Exploring the antifungal potential of Annona muricata leaf extract-loaded hydrogel in treating vulvovaginal candidiasis.

Vulvovaginal candidiasis, mostly caused by Candida albicans, remains a prevalent concern in women's health. Annona muricata L. (Annonaceae), a plant native from Brazil, is well-known for its therapeutic potential, including antitumor, anti-inflammatory, and antimicrobial properties. This study presents an innovative hydrogel formulation containing the ethanolic extract from A. muricata leaves designed to control C. albicans in an in vivo model of vulvovaginal candidiasis. Here, we report the development, thermal, physicochemical and rheological characterization of a Carbopol®-based hydrogel containing A. muricata extract. Furthermore, we evaluated its activity in a vulvovaginal candidiasis in vivo model. Thermal analyses indicated that the addition of the extract increased the polymer-polymer and polymer-solvent interactions.Rheological analysis showed a decrease in the viscosity and elasticity of the formulation as the A. muricata extract concentration increased, suggesting a liquid-like behavior. After treatment with the Carbopol®-based hydrogel with A. muricata, our in vivo results showed a significant reduction in vulvovaginal fungal burden and infection, as well as a reduction in mucosal inflammation. The current research opens up possibilities for the application of the Carbopol®-based hydrogel with A. muricata as a natural therapeutic option for the treatment of vulvovaginal candidiasis.

Correction: Implementation of digital health in rural populations with chronic musculoskeletal conditions: A scoping review protocol.

[This corrects the article DOI: 10.1371/journal.pone.0291638.].

Low Fat Yoghurts Produced with Different Protein Levels and Alternative Natural Sweeteners.

The food industry is looking for substitutes for sucrose in food items due to the excessive consumption of products with added sugar and the demand for healthier products. Alternative natural sweeteners can help achieve this goal. Different types of low-fat yoghurts (1% fat), with low-protein and high-protein levels (3% and 4.5-6.5% protein, respectively), were produced using alternative natural sweeteners. The low-protein yoghurts were made with stevia (0.03% w/w) or agave syrup (4.5% w/w). The high-protein yoghurts were made with stevia (0.04% w/w), xylitol (6% w/w) or honey (6% w/w). Sucrose (6% w/w) was used as a control in both trials. pH and titratable acidity, CIEL*a*b* color parameters, syneresis index, rheology and the texture profile of the low-fat yoghurts were evaluated over refrigerated storage. All products underwent sensory evaluation by an untrained panel. The high-protein yoghurts were found to be more acidic (>1% as lactic acid), had a lower syneresis index (between 2.1 and 16.2%) and a better consistency (stronger gel structure) than the low-protein yoghurts. In terms of rheological parameters, stevia-sweetened yoghurts scored higher than the other sweetened yoghurts, showing a better gel structure. The different sweeteners tested did not significantly affect the sensory properties of the yoghurts, although the high-protein yoghurts scored higher for most of the attributes evaluated. Overall, consumers preferred stevia-sweetened yoghurts to yoghurts sweetened with sucrose or agave for the low-protein yoghurts. Of the tested formulations, those containing high protein with the alternative natural sweetener xylitol received higher scores in all attributes. These results reveal the potential of the tested natural sweeteners as sucrose substitutes, while contributing to improving the nutritional value of yoghurts.

Implementation of digital health in rural populations with chronic musculoskeletal conditions: A scoping review protocol.

Musculoskeletal conditions are a major source of disability worldwide, and its burden have been rising in the last decades. Rural areas, in particular, are associated with higher prevalence of these conditions as well as higher levels of disability, which is likely related to other determinants that affect these communities. Although digital health has been identified as a potential solution to mitigate the impact of these determinants, it is also known that these populations may face barriers that limit the implementation of these interventions. Therefore, the aim of this scoping review is to comprehensively map the evidence regarding the implementation of digital health interventions in rural populations with chronic musculoskeletal conditions. We will include studies published from the year 2000; that report the use of digital interventions that promote prevention, treatment or monitoring of any chronic musculoskeletal condition or chronic pain from musculoskeletal origin, in patients that live in rural areas. This protocol follows the methodological framework for scoping reviews proposed by Arksey and O'Malley, as well as the Joana Briggs Institute (JBI) approach. We will conduct the search on Medline (PubMed), EMBASE, Web of Science and Scopus, as well as grey literature databases. Two independent reviewers will screen titles and abstracts followed by a full-text review to assess the eligibility of the articles. Data extracted will include the identification of the digital interventions used, barriers and enablers identified by the patients or healthcare providers, the patient-level outcomes measured, and the implementation strategies and outcomes reported. By mapping the evidence on the implementation of digital health interventions in rural communities with musculoskeletal conditions, this scoping review will enhance our understanding of their applicability in real-world settings.

