Atividade antibacteriana das plantas copaifera duckei dwyer e pseudobombax marginatum e sua interação com a superfície celular bacteriana

A resistência microbiana a antimicrobianos tem favorecido a busca por substâncias bioativas provenientes de plantas usadas na medicina popular, com o intuito de se obter novos fármacos com atividade antimicrobiana. Neste estudo, foi proposta a investigação da atividade antibacteriana do óleo-resina de Copaifera duckei e de diferentes extratos da casca de Pseudobombax marginatum, e seus possíveis mecanismos de ação. O potencial inibitório antibacteriano foi avaliado utilizando-se os métodos de difusão e diluição em ágar, e a bioautografia. O mecanismo de ação foi analisado por microscopia eletrônica, no qual se observou alterações na ultraestrutura bacteriana, e por eletroforese em SDS-PAGE, que determinou ação sobre as proteínas das superfícies celulares. A análise química foi realizada pelas técnicas de Espectrometria de massas acoplada ao Cromatógrafo a gás- EM/CG (C. duckei) e Cromatografia Líquida de Alta Eficiência- CLAE (P. marginatum). Entre as bactérias estudadas, B. cereus foi a mais suscetível às plantas em estudo, com concentrações inibitórias mínimas (CIMs) correspondentes a 0,3125 mg/mL para o óleo-resina de copaíba, e 0,5 mg/mL para extrato hidroalcoólico (1:1) e 0,512 mg/mL para a fração butanólica da casca P. marginatum, nos quais pôde-se observar alterações na parede celular do B. cereus, com remoção da camada S, espessamento da parede celular e formação de diversos septos nos centros de divisão celular. A análise química por EM/CG mostrou compostos terpênicos no óleo-resina de C. duckei, tendo como composto majoritário o β-bisaboleno, e a análise por CLAE mostrou a presença de compostos derivados da catequina na casca do P. marginatum. Desta forma, as plantas em estudo mostram um potencial antibacteriano considerável, podendo contribuir tanto na terapia antimicrobiana como na área de alimentos, tendo como um de seus prováveis sítios de ação a parede celular bacteriana

Avaliação de efeitos biológicos de antiinflamatórios derivados do ácido propiônico através de modelos experimentais em nível molecular e celular

Derivatives of propionic acid NSAIDs are irreversible inhibitors of cyclooxygenase enzyme widely used. The aim of this study was to evaluate, through different experimental models, biological effects of derivatives of propionic acid (fenoprofen, naproxen, ibuprofen and ketoprofen) in cellular and molecular level. The labeling of blood constituents with technetium-99m (99mTc) and morphological analysis of erythrocytes of blood of rats, as well as growth, survival of cultures of Escherichia coli (E. coli) and the assessment of bacterial plasmid electrophoretic profiles were models used for experimental evaluation of possible biological effects of antiinflammatory drugs. The results show that, in general, anti-inflammatory drugs evaluated were not able to alter the labeling of blood constituents with 99mTc, the morphology of red blood cells from blood of rats, as well as the growth of cultures of E. coli and the electrophoretic profile of plasmid DNA. However, naproxen appears to cytotoxic effect on bacterial cultures, plasmids and genotoxic effects in reducing the action of stannous chloride in cultures of E. coli. The use of experimental fast performance and low cost was important for assessment of biological effects, contributing to a better understanding of the properties of propionic acid derivatives studied. anti-inflammatory, blood constituents, technetium-99m, stannous chloride, Escherichia coli; DNA

Resposta celular Th1 /Th2 na doença periodontal experimental, em ratos: um estudo imuno-histoquímico

Host response plays a major role in the pathogenesis of periodontal disease. Mediators such as inflammatory cytokines which are secreted during the immune response to bacterial challenges have ambiguous functions that may or may not lead to protection of the attacked tissue. In this context, experimental evidence suggests that T-helper 1 (Th-1) and T-helper 2 (Th-2) mediated responses are potentially important during the disease process. The aims of this study therefore were to further clarify the role played by Th2 cells during different time points of the active phase of periodontal disease, as well as, to investigate whether there was any evidence of a Th1 response in the periodontal disease microenvironment. Experimental periodontitis was induced in 30 Wistar male rats by placing cotton ligatures around the mandibular first molars. The rats were then randomly divided into two groups. Group1 (G1=15) and Group 2 (G2=15). In G1 the ligatures were maintained for 2 days, whereas in G2 the ligatures were left for 15 days, a time point that corresponds to the advanced stage of periodontal disease The contra-lateral teeth served as controls (no ligatures). Immunohistochemical investigation for the presence in gingival tissue of Th2 specific transcription factor (GATA3) and the subunit of the IFN-γ receptor was carried out after the disease induction period. Light microscopy analysis revealed a decrease in the expression of GATA-3 as bone loss progressed. On the other hand, although IFN-γ R1 was detected at an early stage of the active phase of disease its expression remained unaltered during the remaining period of the study. These results indicate that the Th2 response have a protective role during the pathogenesis of periodontal disease and that the progression of the periodontal disease is related with the unbalance of the responses Th1/Th2

