High-protein diet associated with resistance training reduces cardiac TNF-α levels and up-regulates MMP-2 activity in rats.

The consumption of high-protein diets (HPD) is associated with resistance training (RT) due to effects on metabolism. However, little is known about these effects on cardiac tissue. This study aimed to investigate effects of HPD and RT on cardiac biomarkers. 18 rats were divided into normo-protein (NPD), and HPD groups: NPD-Control, NPD-RT, HPD-Control, and HPD-RT. Interleukin-6 (IL-6), tumour necrosis factor alpha (TNF-a), nitric oxide (NO), activity of metalloproteinase-2 (MMP-2), and vascular factor (VEGF) were analysed. RT was effective in regulating body weight, increasing strength, and reducing food consumption (p < .05). HPD induces higher levels of interleukin 6 (p = .0169), and lowers NO (p < .0001). When associated with RT, the HPD decreases levels of tumour necrosis factor alpha, while enhances NO, and MMP activity (p < .05). The association of RT with HDP decreases inflammatory parameters and indicates an enhancement in the molecular parameters of cardiac tissue.

Host defense peptides clavanins A and MO reduce in vitro osteoclastogenesis

Aim: Several systemic diseases, such as periodontitis and apical periodontitis, can cause extensive bone resorption. Host defense peptides may have the potential for the development of novel therapies for the bone resorption process. This study evaluated the potential of host defense peptides clavanins A, MO, and LL-37 in in vitro osteoclastogenesis. Methods: RAW 264.7 cultures were stimulated with recombinant of receptor activator of nuclear factor kappa B ligand in the presence of different tested concentrations of host defense peptides, besides calcium hydroxide and doxycycline. Cellular viability, nitric oxide production, and a number of differentiated osteoclast-like cells were also evaluated. Results: Results showed that none of the substances were cytotoxic, except for 128 µg.mL-1 of doxycycline after 3 days. Host defense peptides, calcium hydroxide, and doxycycline did not interfere in nitric oxide production or downregulated it. An exception was observed in the presence of 2 µg.mL-1 of doxycycline, in which nitric oxide production was up-regulated. All host defense peptides were capable of reducing osteoclast-like cell differentiation. Conclusion: Host defense peptides clavanins A and MO demonstrated to be potential suppressors of osteoclastogenesis in vitro without interfering in cellular viability and nitric oxide production. These promising results need to be further analyzed in in vivo models of bone resorption

Concentrated MTA Repair HP reduced biofilm and can cause reparative action at a distance.

AIM: To evaluate in vitro whether MTA Repair HP can induce repair processes at a distance, including its effects on biofilm, cell viability, migration, production of TGF-ß, phosphate and ALP, evaluated through MTA diluted extracts. METHODOLOGY: Initially, antibacterial tests were performed with the bacterium Streptococcus mutans (ATCC 25175) in the presence of MTA extracts (dilutions of 1:1, 1:2 and 1:4). Growth inhibition assay by microdilution in broth, antibiofilm plate assay of young biofilm and antibiofilm assay in confocal microscopy of mature biofilm were carried out. Then, pulp cells were stimulated in the presence of several MTA dilutions, and cell viability (MTT assay), proliferation and migration capacity (scratch assay) were evaluated. To evaluate the capacity of 1:1, 1:2 and 1:4 dilutions of MTA Repair HP to promote the production of important agents of odontogenic differentiation and mineralization, ALP activity, TGF-ß secretion and phosphate quantification were measured. Statistical differences were verified using one-way and two-way anova and Tukey's post-tests. RESULTS: The test dilutions of MTA Repair HP did not inhibit planktonic S. mutans growth but were able to reduce young and mature S. mutans biofilm (p < 0.001). In addition, none of the MTA Repair HP dilutions was cytotoxic for pulp cells. The 1:2 and 1:4 dilutions of MTA Repair HP induced migration and proliferation of pulp cells (p < 0.05). ALP activity and TGF-ß secretion were independent of the tested dilution (p < 0.001). Diluted 1:4 MTA Repair HP produced less phosphate than the more concentrated 1:1 and 1:2 MTA dilutions (p < 0.001). CONCLUSIONS: Undiluted MTA Repair HP reduced S. mutans biofilm, when compared to 1:2 and 1:4 MTA dilutions. Furthermore, none of the tested dilutions was cytotoxic to pulp cells. MTA Repair HP promoted cell migration and proliferation at a distance, assessed through the dilution of the MTA. Even from a distance, MTA Repair HP has the ability to participate in some events related to repair, such as migration, proliferation and TGF production.

