Silver Nanoparticle-Embedded Carbon Nitride: Antifungal Activity on Candida albicans and Toxicity toward Animal Cells.

The development of engineered nanomaterials has been considered a promising strategy to control oral infections. In this study, silver-embedded carbon nitrides (Ag@g-CN) were synthesized and tested against Candida albicans, investigating their antifungal action and biocompatibility in animal cells. Ag@g-CN was synthesized by a simple one-pot thermal polymerization technique and characterized by various analytical techniques. X-ray diffraction (XRD) analysis revealed slight alterations in the crystal structure of g-CN upon the incorporation of Ag. Fourier transform infrared (FT-IR) spectroscopy confirmed the presence of Ag-N bonds, indicating successful silver incorporation and potential interactions with g-CN's amino groups. UV-vis spectroscopy demonstrated a red shift in the absorption edge of Ag@g-CN compared with g-CN, attributed to the surface plasmon resonance effect of silver nanoparticles. Field emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM) confirmed the 2D layered sheet like morphology of both materials. The Ag 3d peaks found in X-ray photoelectron spectroscopy (XPS) confirmed the presence of metallic Ag0 nanoparticles in Ag@g-CN. The Ag@g-CN materials exhibited high antifungal activity against reference and oral clinical strains of C. albicans, with minimal inhibitory concentration (MIC) ranges between 16-256 µg/mL. The mechanism of Ag@g-CN on C. albicans was attributed to the disruption of the membrane integrity and disturbance of the biofilm. In addition, the Ag@g-CN material showed good biocompatibility in the fibroblastic cell line and in Galleria mellonella, with no apparent cytotoxicity observed at a concentration up to 1000 µg/mL. These findings demonstrate the potential of the Ag@g-CN material as an effective and safe antifungal agent for the treatment of oral fungal infections.

Inducing molecular orientation in solution-processed thin films of fluorene-bithiophene-based copolymer: thermal annealing vs. solvent additive.

A deep understanding of the factors influencing the morphology of thin films based on conjugated polymers is essential to boost their performance in optoelectronic devices. Herein, we investigated the electronic structure and morphology of thin films of the copolymer poly(9,9-dioctyl-fluorenyl-co-bithiophene) (F8T2) in its pristine form as well as samples processed with the solvent additive 1,8-diiodooctane (DIO) or post-processed through thermal annealing treatment. Measurements were carried out using angle-resolved S K-edge NEXAFS (near-edge X-ray absorption fine structure) in total electron yield (TEY) and fluorescence yield (FY) detection modes. Two main transitions were observed at the S 1s NEXAFS spectra: S 1s → π* and S 1s → σ* (S-C). The observed dichroism pointed to a face-on orientation of the conjugated backbone, which was significantly increased for F8T2 films processed with DIO. Resonant Auger decay spectra were obtained and analyzed using the core-hole clock (CHC) method. An enhancement in the charge transfer process was observed for thermally annealed films, especially for samples processed with DIO, corresponding to an increase in film ordering. Furthermore, the investigated films were characterized using X-ray photoelectron spectroscopy, attesting to the presence of the thiophene unit in the samples and demonstrating that some of its sulfur atoms were positively polarized in the F8T2 films. All these experimental findings were compared with molecular dynamics (MD) simulations of film evaporation with and without DIO. The use of MD, together with mathematical modeling, was able to explain the major effects found in the experiments, including the polarization of sulfur atoms. The simultaneous use of powerful spectroscopic techniques and theoretical methods shed light on key aspects linking film morphology with fabrication procedures.

Blocking the CTLA-4 and PD-1 pathways during pulmonary paracoccidioidomycosis improves immunity, reduces disease severity, and increases the survival of infected mice.

Immune checkpoint pathways, i.e., coinhibitory pathways expressed as feedback following immune activation, are crucial for controlling an excessive immune response. Cytotoxic T lymphocyte antigen-4 (CTLA-4) and programmed cell death protein-1 (PD-1) are the central classical checkpoint inhibitory (CPI) molecules used for the control of neoplasms and some infectious diseases, including some fungal infections. As the immunosuppression of severe paracoccidioidomycosis (PCM), a chronic granulomatous fungal disease, was shown to be associated with the expression of coinhibitory molecules, we hypothesized that the inhibition of CTLA-4 and PD-1 could have a beneficial effect on pulmonary PCM. To this end, C57BL/6 mice were infected with Paracoccidioides brasiliensis yeasts and treated with monoclonal antibodies (mAbs) α-CTLA-4, α-PD-1, control IgG, or PBS. We verified that blockade of CTLA-4 and PD-1 reduced the fungal load in the lungs and fungal dissemination to the liver and spleen and decreased the size of pulmonary lesions, resulting in increased survival of mice. Compared with PBS-treated infected mice, significantly increased levels of many pro- and anti-inflammatory cytokines were observed in the lungs of α-CTLA-4-treated mice, but a drastic reduction in the liver was observed following PD-1 blockade. In the lungs of α-CPI and IgG-treated mice, there were no changes in the frequency of inflammatory leukocytes, but a significant reduction in the total number of these cells was observed. Compared with PBS-treated controls, α-CPI- and IgG-treated mice exhibited reduced pulmonary infiltration of several myeloid cell subpopulations and decreased expression of costimulatory molecules. In addition, a decreased number of CD4+ and CD8+ T cells but sustained numbers of Th1, Th2, and Th17 T cells were detected. An expressive reduction in several Treg subpopulations and their maturation and suppressive molecules, in addition to reduced numbers of Treg, TCD4+, and TCD8+ cells expressing costimulatory and coinhibitory molecules of immunity, were also detected. The novel cellular and humoral profiles established in the lungs of α-CTLA-4 and α-PD-1-treated mice but not in control IgG-treated mice were more efficient at controlling fungal growth and dissemination without causing increased tissue pathology due to excessive inflammation. This is the first study demonstrating the efficacy of CPI blockade in the treatment of pulmonary PCM, and further studies combining the use of immunotherapy with antifungal drugs are encouraged.