Estudo da detecção do DNA do papiloma vírus humano (HPV) e da expressão imuno-histoquímica de proteína do ciclo celular no carcinoma epidermóide oral

Oral squamous cell carcinoma (OSCC) is the most common malignancy in oral cavity and human papillomavirus (HPV) may have an important role in its development. The aim of this experiment was to investigate the HPV DNA and viral types in 90 cases of OSCC. Moreover, a comparative analysis between the cases of OSSC with and without HPV DNA was performed by using cell cycle markers p21 and pRb in order to detect a possible correlation of these proteins and HPV infection. DNA was extracted from paraffin embedded tissue and amplified by PCR (polymerase chain reaction) with primers PCO3+ e PCO4+ for a fragment of human β-globin gene. After this procedure, PCR for HPV DNA detection was realized using a pair of generic primers GP5+ e GP6+. Immunohistochemical study was performed by streptoavidin-biotin technique and antibodies against p21 and pRb proteins were employed. Eighty-eight cases were positive for human β-globin gene and HPV DNA was found in 26 (29.5%) of then. It could not be detected significant correlation between HPV and age, sex and anatomical sites of the lesion. The most prevalent viral type was HPV 18 (80.8%). Regarding the immunohistochemical analysis, it was detected significant association between HPV presence and pRb immunoexpression (p=0,044), nevertheless, the same was not observed in relation to p21 protein (p =0,416). It can be concluded that the low detection of HPV DNA in OSCC by the present experiment suggests a possible role of the virus in the development and progression in just a subset of this disease

Caracaterização do núcleo pré-geniculado do sagüi (Callithrix jacchus) :projeção retiniana, neuroquímica e atividade celular (expressão de FOS)

In rodents, the suprachiasmatic nucleus (SCN) and the intergeniculate leaflet (IGL) are the main components of the circadian system. The SCN is considerate the site of an endogenous biological clock because can to generate rhythm and to synchronize to the environmental cues (zeitgebers) and IGL has been related as one of the main areas that modulate the action of SCN. Both receive projections of ganglion cells of retina and this projection to SCN is called retinohypothalamic tract (RHT). Moreover, the IGL is connected with SCN through of geniculohypothalamic tract (GHT). In primates (include humans) was not still demonstrated the presence of a homologous structure to the IGL. It is believed that the pregeniculate nucleus (PGN) can be the answer, but nothing it was still proven. Trying to answer that question, the objective of our study is to do a comparative analysis among PGN and IGL through of techniques immunohystochemicals, neural tracers and FOS expression after dark pulses. For this, we used as experimental model a primate of the new world, the common marmoset (Callithrix jacchus). Ours results may contribute to the elucidation of this lacuna in the circadian system once that the IGL is responsible for the transmission of nonphotic information to SCN and participate in the integration between photic and nonphotic stimulus to adjust the function of the SCN. In this way to find a same structure in primates represent an important achieve in the understanding of the biological rhythms in those animals

Os efeitos das queimadas na Amazônia em nível celular e molecular

The Amazon holds over half of the planet's remaining tropical forests and comprises the largest biodiversity in the world, accounting for approximately 60 % of the Brazilian territory. However, deforestation fires in the region causes serious problems to exposed human. The aim of this study was to evaluate the chemical compounds as well as the cellular and molecular effects after exposure to organic material extracted from particulate matter less than 10 µm (PM10) in the Amazon region. As for the chemical composition, n-alkanes analysis showed a prevalence of anthropogenic influence during the fires in the region. In addition, there was a predominance of monosaccharides from biomass burning markers. Also, the Polycyclic Aromatic Hydrocarbons (PAH) and their derivatives have also been identified in samples collected in the Amazon. By using the PAH concentrations was possible to calculate the BaP-equivalent and it was found that the dibenz(a) anthracene contributes with 83% to potential carcinogenic risk. As for the potential mutagenic risk, the benzo (a) pyrene is the HPA that has a major contribution in this analysis. It may be noted that the retene was the most abundant PAH. This compound was genotoxic and cause death by necrosis in the human lung cells. In biological tests, the data showed that organic PM10 is capable of causing genetic damage in both plant cells and in human lung cells. This damage cause an arrest in the G1 phase of the cell cycle exposed, increasing the expression of p53 and p21. Additionally, the PM10 caused cell death by apoptosis, increasing the foci of histone - H2AX. Given these results, it is important to emphasize the reduction and better control of biomass burning in the Amazon region thus improving the quality of health of the population being exposed. As clearly stated recently by the World Health Organization, the reduction of air pollution could save millions of lives annually.