Proteomic analysis of human dental pulp in different clinical diagnosis.

OBJECTIVES: The present study aimed to identify proteins obtained from pulp tissue and correlate with each clinical diagnosis (healthy pulp, inflamed pulp, and necrotic pulp). MATERIALS AND METHODS: A total of forty-five molars were used. Three biological replicas were evaluated. Lysis and sonication were used for protein extraction. Protein quantification was assessed by using the Bradford technique, and shotgun proteome analysis was performed by nanoUPLC-MSE using a Synapt G2 mass spectrometer. Mass spectra data were processed using the Waters PLGS software, and protein identification was done using the human Uniprot database appended to the PLGS search engine. RESULTS: A total of 123 different proteins were identified in all evaluated pulp conditions. Among these, 66 proteins were observed for healthy pulp, 66 for inflamed pulp, and 91 for necrotic pulp. Most protein identification was related to immune response, multi-organism process, platelet activation, and stress in inflamed pulp samples compared to healthy pulp. Proteins related to cellular component organization or biogenesis, developmental process, growth, immune response, multi-organism process, response to stimulus, signaling, stress, and transport were identified in cases of apical periodontitis compared to inflamed pulp. CONCLUSIONS: The progression of the disease to inflamed pulp promoted a high abundance of proteins related to the immune system and stress. Comparing the necrotic pulp with inflamed pulp conditions, a high abundance of proteins was noticed related to metabolism, transport, and response between organisms. CLINICAL RELEVANCE: This finding may assist in future studies of new markers, understanding of tissue engineering, and development of future products.

Systemic conditions of diabetic patients diagnosed with apical periodontitis / Condições sistêmicas de pacientes diabéticos diagnosticados com periodontite apical

ABSTRACT Objective: This study aimed to evaluate the association between glycemic control status in type 2 diabetes mellitus (T2DM) patients and apical periodontitis. Methods: Twenty-seven patients were involved in this study. The survey was based on anamnesis, intra and extra oral examination and radiographic evaluation. Diabetes mellitus information involved type of diabetes and blood glucose analysis. Patients were divided according to their metabolic control status (glycemic controlled and poorly controlled T2DM patients). Results: A higher fasting blood glucose level (p = 0.004) and a higher percentage of HbA1c (p = 0.0001) were demonstrated in poorly controlled T2DM patients when compared to glycemic controlled T2DM. However, the frequency of apical periodontitis and the elapsed time since diabetes mellitus diagnosis were higher in controlled T2DM patients, reaching 64%. Nevertheless, controlled T2DM patients presented a higher number of apical periodontitis cases (p < 0.05). Findings support that controlled patients T2DM presented higher presence of apical periodontitis than poorly controlled T2DM ones. In these patients, the time elapsed since the diagnosis was higher, which may have provided a longer period of oscillation and/or uncontrolled metabolism. Conclusions: Therefore, it might contribute to the development and maintenance of apical periodontitis in glycemic controlled patients of this study.

RESUMO Objetivo: Este estudo objetivou avaliar a associação entre o estado de controle glicêmico em pacientes com diabetes mellitus tipo 2 (DM2) e a periodontite apical. Métodos: Vinte e sete pacientes foram envolvidos neste estudo. A pesquisa baseou-se na anamnese, exame intra e extraoral e avaliação radiográfica. As informações sobre o diabetes mellitus envolveram o tipo de diabetes e a análise da glicose sanguínea. Os pacientes foram divididos de acordo com seu estado de controle metabólico (pacientes com DM2 com controle glicêmico e pacientes com DM2 mal controlados). Resultados: Um maior nível de glicose em jejum (p = 0,004) e uma maior porcentagem de HbA1c (p = 0,0001) foram demonstrados em pacientes com DM2 mal controlada quando comparados com DM2 com controle glicêmico. Porém, a frequência de periodontite apical e o tempo decorrido desde o diagnóstico de diabetes mellitus foram maiores nos pacientes com DM2 controlado, chegando a 64%. No entanto, os pacientes com DM2 controlada apresentaram um maior número de casos de periodontite apical (p < 0,05). Os achados suportam que pacientes controlados com DM2 apresentam maior presença de periodontite apical do que pacientes com DM2 mal controlada. Nesses pacientes, o tempo decorrido desde o diagnóstico foi maior, o que pode ter proporcionado um período maior de oscilação e/ou metabolismo descontrolado. Conclusão: Portanto, pode contribuir para o desenvolvimento e manutenção da periodontite apical nos pacientes com controle glicêmico deste estudo.