Extracellular vesicles from virulent P. brasiliensis induce TLR4 and dectin-1 expression in innate cells and promote enhanced Th1/Th17 response.

Extracellular vesicles (EVs) are membrane-enclosed nanoparticles that transport several biomolecules and are involved in important mechanisms and functions related to the pathophysiology of fungal diseases. EVs from Paracoccidioides brasiliensis, the main causative agent of Paracoccidioidomycosis (PCM), modulate the immune response of macrophages. In this study, we assessed the EVs proteome from a virulent P. brasiliensis isolated from granulomatous lesions and compared their immunomodulatory ability with EVs isolated from the fungus before the animal passage (control EVs) when challenging macrophages and dendritic cells (DCs). Proteome showed that virulent EVs have a higher abundance of virulence factors such as GP43, protein 14-3-3, GAPDH, as well as virulence factors never described in PCM, such as aspartyl aminopeptidase and a SidJ analogue compared with control EVs. Virulent extracellular vesicles induced higher expression of TLR4 and Dectin-1 than control EVs in macrophages and dendritic cells (DCs). In opposition, a lower TLR2 expression was induced by virulent EVs. Additionally, virulent EVs induced lower expression of CD80, CD86 and TNF-α, but promoted a higher expression of IL-6 and IL-10, suggesting that EVs isolated from virulent P. brasiliensis-yeast promote a milder DCs and macrophage maturation. Herein, we showed that EVs from virulent fungi stimulated a higher frequency of Th1/Tc1, Th17, and Treg cells, which gives new insights into fungal extracellular vesicles. Taken together, our results suggest that P. brasiliensis utilizes its EVs as virulence bags that manipulate the immune system in its favour, creating a milder immune response and helping with fungal evasion from the immune system.

Transcriptional profiling of a fungal granuloma reveals a low metabolic activity of Paracoccidioides brasiliensis yeasts and an actively regulated host immune response.

Granulomas are important immunological structures in the host defense against the fungus Paracoccidioides brasiliensis, the main etiologic agent of Paracoccidioidomycosis (PCM), a granulomatous systemic mycosis endemic in Latin America. We have performed transcriptional and proteomic studies of yeasts present in the pulmonary granulomas of PCM aiming to identify relevant genes and proteins that act under stressing conditions. C57BL/6 mice were infected with 1x106 yeasts and after 8- and 12-weeks of infection, granulomatous lesions were obtained for extraction of fungal and murine RNAs and fungal proteins. Dual transcriptional profiling was done comparing lung cells and P. brasiliensis yeasts from granulomas with uninfected lung cells and the original yeast suspension used in the infection, respectively. Mouse transcripts indicated a lung malfunction, with low expression of genes related to muscle contraction and organization. In addition, an increased expression of transcripts related to the activity of neutrophils, eosinophils, macrophages, lymphocytes as well as an elevated expression of IL-1ß, TNF-α, IFN-γ, IL-17 transcripts were observed. The increased expression of transcripts for CTLA-4, PD-1 and arginase-1, provided evidence of immune regulatory mechanisms within the granulomatous lesions. Also, our results indicate iron as a key element for the granuloma to function, where a high number of transcripts related to fungal siderophores for iron uptake was observed, a mechanism of fungal virulence not previously described in granulomas. Furthermore, transcriptomics and proteomics analyzes indicated a low fungal activity within the granuloma, as demonstrated by the decreased expression of genes and proteins related to energy metabolism and cell cycle.

The immunosuppressive activity of myeloid-derived suppressor cells in murine Paracoccidioidomycosis relies on Indoleamine 2,3-dioxygenase activity and Dectin-1 and TLRs signaling.

