Resistin Modulates the Functional Activity of Colostral Macrophages from Mothers with Obesity and Diabetes.

BACKGROUND: Obesity and diabetes are major public health problems. Resistin is an adipokine that links the two diseases. There are few reports regarding colostrum cells and resistin from mothers with obesity and diabetes. Thus, this study aimed to determine the functional activity of macrophages present in the breast milk and colostrum of diabetic mothers with obesity and the effects of resistin on these cells. METHODS: The women were divided according to BMI and glycemic status into normal weight non-diabetic, obese non-diabetic, normal weight type 2 diabetic, or obese type 2 diabetic groups. ELISA determined the resistin in colostrum. The cell subsets and apoptosis were determined by flow cytometry and the functional activity of cells by fluorescence microscopy. RESULTS: The resistin levels were higher in the colostrum from diabetic mothers with obesity. The frequencies of CD14+ cells and cells expressing CD95+, independent of resistin treatment, were higher in the colostrum from diabetic mothers with obesity. The frequency of cells expressing CD14+CD95+ was higher in cells not treated with resistin in the colostrum from diabetic mothers with obesity. Apoptosis, irrespective of the presence of resistin, increased, whereas microbicidal activity decreased in cells from diabetic mothers with obesity. CONCLUSION: The data suggest that hyperglycemia associated with low-grade inflammation caused by obesity affects the percentage of cells expressing CD14+CD95+, death by apoptosis, and microbicidal indices; meanwhile, resistin restored the microbicidal activity of colostrum cells.

Multiomic Approaches Reveal Hormonal Modulation and Nitrogen Uptake and Assimilation in the Initial Growth of Maize Inoculated with Herbaspirillum seropedicae.

Herbaspirillum seropedicae is an endophytic bacterium that can fix nitrogen and synthesize phytohormones, which can lead to a plant growth-promoting effect when used as a microbial inoculant. Studies focused on mechanisms of action are crucial for a better understanding of the bacteria-plant interaction and optimization of plant growth-promoting response. This work aims to understand the underlined mechanisms responsible for the early stimulatory growth effects of H. seropedicae inoculation in maize. To perform these studies, we combined transcriptomic and proteomic approaches with physiological analysis. The results obtained eight days after inoculation (d.a.i) showed increased root biomass (233 and 253%) and shoot biomass (249 and 264%), respectively, for the fresh and dry mass of maize-inoculated seedlings and increased green content and development. Omics data analysis, before a positive biostimulation phenotype (5 d.a.i.) revealed that inoculation increases N-uptake and N-assimilation machinery through differentially expressed nitrate transporters and amino acid pathways, as well carbon/nitrogen metabolism integration by the tricarboxylic acid cycle and the polyamine pathway. Additionally, phytohormone levels of root and shoot tissues increased in bacterium-inoculated-maize plants, leading to feedback regulation by the ubiquitin-proteasome system. The early biostimulatory effect of H. seropedicae partially results from hormonal modulation coupled with efficient nutrient uptake-assimilation and a boost in primary anabolic metabolism of carbon-nitrogen integrative pathways.

Necessidades sentidas pelas mulheres no período puerperal / Needs felt by women in the puerperal period

Objetivo: conhecer as necessidades sentidas pelas mulheres no período puerperal. Método: tratase de estudo qualitativo, exploratório, descritivo, com 20 mulheres. Coletaram-se os dados por meio de entrevistas semiestruturadas e, posteriormente, analisadas e categorizadas conforme a técnica de Análise de Conteúdo na modalidade Análise Temática. Resultados: emergiu-se a temática "Necessidades sentidas pelas mulheres no período puerperal", que se constitui pelas subcategorias "Puerpério imediato", "Puerpério remoto". Conclusão: mostra-se que as mulheres sentem necessidades distintas ao longo do período puerperal suscitando a atuação dos profissionais desde o pré-natal para preparar a mulher para as situações que serão vivenciadas no puerpério, bem como o trabalho com a família no fortalecimento das relações e no preparo da rede de apoio para a chegada do novo membro.(AU)

Functional characterization of a serine protease inhibitor modulated in the infection of the Aedes aegypti with dengue virus.