Enterococcus faecalis and Staphylococcus aureusstimulate nitric oxide production in macrophages and fibroblasts in vitro

Aim: Nitric oxide (NO) is an important mediator related to damage of the pulp tissue and at the same time to regenerative pulp processes. However, it is not clear how common endodontic microorganisms can regulate this mediator. This study aimed to investigate NO production by macrophages and fibroblasts against Enterococcus faecalis- and Staphylococcus aureus-antigens. Methods: RAW 264.7 macrophages and L929 fibroblast cell lines were stimulated with different heat-killed (HK) antigen concentrations (105-108 colony forming units - CFU) from E. faecalis and S. aureus with or without interferon-gamma (IFN-γ). Cell viability by MTT colorimetric assay and NO production from the culture supernatants were evaluated after 72 h. Results: Data here reported demonstrated that none of the antigen concentrations decreased cell viability in macrophages and fibroblasts. The presence of HK-S. aureus and HK-E. faecalis antigen- stimulated NO production with or without IFN-γ on RAW 264.7. The HK-S. aureus antigen stimulated NO production in L929 fibroblasts with or without IFN-γ, and the highest concentration of HK-E. faecalis with IFN-γ also stimulated NO production by these cells. Conclusion: The amount of NO produced by macrophages and fibroblasts may be involved in the concentration and type of prevalent endodontic microorganisms, generating new answers for the understanding of pulpal revascularization/regeneration processes

Efeito remineralizador da caseína fosfato de cálcio fosfopeptídeo amorfo em dentes decíduos e permanentes in vivo / Remineralizing effect of casein fosfopeptideo amorphous calcium phosphate in primary and permanent teeth in vivo

O presente estudo in vivo avaliou o efeito remineralizador da caseína fosfato de cálcio fosfopeptídeo amorfo (CPP-ACP) tooth mousse em lesões de mancha branca em dentes decíduos e permanentes. Dezoito crianças de 4 a 8 anos apresentaram 81 lesões de mancha branca, as quais foram divididas de maneira randômica. Estas foram tratadas com verniz de flúor 2,26% (controle) e CPP-ACP mousse (teste), duas vezes por semana, durante seis e quatro semanas, respectivamente. Um examinador cego avaliou as lesões de mancha branca no período inicial (T0) e após 3 (T1) e 12 (T2) semanas de tratamento. Os dados de índice de placa visível, índice de sangramento gengival, índice ICDAS foram coletados. Em adição, foram realizadas tomadas fotográficas das lesões em todos os períodos analisados. Os dados foram analisados por testes qui-quadrado e t de Student. Resultados não demonstraram diferença entre os grupos controle e teste durante os períodos, a partir do índice ICDAS (p>0,05). No entanto, a partir do mesmo índice, o grupo teste demonstrou diferença significante entre os períodos analisados (p<0,05). A área das manchas brancas, em pixels, dos grupos controle e teste não demonstrou diferença estatística de cada grupo, em qualquer dos períodos analisados (p>0,05). Dessa forma, concluiu- se que a terapia de remineralização de lesões de manchas brancas foi efetiva em ambos os grupos. No entanto, devido ao número amostral reduzido e o curto período de análise do presente estudo, são necessários maiores esclarecimentos em relação a este efeito.

This in vivo study was conducted to evaluate the remineralization effects of CPP-ACP Tooth Mousse on white-spot lesion in primary and permanent teeth. Eighteen children (age from 4-8) exhibiting 81 white-spot lesions sites were randomly divided into two different treatments protocols: test group received applications of the remineralizing mousse twice a week for six weeks; control group received applications of a 2.26% fluoridated coating once a week for four weeks. A blind examiner evaluated the children at baseline, 3 and 12 weeks after the first visit, recorded visible plaque, gingival bleeding, ICDAS index and photographed the lesions. Data was analyzed with chi-square and Student t-test. According to ICDAS index, the control group was not statistically different from the treated group (p>0.05), in each time. However, results from treated group demonstrated that ICDAS index was statistically different from times (p<0.05). The area of the white spots (in pixels) of samples from control and treated groups were not statistically different from each group in any time (p>0.05). It was concluded that the remineralizing therapy of white spot lesions was efficient in the two groups. Nevertheless, due to the small sample size and the brief time for conducting the study, new research is necessary to increase the findings.