Paracoccidioidomycosis (PCM) is a systemic mycosis with a high incidence in Latin America. Prior studies have demonstrated the significance of the enzyme Indoleamine 2,3-dioxygenase (IDO-1) in the immune regulation of PCM as well as the vital role of myeloid-derived suppressor cells (MDSCs) in moderating PCM severity. Additionally, Dectin-1 and Toll-Like Receptors (TLRs) signaling in cancer, infection, and autoimmune diseases have been shown to impact MDSC-IDO-1+ activity. To expand our understanding of MDSCs and the role of IDO-1 and pattern recognition receptors (PRRs) signaling in PCM, we generated MDSCs in vitro and administered an IDO-1 inhibitor before challenging the cells with Paracoccidioides brasiliensis yeasts. By co-culturing MDSCs with lymphocytes, we assessed T-cell proliferation to examine the influence of IDO-1 on MDSC activity. Moreover, we utilized specific antibodies and MDSCs from Dectin-1, TLR4, and TLR2 knockout mice to evaluate the effect of these PRRs on IDO-1 production by MDSCs. We confirmed the importance of these in vitro findings by assessing MDSC-IDO-1+ in the lungs of mice following the fungal infection. Taken together, our data show that IDO-1 expression by MDSCs is crucial for the control of T-cell proliferation, and the production of this enzyme is partially dependent on Dectin-1, TLR2, and TLR4 signaling during murine PCM.

Myeloid-derived suppressor cells are associated with impaired Th1 and Th17 responses and severe pulmonary paracoccidioidomycosis which is reversed by anti-Gr1 therapy.

Previous studies on paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in Latin America, revealed that host immunity is tightly regulated by several suppressive mechanisms mediated by tolerogenic plasmacytoid dendritic cells, the enzyme 2,3 indoleamine dioxygenase (IDO-1), and regulatory T-cells (Tregs). IDO-1 orchestrates local and systemic immunosuppressive effects through the recruitment and activation of myeloid-derived suppressor cells (MDSCs), a heterogeneous population of myeloid cells possessing a potent ability to suppress T-cell responses. However, the involvement of MDSCs in PCM remains uninvestigated. The presence, phenotype, and immunosuppressive activity of MDSCs were evaluated at 96 h, 2 weeks, and 8 weeks of pulmonary infection in C57BL/6 mice. Disease severity and immune responses were assessed in MDSC-depleted and nondepleted mice using an anti-Gr1 antibody. Both monocytic-like MDSCs (M-MDSCs) and polymorphonuclear-like MDSCs (PMN-MDSCs) massively infiltrated the lungs during Paracoccidioides brasiliensis infection. Partial reduction of MDSC frequency led to a robust Th1/Th17 lymphocyte response, resulting in regressive disease with a reduced fungal burden on target organs, diminishing lung pathology, and reducing mortality ratio compared with control IgG2b-treated mice. The suppressive activity of MDSCs on CD4 and CD8 T-lymphocytes and Th1/Th17 cells was also demonstrated in vitro using coculture experiments. Conversely, adoptive transfer of MDSCs to recipient P. brasiliensis-infected mice resulted in a more severe disease. Taken together, our data showed that the increased influx of MDSCs into the lungs was linked to more severe disease and impaired Th1 and Th17 protective responses. However, protective immunity was rescued by anti-Gr1 treatment, resulting in a less severe disease and controlled tissue pathology. In conclusion, MDSCs have emerged as potential target cells for the adjuvant therapy of PCM.

Análise das intercorrências no transporte inter-hospitalar da pessoa em situação crítica / Analysis of intercurrences in the inter-hospital transport of critically ill patients

O transporte inter-hospitalar da pessoa em situação crítica é um procedimento frequente face à necessidade de meios complementares de diagnóstico e terapêutica com elevado nível de diferenciação. Embora represente um risco adicional para o doente, o planeamento adequado é fundamental para a diminuição da ocorrência de complicações. Objetivos: Analisar as intercorrências no transporte inter-hospitalar da pessoa em situação critica do Serviço de Urgência Geral de um Centro Hospitalar da região norte de Portugal de julho de 2020 a março de 2021. Métodos: Estudo transversal retrospetivo, de natureza quantitativa. Realizado em 48 doentes provenientes do Serviço de Urgência Geral de um Centro Hospitalar da região norte de Portugal, sujeitos a transporte inter-hospitalar, por via terrestre, entre julho de 2020 a março de 2021. A recolha de dados decorreu no mês de julho e agosto de 2021. Como instrumento de recolha de dados foi elaborada uma grelha de registos, tendo por base a folha de registo de transporte inter-hospitalar. O tratamento de dados foi no programa Statistical Package for the Social Sciences (SPSS) versão 25.0., com recurso às estatísticas, descritiva e inferencial. O estudo obteve parecer favorável da Comissão de ética R.2644 de 25.06.2021. Resultados: Amostra maioritariamente do sexo masculino (60,4%), no grupo etário entre os 18-64 anos (39,6%), com uma média de idades de 66,75±20,26 anos. A duração média do tempo de transporte foi de 64,11±08,18 minutos. A especialidade de origem mais frequente foi a neurologia (64,4%), a de destino a neurocirurgia (52,1%), com o diagnóstico do sistema nervoso (54,2%). Escala de coma de Glasgow com 15 pontos (60,4%) e com ventilação espontânea (47,9%). Registaram-se 79,2% de intercorrências, sendo que destas 80,9% foram clínicas e a mais frequente foi a tensão arterial ≥ 140mmHg (47,9%), no sexo masculino (63,2%), com idade entre os 75-84 anos (100%), com o diagnóstico do sistema nervoso (84,6%) e a especialidade de origem a neurologia. Verificou-se diferença, estatística significativa, entre as intercorrências clínicas e a especialidade de origem (p=0,033). As intercorrências não clínicas foram de (19,1%) e a mais frequente foi a exteriorização do cateter venoso periférico (16,7%) e no grupo etário superior 85 anos (45,5%) verificando-se uma associação, estatisticamente significativa entre as intercorrências não clinicas e o grupo etário (p=0,007). Conclusão: Verificou-se maior frequência de intercorrências clínicas, no sexo masculino, com idade entre os 75-84 anos e com alterações neurológicas, associando-se com a especialidade de origem. As não clínicas foram mais frequentes nos mais idosos. Sugerimos uma monitorização, dos parâmetros vitais mais frequente ou mesmo invasiva neste grupo etário, bem como vigilância do estado de consciência destes doentes. A realização de outros estudos com amostras maiores e preferencialmente prospetivos.

