Study of Melipona quadrifasciata brain under operant learning using proteomic and phosphoproteomic analysis.

Learning to anticipate events based on the predictive relationship between an action and an outcome (operant conditioning) is a form of associative learning shared by humans and most of other living beings, including invertebrates. Several behavioral studies on the mechanisms of operant conditioning have included Melipona quadrifasciata, a honey bee that is easily manipulated due to lack of sting. In this work, brain proteomes of Melipona bees trained using operant conditioning and untrained (control) bees were compared by two-dimensional gel electrophoresis analysis within pI range of 3-10 and 4-7; in order to find proteins specifically related to this type of associative learning.One protein was detected with differential protein abundance in the brains of trained bees, when compared to not trained ones, through computational gel imaging and statistical analysis. This protein was identified by peptide mass fingerprinting and MS/MS peptide fragmentation using a MALDI-TOF/TOF mass spectrometer as one isoform of arginine kinase monomer, apparently dephosphorylated. Brain protein maps were obtained by 2-DE (Two-dimensional gel electrophoresis) from a total proteins and phosphoproteins extract of the bee Melipona quadrifasciata. One isoform of arginine kinase, probably a dephosphorylated isoform, was significantly more abundant in the brain of trained bees using operant conditioning. Arginine kinase has been reported as an important enzyme of the energy releasing process in the visual system of the bee, but it may carry out additional and unexpected functions in the bee brain for learning process.

Comparative proteomic analysis of kinesin-8B deficient Plasmodium berghei during gametogenesis.

Plasmodium blood stages, responsible for human to vector transmission, termed gametocytes, are the precursor cells that develop into gametes in the mosquito. Male gametogenesis works as a bottleneck for the parasite life cycle, where, during a peculiar and rapid exflagellation, a male gametocyte produces 8 intracellular axonemes that generate by budding 8 motile gametes. Understanding the molecular mechanisms of gametogenesis is key to design strategies for controlling malaria transmission. In the rodent P. berghei, the microtubule-based motor kinesin-8B (PbKIN8B) is essential for flagellum assembly during male gametogenesis and its gene disruption impacts on completion of the parasitic life cycle. In efforts to improve our knowledge about male gametogenesis, we performed an iTRAQ-based quantitative proteomic comparison of P. berghei mutants with disrupted kinesin-8B gene (ΔPbkin8B) and wild type parasites. During the 15 min of gametogenesis, ΔPbkin8B parasites exhibited important motor protein dysregulation that suggests an essential role of PbKIN8B for the correct interaction or integration of axonemal proteins within the growing axoneme. The energy metabolism of ΔPbkin8B mutants was further affected, as well as the response to stress proteins, protein synthesis, as well as chromatin organisation and DNA processes, although endomitoses seemed to occur. SIGNIFICANCE: Malaria continues to be a global scourge, mainly in subtropical and tropical areas. The disease is caused by parasites from the Plasmodium genus. Plasmodium life cycle alternates between female Anopheles mosquitoes and vertebrate hosts through bites. Gametocytes are the parasite blood forms responsible for transmission from vertebrates to vectors. Inside the mosquito midgut, after stimulation, male and female gametocytes transform into gametes resulting in fertilization. During male gametogenesis, one gametocyte generates eight intracytoplasmic axonemes that generate, by budding, flagellated motile gametes involving a process termed exflagellation. Sexual development has a central role in ensuring malaria transmission. However, molecular data on male gametogenesis and particularly on intracytoplasmic axoneme assembly are still lacking. Since rodent malaria parasites permit the combination of in vivo and in vitro experiments and reverse genetic studies, our group investigated the molecular events in rodent P. berghei gametogenesis. The P. berghei motor ATPase kinesin-8B is proposed as an important component for male gametogenesis. We generated Pbkin8B gene-disrupted gametocytes (ΔPbkin8B) that were morphologically similar to the wild- type (WT) parasites. However, in mutants, male gametogenesis is impaired, male gametocytes are disabled in their ability to assemble axonemes and to exflagellate to release gametes, reducing fertilization drastically. Using a comparative quantitative proteomic analysis, we associated the nonfunctional axoneme of the mutants with the abnormal differential expression of proteins essential to axoneme organisation and stability. We also observed a differential dysregulation of proteins involved in protein biosynthesis and degradation, chromatin organisation and DNA processes in ΔPbkin8B parasites, although DNA condensation, mitotic spindle formation and endomitoses seem to occur. This is the first functional proteomic study of a kinesin gene-disrupted Plasmodium parasite providing new insights into Plasmodium male gametogenesis.