Immune Response Profile against Persistent Endodontic Pathogens Candida albicans and Enterococcus faecalis In Vitro.

INTRODUCTION: Persistent microorganisms such as Candida albicans and Enterococcus faecalis might be directly related to endodontic treatment failure. The host response to these microorganisms impairs the reestablishment of intraradicular and periradicular health. METHODS: The present investigation evaluated the expression of inflammatory mediators produced by RAW 264.7 cells in the presence of heat-killed antigens (HK) C. albicans and E. faecalis. Cultures of RAW cells were stimulated with both antigens in the presence or absence of recombinant interferon (rIFN)-γ. Parameters of cell viability, production of nitric oxide (NO), as well as the synthesis of interleukin (IL)-1α, IL-6, IL-10, IL-12, monocyte chemotactic protein-1, and tumor necrosis factor (TNF)-α were analyzed. RESULTS: Results demonstrated that cell viability was especially reduced in antigens and rIFN-γ-stimulated groups. Groups stimulated with HK C. albicans upregulated IL-10 production. Otherwise, the addition of rIFN-γ to HK C. albicans upregulated TNF-α and NO production. Groups stimulated with HK E. faecalis upregulated TNF-α production. HK E. faecalis and rIFN-γ upregulated TNF-α and NO synthesis. The production of other cytokines remained unchanged by all stimuli. CONCLUSIONS: Knowledge regarding the host mechanism of response to microorganisms that perpetuate endodontic infection and the periradicular lesions can contribute to optimization of endodontic therapy. The mentioned inflammatory mediators and virulence factors involved in endodontic failure might guide lesion progression and also be targets in the development of disinfectant and immunomodulatory agents.

Antimicrobial peptide-based treatment for endodontic infections--biotechnological innovation in endodontics.

The presence/persistence of microorganisms in the pulp and periapical area corresponds to the maintenance of an exacerbated immune response that leads to the start of periradicular bone resorption and its perpetuation. In endodontic treatment, the available intracanal medications do not have all the desirable properties in the context of endodontic infection and apical periodontitis; they need to include not only strong antimicrobial performance but also an immunomodulatory and reparative activity, without host damage. In addition, there are various levels of resistance to root canal medications. Thus, antimicrobial agents that effectively eliminate resistant species in root canals could potentially improve endodontic treatment. In the emergence of new therapies, an increasing number of studies on antimicrobial peptides (AMPs) have been seen over the past few years. AMPs are defense biomolecules produced in response to infection, and they have a wide spectrum of action against many oral microorganisms. There are some studies that correlate peptides and oral infections, including oral peptides, neuropeptides, and bacterial, fish, bovine and synthetic peptides. So far, there are around 120 published studies correlating endodontic microbiota with AMPs but, according to our knowledge, there are no registered patents in the American patent database. There are a considerable number of AMPs that exhibit excellent antimicrobial activity against endodontic microbiota at a small inhibitory concentration and modulate an exacerbated immune response, down-regulating bone resorption. All these reasons indicate the antimicrobial peptide-based endodontic treatment as an emerging and promising option.

Shedding some light over the floral metabolism by arum lily (Zantedeschia aethiopica) spathe de novo transcriptome assembly.

Zantedeschia aethiopica is an evergreen perennial plant cultivated worldwide and commonly used for ornamental and medicinal purposes including the treatment of bacterial infections. However, the current understanding of molecular and physiological mechanisms in this plant is limited, in comparison to other non-model plants. In order to improve understanding of the biology of this botanical species, RNA-Seq technology was used for transcriptome assembly and characterization. Following Z. aethiopica spathe tissue RNA extraction, high-throughput RNA sequencing was performed with the aim of obtaining both abundant and rare transcript data. Functional profiling based on KEGG Orthology (KO) analysis highlighted contigs that were involved predominantly in genetic information (37%) and metabolism (34%) processes. Predicted proteins involved in the plant circadian system, hormone signal transduction, secondary metabolism and basal immunity are described here. In silico screening of the transcriptome data set for antimicrobial peptide (AMP) -encoding sequences was also carried out and three lipid transfer proteins (LTP) were identified as potential AMPs involved in plant defense. Spathe predicted protein maps were drawn, and suggested that major plant efforts are expended in guaranteeing the maintenance of cell homeostasis, characterized by high investment in carbohydrate, amino acid and energy metabolism as well as in genetic information.