Cryptocarya moschata extract decreases single and mixed biofilms on acrylic resins.

OBJECTIVE: This study proposed to assess the effect of Cryptocarya moschata extract on single and mixed biofilms formed on denture base and reline acrylic resin. MATERIALS AND METHODS: Single and mixed biofilms of Candida albicans and Streptococcus mutans were formed on the samples and treated with C. moschata extract; Nystatin solution at 100,000 IU/mL or Penicillin antibiotic solution at 100,000 IU/mL; or PBS solution. Antimicrobial activity was analyzed by counting colony-forming units, metabolism assay, assessment of protein components of the biofilm matrix, and of cell viability using confocal laser scanning microscopy (CLSM). Data were submitted to ANOVA and Tukey's post-test (α = 0.05). RESULTS: Cryptocarya moschata extract reduced cell viability of C. albicans and S. mutans single and mixed biofilms formed on samples. For all types of biofilms in the C. moschata group, there was a log reduction of the biofilm, proven by the Alamar Blue assay. Analyzing the extracellular matrix protein components, groups treated with the extract exhibited a lower level of fluorescence compared to the PBS groups. Reduction in thickness biofilm and viable cells was perceptible in the C. moschata group when assessing through CLSM. CONCLUSION: Cryptocarya moschata extract reduced the single and mixed biofilms of C. albicans and S. mutans on acrylic resins.

Streptococcus mutans supernatant affects the virulence of Candida albicans.

Candida albicans causes a variety of clinical manifestations through multiple virulence factors that act simultaneously to overcome the immune system and invade the host tissues. Owing to the limited number of antifungal agents available, new candidiasis therapeutic strategies are required. Previous studies have demonstrated that the metabolites produced by Streptococcus mutans lead to a decrease in the number of Candida cells. Here, for the first time, we evaluated whether the C. albicans cells that survived the pretreatment with S. mutans supernatant can modify their virulence factors and their capability to infect Galleria mellonella larvae. Streptococcus mutans supernatant (SM-S) was obtained by filtering the culture supernatant of this bacterium. Then, C. albicans cells were pretreated with SM-S for 24 h, and the surviving cells were evaluated using in vitro and in vivo assays. The C. albicans pretreated with SM-S showed a significant inhibition of hyphal growth, an altered adhesion pattern, and an impaired capability to form biofilms; however, its proteolytic activity was not affected. In the in vivo assays, C. albicans cells previously exposed to SM-S exhibited a reduced ability to infect G. mellonella and a higher amount of circulating hemocytes. Thus, SM-S could inhibit important virulence factors of C. albicans, which may contribute to the development of new candidiasis therapeutic strategies.

Very low-calorie ketogenic diet in the treatment of adaptive thermogenesis: A case report.

OBJECTIVES: The management of the phenomenon of adaptive thermogenesis poses a challenge to the successful treatment of overweight/obesity with a nutritional intervention that minimizes the loss of muscle mass, with little cognitive restraint use and disorganization of eating behavior. On the other hand, it creates a significant calorie deficit for the reduction of body fat. The aim of this case report was to discuss the effects of a very low-calorie ketogenic diet in a woman with obesity and low resting metabolic rate. CASE DESCRIPTION: A 36-y-old white woman with a history of obesity and bulimia nervosa who has had difficulty losing and maintaining weight despite numerous dietary and pharmacologic treatments. RESULTS: There was a loss of 12 kg in 115 d, reaching 13.4 kg, with 11.4 kg of fat mass (FM). The resting metabolic rate showed an increase of 79% in relation to the initial rate, reaching normal levels for the predictive equations and maintaining this level in the first-year follow-up. Additionally, improvement of metabolic laboratory parameters and eating behavior traits were described. CONCLUSIONS: In this specific case of bulimia nervosa resulting in hypometabolism (low resting metabolic rate/fat-free mass) and obesity, the very low-calorie ketogenic diet intervention has demonstrated a possibility of weight loss with little cognitive restraint use, thereby increasing resting metabolic rate in the short and medium terms, ultimately promoting a negative energy balance. In relation to the numeric results, it seems positive; however, more research is necessary to evaluate the effects on the overall relationship with food and its long-term repercussions.