The inter-hospital transport of the critically ill person is a frequent procedure given the need for complementary means of diagnosis and therapy with a high level of differentiation. Although it represents an additional risk for the patient, proper planning is essential to reduce the occurrence of complications. Objectives: To analyze the intercurrences in the inter-hospital transport of the person in a critical situation of the General Emergency Service of a Hospital Center in the northern region of Portugal from July 2020 to March 2021. Methods: Retrospective cross-sectional study of a quantitative nature. Performed on 48 patients from the General Emergency Department of a Hospital Center in the northern region of Portugal, subject to inter-hospital transport, by land, between July 2020 and March 2021. Data collection took place in July and August 2021. As a data collection instrument, a grid of records was prepared, based on the inter-hospital transport record sheet. Data processing was done using the Statistical Package for the Social Sciences (SPSS) version 25.0., using descriptive and inferential statistics. The study obtained a favorable opinion from the Ethics Committee R.2644 de 25.06.2021. Results: The sample was mostly male (60.4%), in the age group between 18-64 years (39.6%), with a mean age of 66.75±20.26 years. The mean duration of transport time was 64.11±08.18 minutes. The most frequent specialty of origin was neurology (64.4%), that of destination neurosurgery (52.1%), with the diagnosis of the nervous system (54.2%). Glasgow Coma Scale with 15 points (60.4%) and with spontaneous ventilation (47.9%). There were 79.2% of complications, of which 80.9% were clinical and the most frequent was blood pressure ≥ 140mmHg (47.9%), in males (63.2%) and aged between 75-84 years (100%), with the diagnosis of the nervous system (84.6%) and the specialty of origin neurology. There was a statistically significant difference between clinical complications and the specialty of origin (p=0.033). Non-clinical complications were (19.1%) and the most frequent was the externalization of the peripheral venous catheter (16.7%) and in the age group over 85 years (45.5%) there was a statistically significant association between non-clinical complications and age group (p=0.007). Conclusion: There was a higher frequency of clinical complications, in males, aged between 75- 84 years and with neurological alterations, associated with the specialty of origin. Non-clinical ones were more frequent in the elderly. We suggest a more frequent or even invasive monitoring of vital parameters in this age group, as well as monitoring the state of consciousness of these patients. Carrying out other studies with larger and preferably prospective samples.

Beneficial Effects of Flaxseed and/or Mulberry Extracts Supplementation in Ovariectomized Wistar Rats.

Low endogenous estrogen action causes several injuries. Medicinal plants, such as flaxseed and mulberry, contain substances that have been shown to be effective to the organism. The aim was to verify the effects of flaxseed and/or mulberry extracts on ovariectomized Wistar rats. The animals received supplements of extracts and estrogen or saline by gavage for 60 days and were weighed weekly. Vaginal wash, blood, pituitary, uterus, liver, and kidneys were collected. Phenolic compounds and the antioxidant activity of the extracts, lipid profile, uric acid, liver enzymes, and pituitary weight were measured. Histomorphometric for uterine wall and histopathological analyses for liver and kidney were performed. Flaxseed and mulberry extracts showed great antioxidant activity and large amounts of phenolic compounds. The treatment with extracts had less weight gain, increased pituitary weight, the predominance of vaginal epithelial cells, and reduced TC, LDL-c and lipase activity, similar to estrogen animals. Estrogen or flaxseed + mulberry animals reduced VLDL-c and TAG. HDL-c, uric acid, and liver enzymes did not differ. Estrogen or extracts demonstrated trophic action on the endometrial thickness and have not shown hepatotoxicity or nephrotoxicity. We suggested the beneficial effects of flaxseed and mulberry extract as an alternative to reduce and/or prevent the negative effects caused by low estrogenic action.

A New Crystalline Ketoprofen Sodium Salt: Solid-State Characterization, Solubility, and Stability.