Mitracarpus frigidus reduces lipid metabolism and PGE2 levels in inflammatory cells.

OBJECTIVES: To evaluate the ability of the aqueous extract of Mitracarpus frigidus (MFAq) to inhibit lipid body formation and inflammatory mediator production in macrophages stimulated with lipopolysaccharide (LPS) and interferon gamma (IFN-γ). METHODS: MFAq was chemically characterized by ultrafast liquid chromatography/quadruple time-of-flight tandem mass spectrometry. The macrophages obtained from mice were incubated with MFAq. Cell viability and membrane integrity were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and propidium iodide assays, respectively. Moreover, NO, reactive oxygen species (ROS), transforming growth factor beta (TGF-ß), prostaglandin E2 (PGE2) levels and lipid bodies (LBs) were examined in macrophages that were stimulated with LPS and IFN-γ and treated with MFAq. Finally, molecular docking analysis was conducted to investigate the interaction of MFAq with the cyclooxygenase 2 (COX-2) enzyme. KEY FINDINGS: Chlorogenic acid, clarinoside, harounoside, rutin, kaempferol-3O-rutinoside and 2-azaanthraquinone were identified in MFAq. MFAq significantly inhibited NO, ROS and LBs, and did not affect the membrane integrity of macrophages. MFAq-treated cells showed significantly lower levels of TGF-ß and PGE2. Molecular docking demonstrated that the compounds found in MFAq are able to inhibit COX-2 by binding to important residues in the catalytic site. CONCLUSIONS: MFAq interferes with lipid metabolism in stimulated macrophages, leading to the reduction of important inflammatory mediators. Furthermore, MFAq can directly inhibit the COX-2 enzyme or inhibit its expression owing to its ability to reduce NO production.

Antibiofilm potential of Annona muricata L. ethanolic extract against multi-drug resistant Candida albicans.

ETNOPHARMACOLOGICAL RELEVANCE: Traditional uses of Annona muricata L. (soursop) include treatment for cancer, fungal infections, and inflammatory diseases. Its phytoconstituents, mainly acetogenins and alkaloids, are associated with therapeutic activity and clinical application is currently under investigation. However, the application of phytotherapy to treat diseases caused by fungal biofilms, such as vulvovaginal candidiasis (VVC), is still limited. AIM OF THE STUDY: To investigate the activity of the ethanolic extract of A. muricata leaves (AML) against biofilms formed by multiresistant Candida albicans (ATCC® 10231) both in vitro and in a VVC experimental model. MATERIAL AND METHODS: C. albicans biofilms were grown and their adhesion, proliferation, development, and matrix composition studied by spectrophotometry, scanning electron microscopy (SEM), whole slide imaging (WSI), and biochemical assays without or with AML treatment. In parallel, in vivo experiments were conducted using a murine model of infection treated with different concentrations of the extract and nystatin. Fungal burden and histological changes were investigated. RESULTS: The proliferation and adhesion of C. albicans biofilms were significantly reduced as confirmed by SEM and WSI quantitative analyses. Furthermore, the concentration of carbohydrates, proteins and DNA was reduced in the biofilm matrix. In vivo assays demonstrated that AML was able to reduce the fungal burden and the inflammatory process. CONCLUSIONS: The findings further emphasized the therapeutic and scientific potential of AML, thus encouraging its future use in the treatment of VVC.

Isolation and Chemical Characterization of Antifungal, Antioxidant, and Anti-Inflammatory Compounds from Centrosema coriaceum using GC/MS, UFLC-QTOF-MS, and FACE.

