A Saúde Bucal e a Prevenção da Pneumonia na UTI: Revisão Literatura / Bucal Health and Prevention of Pneumonia in the ICU: Literature Review / Salud Bucodental y Neumonía en la UCI: Revisión Bibliográfica

Objetivo: Analisar os fatores que interferem na saúde bucal na prevenção da pneumonia, cujos procedimentos metodológicos se fundamentaram a partir da literatura na modalidade integrativa. Método: Estudo do tipo Revisão de literatura nas bases de dados Latino-Americana e do Caribe em Ciências da Saúde (LILACS), Base de dados de Enfermagem (BDENF), Medical Literature Analysis and Retrieval System Online (MEDLINE), Scientific Electronic Library Online (SciELo). Resultados:Foram encontrados resultados mais relevantes que apesar dos profissionais de enfermagem afirmarem que possuem conhecimento suficiente sobre como realizar a higiene oral em pacientes, a aplicação do protocolo adequado diminui drasticamente nos casos de pneumonia associado a ventilação mecânica. Conclusão: Conclui-se que os protocolos de atividades voltadas para assistência à saúde dos pacientes com pneumonia precisam ser intensificados, sobretudo pelos profissionais de enfermagem

Objective: To analyze the factors that interfere with oralhealth in the prevention of pneumonia, whose methodological procedures were based on the literature in the integrative modality. Method:Literature review study in the Latin American and Caribbean Health Sciences databases (LILACS), Nursing Database (BDENF), Medical Literature Analysis and Retrieval System Online (MEDLINE), Scientific Electronic Library Online (SciELo). Results:More relevant results were found that although nursing professionals claim to have sufficient knowledge about how to perform oralhygiene in patients, the application of the appropriate protocol drastically decreases in cases of pneumonia associated with mechanical ventilation. Conclusion:It is concluded that the protocols of activities aimed at health care for patients with pneumonia need to be intensified, especially by nursing professionals.

Objetivo: Analizar los factores que interfieren en la salud bucal en la prevención de la neumonía, cuyos procedimientos metodológicos se basaron en la literatura en la modalidad integrativa. Método:Revisión de la literatura en las bases de datos de Ciencias de la Salud de América Latina y el Caribe (LILACS), Base de Datos de Enfermería (BDENF), Sistema de Análisis y Recuperación de Literatura Médica en Línea (MEDLINE), Scientific Electronic Library Online (SciELo). Resultados: Se encontraron resultados más relevantes que aunque los profesionales de enfermería afirman tener conocimientos suficientes sobre cómo realizar la higiene bucal en los pacientes, la aplicación del protocolo adecuado disminuye drásticamente en los casos de neumonía asociada al ventilador. Conclusión:Se concluye que los protocolos de actividades dirigidas al cuidado de la salud de pacientes con neumonía necesitan ser intensificados, especialmente por los profesionales de enfermería

Calotropis procera (Aiton) Dryand (Apocynaceae): State of the art of its uses and Applications.

Calotropis procera (Aiton) Dryand (Apocynaceae), popularly known as milkweed, has been traditionally used to treat diseases particularly associated with gastric disorders, skin disease and inflammatory processes. The present study aimed to review the current scientific evidence regarding the pharmacological effects of C. procera extracted phytochemicals and possible research opportunities as complementary and alternative medicine. Scientific publications were searched in various electronic databases (PubMed, Scopus, Web of Science, Google Scholar, Springer, Wiley, and Mendeley) using the following search terms: Calotropis procera, medicinal plants, toxicity, phytochemical characterization, and biological effects. Collected data showed that cardenolides, steroid glycoside and flavonoids are the main classes of phytochemicals identified in C. procera latex and leaves. In addition, lignans, terpenes, coumarins, and phenolic acids have been reported. These metabolites have been correlated with their biological activities, including mainly antioxidant, anti-inflammatory, antitumoral, hypoglycemic, gastric protective, anti-microbial, insecticide, anti-fungal, anti-parasitic, among others. However, some of the studies were carried out with only a single dose or with a high dose not achievable under physiological conditions. Therefore, the validity of C. procera biological activity may be questionable. Not less important to highlight are the risks associated with its use and the possibility of accumulation of heavy metals that can be toxic. Furthermore, there are no clinical trials with C. procera to date. In conclusion, the need of bioassayguided isolation of bioactive compounds, bioavailability and efficacy, as well as pharmacological and toxicity studies, are needed using in vivo models and clinical trials in order to support the traditionally claimed health benefits.