During feeding with blood meal, female Aedes aegypti can transmit infectious agents, such as dengue, yellow fever, chikungunya and Zika viruses. Dengue virus causes human mortality in tropical regions of the world, and there is no specific treatment or vaccine with maximum efficiency being used for these infections. In the vector-virus interaction, the production of several molecules is modulated by both mosquitoes and invading agents. However, little information is available about these molecules in the Ae. aegypti mosquito during dengue infection. Inhibitors of the pacifastin family have been described to participate in the immune response of insects and Pac2 is the only gene of this family present in Ae. aegypti being then chosen for investigation. Pac2 was expressed in E. coli, purified and analyzed by mass spectrometry and SDS-PAGE. The Pac2 transcript was detected by qPCR, and its protein levels were assessed by Western blotting. The inhibitory activity of Pac2 was measured using its Ki, IC50 and zymography. Mosquito infections with DENV were introduced with the Brazilian ACS-46 DENV-2 strain propagated in C6/36 cells. In the present work, we showed that it is possibly involved in the interaction of the mosquitoes with the dengue virus. The Pac2 transcript was detected in larvae and in both the salivary gland and midgut of Ae. aegypti females, while the native protein was identified in females 3 h post-blood meal. Pac2 is a strong inhibitor of trypsin-like and thrombin-like proteases, which are present in 4th instar larvae midgut and females 24 h after blood meal. During DENV infection, up regulation of Pac2 expression occurs in the salivary gland and midgut. Pac2 is the first Pacifastin inhibitor member described in mosquitoes. Our results suggest that Pac2 acts on mosquito serine proteases, mainly the trypsin-like type, and is under transcriptional control by virus infection signals to allow its survival in the vector or by the mosquito as a defense mechanism against virus infection.

Gestantes e puérperas soropositivas para o HIV e suas interfaces de cuidado / HIV-positive pregnant and puerperal women and their interfaces of care

Objetivo: compreender a percepção de ser gestante/puérpera soropositiva para o HIV. Método: estudo qualitativo, descritivo-exploratório, realizado com uma gestante e duas puérperas internadas soropositivas para o HIV. Os dados foram obtidos por meio entrevista com questões semiestruturadas. A análise das informações ocorreu a partir da técnica de análise de conteúdo na modalidade análise temática. Resultados: da análise, emergiram duas categorias > e >. O desfecho dos cuidados em saúde tem relação direta com a assistência profissional, em que práticas humanizadas, pautadas numa relação empática de apoio e acolhimento, mostram-se eficazes para o desenvolvimento do autocuidado e cuidado do outro. Conclusão: considera-se necessário criar ações intersetoriais que repercutam na assistência prestada às portadoras do HIV, sensibilizando os profissionais para acolher este público, em todos os níveis atenção.

Insatisfação e checagem corporal e comportamento alimentar em estudantes de Educação Física, Nutrição e Estética / Body dissatisfaction and body-checking and eating behavior in undergraduate in Physical Education, Nutrition and Aesthetics

Esse estudo objetivou verificar a correlação da checagem e da insatisfação corporal com o comportamento alimentar em acadêmicos dos cursos de Educação Física, Nutrição e Estética, assim como comparar as variáveis entre ambos os sexos. No total, participaram 207 acadêmicos, com idade entre 19 a 45 anos. Para avaliar a insatisfação com a imagem corporal em ambos os sexos foi utilizado o Body Shape Questionnaire (BSQ). Para a verificação do comportamento de checagem corporal foi utilizado o Body Checking Questionnaire (BCQ) em mulheres e o Male Body Checking Questionnaire (MBCQ) em homens. A fim de avaliar os comportamentos de risco para transtornos alimentares foi utilizado o Eating Attitudes Test-26 (EAT-26). Além disso, foram coletados dados de massa corporal e estatura dos voluntários, através de um questionário sociodemográfico. A partir dos achados, na Educação Física as mulheres apresentaram maior média para insatisfação corporal (81,67 pontos) e para os comportamentos alimentares inadequados (13,13 pontos) quando comparadas aos homens (49,46 pontos e 6,49 pontos, respectivamente). No curso de Nutrição, o IMC (25,86) e a checagem corporal (1,29) foram estatisticamente maiores em homens quando comprados às mulheres do mesmo curso. Conclui-se que, a partir das análises de regressão, o comportamento alimentar foi correlacionado com a insatisfação corporal para os cursos de Educação Física e Estética. Nas estudantes de Nutrição, a insatisfação e a checagem corporal influenciaram o comportamento alimentar....(Au)