Craving for carbs: food craving and disordered eating in low-carb dieters and its association with intermittent fasting.

Studies point to positive outcomes in a diet with reduction of carbohydrates and that the associated practice of intermittent fasting (IF) might increase weight loss. Although dieting might be related to disordered eating, little evidence is available about the role of restrictive carbohydrates diets on disordered eating. This study aimed to explore if doing low-carb (LC) diets was related to disordered eating and if IF would increase these symptoms. The sample comprised university students (n = 682), with a mean age of 22 years old and average BMI of 23.6 kg/m2 (SD = 4.3). Twenty-seven percent (n = 188) of respondents reported doing LC diet in the last three months. Of those, 31% (n = 58) reported doing LC diet combined with periods of IF. Mean scores were compared using parametric tests, and effects size and correlations between variables were calculated. Dieters showed higher levels of binge eating, food cravings, cognitive restraint, cognitive restraint toward carbohydrates when compared to non-dieters. The association of LC and IF was related to an increase in disordered eating, especially binge eating and food cravings, specifically 'Lack of control', 'Thoughts or preoccupation with food,' and 'Guilt from cravings and/or for giving in to them'. These results provide evidence that restrictive carbohydrate diets and IF may increase cognitive restraint and, consequently, food cravings.Level III: Evidence obtained from cohort or case-control analytic studies.

Antiviral activity of medicinal plant-derived products against SARS-CoV-2.

This review presents information from several studies that have demonstrated the antiviral activity of extracts (Andrographis paniculata, Artemisia annua, Artemisia afra, Cannabis sativa, Curcuma longa, Echinacea purpurea, Olea europaea, Piper nigrum, and Punica granatum) and phytocompounds derived from medicinal plants (artemisinins, glycyrrhizin, and phenolic compounds) against SARS-CoV-2. A brief background of the plant products studied, the methodology used to evaluate the antiviral activity, the main findings from the research, and the possible mechanisms of action are presented. These plant products have been shown to impede the adsorption of SARS-CoV-2 to the host cell, and prevent multiplication of the virus post its entry into the host cell. In addition to antiviral activity, the plant products have also been demonstrated to exert an immunomodulatory effect by controlling the excessive release of cytokines, which is commonly associated with SARS-CoV-2 infections.

Curcuma longa L. helps macrophages to control opportunistic micro-organisms during host-microbe interactions.

Aim: Plant products have been evaluated to control opportunistic micro-organisms, as well as fortify immune system cells. Thus, Curcuma longa L. (turmeric) extract was evaluated in interactions of murine macrophages (RAW 264.7) with Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans, in order to establish cooperation with defense cells. Materials & methods: Effects of minimal inhibitory concentrations (MIC) of the plant extract were analyzed on phagocytosis, cell viability of RAW 264.7 and production of inflammation-related molecules (IL-1ß, TNF-α, IL-10 and NO). Results: The plant extract was cytocompatible and promoted significant reductions of micro-organisms, and synthesis of inflammation-related molecules, during interactions. Conclusion:C. longa L. extract showed significant antimicrobial response and cooperation with macrophages, by fighting bacteria and yeasts during host-microbe interactions.

Curcuma longa L. (turmeric), Rosmarinus officinalis L. (rosemary), and Thymus vulgaris L. (thyme) extracts aid murine macrophages (RAW 264.7) to fight Streptococcus mutans during in vitro infection.