Anthocyanins Reduce Cell Invasion and Migration through Akt/mTOR Downregulation and Apoptosis Activation in Triple-Negative Breast Cancer Cells: A Systematic Review and Meta-Analysis.

BACKGROUND: Studies have suggested the chemopreventive effects of anthocyanins on breast cancer carcinogenesis. This systematic review and meta-analysis aimed to evaluate the effect of anthocyanins on triple-negative breast cancer cells (TNBC) cultured in vitro. METHODS: We searched for all relevant studies that evaluated the mechanisms of migration, invasion, Akt/mTOR and MAPK pathways, and apoptosis, using PubMed and Scopus. Means and standard deviation were used, and a randomized effects model was applied, with a confidence interval of 95%. Statistical heterogeneity between studies was assessed using the Chi2 test and I2 statistics. All analyses were performed using RevMan software (version 5.4). RESULTS: Eleven studies were included in the systematic review and ten in the meta-analysis, where the roles of anthocyanin-enriched extract or cyanidin-3-O-glucoside (C-3-O-G) on MDA-MB-231 and MDA-MB-453 cells were investigated. DISCUSSION: There was a significant reduction in invasion (mean difference: -98.64; 95% CI: -153.98, -43.3; p ˂ 0.00001) and migration (mean difference: -90.13; 95% CI: -130.57, -49.68; p ˂ 0.00001) in TNBC cells after anthocyanins treatment. Anthocyanins also downregulated Akt (mean difference: -0.63; 95% CI: -0.70, -0.57; p ˂ 0.00001) and mTOR (mean difference: -0.93; 95% CI: -1.58, -0.29; p = 0.005), while JNK (mean difference: -0.06; 95% CI: -1.21, 1.09; p = 0.92) and p38 (mean difference: 0.05; 95% CI: -1.32, 1.41; p = 0.95) were not modulated. There was also an increase in cleaved caspase-3 (mean difference: 1.13; 95% CI: 0.11, 2.16; p = 0.03), cleaved caspase-8 (mean difference: 1.64; 95% CI: 0.05, 3.22; p = 0.04), and cleaved PARP (mean difference: 0.93; 95% CI: 0.54, 1.32). Although the difference between control and anthocyanin groups was not significant regarding apoptosis rate (mean difference: 3.63; 95% CI: -2.88, 10.14; p = 0.27), the analysis between subgroups showed that anthocyanins are more favorable in inducing overall apoptosis (p ˂ 0.00001). CONCLUSION: The results show that anthocyanins hold promise in fighting against TNBC, but their effects should not be generalized. In addition, further primary studies should be conducted so that more accurate conclusions can be drawn.

Dark Sweet Cherry (Prunus avium) Anthocyanins Suppressed ERK1/2-Akt/mTOR Cell Signaling and Oxidative Stress: Implications for TNBC Growth and Invasion.

This study aimed to assess dark sweet cherry (DSC) total polyphenols (WE) and anthocyanins (ACN) against metastatic breast cancer (BC). The WE and ACN anticancer activity and underlying mechanisms were assessed in vitro using 4T1 BC cells. A pilot study using a BALB/C mouse syngeneic model bearing 4T1 tumors assessed the anti-metastatic potential of ACN in vivo. ACN inhibited cell viability with higher potency than WE and reduced reactive oxygen species (ROS) (IC50 = 58.6 µg cyanidin 3-glucoside equivalent (C3G)/mL or 122 µM). ACN induced p38 stress-related intrinsic apoptosis, leading to caspase-3 cleavage and total PARP decrease. ACN suppressed ERK1/2 and Akt/mTOR signaling pathways, which are abnormally activated in BC and promote motility and invasion. This was consistent with suppression of VCAM-1 mRNA, Scr phosphorylation and 88.6% reduction of cells migrating to wounded area. The pilot in vivo results supported the ACN-mediated suppression of angiogenesis in tumors and lungs. ACN also lowered Cenpf mRNA in lungs, associated with lung metastasis lesions and poor survival. Results demonstrated the dual Akt-ERK inhibitory role of ACN and suppression of their downstream pro-invasive targets. These results encourage a larger scale in vivo study to confirm that ACN may help to fight BC invasion and metastasis.