In recent years, natural products with biological activities have been increasingly researched. The elucidation of phytoconstituents is necessary for the development of drugs as a natural alternative for the treatment of various diseases. The work aimed to evaluate in vitro and in silico bioactivities of hexane (CCHE) and methanol (CCME) fractions of ethanolic extract from Centrosema coriaceum Benth (Fabaceae) leaves and elucidate their phytoconstituents. CCHE and CCME showed antifungal activity for Candida glabrata (MIC of 1000 µg/mL) with fungistatic effect and action in cell envelope by sorbitol and ergosterol assays. CCHE and CCME presented promising antioxidant activity against the DPPH radical with IC50 of 13.61±0.50 and 6.31±0.40 µg/mL, respectively, and relative antioxidant activity (RAA%) of 45.77±3.61/ 28.53±2.25 % for CCHE and 82.18±2.25/51.99±3.23 % for CCME when compared to rutin and quercetin, respectively. Moreover, these fractions demonstrated promising results for the inhibition of lipid peroxidation by ß-carotene/linoleic acid assay. For anti-inflammatory and cytotoxicity activities, CCHE and CCME significantly inhibited the production of nitric oxide and TNF-α, without toxicity on murine intraperitoneal macrophages, respectively. Esters, alkanes, steroids, tocopherols, and terpenes were identified in CCHE by GC/MS. Flavonoids, phenolic acids, and disaccharides were detected in CCME by UFLC-QTOF-MS and FACE. Furthermore, rutin was purified from CCME. In silico predictions evidenced that compounds present in both fractions have high affinity to the fungal membrane besides antioxidant and anti-inflammatory activities. Based on these observations, CCHE and CCME have a noteworthy potential for the design of novel antifungal and anti-inflammatory agents that should be explored in future studies.

Antifungal Annona muricata L. (soursop) extract targets the cell envelope of multi-drug resistant Candida albicans.

ETNOPHARMACOLOGICAL RELEVANCE: Annona muricata L. (soursop) is traditionally used in the treatment of inflammatory diseases, cancer, and infections caused by fungi. The therapeutic activity explored by its medicinal use is generally associated with its phytoconstituents, such as acetogenins and alkaloids. However, its potential antifungal bioactivity as well as its mechanism of action remains to be established. AIM OF THE STUDY: To evaluate the antifungal activity of the ethanolic extract of A. muricata leaves against multidrug-resistant Candida albicans (ATCC® 10231). MATERIAL AND METHODS: Phytoconstituents were detected by UFLC-QTOF-MS. The minimum inhibitory concentration was determined, followed by the determination of the minimum fungicidal concentration. For planktonic cells, the growth curve and cell density were evaluated. Studies to understand the mechanism of action on the cell envelope involved crystal violet permeability, membrane extravasation, sorbitol protection, exogenous ergosterol binding assay, metabolic activity, and cell viability. Furthermore, mitochondrial membrane potential was assessed. RESULTS: Our analyses demonstrated a significant inhibitory effect of A. muricata, with the ability to reduce fungal growth by 58% and cell density by 65%. The extract affected both the fungal plasma membrane and cell wall integrity, with significant reduction of the cell viability. Depolarization of the fungal mitochondrial membrane was observed after treatment with A. muricata. Rutin, xi-anomuricine, kaempferol-3O-rutinoside, nornuciferine, xylopine, atherosperminine, caffeic acid, asimilobine, s-norcorydine, loliolide, annohexocin, annomuricin, annopentocin, and sucrose were identified as extract bioactive components. CONCLUSIONS: Our findings show that the A. muricata extract is a source of chemical diversity, which acts as a potential antifungal agent with promising application to the therapy of infections caused by C. albicans.

Evaluation of a Functional Craft Wheat Beer Fermented with Saccharomyces cerevisiae UFMG A-905 to treat Salmonella Typhimurium infection in mice.

Functional foods containing probiotics are generally administered as dairy products. Non-dairy beverages are another possibility, but probiotic functionality must be confirmed in such vehicles. In the present study, a craft wheat beer brewed with the probiotic yeast Saccharomyces cerevisiae UFMG A-905 (905) was evaluated in a murine model of Salmonella Typhimurium infection. Unfiltered or filtered beer brewed with 905, a commercial wheat beer used as a negative control, or saline were administered orally to mice before and during oral S. Typhimurium challenge. High fecal levels of yeast were only counted in mice treated with the unfiltered 905 beer, which also had reduced mortality and body weight loss due to S. Typhimurium infection. Increased levels of intestinal IgA, translocation to liver and spleen, liver and intestinal lesions, pro-inflammatory cytokines in liver and ileum, and hepatic and intestinal myeloperoxidase and eosinophilic peroxidase activities were observed in animals infected with S. Typhimurium. All these parameters were reduced by the treatment with unfiltered 905 beer. In conclusion, the results show that a craft wheat beer brewed with S. cerevisiae UFMG A-905 maintained the probiotic properties of this yeast when administered orally to mice challenged with S. Typhimurium.