Finding an effective alternative way to aid defense cells to fight Streptococcus mutans was the main goal of this study. The effect of plant extracts from Curcuma longa L. (turmeric), Rosmarinus officinalis L. (rosemary), and Thymus vulgaris L. (thyme) was evaluated on murine macrophages (RAW 264.7) infected by S. mutans. Minimum inhibitory concentration (MIC) of the extracts was determined. Macrophages were infected by S. mutans and treated with each extract. From the supernatants, it was measured nitric oxide (NO) level. Posteriorly, RAW 264.7 were lysed to expose living and phagocytosed bacteria. Cytotoxicity was checked by lysosomal activity analysis, using neutral red assay. Each extract helped RAW 264.7 to eliminate S. mutans during infection, as observed by a significant bacterial reduction. Significant cell viability was also found. Besides, an increased production of NO was verified using R. officinalis L. and T. vulgaris L. extracts. The evaluated extracts demonstrated an effective action to assist RAW 264.7 to fight S. mutans during infection.

Titanium alloys: in vitro biological analyzes on biofilm formation, biocompatibility, cell differentiation to induce bone formation, and immunological response.

Biological effects of titanium (Ti) alloys were analyzed on biofilms of Candida albicans, Enterococcus faecalis, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus mutans, and Streptococcus sanguinis, as well as on osteoblast-like cells (MG63) and murine macrophages (RAW 264.7). Standard samples composed of aluminum and vanadium (Ti-6Al-4V), and sample containing niobium (Ti-35Nb) and zirconium (Ti-13Nb-13Zr) were analyzed. Monomicrobial biofilms were formed on the Ti alloys. MG63 cells were grown with the alloys and the biocompatibility (MTT), total protein (TP) level, alkaline phosphatase (ALP) activity, and mineralization nodules (MN) formation were verified. Levels of interleukins (IL-1ß and IL-17), tumor necrosis factor alpha (TNF-α), and oxide nitric (NO) were checked, from RAW 264.7 cells supernatants. Data were statically analyzed by one-way analysis of variance (ANOVA) and Tukey's test, or T-test (P ≤ 0.05). Concerning the biofilm formation, Ti-13Nb-13Zr alloy showed the best inhibitory effect on E. faecalis, P. aeruginosa, and S. aureus. And, it also acted similarly to the Ti-6Al-4V alloy on C. albicans and Streptococcus spp. Both alloys were biocompatible and similar to the Ti-6Al-4V alloy. Additionally, Ti-13Nb-13Zr alloy was more effective for cell differentiation, as observed in the assays of ALP and MN. Regarding the stimulation for release of IL-1ß and TNF-α, Ti-35Nb and Ti-13Nb-13Zr alloys inhibited similarly the synthesis of these molecules. However, both alloys stimulated the production of IL-17. Additionally, all Ti alloys showed the same effect for NO generation. Thus, Ti-13Nb-13Zr alloy was the most effective for inhibition of biofilm formation, cell differentiation, and stimulation for release of immune mediators.

Antimicrobial activity of noncytotoxic concentrations of Salvia officinalis extract against bacterial and fungal species from the oral cavity.

The use of medicinal plants can be an alternative method for the control of microorganisms responsible for human infections. This study evaluated the antimicrobial activity of Salvia officinalis Linnaeus (sage) extract on clinical samples isolated from the oral cavity and reference strains of Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans, Candida albicans, Candida tropicalis, and Candida glabrata. In addition, testing assessed the cytotoxic effect of S officinalis on murine macrophages (RAW 264.7). Minimum inhibitory, minimum bactericidal, and minimum fungicidal concentrations of S officinalis extract were determined by broth microdilution method in 60 microbial samples. The cytotoxicity was checked by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The quantities of the proinflammatory cytokines interleukin 1ß (IL-1ß) and tumor necrosis factor α (TNF-α) produced by RAW 264.7 were analyzed by an enzyme-linked immunosorbent assay. An S officinalis concentration of 50.0 mg/mL was effective against all microorganisms. Regarding cytotoxicity, the groups treated with 50.0-, 25.0-, and 12.5-mg/mL concentrations of S officinalis presented cell viability statistically similar to that of the control group, which was 100% viable. The production of IL-1ß and TNF-α was inhibited at a 50.0-mg/mL concentration of S officinalis. Thus, S officinalis extract presented antimicrobial activity on all isolates of Staphylococcus spp, S mutans, and Candida spp. No cytotoxic effect was observed, as demonstrated by the survival of RAW 264.7 and inhibition of IL-1ß and of TNF-α.