Calotropis Procera Induced Caspase-Dependent Apoptosis and Impaired Akt/mTOR Signaling in 4T1 Breast Cancer Cells.

INTRODUCTION: Calotropis procera (Aiton) Dryand (Apocynaceae) is an herb that has been commonly used in folk medicine to treat various diseases for more than 1500 years. AIMS: Our goal was to investigate the anti-metastatic effects of phenolics extracted from C. procera (CphE) against 4T1 breast cancer cells and in BALB/c mice. METHODS: 4T1 cells were treated with CphE and quercetin (positive control) at concentrations that inhibited cell viability by 50% (IC50). Levels of reactive oxygen species (ROS), wound healing, and protein expressions were determined following standard protocols. For the in vivo pilot study, the syngeneic BALB/c mouse model was used. 4T1 cells were injected into mammary fat pads. Tumors were allowed to grow for 9 days before gavage treatment with CphE (150 mg GAE/kg/day) or PBS (controls) for one week. Excised tumors, liver, and lungs were analyzed for gene and protein expression and histology. RESULTS: In vitro results showed that CphE suppressed cell viability through apoptosis induction, via caspase-3 cleavage and total PARP reduction. CphE also scavenged ROS and suppressed Akt, mTOR, ERK1/2, CREB, and Src activation contributing to cell motility inhibition. CphE reduced IR, PTEN, TSC2, p70S6, and RPS6, protein levels, which are proteins involved in the PI3K/Akt/mTOR pathway, suggesting this pathway as CphE primary target. In vivo results showed downregulation of ERK1/2 activation by phosphorylation in tumor tissues, accompanied by angiogenesis reduction in tumor and lung tissues. A reduction of Cenpf mRNA levels in liver and lung tissues strongly suggested anti-invasive cancer activity of CphE. CONCLUSION: CphE inhibited 4T1 cell signal pathways that play a key role in cell growth and invasion. The potential for in vitro results to be translated in vivo was confirmed. A complete animal study is a guarantee to confirm the CphE anticancer and antimetastatic activity in vivo.

Priming of defense-related genes in Brassica oleracea var. capitata using concentrated metabolites produced by Rhizobium tropici CIAT 899.

To verify the potential of metabolites extracted from Rhizobium tropici to trigger the priming of defense responses in cruciferous plants, we analyzed the expression of defense-related genes by qRT-PCR. Brassica oleracea var. capitata, susceptible to Xanthomonas campestris pv. campestris, were grown in greenhouse conditions. At 18 days after sowing, plants were inoculated with 1 mL of 1% concentrated metabolites produced by R. tropici (CM-RT) in the root. In a second experiment, leaves were sprayed with 1 mL of a solution containing 1% CM-RT. Aerial and root tissue were collected separately at 0 (non-treated control condition), 24, and 48 h after application, submitted to RNA extraction and gene expression analysis by qRT-PCR. The results showed that, after root treatment with CM-RT, most evaluated genes were upregulated at 24 h after application and downregulated at 48 h after application in roots, while in leaves, genes were downregulated both at 24 and 48 h after application. On the other hand, leaf treatment with CM-RT showed that most evaluated genes in leaves and roots were upregulated at 24 and 48 h after application. These results indicate that the effect of CM-RT applied in roots seems restricted to the applied region and is not sustained, while the application in leaves results in a more systemic response and maintenance of the effect of CM-RT for a longer period. The results obtained in this study emphasize the biotechnological potential of using metabolites of R. tropici as an elicitor of active defense responses in plants.