Pomegranate Peels and Seeds as a Source of Phenolic Compounds: Effect of Cultivar, By-Product, and Extraction Solvent.

The nutraceutical properties of Punica granatum L. are not restricted to the edible portion of the fruit but also to the peels and seeds, flowers, leaves, and tree bark. The recovery and valorization of the peel and seeds (ca. 50% of the whole fruit), besides the positive environmental impact, can be viewed as a source of natural bioactive compounds. Thus, the bioactive properties of extracts of pomegranate peel and seeds from Acco and Wonderful known cultivars, as well as of the novel Big Full cultivar, were evaluated. The dried and ground pomegranate by-products were submitted to a conventional solid/liquid extraction with ethanol/water mixtures (0%, 25%, 50%, and 75% of EtOH, v/v). The obtained extracts were characterized in terms of total phenolic compounds (TPC), total flavonoids (TF), and antioxidant activity (AA), determined by the DPPH radical scavenging activity and expressed as IC50 (half maximum inhibitory concentration). With the exception of the Acco cultivar, the extraction yield (EY) was higher for peels, whose extracts showed higher TPC, TF, and IC50 (lower AA). The extracts obtained from the by-products of the Big Full cultivar had a statistically higher overall bioactive potential (TPC: 0.36 mg GAE/mg extract; TF: 0.031 mg CATE/mg extract; IC50: 0.51 mg/mL) compared to the other two studied cultivars. Furthermore, the EY was enhanced by solvents richer in ethanol (50-75%), allowing obtaining extracts richer in TPC and TF with higher AA. Finally, it was shown that EY combined with bioactive data allowed a satisfactory principal component unsupervised differentiation of the pomegranate extracts according to the type of by-product used.

Effect of Extraction Method on the Bioactive Composition, Antimicrobial Activity and Phytotoxicity of Pomegranate By-Products.

Pomegranate by-products can be an asset to the food industry due to the richness in bioactive and antimicrobial compounds. This work studied the influence of conventional solvent and sonication-assisted extraction methods on the bioactive profile, antimicrobial properties, and phytotoxicity effect of the peels and seeds extracts from Acco, Big Full, and Wonderful pomegranate cultivars. The bioactive composition of the extracts was evaluated for the content of total phenolics, total flavonoids, and antioxidant activity (expressed as the half-maximal inhibitory concentration-IC50) by spectrophotometric methods, while the tannins were determined by titration and the anthocyanins were estimated by the pH-differential method. For the evaluation of the antimicrobial activity, the disk diffusion method of Kirby-Bauer was adapted through inhibition halos against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, and Yarrowia lipolytica. The extracts' phytotoxicity was evaluated in vitro on garden-cress seeds. Extracts from conventional extraction were richer in total phenolics, expressed as gallic acid equivalents (0.16-0.73 mg GAE/mg extract), while those from sonication-assisted extraction had higher contents of total flavonoids, expressed as catechin equivalents (0.019-0.068 mg CATE/mg extract); anthocyanins, expressed as cyanidin-3-glucoside (0.06-0.60 µg C3G/mg, dry basis); and antioxidant activity (IC50, 0.01-0.20 mg/mL). All extracts were more effective against Gram-positive bacteria and yeasts than Gram-negative bacteria. In general, the sonication-assisted extracts led to higher inhibition halos (8.7 to 11.4 mm). All extracts presented phytotoxicity against garden-cress seeds in the tested concentrations. Only the lowest concentration (0.003 mg/mL) enabled the germination of seeds and root growth, and the sonication-assisted extracts showed the highest Munoo-Liisa vitality index (51.3%). Overall, sonication-assisted extraction obtained extracts with greater bioactive and antimicrobial potential and less phytotoxicity.