Exploring the molecular complexity of Triatoma dimidiata sialome.

Triatoma dimidiata, a Chagas disease vector widely distributed along Central America, has great capability for domestic adaptation as the majority of specimens caught inside human dwellings or in peridomestic areas fed human blood. Exploring the salivary compounds that overcome host haemostatic and immune responses is of great scientific interest. Here, we provide a deeper insight into its salivary gland molecules. We used high-throughput RNA sequencing to examine in depth the T. dimidiata salivary gland transcriptome. From >51 million reads assembled, 92.21% are related to putative secreted proteins. Lipocalin is the most abundant gene family, confirming it is an expanded family in Triatoma genus salivary repertoire. Other putatively secreted members include phosphatases, odorant binding protein, hemolysin, proteases, protease inhibitors, antigen-5 and antimicrobial peptides. This work expands the previous set of functionally annotated sequences from T. dimidiata salivary glands available in NCBI from 388 to 3815. Additionally, we complemented the salivary analysis through proteomics (available data via ProteomeXchange with identifier PXD008510), disclosing the set complexity of 119 secreted proteins and validating the transcriptomic results. Our large-scale approach enriches the pharmacologically active molecules database and improves our knowledge about the complexity of salivary compounds from haematophagous vectors and their biological interactions. SIGNIFICANCE: Several haematophagous triatomine species can transmit Trypanosoma cruzi, the etiological agent of Chagas disease. Due to the reemergence of this disease, new drugs for its prevention and treatment are considered priorities. For this reason, the knowledge of vector saliva emerges as relevant biological finding, contributing to the design of different strategies for vector control and disease transmission. Here we report the transcriptomic and proteomic compositions of the salivary glands (sialome) of the reduviid bug Triatoma dimidiata, a relevant Chagas disease vector in Central America. Our results are robust and disclosed unprecedented insights into the notable diversity of its salivary glands content, revealing relevant anti-haemostatic salivary gene families. Our work expands almost ten times the previous set of functionally annotated sequences from T. dimidiata salivary glands available in NCBI. Moreover, using an integrated transcriptomic and proteomic approach, we showed a correlation pattern of transcription and translation processes for the main gene families found, an important contribution to the research of triatomine sialomes. Furthermore, data generated here reinforces the secreted proteins encountered can greatly contribute for haematophagic habit, Trypanosoma cruzi transmission and development of therapeutic agent studies.

A proteomic approach to compare saliva from individuals with and without oral leukoplakia.

BACKGROUND: Oral leukoplakia is the most common potentially malignant disorder in the oral cavity and can precede carcinoma. This study aimed to identify possible oral leukoplakia salivary biomarkers. METHODS: Unstimulated saliva was collected from participants and protein concentration was determined. Proteins were then precipitated with cold acetone and separated using 2DE over a pH range of 3-10. Spot demarcation and matching were performed and protein identification was done through MS analysis. Oral leukoplakia tissues were submitted to immunohistochemistry analysis for keratin 10 (CK10). A complementary analysis of oral leukoplakias that were not included previously was performed in addition. RESULTS: 226±10 spots were identified in oral leukoplakia 2DE gels, and 262±12 spots were identified in volunteers. Twenty-two spots were highly abundant in oral leukoplakias or not detected in the control group, such as apolipoprotein A1, alpha amylase, cystatins, keratin 10, and lysozyme precursor. All were identified. All oral leukoplakia cases were immunopositive for CK10, mainly in the superficial epithelial layers. CONCLUSIONS: The 2DE salivary protein profiles of individuals with and without oral leukoplakia were observably different. CK10 appears to be an interesting protein and should be further studied in oral carcinogenesis. SIGNIFICANCE: MS-based proteomics enables large-scale analysis of proteins. Proteomics can provide detailed descriptions of proteomes of cells and tissues, including body fluids, and appears as a powerful tool to study human disorders. Saliva is readily accessible through non invasive collection and can mirror diverse disease states. Saliva from both diseased and healthy subjects can be analyzed through 2DE and differences between groups could be found. Routine immunohistochemistry analysis confirmed one of these findings, with CK10 being positive tissues from individuals with oral leukoplakia. Therefore, the present study allows insights into development of an important potential oral cancer precursor, named oral leukoplakia. However, the results can be extrapolated and tested in other precancer states, such as proliferative verrucous leukoplakia, patients at risk of oral cancer due to lifestyle behavior and/or cancer history in the family or even those who are under surveillance after a treated primary oral cancer.