Ketoprofen (KTP) is an Active Pharmaceutical Ingredient (API) that has low solubility in aqueous solvents. The use of KTP salts has attracted attention due to its improvements in terms of solubility, tolerability, higher rate and extent of absorption, and faster onset of the therapeutic effect. In this work, a crystalline KTP sodium salt (coded as KTP-Na) was successfully obtained and widely characterized by X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), solubility and accelerated stability studies. XRD results showed that KTP-Na is not yet reported in the literature. Moreover, FTIR, DSC and TGA were useful for differentiation of KTP-Na from the KTP commercialized form (coded as KTP-R1). The solubility of KTP-Na in water was about 80 times greater than the KTP-R1. However, KTP-Na showed lower physical stability in storage conditions at 40 ± 2°C/ 75% ± 5% RH when compared to KTP-R1, which was shown to be related to a high hygroscopicity of KTP-Na. Therefore, due to its higher solubility, KTP-Na may be a viable alternative for use in solid dosage forms. However, the presence of moisture must be strictly controlled to avoid water absorption and consequent amorphization.

Myocardial function reclassification: Echocardiographic strain patterns in patients with chronic Chagas cardiomyopathy and intraventricular dyssynchrony.

BACKGROUND: We aimed to identify, among Chronic Chagas Cardiomyopathy (CCC) patients with left ventricular dysfunction (LVD) and non-left bundle branch block (non-LBBB), subgroups with different functional and mechanical patterns of global longitudinal strain (GLS) and intraventricular dyssynchrony (IVD) at rest and after exercise stress test, and reclassify them using a new echocardiographic approach. METHODOLOGY: In this single-center cross-sectional study, 40 patients with CCC, left ventricular ejection fraction (LVEF) ≤ 35% and non-LBBB underwent rest echocardiography and then treadmill exercise stress echocardiography with GLS and IVD analysis. The sample was divided into four groups, based on GLS and IVD significant variation between rest and exercise: GLS + IVD+ (9 patients); GLS + IVD- (9 patients); GLS-IVD+ (10 patients); GLS-IVD- (10 patients). RESULTS: At rest, median LVEF was 28% (21.3%-33%) and GLS (-7% (-5%/-9.3%), were not different among groups. The average response of GLS was an increase of 0.74% over rest values, and the average response of IVD was a decrease of 6.9 ms. Group GLS-IVD+ presented more dyssynchrony at rest (p = 0.01). Left atrial (LA) volume (higher in GLS-IVD-) (p = 0.022) and TAPSE (higher in GLS + IVD+) (p = 0.015) were also different among groups at baseline. Of the 40 patients evaluated, 27 (67.5%) had very severe LVD (GLS < -8%). In addition, among these patients, 11 patients had contractile reserve after undergoing stress echocardiography. CONCLUSIONS: In patients with CCC, severe LVD and non-LBBB, the evaluation of GLS and IVD between rest and exercise was able to reclassify myocardial function and to identify subgroups with contractile reserve and significant dyssynchronopathy.

Dynamic cross correlation analysis of Thermus thermophilus alkaline phosphatase and determinants of thermostability.

BACKGROUND: Understanding the determinants of protein thermostability is very important both from the theoretical and applied perspective. One emerging view in thermostable enzymes seems to indicate that a salt bridge/charged residue network plays a fundamental role in their thermostability. METHODS: The structure of alkaline phosphatase (AP) from Thermus thermophilus HB8 was solved by X-ray crystallography at 2.1 Å resolution. The obtained structure was further analyzed by molecular dynamics studies at different temperatures (303 K, 333 K and 363 K) and compared to homologous proteins from the cold-adapted organisms Shewanella sp. and Vibrio strain G15-21. To analyze differences in measures of dynamic variation, several data reduction techniques like principal component analysis (PCA), residue interaction network (RIN) analysis and rotamer analysis were used. Using hierarchical clustering, the obtained results were combined to determine residues showing high degree dynamical variations due to temperature jumps. Furthermore, dynamic cross correlation (DCC) analysis was carried out to characterize networks of charged residues. RESULTS: Top clustered residues showed a higher propensity for thermostabilizing mutations, indicating evolutionary pressure acting on thermophilic organisms. The description of rotamer distributions by Gini coefficients and Kullback-Leibler (KL) divergence both revealed significant correlations with temperature. DCC analysis revealed a significant trend to de-correlation of the movement of charged residues at higher temperatures. SIGNIFICANCE: The de-correlation of charged residues detected in Thermus thermophilus AP, highlights the importance of dynamic electrostatic network interactions for the thermostability of this enzyme.

The influence of phytoestrogens on different physiological and pathological processes: An overview.

Functional foods have nutritional properties and organic functions, which are beneficial to health. Certain types of functional food components are so-called phytoestrogens, non-steroidal compounds derived from the metabolism of precursors contained in plants, which originate secondary metabotypes known to induce biological responses and by mimicry or modulating the action of endogenous estrogen. These molecules are involved in several physiological and pathological processes related to reproduction, bone remodeling, skin, cardiovascular, nervous, immune systems, and metabolism. This review aimed to present an overview of phytoestrogens regarding their chemical structure, actions, and effects in the organism given several pathologies. Several studies have demonstrated beneficial phytoestrogen actions, such as lipid profile improvement, cognitive function, menopause, oxidative stress, among others. Phytoestrogens effects are not completely elucidated, being necessary future research to understand the exact action mechanisms, whether they are via estrogen receptor or whether other hidden mechanisms produce these effects. Thus, this review makes a general approach to the phytoestrogen actions, beneficial effects, risk and limitations. However, the complexities of biological effects after ingestion of phytoestrogens and the differences in their metabolism and bioavailability indicate that interpretation of either risk or benefits needs to be made with caution.

