Cunninghamella echinulata DSM1905 biofilm-based L-asparaginase production in pneumatically-driven bioreactors.

We evaluated by comparing the performance of three pneumatically-driven bioreactors in the production of L-asparaginase (L-ASNase), an enzyme used to treat leukaemia and lymphoma. A two-step screening process was conducted to detect Cunninghamella spp. strains producing L-ASNase. Cunninghamella echinulata DSM1905 produced the highest levels of L-ASNase during screening assays. Subsequently, fermentations were performed in bubble column (BCR), airlift (ALR), and hybrid fixed-bed airlift (FB-ALR) bioreactors to determine the best upstream bioprocess. Mycelial biomass production was higher in BCR than in ALR and FB-ALR (p ≤ 0.0322). The activity of L-ASNase produced in FB-ALR, in which the fungus grew as a consistent biofilm, was significantly higher (p ≤ 0.022) than that from ALR, which was higher than that of BCR (p = 0.036). The specific activity of ALR and FB-ALR presented no differences (p = 0.073), but it was higher than that of BCR (p ≤ 0.032). In conclusion, C. echinulata DSM1905, grown under the biofilm phenotype, produced the highest levels of L-ASNase, and FB-ALR was the best upstream system for enzyme production.

Analysis of osseointegration of implants with macrogeometries with healing chambers: a randomized clinical trial.

BACKGROUND: To verify the influence of macrogeometry with healing chambers on the osseointegration of dental implants by analyzing implant stability quotient (ISQ) and evaluate the correlation between insertion torque and ISQ insertion with different macrogeometries. METHODS: In total, 26 implants were installed in the posterior mandible of eight patients with sufficient bone height for the installation of implants measuring 3.5 mm in diameter and 9.0 mm in length. The implants were categorized according to two types of macrogeometry: a test group (GT) with 13 conical implants with healing chambers and a control group (GC) with 13 conical implants with conventional threads. To insert the implants, a bone drilling protocol was used up to a diameter of 3 mm with the last helical bur. The insertion torque of the implants was evaluated, followed by the measurement of ISQ at 0 (T-0), 7 (T-7), 14 (T-14), 21 (T-21), 28 (T-28), and 42 (T-42) days. RESULTS: The mean insertion torque was 43 Ncm in both groups, without a significant difference. Moreover, no significant difference in the ISQ values was found between the groups at different time points (p > 0.05), except at T-7 (GT = 69.87±1.89 and GC = 66.48±4.49; p = 0.01). Although there was no significant difference, ISQ median values were higher in the GT group than GC group at 28 days (GT = 67.98 and GC = 63.46; p = 0.05) and 42 days (GT = 66.12 and GC = 60.33; p = 0.09). No correlation was found between the insertion torque and ISQ insertion (p > 0.05). CONCLUSION: Furthermore, implants with a 3.5 mm diameter macrogeometry, with or without healing chambers, inserted with a drilling protocol up to 3 mm in diameter of the last helical bur, led to a similar secondary stability, with no difference in ISQ values. Although, implants with healing chamber demonstrates ascending values in the graph of ISQ, having a trend of faster osseointegration than implants without healing chambers. Both macrogeometries provide a similar primary stability to implants. TRIAL REGISTRATION: This study was registered retrospectively in ReBec (brazilian registry of clinical trials) under the number RBR-96n5×69, on the date of 19/06/2023.

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.

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.

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.

Correction: Anabolic steroids among resistance training practitioners.

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

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).

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.

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.

Cytopathological Changes Induced by the Crack Use in Oral Mucosa.

AIMS: To evaluate cytological alterations, inflammation, and microbial charge of the oral mucosa epithelium in crack users in in terms of the amount and duration of use. METHODS: Two hundred thirty four crack users (case group) and 120 non-users (control group) participated in this study. Clinically healthy epithelial cells were collected from the posterior mouth floor, using the conventional exfoliative cytology. Some of the aspects evaluated were as follows: Papanicolaou classification, nuclear area (NA), cytoplasmic area (CA), nuclear/cytoplasmic area ratio (NA/CA), inflammation, microbial charge, keratinization, enucleated superficial cells, and binucleation. RESULTS: The average time of crack consumption was 9.8 years (±7.1) and the average quantity of use was 13.97 g/week (±18.5). The average NA values and NA/CA ratio were increased and CA values were decreased in the case group compared to those in the controls (p < 0.05). Papanicolaou class II, intense inflammation, and intense microbial charge were more prevalent in the case group than in the controls (p < 0.05). There was a significant association between high quantity of smoked crack rocks per week and increased CA values, absence of keratinization, and presence of enucleated superficial cells (p < 0.05). CONCLUSION: Crack use seemed to induce inflammatory alterations and early indicators of malignant transformation on the oral mucosa epithelium.

The effect of two drug delivery systems in ropivacaine cytotoxicity and cytokine release by human keratinocytes and fibroblasts.

OBJECTIVES: Modified drug delivery systems have been developed to improve pharmacological properties of local anaesthetics. However, the inflammatory potential of these formulations was not investigated. This study compared the in-vitro effects of ropivacaine (ropi) in plain, liposomal (MLV) or 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) formulations on cell viability, apoptosis and cytokine (IL-1α, TNF-α, IL-6 and IL-10) release. METHODS: Human immortalized keratinocytes (HaCaT) and human immortalized gingival fibroblasts (HGF) were exposed to 1-100 µm ropi concentrations. The cell viability was measured by XTT and LIVE/DEAD assay. Apoptosis was performed by flow cytometry, and cytokine release was measured by ELISA assay. KEY FINDINGS: Human immortalized keratinocyte viability was reduced by ropi and both drug delivery systems. However, none of the formulations induced apoptosis. Results showed a differential regulation of IL-1α TNF-α, IL-6 and IL-10 by HaCaT and HGF. Ropi-HP-ß-CD increased twofold the IL-6 release by HGF in comparison with the control, while 100 µm ropi-MLV led to an increased release of all pro-inflammatory cytokines by HGF. CONCLUSION: The loss in cell viability was not related to cellular apoptosis. Ropi complexed with HP-ß-CD showed a similar cytokine release pattern when compared to the plain formulation. Thus, the HP-ß-CD form was a better drug carrier than the MLV form for ropivacaine drug delivery.

Growing Candida albicans Biofilms on Paper Support and Dynamic Conditions.

A stainless steel paper-embedded biofilm reactor (PEBR) was developed for Candida spp. growth, permitting confluent distribution of nutrients by capillary diffusion through ordinary laboratory filter paper. Antibiogram disks were distributed along the filter paper rim, and the PEBR received 0.1 or 0.01 % crystal violet (CV) at 200 µL min(-1) and at 37 °C, for 48 h. CV was recovered from the disks and measured at 540 nm. Candida albicans SC5314 cells were applied onto antibiogram disks. The bioreactor was assembled, and YEPD broth was admitted (200 µL min(-1)) at 37 °C, for 72 h. Biofilm growth was estimated via the MTT reduction test. Controls were disks that received the same treatments, except for the fungus. The PEBR was considered high-throughput table, low-cost, and feasible to grow C. albicans biofilms.