O efeito do tratamento periodontal na proteína c-reativa em pacientes com indicação de cirurgia de revascularização: resultados preliminares / The effect of periodontal treatment in c-reactive protein in patients with indications of surgery revascularization: preliminary results

O objetivo do presente estudo foi examinar o efeito do tratamento periodontal não-cirúrgico sobre a proteína C-reativa, em pacientes com indicação de cirurgia de revascularização no Instituto Nacional de Cardiologia. Foram avaliados 22 pacientes portadores de periodontite crônica, com idade acima de 35 anos (média de idade 59,45 anos) separados em 2 grupos. O grupo I, estudo, (n = 11) recebeu tratamento periodontal anterior ao procedimento cirúrgico, e o grupo II, controle, (n = 11) não realizou tratamento periodontal. Os parâmetros periodontais avaliados foram: profundidade de bolsa à sondagem e nível de inserção clínica. Foram coletadas amostras de sangue de todos os pacientes para análise da proteína C-reativa. Os parâmetros foram avaliados no dia 0, ao início do tratamento periodontal e repetida aos 90 dias, junto com a coleta sanguínea. O tratamento periodontal não-cirúrgico foi eficaz no controle da doença periodontal e aumentou os níveis séricos de proteína C-reativa.

The aim of this study was to examine the effect of periodontal treatment on nonsurgical blood components like C-reactive protein in patients referred for bypass surgery at the Instituto Nacional de Cardiologia. We evaluate 22 patients with chronic periodontitis, aged above 35 years (mean age 59.45 years) divided into 2 groups. The group I, study (n=11), received periodontal treatment prior to cardiovascular surgvyery, and group II, control (n=11) did not have periodontal treatment. The periodontal parameters evaluated were: probing depths and clinical attachment level. Blood samples were collected from all patients for analysis of C-reactive protein. The parameters were evaluated at baseline, in the beginning of periodontal treatment and were repeated after 90 days, along with blood collection. The non-surgical periodontal treatment was effective in controlling periodontal disease and increased serum levels of C-reactive protein.

[Oral infection control in hospitalized patients: an approach to cardiologist and intensive care units doctors]. / Controle de infecção oral em pacientes internados: uma abordagem direcionada aos médicos intensivistas e cardiologistas.

This paper aims to find the current level of periodontal med-care knowledge, as well as the possible existence of some oral infection control protocol regarding hospitalized patients. Our sample gathered 110 cardiologists and intensive care units doctors selected from medical teams of five Rio de Janeiro hospitals. Preliminary numbers: 75.4% said to have heard something about Periodontal Medicine, although only 30% out of this group admitted to have read something concerning such subject. On the other side, only 2.7% of the sample informed to do consistent information searching along their patients anamnese, while 58.2% out of this group admitted such procedure conditional to the patient's general state at the due moment. Through such numbers, we tend to come up to the conclusion that, be it either through direct or indirect Periodontal Medicine technical information (and consequently with regards to the absolut importance of preservation and control of oral biofilm and its impact on one's systemic health), the matter has been dimly spread among medical groups. The search also revealed the probability that Rio de Janeiro hospitals lack either units or agents designed for prevention and control of oral infection; consequently, such organizations do not have any kind of protocols, reliable or not, concerned to oral infections.

Controle de infecção oral em pacientes internados: uma abordagem direcionada aos médicos intensivistas e cardiologistas / Oral infection control in hospitalized patients: an approach to cardiologist and intensive care units doctors

O objetivo do presente estudo foi verificar o grau de conhecimento médico sobre medicina periodontal e verificar a existência de algum protocolo de controle de infecção da cavidade oral em pacientes internados em hospitais. Para tal, 110 médicos cardiologistas e intensivistas lotados em cinco hospitais no município do Rio de Janeiro foram entrevistados. Dentre os indivíduos, 75,4 por cento afirmaram ter conhecimento sobre o termo medicina periodontal; entretanto, apenas 30 por cento declararam já ter lido algo a respeito. Apenas 2,7 por cento dos médicos possuem o hábito de coletar informações sobre a história odontológica de seus pacientes e 58,2 por cento afirmaram que essa conduta é condicional ao quadro apresentado pelo paciente. Com base nos dados obtidos, pode-se concluir que o conhecimento sobre medicina periodontal e, consequentemente, sobre a importância do controle do biofilme oral na manutenção da saúde sistêmica, apresenta-se pouco difundido entre a classe médica. Verificou-se não haver setor ou pessoa responsável pelo controle de infecção oral dentro dos hospitais avaliados e, consequentemente, a não existência de qualquer protocolo, eficaz ou não, de controle de infecção oral nessas unidades.