Impact of sesquiterpene lactones on the skin and skin-related cells? A systematic review of in vitro and in vivo evidence.

Although anti-inflammatory properties are attributed to sesquiterpene lactones (SL), cutaneous hypersensitivity reactions are proposed as limitations for SL-based therapies. Thus, the impact of SL on the skin and skin-related cells was systematically reviewed. Studies indexed in electronic databases were screened from the PRISMA strategy. The risk of bias in all studies was verified from the SYRCLE's tool. Thirty original studies were recovered and analyzed. Mice and guinea pig, keratinocytes and fibroblasts were predominantly investigated from in vivo and in vitro studies, respectively. In vivo studies indicated that most SL induced contact dermatitis associated with edema, erythema, and inflammatory infiltrate. Conversely, in vitro evidence was consistent with a dose-dependent anti-inflammatory effect of SL in response to reduced cytokines, 5-LOX, and COX-2 levels or activity in keratinocytes, fibroblasts, macrophages and dendritic cells; which are events potentially triggered by downregulation of gene expression and/or inhibition of the NF-κB signaling pathway. In vivo studies presented uncertain to high-risk of bias mainly associated with underreporting of randomization and experimental blinding. The current evidence supports potent cutaneous immunomodulatory properties of SL. Although in vitro and in vivo studies indicate opposite anti- or proinflammatory effects, this contradiction exhibits a dose-dependent component. In addition, the anti-inflammatory pathways activated by SL are better understood from in vitro evidence. However, additional studies are required to elucidating specific anti-inflammatory and proinflammatory mechanisms triggered by SL in vivo. Thus, controlling the sources of bias described in this review can contribute to improving the quality of the evidence in further investigations.

Tagitinin F has anti-inflammatory, anti-nociceptive and anti-matrix metalloproteinase properties: An in silico, in vitro and in vivo study.

Sesquiterpene lactones (SL) are natural bioactive molecules indicated as potential scaffolds for anti-inflammatory and analgesic drug design. However, their anti-inflammatory applicability remains underestimated since the impact of SL on inflammatory nociception and tissue repair are overlooked. Thus, we used an integrated in silico, in vitro and in vivo framework to investigate the impact of tagitinin F (TAG-F) on lipopolysaccharide (LPS)-challenged macrophages, excisional skin wounds, and carrageenan-induced paw edema and mechanical hyperalgesia in mice. RAW 264.7 macrophages in culture were challenged with LPS and treated with TAG-F (5, 10, 50 and 100 µM). The paw of BALB/c mice was injected with carrageenan and treated with 0.5% and 1% TAG-F. Excisional wounds were also produced in BALB/c mice and treated with 0.5% and 1% TAG-F. Our results indicated a consistent concentration-dependent downregulation in 5-lipoxygenase, cyclooxygenase 1 and 2 (COX-1 and COX-2), matrix metalloproteinase 1 and 2 (MMP-1 and MMP-2) activities; as well as attenuation in prostaglandin E2 (PGE2), leukotriene B4 (LTB4) and tumor necrosis factor-α (TNF-α) production in both in vitro and in vivo models. In vivo, TAG-F also attenuated carrageenan-induced paw edema and mechanical hyperalgesia in mice. From the excisional skin wound, TAG-F was still effective in reducing neutrophils and macrophages infiltration and stimulating collagen deposition in the scar tissue, accelerating tissue maturation. Together, our findings indicate that the anti-inflammatory effect of TAG-F is more comprehensive than previously suggested, exerting a significant impact on the control of edema, inflammatory pain and modulating central metabolic processes linked to skin wound healing.

Influence of mentalis muscle relaxation on oronasal breathing / A repercussão do relaxamento do músculo mentual na respiração oronasal

ABSTRACT Purpose: to investigate the existence of changes in the electromyographic patterns of the mentalis and inferior orbicularis oris muscles in oronasal breathers, submitted to massage therapy on the mentalis muscle. Methods: a controlled blind placebo experiment, with a sample of 19 oronasal breathers (1 man and 18 women), mean age (standard deviation) 22.3 (2.63) years, randomly divided into control and experimental groups, respectively with 7 and 12 volunteers. The experimental group alone underwent myotherapy with massages for 3 months, while electromyographic data were collected from both groups at the beginning and end of the treatment, both at rest and when swallowing water. The analysis of variance was conducted to test the existence of differences between the means; the 5% significance level was used. Results: the analysis of variance revealed signs of interaction between the group and phase effects when analyzing the root mean square values of both the inferior orbicularis oris and the mentalis muscles. As expected, no signs of significant differences were found between the means of the phases in the control group. On the other hand, signs of significant difference were found in the experimental group, with reduced root mean square values in both muscles. The inferior orbicularis oris muscle, which in the pre-phase had a mean (standard deviation) of 202.10 (161.47) µV, had, in the post-phase, values of 131.49 (159.18) µV. The mentalis muscle, in its turn, had in the pre- and post-phase, respectively, a mean (standard deviation) of 199.31 (279.77) µV and 114.58 (253.56) µV. Conclusion: given that no effect was detected in the control group, the decrease in the root mean square values of the mentalis and inferior orbicularis oris muscles in oronasal breathers was attributed to the massage therapy on the mentalis muscle.