Avaliação do efeito da inibição do reparo de sítios abásicos na resposta inflamatória celular

Base excision repair (BER) proteins has been associated with functions beyond DNA repair. Apurynic/apyrimidinic endonuclease 1 (APE1) is a multifunctional protein involved in a plethora of cellular activities, such as redox activation of transcription factors, RNA processing and DNA repair. Some studies have described the action of the protein 8-oxoguanine (OGG1) in correcting oxidized lesions in promoters as a step in the transcription of pro-inflammatory cytokines. Despite being especially important in redox activation of transcription factors such as nuclear factor κB (NF-κB) and AP- 1, the repair activity of APE1 has not yet been associated with the inflammatory response. In this study, experimental and bioinformatic analysis approaches have been used to investigate the relationship between inhibition of the repair of abasic sites in DNA by MX, a synthetic molecule designed to inhibt the repair activity of APE1, and the modulation of the inflammatory response. The results showed that treatment of monocytes with lipopolysaccharide (LPS) and MX reduced the expression of cytokines, chemokines and toll-like receptors, and negatively regulated biological immune processes, as macrophages activation, and NF-κB and tumor necrosis factor (TNF-α) and interferon pathways, without inducing cell death. The transcriptomic analysis suggests that LPS/MX treatment induces mitochondrial dysfunction, endoplasmic reticulum stress and activation of autophagy pathways, probably activated by impairment of cellular energy and/or the accumulation of nuclear and mitochondria DNA damage. Additionally, it is proposed that the repair activity of APE1 is required for transcription of inflammatory genes by interaction with abasic sites at specific promoters and recruitment of transcriptional complexes during inflammatory signaling. This work presents a new perspective on the interactions between the BER activity and the modulation of inflammatory response, and suggests a new activity for APE1 protein as modulator of the immune response in a redox-independent manner.

Expressão de MMPs, marcadores angiogênicos e proliferação celular em tumores odontogênicos

INTRODUCTION AND OBJECTIVE: The study of biological behavior of odontogenic lesions is essential to the establishment of appropriate therapeutic approach and prognosis. The production of extracellular matrix metalloproteinases (MMPs), angiogenesis and cell proliferation contribute to tumor growth. This paper aims to review the literature on odontogenic tumors (OT) selected according to the new World Health Organization classification (WHO- 2005) by evaluating the expression of MMPs, angiogenic and cell proliferation. Furthermore, it aims to verify the relation between these markers and the biological behavior of these lesions. RESULTS: it was found that MMPs -1, -2, -7, -9 and -26 had a higher expression in both epithelial component and stroma, and 13 particularly in the stroma. Increased angiogenesis was observed in more aggressive OT. CD105 expression was higher in keratocystic odontogenic tumour (KOT) and CD34 in solid ameloblastomas (SA). It was observed a higher expression of Ki-67 and p53 in SA and KOT and a low cell proliferation rate in the adenomatoid odontogenic tumour (AOT). CONCLUSION: These results show that MMPs are involved in invasion and recurrence of some odontogenic lesions and are associated with the biological behavior of these tumors. Angiogenesis is critical to provide support to cell proliferation and these concomitant events are correlated with different levels of biological behavior in OT when compared to odontogenic cysts, hence the use of angiogenic inhibitors may be a potential therapeutic approach in these lesions.