This study aimed to verify the correlation between body-checking and body dissatisfaction with eating behavior in undergraduate in Physical Education, Nutrition and Aesthetics, as well as to compare the variables between both sexes. In total, 207 students took part, aged between 19-45 years old. To assess the body image dissatisfaction in both sexes was used the Body Shape Questionnaire (BSQ). To verify body-checking behaviors was used the Body Checking Questionnaire (BCQ) for women and the Male Body Checking Questionnaire (MBCQ) for men. In order to assess risk behaviors for eating disorders was used the Eating Attitudes Test-26 (EAT-26). In addition, we collected body mass and height of the volunteers through a sociodemographic questionnaire. From the findings, Physical Education women had a higher average for body dissatisfaction (81.67 points) and inappropriate eating behaviors (13.13 points) than men (49.46 points and 6.49 points respectively). In the course of Nutrition, the BMI (25.86) and body-checking (1.29) were statistically higher in men when compared to women in the same course. We concluded that, in regression analyzes, eating behavior was correlated with body dissatisfaction in the courses of Physical Education and Aesthetics. In the Nutrition's female students, the dissatisfaction and body-checking influenced eating behavior....(AU)

BmTI-A, a Kunitz type inhibitor from Rhipicephalus microplus able to interfere in vessel formation.

Rhipicephalus microplus is an ectoparasite responsible for transmissions of babesiosis and anaplasmosis causing large losses to livestock production. To survive R. microplus tick produces several active molecules, such as protease inhibitors. This ectoparasite has been described as a rich source of serine protease inhibitors most of them are Kunitz-BPTI members named BmTIs which have no clear function yet. In the present work, we described the expression and functional characterization of rBmTI-A which showed to be similar to the native BmTI-A, a double-headed Kunitz-BPTI inhibitor, capable to inhibit trypsin, human neutrophil elastase (HNE), human plasma kalikrein (HuPK) and human plasmin. rBmTI-A was able to cause a decrease of HUVEC cell viability. Besides, the rBmTI-A showed to be a potent inhibitor of "in vitro" vessel formation. Our results suggested that BmTI-A may participate in the blood acquisition process interfering in the vessel formation during the tick parasite life stage, around 20 days. In conclusion, BmTI-A is a promising molecule to be used in the drug design and development of new method of R. microplus control.

Educação sexual para adolescentes: aliança entre escola e enfermagem/saúde / Sex education for teens: alliance between school and nursing/ healthcare

Este estudo objetivou destacar a importância da educação sexual na escola, a partir da aliança entre escola e enfermagem/saúde. Trata-se de um estudo qualitativo de caráter exploratório e descritivo, desenvolvido com cinco adolescentes, cujos dados foram coletados por meio de entrevistas semi estruturadas. A partir da organização dos dados emergiram as seguintes temáticas: a escola como espaço para a educação sexual de adolescentes; escola e enfermagem/saúde: aliança necessária para a educação sexual de adolescentes. Os resultados evidenciam que a escola constitui-se em um importante espaço para a educação sexual, no entanto, os profissionais desse setor precisam atentar para aspectos individuais e contextuais de cada adolescente. Destaca-se também a necessidade de alianças entre educação e saúde, uma vez que possibilitam o desenvolvimento de competências entre os dois serviços...

This study aimed to highlight the importance of sex education in the school, through the alliance between school and nursing/healthcare. This is a qualitative study of exploratory and descriptive nature, developed with five teenagers, whose data was collected through semi-structured interviews. From the organization of the data, the following themes emerged: the school as a place for sex education for teenagers; school and nursing/healthcare: alliance necessary for the sexual education of teenagers.The results show that the school is an important site for sex education; however, the professionals in this area shall be attentive to individual and contextual aspects of each adolescent. Also important is the need for alliances between education and healthcare, since they would allow the development of responsibilities between the two services...