Effectiveness of the Semi-Adjustable Articulator Compared to Clinical Examination

ABSTRACT: This work aims to verify the effectiveness of the semi-adjustable articulator (SAA) when compared to a clinical examination of samples in individuals comparing the number of occlusal interferences. The research was carried out at FAESA (Integrated University of São Pedro), in which two independent researchers compared occlusal interferences in protrusion and laterality movements in the clinical examination and in models mounted on a semi-adjustable articulator, with carbon from Baush, in 77 patients who attended to the graduation clinic. Data were tabulated and evaluated by the Mann- Whitney statistical test and data normality was verified by the Kolmogorov-Smirnov test. When analyzing the interference of protrusion on central incisors, it was found that the mean was higher for the SAA on clinical examination. As for the right laterality in the canine guides, it was observed that the clinical examination had a higher interference average in relation to the SAA. The same was true for left laterality, where the mean interference was also higher for the clinical examination. That is, by increasing the values of occlusions in the SAA, the values of occlusions in the clinical examination also increase. As central incisor protrusions between the SAA and clinical examination were moderately positive, there was a strong positive relationship for the right sides and a moderate positive relationship for the left sides. It is concluded that a semi-adjustable articulator is an effective tool for the diagnosis and planning of the dentist, due to its ability to simulate mandibular movements. However, such a tool requires prior knowledge for an ideal assembly for treatment success.

RESUMEN: Este trabajo tuvo como objetivo verificar la efectividad del articulador semi-ajustable (SAA) cuando se compara con un examen clínico de muestras en individuos de acuerdo al número de interferencias oclusales. La investigación se llevó a cabo en FAESA (Universidad Integrada de São Pedro), en la que dos investigadores independientes compararon las interferencias oclusales en los movimientos de protrusión y lateralidad en el examen clínico y en modelos montados en un articulador semiajustable, con carbón de Baush, en 77 pacientes que acudieron a la clínica de graduación. Los datos se tabularon y evaluaron mediante la prueba estadística de Mann-Whitney y la normalidad de los datos se verificó mediante la prueba de Kolmogorov-Smirnov. Al analizar la interferencia de la protrusión en los incisivos centrales, se encontró que la media era mayor para el SAA en el examen clínico. En cuanto a la lateralidad derecha en las guías caninas, se observó que el examen clínico tuvo un promedio de interferencia más alto en relación al SAA. Lo mismo ocurrió con la lateralidad izquierda, donde la interferencia media también fue mayor para el examen clínico. Es decir, al aumentar los valores de oclusiones en el SAA, también aumentan los valores de oclusiones en el examen clínico. Como las protuberancias de los incisivos centrales entre el SAA y el examen clínico fueron moderadamente positivas, hubo una fuerte relación positiva para los lados derechos y una relación positiva moderada para los lados izquierdos. Se concluye que un articulador semi-ajustable es una herramienta eficaz para el diagnóstico y planificación del odontólogo, debido a su capacidad para simular movimientos mandibulares. Sin embargo, tal herramienta requiere conocimientos previos para un montaje ideal para el éxito del tratamiento.

Immunophenotyping of progenitor cells from articular cartilage of New Zealand Rabbits (Oryctolagus cuniculus).

Mesenchymal stem cell (MSC) have immunomodulatory and anti-inflammatory effects, allowing its application in the therapy of different diseases, including articular cartilage injuries, which induce the establishment of a pro-regenerative microenvironment in the injured tissue. Therefore, our objective was to isolate, characterize and differentiate cartilage cells from different joints of New Zealand rabbit (Oryctolagus cuniculus), in order to verify their potential as MSC for future clinical use. For this, cartilage fragments were isolated from the humerus-radio-ulnar joints, humeral scapula, femoro-tibio-patellar, and lame femoris from rabbits. The results showed that the cells were rounded in the center of the plate and fibroblastoids in the periphery. After thawing, the cells did not change their growth time in culture, nor their morphology. The cells showed labeling for mesenchymal stem cell, cytoskeleton, pluripotency and cell proliferation, but not for hematopoiesis markers (CD105+ and CD34-). We also observed that, when induced, they were able to differentiate into osteogenic, adipogenic, and chondrogenic cells. After application of these cells in nude mice, no tumor growth was observed in spleen, kidney, liver, lung and heart. Therefore, we conclude that cells isolated from the articular cartilage of rabbits present characteristics of MSC with potential for future clinical applications.

