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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.

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.

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.

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).

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.

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. 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. 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. 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.

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.

BackgroundThe 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.ObjectiveThe 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.MethodsWe 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.ResultsThe 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.ConclusionMicrobial 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.

The habit of cigarette smoking is associated with higher oral candidal carriage and possible predisposition to oral candidosis. The effects of exposure to smoke on the virulence properties of oral yeasts remain obscure. Hence, we showed in vitro the effect of cigarette smoke condensate (CSC) on ten clinical isolates of Candida albicans obtained from nonsmoking volunteers, as well the type-strain CBS562. CSC was generated by complete burn of five commercial cigarettes in an in-house smoking machine and used to prepare the culture broth in which the strains were grown. In 24-h intervals (T₂₄, T₄₈, and T₇₂), the cells were harvested, washed, subcultured, and the resultant growth were evaluated for possible variations for secreted aspartyl protease, phospholipase, chondroitinase, and hemolysins, adhesion to acrylic and cell surface hydrophobicity (CSH). The results indicated a temporal increase in the secretion rates of enzymes, particularly when yeast cells were exposed to CSC for 48-72 h (P < 0.05). Similarly, adhesion to acrylic and CSH increased with exposure period (P < 0.05). Based on foregoing, we concluded that CSC may promote significant enhance in the secretion of candidal histolytic enzymes and adherence to denture surfaces, thereby promoting oral yeast carriage and possible infection.

Benzodiazepines (BZDs), the most commonly prescribed psychotropic drugs with anxiolytic action, may cause hyposalivation. It has been previously shown that BZDs can cause hypertrophy and decrease the acini cell number. In this study, we investigated the effects of BZDs and pilocarpine on rat parotid glands, specifically on acinar, ductal, and myoepithelial cells. Ninety male Wistar rats were divided into nine groups. Control groups received a saline solution for 30 days (C30) and 60 days (C60), and pilocarpine (PILO) for 60 days. Experimental groups received lorazepam (L30) and midazolam (M30) for 30 days. Another group (LS60 or MS60) received lorazepam or midazolam for 30 days, respectively, and saline for additional 30 days. Finally, other groups (LP60 or MP60) received either lorazepam or midazolam for 30 days, respectively, and pilocarpine for additional 30 days. The expression of calponin in myoepithelial cells and the proliferating cell nuclear antigen (PCNA) in acinar and ductal cells were evaluated. Animals treated with lorazepam showed an increase in the number of positive staining cells for calponin as compared to control animals (p < 0.05). Midazolam administered with pilocarpine (MP60) induced an increase in the proliferation of acinar and ductal cells and a decrease in the positive staining cells for calponin as compared to midazolam administered with saline (MS60). We found that myoepithelial cells might be more sensitive to the effects of BZD than acinar and ductal cells in rat parotid glands.