Exame de toque retal: a percepção de homens quanto à sua realização / Rectal exam: the perception of men as their realization

Objetivo: compreender a percepção de homens quanto ao exame de toque retal. Metodologia: pesquisa qualitativa com doze homens com 40 anos de idade ou mais que nunca se submeteram ao exame de toque retal. Os dados foram coletados por meio de entrevista, em 2011, contendo a seguinte questão norteadora “Conta pra mim, o que você sabe sobre o exame de toque retal em homens?”. Os dados foram analisados pela técnica de análise do conteúdo. Resultados: emergiram-se três categorias “Realização do exame de toque retal: o que é e quando fazer?”, obtendo-se que os entrevistados desconhecem sobre o exame e quando deve ser realizado; “O que influencia a não realização do exame”, evidenciando a deficiência das informações, falta de solicitação do profissional médico, opção pelo PSA, ausência de sinais e sintomas; “Sentimentos atribuídos ao exame de toque retal”, emergindo sentimentos: vergonha, medo, desconforto, sendo os mais representativos o constrangimento e machismo. Revelou-se ainda sentimento de conformação relacionado ao exame. Conclusão: a ausência de informação de caráter científico é a responsável pela banalização e recusa dos homens, apontando para a necessidade contínua de promover ações educativas sobre a prevenção câncer de próstata.

Objective: to understand the perception of men as the rectal exam. Methodology: qualitative research with twelve men with 40 years old or more who never underwent the rectal exam. The data were collected through interviews, in 2011, containing the following guiding question "tell me, what do you know about the rectal exam in men?". Data were analyzed by the technique of content analysis. Results: three categories emerged "realization of rectal exam: what it is and when to do it?", obtaining respondents unaware about the exam and when should be performed; "What influences not examination", demonstratingthe deficiency of information, lack of medical professional's request, option by the PSA, and the absence of signs and symptoms; Feelings attributed to the rectal exam, "emerging feelings: shame, fear, discomfort, being the most representative the embarrassment and the machismo. It also revealed a feeling of conformation exam-related. Conclusion: the lack of scientific character information is responsible for the trivialization and denial of men, pointing to the continuing need for promoting educational activities on the prevention of prostate cancer.

Production of serine protease inhibitors by mutagenesis and their effects on the mortality of Aedes aegypti L. larvae.

BACKGROUND: Dengue, transmitted primarily by the bites of infected Aedes aegypti L., is transmitted to millions of individuals each year in tropical and subtropical areas. Dengue control strategies are primarily based on controlling the vector, using insecticides, but the appearance of resistance poses new challenges. Recently, highly selective protease inhibitors by phage display were obtained for digestive enzymes of the 4th instar larvae (L4) midgut. These mutants were not confirmed as a larvicide due to the low yield of the expression of these inhibitors. In the present study, chimera molecules were constructed based on the mutations at positions P1-P4' selected previously. The T6, T23 and T149 mutants were mixed with another Kunitz inhibitor, domain 1 of the inhibitor boophilin (D1). METHODS: The chimeras T6/D1, T149/D1 and T23/D1 were expressed at high levels in P. pastoris yeast, purified by ionic exchange chromatography and their homogeneity was analyzed by SDS-PAGE. The chimera inhibitors were assayed against larval trypsin, chymotrypsin and elastase using specific chromogenic substrates. The inhibitors were assayed for their larvicide potential against L4. RESULTS: The chimeras exhibited strong inhibitory activities against the larval digestive enzymes in a dose-dependent manner. T6/D1, T149/D1 and T23/D1 exhibited strong larvicidal activity against L4 of Ae. aegypti with inhibitor concentrations in the µM range. A synergistic increase in mortality was observed when a mixture of the three chimeric inhibitors was tested. CONCLUSIONS: The strategy for constructing the chimeric inhibitors was successful. The chimeras showed strong larvicidal activity against Ae. aegypti. In the future, our findings can be used to design synthetic inhibitors for larvae digestive enzymes as an alternative method to control the dengue vector.

A Kazal-type inhibitor is modulated by Trypanosoma cruzi to control microbiota inside the anterior midgut of Rhodnius prolixus.

The triatomine insect, Rhodnius prolixus, is a vector of Trypanosoma cruzi, a protozoan parasite that causes Chagas disease. The parasite must overcome immune response and microbiota to develop inside the midgut of triatomines. In this study, we expressed, purified and characterized a Kazal-type inhibitor from the midgut of R. prolixus, named RpTI, which may be involved in microbiota - T. cruzi interactions. The qPCR showed that the RpTI transcript was primarily expressed in tissues from the intestinal tract and that it was upregulated in the anterior midgut after T. cruzi infection. A 315-bp cDNA fragment encoding the mature protein was cloned into the pPIC9 vector and expressed in Pichia pastoris system. Recombinant RpTI (rRpTI) was purified on a trypsin-Sepharose column and had a molecular mass of 11.5 kDa as determined by SDS-PAGE analysis. This protein inhibited trypsin (Ki = 0.42 nM), whereas serine proteases from the coagulation cascade were not inhibited. Moreover, trypanocidal assays revealed that rRpTI did not interfere in the viability of T. cruzi trypomastigotes. The RpTI transcript was also knocked down by RNA interference prior to infection of R. prolixus with T. cruzi. The amount of T. cruzi in the anterior midgut was significantly lower in RpTI knockdown insects compared to the non-silenced groups. We also verified that the bacterial load is higher in the anterior midgut of silenced and infected R. prolixus compared to non-silenced and infected insects. Our results suggest that T. cruzi infection increases the expression of RpTI to mediate microbiota modulation and is important for parasite immediately after infection with R. prolixus.

