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Stress, infections, and psychological and social well-being can affect the reproductive system. Activation of the hypothalamic-pituitary-adrenal axis can disrupt ovarian cyclicity. Estrogens can modulate stress responsiveness and mood. Thus, understanding this interaction and how it modulates the menstrual cycle is crucial for women's reproductive health. PURPOSE: The objective of this study was to analyze the influence of a stressor, a period of the Covid-19 pandemic when there were no vaccines available yet, on the psychological state of women aged 18 to 45 years; as well as the influence of mental health on the menstrual cycle, considering the influence of age and hormonal contraceptives. METHOD: Online questionnaire using the Google Forms platform was used. RESULTS: There is a high prevalence of the onset of new psychosocial symptoms. Moreover, most women reported some type of change in their menstrual cycles. The women who were using hormonal contraceptives demonstrated a higher frequency of spotting and menstrual color alterations, while women without hormonal contraceptives demonstrated a higher frequency of cycle duration and menstrual odor alterations. Women without hormonal contraceptives were more susceptible to the development of psychosocial symptoms. Younger adult women were more affected by menstrual changes and psychosocial symptoms. Close to 90% of women who reported several psychosocial symptoms had changes in their menstrual cycles. CONCLUSION: These data suggest the impact of stressors, such as a period of the pandemic, on mental health and menstrual cycles, and younger adult women can be more susceptible. This reflects the relationship between mental and reproductive health.
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BACKGROUND: Paracoccidioidomycosis (PCM), a systemic mycosis in Latin America, is regulated by suppressive mechanisms mediated by tolerogenic plasmacytoid-dendritic-cells and regulatory T-cells. Our recent studies revealed that myeloid-derived suppressor cells (MDSCs), are important mediators in PCM. Their suppressive activity on Th1/Th17 immunity was shown to be mediated by inhibitory effect of IL-10, IDO-1 and PD-L1. Studies revealed the chemotherapeutic drug 5-Fluorouracil (5-FU) as a selective MDSC apoptosis-inducing agent, but its in vivo effect on infectious processes remains poorly investigated. METHODS: MDSCs and other leukocytes were evaluated in the lungs of 5-FU-treated mice after four, six, and eight weeks of P. brasiliensis infection. Disease severity and immunological response were evaluated in MDSCs-depleted. RESULTS: 5-FU treatment caused a significant reduction of pulmonary MDSCs and fungal loads. The specific depletion of MDSCs by 5-FU reduced all pulmonary CD4+ T-cell populations (Th1, Th2, Th17, and Treg) resulting in improved tissue pathology and increased survival rates. Importantly, this reduction was concomitant with increased frequencies of Th1/Th17 cells and the increased levels of Th1/Th2/Th17 cytokines in the lungs and liver of treated mice suggesting an early and efficient protective effect of these cells. Furthermore, the immuneprotection conferred by the specific depletion of MDSCs by 5FU treatment could be reversed by the adoptive transfer of MDSCs. CONCLUSIONS: 5-FU treatment depletes lung-MDSCs of P. brasiliensis-infected mice resulting in enhanced immunity. The protective effect of 5-FU treatment in pulmonary PCM suggests that the specific depletion of MDSCs can be viewed as a potential immunotherapeutic tool for PCM.