Pharmacological investigation of antioxidant and anti-inflammatory activities of aqueous extract from Mitracarpus frigidus (Rubiaceae).

OBJECTIVES: This study aimed to evaluate the potential of aqueous extract from Mitracarpus frigidus aerial parts (MFAq) in the treatment of inflammation and oxidative stress, as well as to characterize its chemical constituents. METHODS: Total phenolic and flavonoid contents were determined, and phytoconstituents were detected by ultra-fast liquid chromatography/quadrupole-time-of-flight tandem mass spectrometry (UFLC-QTOF-MS). The antioxidant activity was evaluated by DPPH, TAC and ß-carotene/linoleic acid assays. In-vitro anti-inflammatory activity, cell viability and cell cycle were performed in J774A.1 cell line. In-vivo anti-inflammatory activity was investigated by two ear oedema assays (croton oil and phenol). KEY FINDINGS: Chlorogenic acid, clarinoside, quercetin-hexosylpentoside, rutin, kaempferol-3-O-rutinoside, kaempferol-rhamnosylhexoside, quercetin-pentosylrhamnosylhexoside, harounoside, 2-azaanthraquinone and sucrose were identified by UFLC-QTOF-MS. MFAq showed antioxidant activity, which was positively correlated to the content of phenolic compounds. MFAq significantly inhibited the production of nitric oxide, did not decrease viability in MTT assay (all concentrations) and showed no changes in membrane permeability and cell cycle of J774A.1 cell line. Furthermore, MFAq showed a reduction in ear oedema in all tested doses. CONCLUSION: MFAq was effective in some antioxidant and inflammatory parameters, in the experimental conditions that were used in the study. This is the first report of chemical composition and bioactivities from this extract.

Combining UFLC-QTOF-MS analysis with biological evaluation of Centrosema coriaceum (Fabaceae) leaves.

Centrosema coriaceum Benth belongs to Fabaceae family and have few studies of biological activity and chemical composition. Thus, the aims of this work were to determine chemical profile of the ethanolic extract of C. coriaceum leaves (CCE) by UFLC-QTOF-MS and to evaluate its in vitro biological potential. CCE showed MIC value of 1000 µg/mL against Candida glabrata (fungistatic effect) and high affinity in cell envelope by increasing cell permeability in nucleotide leakage, sorbitol and ergosterol assays. CCE showed antioxidant activity in all assays performed. For the anti-inflammatory and cytotoxicity activities, CCE, at all tested concentrations, significantly inhibited the production of nitric oxide and did not decrease J774A.1 cell viability below 70%. Finally, rutin, kaempferol-3O-rutinoside, caffeic acid, and sucrose were identified in CCE by UFLC-QTOF-MS. These results suggest, for the first time, that C. coriaceum has interesting antifungal, antioxidant, and anti-inflammatory activities.

Implementação do modelo híbrido da teleaudiologia: aceitação, viabilidade e satisfação em um programa de implante coclear / Implementation of the hybrid teleaudiology model: acceptance, feasibility and satisfaction in a cochlear implant program

RESUMO Objetivo descrever a incorporação da teleaudiologia na rotina de um programa público referência em implante coclear e avaliar a satisfação dos usuários e de seus familiares com a assistência híbrida. Métodos foram desenvolvidas ações que proporcionaram a conexão entre usuários e especialistas, por meio do modelo híbrido da teleaudiologia, combinando práticas remotas (síncronas/ assíncronas) e/ou atendimentos presenciais. Para verificar a satisfação dos pacientes com a teleaudiologia, foi aplicado o Questionário de Satisfação do Cliente. Resultados foram realizados 218 atendimentos remotos e 23 atendimentos presenciais coordenados, entre teleconsultas para orientação e aconselhamento, coaching de pais, terapia para reabilitação auditiva, validação dos resultados, resolução de problemas, ativação e mapeamentos, procedimentos intraoperatórios, teleconsultas com especialistas e entregas de peças de manutenção da tecnologia auditiva. Dos participantes, 36 responderam ao questionário, relataram estar muito satisfeitos com os serviços prestados de maneira geral, ou em grande parte (88,9%) e com a quantidade de atendimentos que receberam (72,2%). Este estudo sugeriu que, em algum momento durante a pandemia, a maioria dos pacientes teve suas necessidades atendidas usando teleaudiologia para cuidados continuados. Conclusão a abordagem de atendimento híbrida em audiologia foi viável, aceita e alcançou satisfação dos usuários.