RESUMO Objetivo: investigar a existência de modificações dos padrões eletromiográficos dos músculos mentual e orbicular inferior da boca em respiradores oronasais submetidas à massoterapia no músculo mentual. Métodos: experimento cego placebo controlado, com amostra de 19 respiradores oronasais, 1 homem e 18 mulheres, com média de idade (desvio-padrão) de 22,3 (2,63) anos, aleatoriamente dividida nos grupos controle e experimental, respectivamente com 7 e 12 voluntários. Apenas no grupo experimental foi aplicada mioterapia por meio de massagens, por três meses e em ambos os grupos foram coletados dados eletromiográficos no início e no final do tempo de tratamento nas condições de repouso e deglutição de água. A análise de variância foi aplicada para testar a existência de diferenças entre as médias e foi adotado o nível de significância de 5%. Resultados: a análise de variância revelou indícios de interação entre os efeitos de grupo e fase quando analisados os valores de Root Mean Square (RMS), tanto do músculo orbicular inferior como do músculo mentual. Como esperado, não foram encontrados indícios de diferenças significantes entre as médias das fases no grupo controle, entretanto, foram encontrados indícios de diferença significante no grupo experimental havendo redução das médias de RMS em ambos os músculos. O orbicular inferior, que na fase pré apresentava média (desvio padrão) de 202,10 (161,47) µV, apresentou valores de 131,49 (159,18) µV na fase pós, enquanto que o músculo mentual apresentou, respectivamente, nas fases pré e pós, média (desvio padrão) de 199,31 (279,77) µV e 114,58 (253,56) µV. Conclusão: tendo em vista a não detecção de efeito no grupo controle, atribui-se à massoterapia no músculo mentual a redução das médias de Root Mean Square dos músculos mentual e orbicular inferior da boca em respiradores oronasais.

Mowing and lowering the marandu palisadegrass sward height at late winter improve the sward structure / A roçada e a redução da altura do capim-marandu no final do inverno melhora a estrutura do pasto

The structure of the sward is important because it influences the responses of plants and animals in grazing conditions. The objective with this work was to evaluate the structural characteristics of the Marandu palisade grass pasture (Brachiaria brizantha syn. Urochloa brizantha) during spring and summer, depending on the condition of the sward at the end of winter. Four grazing conditions at the end of winter were evaluated: short (24.1 cm), short (25.2 cm)/mown (8 cm), tall (49.0 cm) and tall (50.0 cm)/mown (8 cm). The foliar area index was lower in the tall pasture than in the tall/mown pasture. The mass and the volumetric density of dead stem were higher in the tall pasture, intermediate in the short pasture, and lower in mowed pastures. At the beginning of the grazing period, the live stem mass was higher in the tall pasture. At the beginning and middle of the grazing period, the volumetric density of dead leaf lamina was lower in mowed pastures. At the beginning of the pasture period, the short pasture had a higher tiller number (TN). In the middle of the grazing period, the short and short/mown pastures had higher TN. At the end of the pasture period, the lowest TN value occurred in the tall pasture. The tall pasture at the end of winter presents an unfavorable structure to the animal in grazing, while the mowing and reduction in the height of the marandu palisade grass improves the structure of the sward in spring and summer.

A estrutura do pasto é importante porque influencia as respostas de plantas e animais em condições de pastejo. O objetivo com este trabalho foi avaliar as características estruturais do pasto de capim-marandu (Brachiaria brizantha syn. Urochloa brizantha) durante a primavera e o verão, em função da condição do pasto no fim do inverno. Quatro condições de pasto no fim do inverno foram avaliadas: baixo (24,1 cm), baixo (25,2 cm)/roçado (8 cm), alto (49,0 cm) e alto (50,0 cm)/roçado (8 cm). O índice de área foliar foi menor no pasto alto do que no pasto alto/roçado. A massa e a densidade volumétrica de colmo morto foram maiores no pasto alto, intermediárias no pasto baixo, e inferiores nos pastos roçados. No início do período de pastejo, a massa de colmo vivo foi maior no pasto alto. No início e meio do período de pastejo, a densidade volumétrica de lâmina foliar morta foi menor nos pastos roçados. No início do período de pastejo, o pasto baixo apresentou maior número de perfilho (NP). No meio do período de pastejo, os pastos baixo e baixo/roçado tiveram maior NP. No fim do período de pastejo, o menor valor de NP ocorreu no pasto alto. O pasto alto no fim do inverno apresenta estrutura desfavorável ao animal em pastejo, enquanto que a roçada e a redução da altura do capim-marandu melhora a estrutura do pasto na primavera e no verão.

Estradiol and the feeding state modulate the interaction between leptin and the nitrergic system in female rats.

