, such as TP, amylase, calcium, and glucose. CLINICAL SIGNIFICANCE: This article is a valuable addition to the scientific literature, due to its novelty. There are no papers that show salivary alterations related to bariatric surgeries.

The apoptotic effect of midazolam on the salivary gland of rats and the reversal effect by pilocarpine.

To evaluate the apoptosis in parotid glands of rats treated with midazolam associated or not with pilocarpine, 60 Wistar rats were assigned to 6 groups: control groups received saline solution for 30 days (S30) and 60 days (S60) and the other groups received pilocarpine for 60 days (P60), midazolam for 30 days (M30), midazolam for 30 days and 30 days of saline (M30 + S30), and finally midazolam for 30 days and 30 days of midazolam and pilocarpine (M30 + MP30). Histological sections were subjected to the TdT-mediated dUTP-biotin nick and labeling technique. The number of positive and negative cells was quantified, calculating the apoptotic index. ANOVA at 2 criteria and Tukey's test were used. A greater apoptotic index was observed in the M30 (52.79 ± 9.01) and M30 + S30 (62.43 ± 8.52) groups when compared with the S30 (37.94 ± 5.94) and S60 (31.85 ± 9.18) groups, respectively (p < 0.05). There was no difference between M30 + MP30 (30.98 ± 6.19) and S60 (31.85 ± 9.18) groups regarding apoptotic index. Chronic administration of midazolam has been shown to increase the number of apoptotic cells in the parotid glands of rats. However, pilocarpine inhibited this effect, thus inhibiting the apoptosis.

Dynamics of the seasonal airborne propagation of Staphylococcus aureus in academic dental clinics.

OBJECTIVE: Staphylococcus aureus strains can be disseminated during dental treatments and occasionally lead to the contamination and infection of patients and dentists, which is an important public health problem. The dynamics of the airborne propagation and the genetic diversity of S. aureus isolated in an academic dental clinic environment were investigated using isoenzyme typing. Materials and MethodsThe isoenzymes of 44 previously reported isolates were obtained from fresh cultures and extracted using glass beads. Nine isoenzymes were investigated using multilocus enzyme electrophoresis (MLEE). The genetic diversity and relationship among the strains (electrophoretic type - ET) were determined using statistics previously described by Nei25 (1972) and the SAHN grouping method (UPGMA algorithm). RESULTS: Clonal pattern analyses indicated a high level of genetic polymorphism occurring among the 33 ETs, which were grouped into five taxa. Each taxon presented one or more clusters that were moderately related and that contained two or more identical/highly related isolates, revealing seasonal airborne propagation in these dental clinic environments. CONCLUSIONS: These data suggest the occurrence of active microevolutionary processes in S. aureus as well as the possibility of environmental propagation during a 14-month time span. Such findings are important to show that multiuser academic dental clinics can retain certain strains that are spreadable to different niches.

Dynamics of the seasonal airborne propagation of Staphylococcus aureus in academic dental clinics

Abstract Objective Staphylococcus aureus strains can be disseminated during dental treatments and occasionally lead to the contamination and infection of patients and dentists, which is an important public health problem. The dynamics of the airborne propagation and the genetic diversity of S. aureus isolated in an academic dental clinic environment were investigated using isoenzyme typing. Material and Methods The isoenzymes of 44 previously reported isolates were obtained from fresh cultures and extracted using glass beads. Nine isoenzymes were investigated using multilocus enzyme electrophoresis (MLEE). The genetic diversity and relationship among the strains (electrophoretic type - ET) were determined using statistics previously described by Nei25 (1972) and the SAHN grouping method (UPGMA algorithm). Results Clonal pattern analyses indicated a high level of genetic polymorphism occurring among the 33 ETs, which were grouped into five taxa. Each taxon presented one or more clusters that were moderately related and that contained two or more identical/highly related isolates, revealing seasonal airborne propagation in these dental clinic environments. Conclusions These data suggest the occurrence of active microevolutionary processes in S. aureus as well as the possibility of environmental propagation during a 14-month time span. Such findings are important to show that multiuser academic dental clinics can retain certain strains that are spreadable to different niches.