ABSTRACT Purpose This article aims to describe the incorporation of teleaudiology in the routine of a highly regarded, government funded, cochlear implantation program and evaluate the satisfaction of users and their families with hybrid assistance. Methods Actions were developed that provided the connection between users and specialists, through the hybrid model of teleaudiology, combining remote practices (synchronous/asynchronous) with face-to-face care. To check patient satisfaction with teleaudiology, the Customer Satisfaction Questionnaire (CSQ) was applied. Results 218 remote appointments and 23 other presential attendances were coordinated, between teleconsultations, guidance and counseling; parent coaching and hearing rehabilitation validation, therapy; troubleshooting, cochlear implant first activations and mappings, intra-operative measures, teleconsultations with specialists and delivery of repaired electronic devices. Of the participants, 36 patients responded to the questionnaire and reported being very satisfied with the services provided in general or in large part (88.9%) and with the amount of care they received (72.2%).This study suggested that, at some point during the pandemic, most patients had their needs met using teleaudiology long term care. Conclusion The hybrid approach to audiology care was feasible, accepted and achieved user satisfaction.

Spilanthol as a promising antifungal alkylamide for the treatment of vulvovaginal candidiasis.

Spilanthol is a bioactive alkylamide from the native Amazon plant species, Acmella oleracea. However, antifungal activities of spilanthol and its application to the therapeutic treatment of candidiasis remain to be explored. This study sought to evaluate the in vitro and in vivo antifungal activity of spilanthol previously isolated from A. oleracea (spilanthol(AcO)) against Candida albicans ATCC® 10231™, a multidrug-resistant fungal strain. Microdilution methods were used to determine inhibitory and fungicidal concentrations of spilanthol(AcO). In planktonic cultures, the fungal growth kinetics, yeast cell metabolic activity, cell membrane permeability and cell wall integrity were investigated. The effect of spilanthol(AcO) on the proliferation and adhesion of fungal biofilms was evaluated by whole slide imaging and scanning electron microscopy. The biochemical composition of the biofilm matrix was also analyzed. In parallel, spilanthol(AcO) was tested in vivo in an experimental vulvovaginal candidiasis model. Our in vitro analyses in C. albicans planktonic cultures detected a significant inhibitory effect of spilanthol(AcO), which affects both yeast cell membrane and cell wall integrity, interfering with the fungus growth. C. albicans biofilm proliferation and adhesion, as well as, carbohydrates and DNA in biofilm matrix were reduced after spilanthol(AcO) treatment. Moreover, infected rats treated with spilanthol(AcO) showed consistent reduction of both fungal burden and inflammatory processes compared to the untreated animals. Altogether, our findings demonstrated that spilanthol(AcO) is an bioactive compound against planktonic and biofilm forms of a multidrug resistant C. albicans strain. Furthermore, spilanthol(AcO) can be potentially considered for therapeutical treatment of vulvovaginal candidiasis caused by C. albicans. LAY SUMMARY: This study sought to evaluate the antifungal activity of spilanthol against Candida albicans ATCC® 10 231™, a multidrug-resistant fungal strain. Our findings demonstrated that spilanthol(AcO) can be potentially considered for therapeutical treatment of vulvovaginal candidiasis caused by C. albicans.

Pharmacological investigation of antioxidant and anti-inflammatory activities of leaves and branches extracts from Plinia cauliflora (Jaboticaba).