Rmcystatin3, a cysteine protease inhibitor from Rhipicephalus microplus hemocytes involved in immune response.

The Rhipicephalus microplus tick is responsible for losses in the livestock production estimated in 2 billions USD. Despite its economical importance the knowledge in tick's physiology is sparse. In order to contribute to this scenario we describe the characterization of a cysteine proteinase inhibitor named Rmcystatin-3. Purified recombinant Rmcystatin-3 was able to inhibit cathepsin L (Ki = 2.5 nM), BmCl1 (Ki = 1.8 nM) and cathepsin B (Ki = 136 nM). Western blot and quantitative PCR analysis revealed the presence of Rmcystatin-3 in fat body, salivary gland but mainly in hemocytes. The mRNA levels of Rmcystatin-3 during bacterial challenge are drastically down-regulated. In order to define the Rmcystatin-3 possible role in tick immunity, the cystatin gene was knockdown by RNA interference with and without Escherichia coli infection. Our results showed that the Rmcystatin-3 silenced group was more immune competent to control bacterial infection than the group injected with non-related dsRNA. Taking together, our data strongly suggested an important role of Rmcystatin-3 in tick immunity.

RmKK, a tissue kallikrein inhibitor from Rhipicephalus microplus eggs.

Rhipicephalus microplus is an important ectoparasite that is responsible for transmission of anaplasmosis and babesiosis to cattle. Tissue kallikrein inhibitors might play an important role in R. microplus eggs. In the present work, we purified and characterized, a tissue kallikrein inhibitor presents in R. microplus eggs (RmKK), a protein which contains two Kunitz domain in tandem. Purified inhibitor was confirmed by amino terminal determination and its dissociation constant (Ki) for bovine trypsin and porcine pancreatic kallikrein were 0.6 nM and 91.5 nM, respectively. Using a cDNA library from R. microplus midgut, we cloned the cDNA fragment encoding mature RmKK and expressed the protein in Pichia pastoris system. Recombinant RmKK was purified by ion exchange chromatography and presented molecular mass of 16.3 kDa by MALDI-TOF analysis. Moreover, RmKK showed a tight binding inhibition for serine proteases as bovine trypsin (Ki=0.2 nM) and porcine pancreatic kallikrein (PPK) (Ki=300 nM). We performed, for the first time, the characterization of a tissue kallikrein inhibitor presents in R. microplus eggs, which the transcript is produced in the adult female gut. BmKK seems to be the strongest PPK inhibitor among all BmTIs present in the eggs and larvae (Andreotti et al., 2001; Sasaki et al., 2004). This data suggests that BmKK may participate in the development of tick egg and larvae phase.

Selective inhibitors of digestive enzymes from Aedes aegypti larvae identified by phage display.

Dengue is a serious disease transmitted by the mosquito Aedes aegypti during blood meal feeding. It is estimated that the dengue virus is transmitted to millions of individuals each year in tropical and subtropical areas. Dengue control strategies have been based on controlling the vector, Ae. aegypti, using insecticide, but the emergence of resistance poses new challenges. The aim of this study was the identification of specific protease inhibitors of the digestive enzymes from Ae. aegypti larvae, which may serve as a prospective alternative biocontrol method. High affinity protein inhibitors were selected by all of the digestive serine proteases of the 4th instar larval midgut, and the specificity of these inhibitors was characterized. These inhibitors were obtained from a phage library displaying variants of HiTI, a trypsin inhibitor from Haematobia irritans, that are mutated in the reactive loop (P1-P4'). Based on the selected amino acid sequence pattern, seven HiTI inhibitor variants were cloned, expressed and purified. The results indicate that the HiTI variants named T6 (RGGAV) and T128 (WNEGL) were selected by larval trypsin-like (IC(50) of 1.1 nM) and chymotrypsin-like enzymes (IC(50) of 11.6 nM), respectively. The variants T23 (LLGGL) and T149 (GGVWR) inhibited both larval chymotrypsin-like (IC(50) of 4.2 nM and 29.0 nM, respectively) and elastase-like enzymes (IC(50) of 1.2 nM for both). Specific inhibitors were successfully obtained for the digestive enzymes of Ae. aegypti larvae by phage display. Our data also strongly suggest the presence of elastase-like enzymes in Ae. aegypti larvae. The HiTI variants T6 and T23 are good candidates for the development as a larvicide to control the vector.