Expressão de MMPs, marcadores angiogênicos e proliferação celular em tumores odontogênicos

INTRODUCTION AND OBJECTIVE: The study of biological behavior of odontogenic lesions is essential to the establishment of appropriate therapeutic approach and prognosis. The production of extracellular matrix metalloproteinases (MMPs), angiogenesis and cell proliferation contribute to tumor growth. This paper aims to review the literature on odontogenic tumors (OT) selected according to the new World Health Organization classification (WHO- 2005) by evaluating the expression of MMPs, angiogenic and cell proliferation. Furthermore, it aims to verify the relation between these markers and the biological behavior of these lesions. RESULTS: it was found that MMPs -1, -2, -7, -9 and -26 had a higher expression in both epithelial component and stroma, and 13 particularly in the stroma. Increased angiogenesis was observed in more aggressive OT. CD105 expression was higher in keratocystic odontogenic tumour (KOT) and CD34 in solid ameloblastomas (SA). It was observed a higher expression of Ki-67 and p53 in SA and KOT and a low cell proliferation rate in the adenomatoid odontogenic tumour (AOT). CONCLUSION: These results show that MMPs are involved in invasion and recurrence of some odontogenic lesions and are associated with the biological behavior of these tumors. Angiogenesis is critical to provide support to cell proliferation and these concomitant events are correlated with different levels of biological behavior in OT when compared to odontogenic cysts, hence the use of angiogenic inhibitors may be a potential therapeutic approach in these lesions.

Os efeitos das reclamações online na lealdade dos consumidores: um estudo experimental

In an environment of constant change, technological developments, market competition and more informed consumers, the search for a lasting relationship through the conquest of loyalty has become the objective of companies. However, several authors suggest that this loyalty can be affected by negative comments available on the internet. Therefore, this dissertation has as objective to examine if the complaints are available on the internet impact the loyalty to a brand of mobile phone. The research used as the basis the Expanded NCSB model suggest by Johnson et al. (2001), studying five prominent drives of loyalty: image/brand reputation, affective commitment, calculative commitment, perceived value and trust, beyond the satisfaction construct as moderator variable. The research method adopted was the experimental design which included 285 undergraduate students, with the trial which included 285 undergraduate students, with the field study of the mobile industry, specifically, the brands of cell phones. The research approach was quantitative and methods were descriptive statistics, factor analysis, cluster analysis, linear regression and non-parametric test of Wilcoxon for data analysis. Of the 16 hypothesis stemmed from the research model proposed, 12 were confirmed. The results showed that the complaint available on the internet, here represented by the available on the site Reclame Aqui, may impact consumer perceptions about brand loyalty, as well as its antecedents, being that these complaints can affect all the consumers, regardless of historical satisfaction with the brand. It also noted the positive relationship between the independent variables trust, image/brand reputation, perceived value, affective commitment and calculative commitment and the dependent variable - loyalty, even when considering the data obtained after exposure to the complaint. However, no unanimous conclusion that the relationship between these variables was strongest in the group with satisfactory experience. At the first moment of the research, the trust was the most important variable for the formation of loyalty. However, after exposure to treatment, the image/brand reputation, was more relevant. Contributions of the study, limitations and recommendations for future researches are approached in the present investigation

Caracterização físico-química e efeitos de frações polissacarídicas da alga amansia multifida na coagulação, inflamação, radicais livres e viabilidade celular

Galactans are polysaccharides sulfated present in the cell wall of red algae. Carrageenans are galactans well known in the food industry as gelling polysaccharides and for induce inflammatory process in rodents as animal model. The extraction of polysaccharides from A. multifida has been carried out by proteolysis and precipitation in different volumes of acetone, which produced three fractions (F1, F2, and FT). Chemical and physical analyses revealed that these fractions are sulfated galactan predominantly. Results of the antioxidant activity assays showed that all of these fractions have antioxidant activity and that was associated with sulfate content of the analysis of reducing power and total antioxidant capacity. However, these fractions were not effective against lipid peroxidation. The fraction FT presented higher activity on the APTT test at 200 μg (> 240 s). The assessment of the hemolytic activity showed that the FT fraction has the best activity, increasing lyses by the complement system to 42.3% (50 μg) (p< 0,001). The fraction FT showed the best yield, anticoagulant and hemolytic activity between the three fractions and therefore it was choose for the in vivo studies. The Inflammation assessment using the FT fraction (50 mg / kg MB) showed that the cellular migration and the IL-6 production increased 670.1% (p< 0,001) and 531.8% (p< 0,001), respectively. These results confirmed its use as an inflammation inducer in animal model. Cytotoxicity assay results showed that all fractions have toxic effects on 3T3 and HeLa cells after exposition of 48 hours, except when 100 μg for both F1 and FT were used. These results arise the discussion whether these polysaccharides it should be used as additive in foods, cosmetics and medicines.