This paper aims to find the current level of periodontal med-care knowledge, as well as the possible existence of some oral infection control protocol regarding hospitalized patients. Our sample gathered 110 cardiologists and intensive care units doctors selected from medical teams of five Rio de Janeiro hospitals. Preliminary numbers: 75.4 percent said to have heard something about Periodontal Medicine, although only 30 percent out of this group admitted to have read something concerning such subject. On the other side, only 2.7 percent of the sample informed to do consistent information searching along their patients anamnese, while 58.2 percent out of this group admitted such procedure conditional to the patient's general state at the due moment. Through such numbers, we tend to come up to the conclusion that, be it either through direct or indirect Periodontal Medicine technical information (and consequently with regards to the absolut importance of preservation and control of oral biofilm and its impact on one's systemic health), the matter has been dimly spread among medical groups. The search also revealed the probability that Rio de Janeiro hospitals lack either units or agents designed for prevention and control of oral infection; consequently, such organizations do not have any kind of protocols, reliable or not, concerned to oral infections.

Avaliação clínica da hipersensibilidade dentinária após tratamento com dessensibilizantes comerciais / Clinical assessment of dentine hypersensitivity after commercial desensitivity treatment

O objetivo do presente trabalho foi avaliar clinicamente o efeito de dois medicamentos dessensibilizantes de dentina aplicados diretamente na região cervical de dentes com dentina exposta e história clínica de sensibilidade dentinária. Foram selecionados 150 dentes com dentina exposta, ausência de lesões cariosas e queixa de sensibilidade cervical dentinária de um total de 25 pacientes. Os produtos comerciais testados (Dessensiv e Sensikill) e um placebo foram aplicados a cada 7 dias durante quatro semanas. Todos os dentes sensíveis de um mesmo quadrante receberam aplicação tópica do mesmo produto, sorteados aleatoriamente. Em todas as sessões avaliou-se o grau de sensibilidade cervical de cada dente através de estímulos táctil (ponta de uma sonda exploradora) e volátil (jato de arda seringa tríplice). Na semana subsequente a cada aplicação dos produtos (7, 14 e 21 dias) foi calculado o índice de melhora ou piora na sensibilidade dentinária aos estímulos aplicados. Após a 3ª aplicação dos produtos o índice de melhora na sensibilidade ao estímulo táctil foi de 87,2% para os dentes que foram tratados com o produto Dessensiv, 91,9% para o produto Sensikill e 27,5% no grupo placebo. Quanto à sensibilidade ao estímulo volátil, o índice de melhora nos dentes que receberam tratamento dos produtos Dessensiv, Sensikill e placebo foi, respectivamente, de 83,5%, 87,1% e 20,4%. A análise dos resultados permitiu concluir que os produtos comerciais testados são eficazes no tratamento da hipersensibilidade cervical dentinária após três aplicações, não havendo diferença estatisticamente significante entre os dois produtos e ambos foram mais eficazes que o placebo.

The aim of this paper is to make a clinical assessment of the effect of two dentine desensitizing medications applied directly to the cervical region of teeth with exposed dentine and with clinical histories of dentine sensitivity. A total of 150 teeth from 25 patients were selected which had exposed dentine, absence of carious lesions and dentinal cervical sensitivity. The commercial products being tested (Dessensivand Sensikill) and a placebo were applied every seven days for four weeks. All sensitive teeth from the same quadrant were treated topically with the same randomly-chosen product. In all sessions the degree of cervical sensitivity was evaluated for each tooth by means of tactile stimulus (the tip of an exploratory probe) and volatile stimulus (ablast of air from a triple syringe). The rate of improvement or relapse of dentine sensitivity was estimated after each weekly application of products (day 7, 14 and 21). After the third application of the products, however, the rate of improvement in sensitivity to tactile stimuli was 87.2% for teeth treated with Dessensiv, 91.9% for those treated with Sensikill and 27.5% for the placebo group. With regard to sensitivity to volatile stimulus, the rate of improvement for teeth treated with Dessensiv, Sensikill and placebo was, respectively, 83.5%, 87.1% and 20.4%. Upon analyzing the results, one can conclude that the commercial products tested are efficient in the treatment of dentinal cervical hypersensitivity after three applications and that there is no significant statistical difference between the two products, both being more effective than the placebo.