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Background: Paracoccidioidomycosis (PCM) is a systemic endemic fungal disease prevalent in Latin America. Previous studies revealed that host immunity against PCM is tightly regulated by several suppressive mechanisms mediated by tolerogenic plasmacytoid dendritic cells, the enzyme 2,3 indoleamine dioxygenase (IDO-1), regulatory T-cells (Tregs), and through the recruitment and activation of myeloid-derived suppressor cells (MDSCs). We have recently shown that Dectin-1, TLR2, and TLR4 signaling influence the IDO-1-mediated suppression caused by MDSCs. However, the contribution of these receptors in the production of important immunosuppressive molecules used by MDSCs has not yet been explored in pulmonary PCM. Methods: We evaluated the expression of PD-L1, IL-10, as well as nitrotyrosine by MDSCs after anti-Dectin-1, anti-TLR2, and anti-TLR4 antibody treatment followed by P. brasiliensis yeasts challenge in vitro. We also investigated the influence of PD-L1, IL-10, and nitrotyrosine in the suppressive activity of lung-infiltrating MDSCs of C57BL/6-WT, Dectin-1KO, TLR2KO, and TLR4KO mice after in vivo fungal infection. The suppressive activity of MDSCs was evaluated in cocultures of isolated MDSCs with activated T-cells. Results: A reduced expression of IL-10 and nitrotyrosine was observed after in vitro anti-Dectin-1 treatment of MDSCs challenged with fungal cells. This finding was further confirmed in vitro and in vivo by using Dectin-1KO mice. Furthermore, MDSCs derived from Dectin-1KO mice showed a significantly reduced immunosuppressive activity on the proliferation of CD4+ and CD8+ T lymphocytes. Blocking of TLR2 and TLR4 by mAbs and using MDSCs from TLR2KO and TLR4KO mice also reduced the production of suppressive molecules induced by fungal challenge. In vitro, MDSCs from TLR4KO mice presented a reduced suppressive capacity over the proliferation of CD4+ T-cells. Conclusion: We showed that the pathogen recognition receptors (PRRs) Dectin-1, TLR2, and TLR4 contribute to the suppressive activity of MDSCs by inducing the expression of several immunosuppressive molecules such as PD-L1, IL-10, and nitrotyrosine. This is the first demonstration of a complex network of PRRs signaling in the induction of several suppressive molecules by MDSCs and its contribution to the immunosuppressive mechanisms that control immunity and severity of pulmonary PCM.
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Antígeno B7-H1 , Modelos Animais de Doenças , Interleucina-10 , Lectinas Tipo C , Camundongos Endogâmicos C57BL , Células Supressoras Mieloides , Paracoccidioidomicose , Receptor 2 Toll-Like , Receptor 4 Toll-Like , Animais , Camundongos , Interleucina-10/metabolismo , Receptor 2 Toll-Like/metabolismo , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/imunologia , Células Supressoras Mieloides/imunologia , Células Supressoras Mieloides/metabolismo , Receptor 4 Toll-Like/metabolismo , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologia , Lectinas Tipo C/metabolismo , Lectinas Tipo C/genética , Antígeno B7-H1/metabolismo , Antígeno B7-H1/genética , Paracoccidioidomicose/imunologia , Paracoccidioides/imunologia , Tirosina/análogos & derivados , Tirosina/metabolismo , Linfócitos T Reguladores/imunologia , Pulmão/imunologia , Pulmão/microbiologia , Transdução de Sinais , Masculino , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Indolamina-Pirrol 2,3,-Dioxigenase/genética , Camundongos KnockoutRESUMO
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.
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Antifúngicos , Candida albicans , Nanopartículas Metálicas , Prata , Candida albicans/efeitos dos fármacos , Prata/química , Prata/farmacologia , Antifúngicos/farmacologia , Antifúngicos/química , Antifúngicos/síntese química , Nanopartículas Metálicas/química , Nanopartículas Metálicas/toxicidade , Animais , Testes de Sensibilidade Microbiana , Compostos de Nitrogênio/química , Compostos de Nitrogênio/farmacologia , Compostos de Nitrogênio/toxicidade , Camundongos , NitrilasRESUMO
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.
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Paracoccidioidomicose , Camundongos , Animais , Antígeno CTLA-4 , Receptor de Morte Celular Programada 1 , Camundongos Endogâmicos C57BL , Gravidade do Paciente , Imunoglobulina GRESUMO
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.
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Vesículas Extracelulares , Lectinas Tipo C , Paracoccidioides , Paracoccidioidomicose , Animais , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo , Proteoma , Paracoccidioidomicose/microbiologia , Vesículas Extracelulares/metabolismo , Fatores de VirulênciaRESUMO
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.
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Paracoccidioides , Paracoccidioidomicose , Animais , Camundongos , Paracoccidioides/genética , Proteômica , Camundongos Endogâmicos C57BL , Ferro/metabolismo , Imunidade , GranulomaRESUMO
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.
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Células Supressoras Mieloides , Paracoccidioidomicose , Animais , Camundongos , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Camundongos KnockoutRESUMO
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.
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Células Supressoras Mieloides , Paracoccidioidomicose , Camundongos , Animais , Células Th17/patologia , Camundongos Endogâmicos C57BL , PulmãoRESUMO
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.