Indução de morte celular em células de adenocarcinoma cervical humano (HeLa) pela lectina da esponja Cinachyrella apion (CaL)

Cancer is a term used to represent a set of more than 100 diseases, including malignant tumors from different locations. The malignancies are the second leading cause of death in the population, representing approximately 17% of deaths of known cause. Strategies that induce differentiation have had limited success in the treatment of established cancers. In this work, a lectin purified from the marine sponge Cinachyrella apion (CaL) was evaluated due to its hemolytic, cytotoxic and antiproliferative properties, besides the ability to induce cell death via apoptosis in tumor cells. The antiproliferative activity of CaL was tested against cell lines, with the highest inhibition of tumor growth for HeLa, reducing cell growth at a dose dependent manner, with a concentration of 10 μg/mL. The hemolytic activity and toxicity against peripheral blood cells were tested using the concentration of IC50 for both trials and twice the IC50 for analysis in flow cytometry, indicating that CaL is not toxic to these cells. To assess the mechanism of cell death caused by CaL in HeLa cells, we performed flow cytometry and western blotting. The results showed the lectin probably induces cell death by apoptosis activation by pro-apoptotic protein Bax, promoting mitochondrial membrane permeabilization, cell cycle arrest in S phase, with accumulation of cells of approximately 57% in this phase, and acting as both dependent and/or independent of caspases pathway. These results suggest that CaL has the potential to be used as drug treatment against cancer.

Avaliação do efeito do composto tipo heparina isolado do caranguejo Chaceon fenneri na hemostasia e na morte celular

Heparin is a pharmaceutical animal widely used in medicine due to its potent anticoagulant effect. Furthermore, it has the ability to inhibit the proliferation, invasion and adhesion of cancer cells to vascular endothelium. However, its clinical applicability can be compromised by side effects such as bleeding. Thus, the search for natural compounds with low bleeding risk and possible therapeutic applicability has been targeted by several research groups. From this perspective, this study aims to evaluate the hemorrhagic and anticoagulant activities and citotoxic effect for different tumor cell lines (HeLa, B16-F10, HepG2, HS-5,) and fibroblast cells (3T3) of the Heparin-like from the crab Chaceon fenneri (HEP-like). The HEP-like was purified after proteolysis, ion-exchange chromatography, fractionation with acetone and characterized by electrophoresis (agarose gel) and enzymatic degradation. Hep-like showed eletroforetic behavior similar to mammalian heparin, and high trisulfated /Nacetylated disaccharides ratio. In addition, HEP-like presented low in vitro anticoagulant activity using aPTT and a minor hemorrhagic effect when compared to mammalian heparin. Furthermore, the HEP-like showed significant cytotoxic effect (p<0.001) on HeLa, HepG2 and B16-F10 tumor cells with IC50 values of 1000 ug/mL, after incubation for 72 hours. To assess the influence of heparin-like on the cell cycle in HeLa cells, analysis was performed by flow cytometry. The results of this analysis showed that HEP-like influence on the cell cycle increasing S phase and decreasing phase G2. Thus, these properties of HEP-like make these compounds potential therapeutic agents

Concreto celular polimérico: influência na adição de resíduo de poliéster insaturado termofixo

This work addresses the production of lightweight concrete building elements, such as plates, prefabricated slabs for pre-molded and panels of fencing, presenting a singular concrete: the Lightweight Concrete, with special properties such low density and good strength, by means of the joint use of industrial waste of thermosetting unsaturated polyesters and biodegradable foaming agent, named Polymeric Lightweight Concrete. This study covered various features of the materials used in the composition of the Polymeric Lightweight Concrete, using a planning of factorial design 23, aiming at studying of the strength, production, dosage processes, characterization of mechanical properties and microstructural analysis of the transition zone between the light artificial aggregate and the matrix of cement. The results of the mechanical strength tests were analyzed using a computational statistics tool (Statistica software) to understand the behavior and obtain the ideal quantity of each material used in the formula of the Polymeric Lightweight Concrete. The definition of the ideal formula has the purpose of obtaining a material with the lowest possible dry density and resistance to compression in accordance with NBR 12.646/92 (≥ 2.5 MPa after 28 days). In the microstructural characterization by scanning electron microscopy it was observed an influence of the materials in the process of cement hydration, showing good interaction between the wrinkled face of the residue of unsaturated polyesters thermosetting and putty and, consequently, the final strength. The attaining of an ideal formula, given the Brazilian standards, the experimental results obtained in the characterization and comparison of these results with conventional materials, confirmed that the developed Polymeric Lightweight Concrete is suitable for the production of building elements that are advantageous for construction