ETHNOPHARMACOLOGICAL RELEVANCE: Among all native Brazilian plant species, Plinia cauliflora (DC.) Kausel (Jaboticaba), is well known for producing "superfruits", due to their high phenolic content and antioxidant property. The fruit has astringent characteristics, and it is popularly known for the treatment of diarrhea, rash, and intestinal inflammation. However, there are only a few studies on the use of leaves and branches of this species in the literature, mainly to treat oxidative stress and inflammation. AIM OF THE STUDY: The present study aimed to investigate the antioxidant and anti-inflammatory potential of leaves and branches extracts from P. cauliflora. MATERIAL AND METHODS: The phytochemical analysis of P. cauliflora extracts was performed by the total phenolic, flavonoid, and tannin dosage method. Moreover, the compounds were identified by HPLC-MS-Q-TOF. Antioxidant capacity was determined by DPPH, ß-carotene/linoleic acid system, MDA formation, and phosphomolybdenum assays. In vitro and in vivo anti-inflammatory activities of P. cauliflora were evaluated by the reduction of nitric oxide in the J774A.1 cell line and inhibition of ear edema in mice, respectively. RESULTS: The ethanolic extract of the leaves exhibited greater flavonoid content whereas the ethanolic extract of the branches showed higher tannins content. Twenty-two and seventeen compounds were identified by HPLC-MS-Q-TOF in the leaves and branches, respectively, being tellimagrandin I, castalagin, and valoneic acid dilactone reported for the first time in P. cauliflora. The antioxidant potential of extracts was confirmed through different oxidation pathways from oxidizing radicals, which might be related to the presence of phenolic compounds. For the anti-inflammatory assay, the leaves and branches extracts showed promising results, with a reduction of nitric oxide ear edema inhibition around 95% and 80%, respectively. CONCLUSIONS: Herein, the great biological potential of leaves and branches extracts from P. cauliflora was highlighted. These parts of the plant are underused and poorly reported in the literature, especially for the antioxidant and anti-inflammatory activities.

Whole slide imaging is a high-throughput method to assess Candida biofilm formation.

New strategies that enable fast and accurate visualization of Candida biofilms are necessary to better study their structure and response to antifungals agents. Here, we applied whole slide imaging (WSI) to study biofilm formation of Candida species. Three relevant biofilm-forming Candida species (C. albicans ATCC 10231, C. glabrata ATCC 2001, and C. tropicalis ATCC 750) were cultivated on glass coverslips both in presence and absence of widely used antifungals. Accumulated biofilms were stained with fluorescent markers and scanned in both bright-field and fluorescence modes using a WSI digital scanner. WSI enabled clear assessment of both size and structural features of Candida biofilms. Quantitative analyses readily detected reductions in biofilm-covered surface area upon antifungal exposure. Furthermore, we show that the overall biofilm growth can be adequately assessed across both bright-field and fluorescence modes. At the single-cell level, WSI proved adequate, as morphometric parameters evaluated with WSI did not differ significantly from those obtained with scanning electron microscopy, considered as golden standard at single-cell resolution. Thus, WSI allows for reliable visualization of Candida biofilms enabling both large-scale growth assessment and morphometric characterization of single-cell features, making it an important addition to the available microscopic toolset to image and analyse fungal biofilm growth.

Prevalence evolution of SARS-CoV-2 infection in the Municipality of Sao Paulo, 2020 - 2021

ObjectivesTo estimate the evolution of the prevalence of SARS-CoV-2 virus infection among residents aged 18 years or over in the municipality of Sao Paulo. MethodsThis is a population-based household survey conducted every 15 days, between June and September 2020 and January and February 2021. In total, 11 phases were performed. The presence of antibodies against SARS-CoV-2 was identified in venous blood using a lateral flow test, Wondfo Biotech. In the last phase, it was combined with an immunoenzymatic test, Euroimmun. Participants also answered a semi-structured questionnaire on sociodemographic and economic factors and social distancing measures. Prevalence estimates and 95% confidence intervals were estimated according to the region, Human Development Index, sex, age group, ethnicity, education, income and variables associated with risk or prevention of the infection. To compare the frequencies among the categories of each variable, the chi-square test with Rao Scott correction was used, considering a 5% significance level. ResultsIn total, 23,397 individuals were interviewed and had their samples collected. The estimated prevalence of antibodies against SARS-CoV-2 ranged from 9.7% (95%CI: 7.9-11.8%) to 25.0% (95%CI: 21.7-28.7). The prevalence of individuals with antibodies against SARS-CoV-2 was higher among black and pardo people, people with lower schooling, people with lower income and among residents of regions with lower Human Development Index. The lowest prevalences were associated with recommended measures of disease protection. The proportion of asymptomatic infection was 45.1%. ConclusionThe estimated prevalence of SARS-CoV-2 infection was lower than the cumulative incidence variation, except for the last phase of the study. The differences in prevalence estimates observed among subpopulations showed the social inequality as a risk of infection. The lower prevalence observed among those who could follow prevention measures reinforce the need to maintain the social distancing measures as ways to prevent SARS-CoV-2 infection.