Expression and functional characterization of boophilin, a thrombin inhibitor from Rhipicephalus (Boophilus) microplus midgut.

Rhipicephalus (Boophilus) microplus is an ectoparasite responsible for an important decrease in meat, milk and leather production, caused both by cattle blood loss and by the transmission of anaplasmosis and babesiosis. R. microplus is a rich source of serine protease inhibitors, including the trypsin inhibitors BmTI-A and BmTI-6, the subtilisin inhibitor BmSI, and the recently described thrombin inhibitor, boophilin. Boophilin is a double Kunitz-type thrombin inhibitor, with the unusual ability to form a ternary complex with a second (non-thrombin) serine proteinase molecule. The large-scale expression and purification of boophilin and of its isolated N-terminal (D1) domain in Pichia pastoris, its expression profile, and the effect of RNAi-mediated gene silencing in tick egg production are reported. Full-length boophilin and D1 were expressed at 21 and 37.5mg/L of culture, respectively. Purified boophilin inhibited trypsin (K(i) 0.65 nM), neutrophil elastase (K(i) 21 nM) and bovine thrombin (K(i) 57 pM), while D1 inhibited trypsin and neutrophil elastase (K(i) of 2.0 and 129 nM, respectively), but not thrombin. Boophilin gene silencing using RNAi resulted in 20% reduction in egg weight production, suggesting that the expression of boophilin in this life stage would be important but not vital, probably due to functional overlap with other serine proteinase inhibitors in the midgut of R. microplus. Considering our data, Boophilin could be combining with other antigen in a vaccine production for tick control.

Molecular characterization of genes encoding trypsin-like enzymes from Aedes aegypti larvae and identification of digestive enzymes.

Trypsin-like enzymes play an important role in the Aedes aegypti digestive process. The trypsin-like enzymes present in adults were characterized previously, but little is known about trypsins in larvae. In the present work, we identified one of the trypsin enzymes from Ae. aegypti larval midgut using a library of trypsin gene fragments, which was the sequence known as AAEL005607 from the Ae. aegypti genome. Quantitative PCR analysis showed that AAEL005607 was transcribed in all larval instars, but it was not present in adult midgut. In order to confirm transcription data, the trypsin-like enzymes from 4th instar larvae of Ae. aegypti midgut were purified and sequenced. Purified trypsin showed identity with the amino-terminal sequence of AAEL005607, AAEL005609 and AAEL005614. These three trypsins have high amino acids identity, and could all be used as a template for the design of inhibitors. In conclusion, for the first time, digestive enzymes of 4th larval instar of Ae. aegypti were purified and characterized. The knowledge of digestive enzymes present in Ae. aegypti larvae may be helpful in the development of a larvicide.

Boophilus microplus cathepsin L-like (BmCL1) cysteine protease: specificity study using a peptide phage display library.

The tick Rhipicephalus (Boophilus) microplus is one of the most important bovine ectoparasites, a disease vector responsible for losses in meat and milk productions. A cysteine protease similar to cathepsin L, named BmCL1, was previously identified in R. microplus gut, suggesting a role of the enzyme in meal digestion. In this work, BmCL1 was successfully expressed in Pichia pastoris system, yielding 54.8 mg/L of culture and its activity was analyzed by synthetic substrates and against a R. microplus cysteine protease inhibitor, Bmcystatin. After rBmCl1 biochemical characterization it was used in a selection of a peptide phage library to determine rBmCL1 substrate preference. Obtained sequenced clones showed that rBmCL1 has preference for Leu or Arg at P(1) position. The preference for Leu at position P(1) and the activation of BmCL1 after a Leu amino acid residue suggest possible self activation.