The role of dietary polyphenols in osteosarcoma: A possible clue about the molecular mechanisms involved in a process that is just in its infancy.

Osteosarcoma (OS) is a primary malignant bone tumor mainly affecting children, teenagers and young adults, being associated with early metastasis and poor prognosis. The beneficial effects of polyphenols have been investigated in different areas, including their potential to fight OS. Polyphenols are believed to reduce morbidity and/or slow down the development of cancer. This review aimed to assess the effect of polyphenols in OS and investigate their molecular mechanisms. It was observed that the broad spectrum of health-promoting properties of plant polyphenols in OS occurs mainly due to modulation of reactive oxygen species, anti-inflammatory activity, anti-angiogenesis, apoptosis inducer, inhibition of invasion and metastasis. However, it is worth mentioning that although the promising effects of polyphenols in the fight against OS, most of the studies have been performed using in vitro and in vivo animal models. Therefore, studies in humans are needed to validate the effectiveness of polyphenols in OS treatment. PRACTICAL APPLICATIONS: Polyphenols are widely used for various diseases, however, until now, their real role in the treatment of osteosarcoma remains unknown. This review provides a broad spectrum of research conducted with polyphenols and their potential as adjuvant therapy in the treatment of osteosarcoma. However, prior to their clinical application for osteosarcoma treatment, there is a need to isolate and identify specific polyphenolic compounds with high antitumor activity, increase their oral bioavailability, and to investigate their interactions with chemotherapeutic drugs being used in clinical practice.

Evaluation of immunohistopathological profile of tubular and solid canine mammary carcinomas.

Breast cancer is the most common cancer in women, but the incidence of mammary carcinoma in female dogs is even higher than in humans. These two tumors have similarities that can be seen by its biological behavior, molecular genetic alterations, and histology. This suggest that female dogs can be an excellent model for preclinical oncological studies. And the mammary carcinoma most frequently found in this species is the tubular and solid carcinomas. The extracellular matrix (ECM) has an important role in the progression of these tumors. Because of that we proposed to evaluate the ECM components of these carcinomas through histology with specific stains such as Masson's Trichrome, Picrosirius Red and the technique of scanning electron microscopy. With that, we found the presence of collagen fibers in the tubular carcinoma and around its parenchyma. On the other hand, the solid carcinoma presented collagen fibers throughout the parenchyma and around each tumor cell. With the transmission electron microscopy, we observed the presence of mitochondrias and rough endoplasmic reticulum in both tumors. And finally, we evaluated the expression of proteins through the immunohistochemistry, in which we found a high expression of VEGF, PCNA, CK-18 and vimentin in solid carcinoma, and a positive mark in the tubular and solid carcinoma for collagen I, III and fibronectin. Thus, we demonstrated some differences in the ECM of these mammary carcinomas, allowing a better understanding of its histological characteristics, and these data may contribute to future studies about therapies focused on tumors ECM.

Effects of doxorubicin associated with amniotic membrane stem cells in the treatment of canine inflammatory breast carcinoma (IPC-366) cells.