Leptin mediates the interaction between reproductive function and energy balance. However, leptin receptors are not expressed in neurons that produce gonadotropin-releasing hormone (GnRH), likely indicating an indirect action through interneurons. Among likely neurons that modulate the secretion of GnRH are NO (nitric oxide) neurons. We assessed whether estradiol and feeding conditions modulate a possible interaction between leptin and NO in brain areas related to the control of reproductive function. Estradiol-treated and untreated ovariectomized rats were normally fed or fasted for 48 h. Then, saline (control) or leptin (3 µg/1 µl) intracerebroventricular microinjections were administered, and after thirty minutes, the brains collected subsequent to the decapitation or transcardially perfusion. Leptin and estradiol increased NO synthase (nNOS) gene expression (RT-PCR) and content (Western blotting) in the medial preoptic area (MPOA) and medial basal hypothalamus (MBH) only in fasted rats. Leptin increased: 1-phosphorylated-signal transducer and activator of transcription-3(pSTAT3) (immunohistochemistry) in the MPOA and various hypothalamic nuclei [arcuate (ARC); ventromedial (VMH); dorsal/ventral dorsomedial (dDMH/vDMH); premammilar ventral (PMV)], effects potentiated by estradiol/fasting interaction; 2- nNOS/pSTAT3 coexpression in the MPOA only in estradiol-treated, fasted rats; 3- nNOS-immunoreactive cell expression in the VMH, DMH and PMV (areas related to reproductive function control) of estradiol -treated rats. Thus, when leptin is reduced during fasting, leptin replacement effectively increased the expression of nitric oxide, which activated the HPG axis only in the presence of estradiol. Estradiol modulates the nitrergic system, leptin sensitivity and consequently leptin's effects on the nitrergic system in hypothalamus and in particular vDMH and PMV.

Sonothrombolysis Improves Myocardial Dynamics and Microvascular Obstruction Preventing Left Ventricular Remodeling in Patients With ST Elevation Myocardial Infarction.

BACKGROUND: It has recently been demonstrated that high-energy diagnostic transthoracic ultrasound and intravenous microbubbles dissolve thrombi (sonothrombolysis) and increase angiographic recanalization rates in patients with ST-segment-elevation myocardial infarction. We aimed to study the effect of sonothrombolysis on the myocardial dynamics and infarct size obtained by real-time myocardial perfusion echocardiography and their value in preventing left ventricular remodeling. METHODS: One hundred patients with ST-segment-elevation myocardial infarction were randomized to therapy (50 patients treated with sonothrombolysis and percutaneous coronary intervention) or control (50 patients treated with percutaneous coronary intervention only). Left ventricular volumes, ejection fraction, risk area (before treatment), myocardial perfusion defect over time (infarct size), and global longitudinal strain were determined by quantitative real-time myocardial perfusion echocardiography and speckle tracking echocardiography imaging. RESULTS: Risk area was similar in the control and therapy groups (19.2±10.1% versus 20.7±8.9%; P=0.56) before treatment. The therapy group presented a behavior significantly different than control group over time (P<0.001). The perfusion defect was smaller in the therapy at 48 to 72 hours even in the subgroup of patients with no recanalization at first angiography (12.9±6.5% therapy versus 18.8±9.9% control; P=0.015). The left ventricular global longitudinal strain was higher in the therapy than control immediately after percutaneous coronary intervention (14.1±4.1% versus 12.0±3.3%; P=0.012), and this difference was maintained until 6 months (17.1±3.5% versus 13.6±3.6%; P<0.001). The only predictor of left ventricular remodeling was treatment with sonothrombolysis: the control group was more likely to exhibit left ventricular remodeling with an odds ratio of 2.79 ([95% CI, 0.13-6.86]; P=0.026). CONCLUSIONS: Sonothrombolysis reduces microvascular obstruction and improves myocardial dynamics in patients with ST-segment-elevation myocardial infarction and is an independent predictor of left ventricular remodeling over time.

Simulation-based assessment in anesthesia: an international multicentre validation study.

PURPOSE: Simulated clinical events provide a means to evaluate a practitioner's performance in a standardized manner for all candidates that are tested. We sought to provide evidence for the validity of simulation-based assessment tools in simulated pediatric anesthesia emergencies. METHODS: Nine centres in two countries recruited subjects to participate in simulated operating room events. Participants ranged in anesthesia experience from junior residents to staff anesthesiologists. Performances were video recorded for review and scored by specially trained, blinded, expert raters. The rating tools consisted of scenario-specific checklists and a global rating scale that allowed the rater to make a judgement about the subject's performance, and by extension, preparedness for independent practice. The reliability of the tools was classified as "substantial" (intraclass correlation coefficients ranged from 0.84 to 0.96 for the checklists and from 0.85 to 0.94 for the global rating scale). RESULTS: Three-hundred and ninety-one simulation encounters were analysed. Senior trainees and staff significantly out-performed junior trainees (P = 0.04 and P < 0.001 respectively). The effect size of grade (junior vs senior trainee vs staff) on performance was classified as "medium" (partial η2 = 0.06). Performance deficits were observed across all grades of anesthesiologist, particularly in two of the scenarios. CONCLUSIONS: This study supports the validity of our simulation-based anesthesiologist assessment tools in several domains of validity. We also describe some residual challenges regarding the validity of our tools, some notes of caution in terms of the intended consequences of their use, and identify opportunities for further research.