Mitracarpus frigidus (Rubiaceae) inhibits inflammatory and oxidative stress mediators in Salmonella sp. mouse infection.

OBJECTIVES: Evaluation of the in-vivo anti-inflammatory activity of the methanolic extract obtained from the aerial parts of Mitracarpus frigidus (MFM) in the infection caused by two Salmonella strains and its chemical fingerprint by UFLC-quadrupole time of flight-MS. METHODS: The efficacy of MFM was investigated in a classical in-vivo Salmonella infection mouse model. A Salmonella reference strain (ATCC 13311) and a clinical isolate were used to infect mice and then MFM was orally administered during 14 days. At the end of the treatment with MFM, the infection and inflammatory levels were assayed. KEY FINDINGS: MFM treatment showed a significant reduction in mice mortality by Salmonella infection and, also, did not cause alterations in the liver function. Inhibitions of inflammatory and oxidative stress mediators [malondialdehyde (MDA), catalase, and metalloproteinase] were possibly involved in the observed effects. Chlorogenic acid, clarinoside, quercetin-pentosylhexoside, rutin, kaempferol-3O-rutinoside, kaempferol-rhamnosylhexoside and 2-azaanthraquinone were identified in MFM. CONCLUSIONS: MFM was effective in some inflammatory parameters, in the experimental conditions that were used in the study. The results presented in this study and the previous in-vitro anti-Salmonella activity reported by our research group reinforce the importance of MFM studies to considerer it as an alternative treatment for salmonellosis.

Protocolo de profilaxis antibiótica preoperatoria para pacientes ortopédicos del Hospital México: un análisis de cumplimiento / Hospital Mexico’s preoperative antibiotic prophylaxis protocol for orthopedic patients: a compliance analysis

Objetivo: Analizar el cumplimiento en la prescripción de la profilaxisantibiótica prequirúrgica, según el protocolo establecido por el Programade Optimización de Antibióticos y el Servicio de Ortopedia del HospitalMéxico, Caja Costarricense del Seguro Social, ubicado en San José,Costa Rica, de febrero a marzo de 2019. Desde la instauración del protocolo en 2018 no se ha realizado una evaluación de su cumplimiento nise ha determinado la tasa de infección del sitio quirúrgico.Método: Estudio observacional retrospectivo; se identificaron lospacientes hospitalizados en el Servicio de Ortopedia desde el 1 defebrero al 31 de marzo de 2019. Se accedió al expediente digitalde cada paciente, se caracterizó la profilaxis antibiótica prescrita y seanalizó el cumplimiento según el protocolo vigente. Se estimó la tasade infección de sitio quirúrgico. Se realizó un análisis descriptivo de lapoblación.Resultados: El estudio incluyó 110 expedientes clínicos. Los antibióticos más prescritos fueron clindamicina y gentamicina de manera concomitante. El cumplimiento de la prescripción de profilaxis antibiótica oscilóentre 89,1% y 100% para los criterios considerados en el protocolo,exceptuando dosis y antibiótico al alta (14,3 y 65,8%, respectivamente). (AU)

Objective: To analyze compliance with a prescribed pre-surgical antibiotic prophylaxis protocol established by the Antimicrobial StewardshipProgram team and the Orthopedics Department of Hospital Mexico, apublic hospital located in Costa Rica, from February to March 2019. Noassessments of compliance with the protocol had been conducted sinceits introduction in 2018, nor had variations in surgical site infection ratesbeen determined.Method: This is a retrospective observational study that extended fromFebruary 1st to March 31st, 2019. We identified patients hospitalizedduring the study period in the hospital’s Orthopedics Department. Wereviewed each patient’s medical record to record their prescribed antibiotic prophylaxis. Following an analysis of the overall compliance with theprotocol, we made an estimation of the surgical site infection rate as wellas a descriptive analysis of the studied population.Results: The study included 110 clinical records. The most frequentlyprescribed antibiotics were clindamycin and gentamicin. Compliancewith the protocol ranged between 89.1% and 100% across the differentcriteria, except for dosing appropriateness and prescription of antibioticsat discharge (14.3% and 65.8%, respectively). The most common noncompliance factor was gentamicin´s dosing. The surgical site infection ratewas 5%. (AU)