BACKGROUND: Tumours in mammary glands represent the most common neoplasia in bitches, as in humans. This high incidence results in part from the stimulation of sex hormones on these glands. Among mammary tumours, inflammatory carcinoma is the most aggressive, presenting a poor prognosis to surgical treatment and chemotherapy. One of the most widely used chemotherapy drugs for breast cancer treatment is doxorubicin (DOXO). Alternative therapies have been introduced in order to assist in these treatments; studies on treatments using stem cells have emerged, since they have anti-inflammatory and immunomodulatory properties. The aim of this study was to evaluate the effects of DOXO and canine amniotic membrane stem cells (AMCs) on the triple-negative canine inflammatory mammary carcinoma cell line IPC-366. METHODS: Four experimental groups were analysed: a control group without treatment; Group I with DOXO, Group II with AMC and Group III with an association of DOXO and AMCs. We performed the MTT assay with DOXO in order to select the best concentration for the experiments. The growth curve was performed with all groups (I-III) in order to verify the potential of treatments to reduce the growth of IPC-366. For the cell cycle, all groups (I-III) were tested using propidium iodide. While in the flow cytometry, antibodies to progesterone receptor (PR), estrogen receptor (ER), PCNA, VEGF, IL-10 and TGF-ß1 were used. For steroidogenic pathway hormones, an ELISA assay was performed. RESULTS: The results showed that cells treated with 10 µg/mL DOXO showed a 71.64% reduction in cellular growth after 72 h of treatment. Reductions in the expression of VEGF and PCNA-3 were observed by flow cytometry in all treatments when compared to the control. The intracellular levels of ERs were also significantly increased in Group III (4.67% vs. 27.1%). Regarding to the levels of steroid hormones, significant increases in the levels of estradiol (E2) and estrone sulphate (S04E1) were observed in Groups I and III. On the other hand, Group II did not show differences in steroid hormone levels in relation to the control. We conclude that the association of DOXO with AMCs (Group III) promoted a reduction in cell growth and in the expression of proteins related to proliferation and angiogenesis in IPC-366 triple-negative cells. CONCLUSIONS: This treatment promoted ER positive expression, suggesting that the accumulated oestrogen conducted these cells to a synergistic state, rendering these tumour cells responsive to ERs and susceptible to new hormonal cancer therapies.

The Tumor Microenvironment: Focus on Extracellular Matrix.

The extracellular matrix (ECM) regulates the development and maintains tissue homeostasis. The ECM is composed of a complex network of molecules presenting distinct biochemical properties to regulate cell growth, survival, motility, and differentiation. Among their components, proteoglycans (PGs) are considered one of the main components of ECM. Its composition, biomechanics, and anisotropy are exquisitely tuned to reflect the physiological state of the tissue. The loss of ECM's homeostasis is seen as one of the hallmarks of cancer and, typically, defines transitional events in tumor progression and metastasis. In this chapter, we discuss the types of proteoglycans and their roles in cancer. It has been observed that the amount of some ECM components is increased, while others are decreased, depending on the type of tumor. However, both conditions corroborate with tumor progression and malignancy. Therefore, ECM components have an increasingly important role in carcinogenesis and this leads us to believe that their understanding may be a key in the discovery of new anti-tumor therapies. In this book, the main ECM components will be discussed in more detail in each chapter.

Caracterização dos casos confirmados de dengue por meio da técnica de linkage de bancos de dados, para avaliar a circulação viral em Belo Horizonte, 2009-2014 / Características de los casos confirmados de dengue utilizando la técnica de enlace de bases de datos, para evaluar la circulación viral en la ciudad de Belo Horizonte, Brasil, 2009-2014 / Characterization of dengue cases confirmed using the database linkage technique: assessment of virus circulation in Belo Horizonte, Brazil, 2009-2014

Objetivo: descrever a circulação viral de dengue em Belo Horizonte, Brasil, de 2009 a 2014. Métodos: trata-se de estudo de série de casos de dengue infectados por diferentes sorotipos do vírus, identificados por isolamento viral ou RT-PCR; foi realizado linkage das bases de dados do Sistema de Informação de Agravos de Notificação (Sinan), do Sistema de Informações Hospitalares do Sistema Único de Saúde (SIH/SUS) e do Gerenciador de Ambiente Laboratorial (GAL). Resultados: foram relacionados 91,1% dos registros (n=775); entre os casos (n=851), 60,4% (n=514) foram confirmados como DENV-1, 22,1% (n=188) como DENV-4, 9,8% (n=83) como DENV-2 e 7,7% (n=66) como DENV-3; DENV-2 apresentou maior percentual de casos graves (4,5%). Conclusão: DENV-1 prevaleceu e circulou em todos os anos avaliados.