Programa de saúde para o pai adolescente: reflexões sobre sua relevância / Health program for the adolescent parents: reflections on its relevance

Objetivo: refletir sobre a relevância do desenvolvimento de programas de saúde para pais adolescentes, de forma que estes possam compreender melhor a paternidade e sentirem-se seguros para assumir o filho e, quem sabe, a família. Metodologia: pesquisa bibliográfica nas bases de dados PubMed, SCIELO e LILACS, entre os meses de maio e junho de 2009. Foram encontrados 105 artigos, publicados entre os anos de 2000 a 2007, com os descritores: Paternidade, Adolescência, Programa de saúde e Sexualidade. Dentre estes artigos, apenas 16 constituiram-se base referencial para este estudo, por focalizarem, especificamente, o tema escolhido. Resultados: na pesquisa não foram encontrados muitos artigos sobre o tema, pois este ainda é pouco explorado. Além disso, os programas de saúde existentes só incluem os cuidados com a mãe adolescente esquecendo-se do pai. Sabe-se que a participação do pai adolescente no cuidado com o filho desde a gestação proporciona um melhor vínculo deste com o filho e com a companheira. Conclusão: faz-se necessário a existência de redes sociais de apoio capazes de inserir estes pais adolescentes no processo complexo que se inicia ainda na concepção.(AU)

A novel trypsin Kazal-type inhibitor from Aedes aegypti with thrombin coagulant inhibitory activity.

Kazal-type inhibitors play several important roles in invertebrates, such as anticoagulant, vasodilator and antimicrobial activities. Putative Kazal-type inhibitors were described in several insect transcriptomes. In this paper we characterized for the first time a Kazal unique domain trypsin inhibitor from the Aedes aegypti mosquito. Previously, analyses of sialotranscriptome of A. aegypti showed the potential presence of a Kazal-type serine protease inhibitor, in female salivary glands, carcass and also in whole male, which we named AaTI (A. aegypti trypsin inhibitor). AaTI sequence showed amino acid sequence similarity with insect thrombin inhibitors, serine protease inhibitor from Litopenaeus vannamei hemocytes and tryptase inhibitor from leech Hirudo medicinalis (LDTI). In this work we expressed, purified and characterized the recombinant AaTI (rAaTI). Molecular weight of purified rAaTI was 7 kDa rAaTI presented dissociation constant (K(i)) of 0.15 and 3.8 nM toward trypsin and plasmin, respectively, and it weakly inhibited thrombin amidolytic activity. The rAaTI was also able to prolong prothrombin time, activated partial thromboplastin time and thrombin time. AaTI transcription was confirmed in A. aegypti female salivary gland and gut 3 h and 24 h after blood feeding, suggesting that this molecule can act as anticoagulant during the feeding and digestive processes. Its transcription in larvae and pupae suggested that AaTI may also play other functions during the mosquito's development.

A novel trypsin Kazal-type inhibitor from Aedes aegypti with thrombincoagulant inhibitory activity

Kazal-type inhibitors play several important roles in invertebrates, such as anticoagulant, vasodilator andantimicrobial activities. Putative Kazal-type inhibitors were described in several insect transcriptomes. Inthis paper we characterized for the first time a Kazal unique domain trypsin inhibitor from the Aedes aegypti mosquito. Previously, analyses of sialotranscriptome of A. aegypti showed the potential presence of a Kazal-type serine protease inhibitor, in female salivary glands, carcass and also in whole male, which we named AaTI (A. aegypti trypsin inhibitor). AaTI sequence showed amino acid sequence similarity withinsect thrombin inhibitors, serine protease inhibitor from Litopenaeus vannamei hemocytes and tryptaseinhibitor from leech Hirudo medicinalis (LDTI). In this work we expressed, purified and characterized therecombinant AaTI (rAaTI). Molecular weight of purified rAaTI was 7 kDa rAaTI presented dissociation constant (Ki) of 0.15 and 3.8 nM toward trypsin and plasmin, respectively, and it weakly inhibited thrombin amidolytic activity. The rAaTI was also able to prolong prothrombin time, activated partial thromboplastin time and thrombin time. AaTI transcription was confirmed in A. aegypti female salivary gland and gut 3 h and 24 h after blood feeding, suggesting that this molecule can act as anticoagulant during the feeding and digestive processes. Its transcription in larvae and pupae suggested that AaTI may also play other functions during the mosquito’s development.