Rosmarinus officinalis L. (rosemary) as therapeutic and prophylactic agent.

Rosmarinus officinalis L. (rosemary) is a medicinal plant native to the Mediterranean region and cultivated around the world. Besides the therapeutic purpose, it is commonly used as a condiment and food preservative. R. officinalis L. is constituted by bioactive molecules, the phytocompounds, responsible for implement several pharmacological activities, such as anti-inflammatory, antioxidant, antimicrobial, antiproliferative, antitumor and protective, inhibitory and attenuating activities. Thus, in vivo and in vitro studies were presented in this Review, approaching the therapeutic and prophylactic effects of R. officinalis L. on some physiological disorders caused by biochemical, chemical or biological agents. In this way, methodology, mechanisms, results, and conclusions were described. The main objective of this study was showing that plant products could be equivalent to the available medicines.

Mouthwashes: an in vitro study of their action on microbial biofilms and cytotoxicity to gingival fibroblasts.

This study evaluated the in vitro antibiofilm effect of 5 different commercial mouthwashes (Cepacol Traditional, Colgate Plax Fresh Mint, Listerine Cool Mint, Oral-B Complete, and Sensodyne) on Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans, Escherichia coli, and Pseudomonas aeruginosa. The cytotoxic effect of the mouthwashes on gingival fibroblasts was also analyzed. A colorimetric assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used to investigate the viability of biofilms after 48 hours and gingival fibroblasts after 24 hours. The biofilms were exposed to the mouthwashes for 2 different lengths of time: T1, the time recommended by the manufacturer (30 or 60 seconds); and T2, double the recommended time (60 or 120 seconds). All antiseptic mouthwashes caused a significant reduction of biofilm (P < 0.05) as well as a significant reduction of viable gingival fibroblasts (P < 0.05) with both exposure times (T1 and T2). It can be concluded that the commercial mouthwashes demonstrated effective antibiofilm activity; they were more effective on bacteria than on C albicans. A significant cytotoxic effect on gingival fibroblasts was also observed.

Thymus vulgaris L. and thymol assist murine macrophages (RAW 264.7) in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans.

Microorganisms are capable to combat defense cells by means of strategies that contribute to their stabilization and proliferation in invaded tissues. Frequently antimicrobial-resistant strains appear; therefore, alternative methods to control them must be investigated, for example, the use of plant products. The capacity of the thyme extract (Thymus vulgaris L.) and phytocompound thymol in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans in murine macrophages (RAW 264.7) was evaluated. Minimal inhibitory concentrations (MIC) of the plant products were used. The effect of these MIC were analyzed in the assays of phagocytosis and immunoregulation by analysis of the production of cytokines (IL-1ß, TNF-α, and IL-10) and nitric oxide (NO). The plant products effectively assisted the macrophages in the phagocytosis of microorganisms, presenting significant reductions of S. aureus and P. aeruginosa. The macrophages also regulated the production of inflammatory mediators in the infections by S. aureus, P. aeruginosa, and C. albicans. In addition, thyme provided a satisfactory effect in response to the bacterial infections, regarding generation of NO. Thus, the effectiveness of the thyme and thymol to control in vitro infections by S. aureus, P. aeruginosa, and C. albicans was observed. HIGHLIGHTS: Phagocytosis of S. aureus by RAW 264.7 was enhanced with thymol Thyme enhanced the phagocytosis of P. aeruginosa by RAW 264.7 Plant products provided immunoregulation of inflammatory cytokines Production of nitric oxide was improved with the treatments in bacterial infections.

Biological activities of Rosmarinus officinalis L. (rosemary) extract as analyzed in microorganisms and cells.