Objetivo: describir la circulación viral del dengue en Belo Horizonte, Brasil, de 2009 a 2014. Métodos: se trata de estudio de una serie de casos de dengue infectados por diferentes serotipos del virus, identificados por aislamiento viral o RT-PCR; se realizó una conexión con las bases de datos del Sistema de Información para Enfermedades de Notificación (Sinan), el Sistema del Información Hospitalaria del Sistema Único de Salud (SIH/SUS) y el Gestor de Ambiente de Laboratorio (GAL). Resultados: fueron relacionados 91,1% de los registros (n=775); entre los casos (n=851), el 60,4% (n=514) fue confirmado como DENV-1, 22,1% (n=188) como DENV-4, 9,8% (n=83) como DENV-2 y 7,7% (n=66) como DENV-3; DENV-2 tuvo un mayor porcentaje de casos graves (4,5%). Conclusión: DENV-1 prevaleció y circuló en todos los años evaluados.

Objective: to describe dengue virus circulation in Belo Horizonte, Brazil, from 2009 to 2014. Methods: this is a series study of cases of dengue infected by different virus serotypes, identified by virus isolation or RT-PCR; database linkage was performed between the Notifiable Health Conditions Information System (SINAN), the Brazilian National Health System Hospital Information System (SIH/SUS) and the Laboratory Environment Manager (GAL). Results: 91.1% of the records were linked (n=775); among the cases (n=851), 60.4% (n=514) were confirmed as DENV-1, 22.1% (n=188) DENV-4, 9.8% (n=83) as DENV-2, and 7.7% (n=66) as DENV-3; DENV-2 accounted for a higher percentage of severe cases (4.5%). Conclusion: DENV-1 prevailed and circulated in all evaluated years.

Canine amniotic membrane mesenchymal stromal/stem cells: Isolation, characterization and differentiation.

The amniotic membrane can be considered as one of the sources of isolation of these cells, since it is found in the fetal maternal interface and has low immunogenicity. Mesenchymal stromal/stem cells (MSCs) have not been identified in canine amniotic membrane (AMC). Therefore, our objective was to isolate, culture, characterize and differentiate cells derived from canine amniotic membrane (AMC) and to verify its immunological and tumorigenic potential. For this, 12 dogs fetuses of each gestational age 32, 43 and 55 days were used, and the isolation and culture of the AMC were performed. We observed that the cells presented fibroblastoid morphology and high confluence even after freezing. We also observed that, when induced, they were able to differentiate into osteogenic, adipogenic, and chondrogenic cells, as well as being CD34- and CD105+. Regarding the immunological markers, we found that IL-1, IL-2, IL-6, IL-10 and MHC II were not expressed, whereas MHC I was expressed. After application of AMC cells in nude mice we can verify that there was no tumor formation. Based on this, we conclude that canine amniotic membrane is a good and accessible source for obtaining MSCs of low immunogenic and tumorigenic potential for veterinary therapeutic applications.

Baccharis trimera protects against ethanol induced hepatotoxicity in vitro and in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Baccharis trimera has been traditionally used in Brazil to treat liver diseases. AIM OF THE STUDY: To evaluate the protective effect of Baccharis trimera in an ethanol induced hepatotoxicity model. MATERIALS AND METHODS: The antioxidant capacity was evaluated in vitro by the ability to scavenged the DPPH radical, by the quantification of ROS, NO and the transcription factor Nrf2. Hepatotoxicity was induced in animals by administration of absolute ethanol for 2 days (acute) or with ethanol diluted for 28 days (chronic). The biochemical parameters of hepatic function (ALT and AST), renal function (urea and creatinine) and lipid profile (total cholesterol, triglycerides and HDL) were evaluated. In addition to antioxidant defense (SOD, catalase, glutathione), oxidative damage markers (TBARS and carbonylated protein), MMP-2 activity and liver histology. RESULTS: Baccharis trimera promoted a decrease in ROS and NO, and at low concentrations promoted increased transcription of Nrf2. In the acute experiment it promoted increase of HDL, in the activity of SOD and GPx, besides diminishing TBARS and microesteatosis. Already in the chronic experiment B. trimera improved the hepatic and renal profile, decreased triglycerides and MMP-2 activity, in addition to diminishing microesteatosis. CONCLUSION: We believe that B. trimera action is possibly more associated with direct neutralizing effects or inhibition of reactive species production pathways rather than the modulation of the antioxidant enzymes activity. Thus it is possible to infer that the biological effects triggered by adaptive responses are complex and multifactorial depending on the dose, the time and the compounds used.