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Linho , Morus , Animais , Antioxidantes/farmacologia , Suplementos Nutricionais , Estrogênios , Feminino , Humanos , Ovariectomia , Fitoestrógenos/farmacologia , Extratos Vegetais/farmacologia , Ratos , Ratos Wistar , Ácido ÚricoRESUMO
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.
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Cardiomiopatia Chagásica , Disfunção Ventricular Esquerda , Cardiomiopatia Chagásica/diagnóstico por imagem , Estudos Transversais , Ecocardiografia , Humanos , Volume Sistólico , Disfunção Ventricular Esquerda/diagnóstico por imagem , Disfunção Ventricular Esquerda/epidemiologia , Função Ventricular EsquerdaRESUMO
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.
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Cetoprofeno , Varredura Diferencial de Calorimetria , Pós , Sódio , Solubilidade , Espectroscopia de Infravermelho com Transformada de Fourier , Água/química , Difração de Raios XRESUMO
The stockpiled forage canopy consists of tillers at different ages, which have specific development patterns. The objective was to understand the development of Urochloa brizanthacv. Marandu during the stockpiling period, by the morphogenic evaluation of tillers at different ages. The experiment was conducted in a completely randomized design, in a split plot scheme, with four replications. Three tillers ages (young, mature and old) and two periods of stockpiling (initial and final) were evaluated. The leaf appearance and elongation rates were higher in young tillers (0.05 tiller-1day-1and 0.62 cm tiller-1day-1, respectively), compared to old tillers (0.02 tiller leaf-1day-1and 0.20 cm tiller-1day-1, respectively). The final leaf length of the tillers' age groups was the same in the beginning of stockpiling. The number of live leaves was lower in the old tillers (2.4), compared to the young (3.6) and mature (4.1) ones, contrary to the stem length. The number of old tillers (800 tillers m-2) was higher than the young (299 tillers m-2) and mature ones (358 tillers m-²). The participation of different age groups of tillersin the canopy influences the development and structure of marandu palisadegrass.(AU)
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Pastagens/análise , Poaceae/química , MorfogêneseRESUMO
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.
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Fosfatase Alcalina/química , Temperatura Alta , Thermus thermophilus/enzimologia , Sequência de Aminoácidos , Cristalografia por Raios X , Estabilidade Enzimática , Ligação de Hidrogênio , Simulação de Dinâmica Molecular , Conformação Proteica , Homologia de SequênciaRESUMO
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.
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Fitoestrógenos/farmacologia , Antioxidantes/farmacologia , Doenças Cardiovasculares , Cognição , Dieta , Feminino , Humanos , Isoflavonas/metabolismo , Lipídeos/sangue , Menopausa , Fármacos Neuroprotetores/farmacologia , Osteoporose , Estresse Oxidativo , Fitoestrógenos/química , Plantas , Receptores de Estrogênio/metabolismoRESUMO
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.
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Lactonas/administração & dosagem , Sesquiterpenos/administração & dosagem , Pele/efeitos dos fármacos , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/efeitos adversos , Anti-Inflamatórios/farmacologia , Dermatite de Contato/etiologia , Relação Dose-Resposta a Droga , Cobaias , Humanos , Lactonas/efeitos adversos , Lactonas/farmacologia , Camundongos , NF-kappa B/metabolismo , Sesquiterpenos/efeitos adversos , Sesquiterpenos/farmacologia , Pele/patologiaRESUMO
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.
Assuntos
Analgésicos/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Cicatriz/tratamento farmacológico , Edema/tratamento farmacológico , Hiperalgesia/tratamento farmacológico , Sesquiterpenos/uso terapêutico , Analgésicos/farmacologia , Animais , Anti-Inflamatórios/farmacologia , Araquidonato 5-Lipoxigenase/metabolismo , Carragenina , Cicatriz/metabolismo , Colágeno/metabolismo , Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/metabolismo , Dinoprostona/metabolismo , Edema/induzido quimicamente , Leucotrieno B4/metabolismo , Lipopolissacarídeos/farmacologia , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Células RAW 264.7 , Sesquiterpenos/farmacologia , Tato , Fator de Necrose Tumoral alfa/metabolismoRESUMO
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.
Assuntos
Humanos , Masculino , Feminino , Adulto Jovem , Terapia Miofuncional/métodos , Respiração Bucal/reabilitação , Eletromiografia , Forame Mentual , Massagem , Músculos da MastigaçãoRESUMO
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.
Assuntos
Pastagens , BrachiariaRESUMO
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.