R. officinalis L. is an aromatic plant commonly used as condiment and for medicinal purposes. Biological activities of its extract were evaluated in this study, as antimicrobial effect on mono- and polymicrobial biofilms, cytotoxicity, anti-inflammatory capacity, and genotoxicity. Monomicrobial biofilms of Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans and Pseudomonas aeruginosa and polymicrobial biofilms composed of C. albicans with each bacterium were formed in microplates during 48 h and exposed for 5 min to R. officinalis L. extract (200 mg/mL). Its cytotoxic effect was examined on murine macrophages (RAW 264.7), human gingival fibroblasts (FMM-1), human breast carcinoma cells (MCF-7), and cervical carcinoma cells (HeLa) after exposure to different concentrations of the extract, analyzed by MTT, neutral red (NR), and crystal violet (CV) assays. The anti-inflammatory activity was evaluated on RAW 264.7 non-stimulated or stimulated by lipopolysaccharide (LPS) from Escherichia coli and treated with different concentrations of the extract for 24 h. Interleukin-1 beta (IL-1ß) and tumor necrosis factor alpha (TNF-α) were quantified by ELISA. Genotoxicity was verified by the frequency of micronuclei (MN) at 1000 cells after exposure to concentrations of the extract for 24 h. Data were analyzed by T-Test or ANOVA and Tukey Test ( P ≤ 0.05). Thus, significant reductions in colony forming units per milliliter (CFU/mL) were observed in all biofilms. Regarding the cells, it was observed that concentrations ≤ 50 mg/mL provided cell viability of above 50%. Production of proinflammatory cytokines in the treated groups was similar or lower compared to the control group. The MN frequency in the groups exposed to extract was similar or less than the untreated group. It was shown that R. officinalis L. extract was effective on mono- and polymicrobial biofilms; it also provided cell viability of above 50% (at ≤ 50 mg/mL), showed anti-inflammatory effect, and was not genotoxic. Impact statement Rosmarinus officinalis L. extract effectively contributed to in vitro control of important species of microorganisms such as Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans, and Pseudomonas aeruginosa in mono- and polymicrobial biofilms that are responsible for several infections in oral cavity as in other regions of the body. Furthermore, this extract promoted also cell viability above 50% at concentrations ≤ 50 mg/mL, excellent anti-inflammatory effect, showing inhibition or reduction of the synthesis of proinflammatory cytokines, being also non-genotoxic to cell lines studied. Thus, this extract may be a promising therapeutic agent that can be added in some medical and dental formulations such as toothpastes, mouthwashes, irrigating root canals, ointments, soaps, in order to control pathogenic microorganisms and biofilms, with anti-inflammatory effect and absence of cytotoxic and genotoxic.

In vitro effect of caffeic acid phenethyl ester on matrix metalloproteinases (MMP-1 and MMP-9) and their inhibitor (TIMP-1) in lipopolysaccharide-activated human monocytes.

OBJECTIVE: The role of matrix metalloproteinases (MMPs) in tissue degradation has become evident in many diseases and great interest therefore exists in the pharmacological control of the activity of these enzymes. This study evaluated the effect of caffeic acid phenethyl ester (CAPE) on the production of MMPs and their inhibitor (TIMP) in monocytes activated by lipopolysaccharide (LPS). DESIGN: The human monocytic cell line (THP-1) was treated with non-cytotoxic concentrations of CAPE (10 and 60µM) combined with 1µg/mL of LPS. The gene expression of MMP-1, MMP-9 and TIMP-1 was evaluated by quantitative real-time polymerase chain reaction. The protein secretion into the culture medium was assessed via enzyme-linked immunosorbent assay and the gelatinolytic activity of MMP-9 by zymography. RESULTS: CAPE, especially at the highest concentration, down-regulated MMP-1 and MMP-9 gene expression but up-regulated the gene expression of TIMP-1. Furthermore, CAPE reduced the secreted protein level of MMP-1 and MMP-9 as well as the gelatinolytic activity of MMP-9. CONCLUSION: CAPE was able to inhibit the gene expression, production and the activity of MMPs induced by LPS and also increased the gene expression of TIMP-1. The present observations suggest that CAPE exerted a positive effect on the regulatory mechanism between MMPs and TIMP, which is important for the control of different diseases.