Aqueous extract of Baccharis trimera improves redox status and decreases the severity of alcoholic hepatotoxicity

ABSTRACT The metabolism of ethanol occurs mainly in the liver, promoting increase of reactive oxygen species and nitrogen, leading to redox imbalance. Therefore, antioxidants can be seen as an alternative to reestablish the oxidizing/reducing equilibrium. The aim of this study was to evaluate the antioxidant and hepatoprotective effect of aqueous extract of Baccharis trimera (Less.) DC., Asteraceae, in a model of hepatotoxicity induced by ethanol. The extract was characterized and in vitro tests were conducted in HepG2 cells. It was evaluated the cells viability exposed to aqueous extract for 24 h, ability to scavenging the radical DPPH, besides the production of reactive oxygen species and nitric oxide, and the influence on the transcriptional activity of transcription factor Nrf2 (12 and 24 h) after exposure to 200 mM ethanol. The results showed that aqueous extract was non-cytotoxic in any concentration tested; moreover, it was observed a decrease in ROS and NO production, also promoting the transcriptional activity of Nrf2. In vivo, we pretreatment male rats Fisher with 600 mg/kg of aqueous extract and 1 h later 5 ml/kg of absolute ethanol was administrated. After two days of treatment, the animals were euthanized and lipid profile, hepatic and renal functions, antioxidant status and oxidative damage were evaluated. The treatment with extract improved liver function and lipid profile, reflecting the reduction of lipid microvesicules in the liver. It also promoted an increase of glutathione peroxidase activity, decrease of oxidative damage and MMP-2 activity. These results, analyzed together, suggest the hepatoprotective effect of B. trimera aqueous extract.

Baccharis trimera inhibits reactive oxygen species production through PKC and down-regulation p47 phox phosphorylation of NADPH oxidase in SK Hep-1 cells.

Baccharis trimera, popularly known as "carqueja", is a native South-American plant possessing a high concentration of polyphenolic compounds and therefore high antioxidant potential. Despite the antioxidant potential described for B. trimera, there are no reports concerning the signaling pathways involved in this process. So, the aim of the present study was to assess the influence of B. trimera on the modulation of PKC signaling pathway and to characterize the effect of the nicotinamide adenine dinucleotide phosphate oxidase enzyme (NOX) on the generation of reactive oxygen species in SK Hep-1 cells. SK-Hep 1 cells were treated with B. trimera, quercetin, or rutin and then stimulated or not with PMA/ionomycin and labeled with carboxy H2DCFDA for detection of reactive oxygen species by flow cytometer. The PKC expression by Western blot and enzyme activity was performed to evaluate the influence of B. trimera and quercetin on PKC signaling pathway. p47 phox and p47 phox phosphorylated expression was performed by Western blot to evaluate the influence of B. trimera on p47 phox phosphorylation. The results showed that cells stimulated with PMA/ionomycin (activators of PKC) showed significantly increased reactive oxygen species production, and this production returned to baseline levels after treatment with DPI (NOX inhibitor). Both B. trimera and quercetin modulated reactive oxygen species production through the inhibition of PKC protein expression and enzymatic activity, also with inhibition of p47 phox phosphorylation. Taken together, these results suggest that B. trimera has a potential mechanism for inhibiting reactive oxygen species production through the PKC signaling pathway and inhibition subunit p47 phox phosphorylation of nicotinamide adenine dinucleotide phosphate oxidase.