Avaliação da existência de controle de infecção oral nos pacientes internados em hospitais do estado do Rio de Janeiro / Evaluating the existence of oral infection control in patients admitted in state hospitals in Rio de Janeiro

O objetivo do presente estudo foi verificar a existência de um protocolo de controle de infecção oral nos hospitais do Estado do Rio de Janeiro. Foram avaliados 62 hospitais da rede pública e privada. Quando questionados se existia na instituição algum procedimento destinado aos pacientes internados (no caso de UTIs), 39 por cento dos hospitais pesquisados responderam positivamente. Das instituições pesquisadas, apenas 15 por cento possuem um protocolo regular para o controle de placa nos pacientes internados. Pesquisas têm demonstrado a inter-relação da doença periodontal com doenças sistêmicas, como doenças cardiovasculares, osteoporose, nascimento de bebês de baixo peso e parto prematuro, diabetes e doenças respiratórias. A cavidade oral tem sido considerada como um potente reservatório de patógenos respiratórios. Diante destes fatos, evidencia-se a importância da higiene bucal como um meio de se prevenir patologias diversas. Desta forma, existe a necessidade de se criar um protocolo de controle de infecção da cavidade oral para contribuir para a redução da mortalidade de pacientes internados e propor medidas preventivas para este fim.

The aim of the present study is to ascertain whether an oral infection control protocol is being used in Rio de Janeiro State hospitals. Sixty two hospitals, both public and private have been surveyed. When asked whether any procedure was being adopted with admitted patients (in case of ICU patients), 39 percent of the hospitals responded positively. Among all institutions surveyed, only 15 percent conduct a regular plaque control protocol in admitted patients. Studies have shown that there is an inter-relationship between periodontal disease and systemic diseases, such as cardiovascular disease, osteoporosis, preterm births and low birthweight, diabetes and respiratory diseases. The oral cavity is considered a potent reservoir of respiratory pathogens. Based on these findings, it has become evident that oral hygiene is an important means of preventing a number of diseases. Thus the need arises not only to create an oral cavity infection control protocol that will contribute to reduce mortality in admitted patients but also to propose preventive measures towards this end.

[Evaluating the existence of oral infection control in patients admitted in state hospitals in Rio de Janeiro]. / Avaliação da existência de controle de infecção oral nos pacientes internados em hospitais do estado do Rio de Janeiro.

The aim of the present study is to ascertain whether an oral infection control protocol is being used in Rio de Janeiro State hospitals. Sixty two hospitals, both public and private have been surveyed. When asked whether any procedure was being adopted with admitted patients (in case of ICU patients), 39% of the hospitals responded positively. Among all institutions surveyed, only 15% conduct a regular plaque control protocol in admitted patients. Studies have shown that there is an inter-relationship between periodontal disease and systemic diseases, such as cardiovascular disease, osteoporosis, preterm births and low birthweight, diabetes and respiratory diseases. The oral cavity is considered a potent reservoir of respiratory pathogens. Based on these findings, it has become evident that oral hygiene is an important means of preventing a number of diseases. Thus the need arises not only to create an oral cavity infection control protocol that will contribute to reduce mortality in admitted patients but also to propose preventive measures towards this end.

Baixa adesão dos pacientes hipertensos a estratégia de tratamento e acompanhamento atraves do cuidado continuado

O objetivo deste estudo foi desenvolver um projeto de intervenção visando atuar sobre a adesão ao tratamento medicamentoso ou não. Foi feita uma revisão narrativa de literatura, nas bases de dados BVS, Portal CAPES e COCHRANE, usando os descritores hipertensão, cooperação do paciente, adesão à medicação e cuidado periódico. Após a revisão foi elaborado um projeto de intervenção tendo como referência o Planejamento Estratégico Situacional, descrito no módulo de Planejamento e Avaliação em Saúde do Curso de Especialização em Saúde da Família do Núcleo de Estudos em Saúde Coletiva. Espera-se com este projeto estimular o conhecimento dos pacientes sobre o autocuidado e o cuidado continuado e estimular a adesão aos tratamentos medicamentoso e não-medicamentoso, com redução da morbimortalidade e melhora da qualidade de vida.