Utilização da lipoproteína de baixa densidade, em diferentes concentrações, em meio diluidor para criopreservação do sêmen ovino / Use of low density lipoprotein in different concentrations in diluent medium for cryopreservation of ovine semen

A utilização da gema de ovo dificulta a padronização de meios diluidores e apresenta riscos biológicos. Assim, este estudo avaliou diferentes concentrações da lipoproteína de baixa densidade (LDL), em substituição à gema de ovo, para a confecção de diluentes para criopreservação espermática em ovinos. Foram utilizados um diluidor controle (CTR= 20% de gema de ovo) e cinco tratamentos, substituindo-se a gema pelas diferentes proporções de LDL (T1=6%; T2=8%; T3=12%; T4=16%; T5=20%), todos à base de TRIS-glicerol. Para o estudo, utilizaram-se dois ejaculados, de seis reprodutores da raça Santa Inês. Sessenta dias após a criopreservação, as amostras foram descongeladas e avaliadas subjetivamente quanto à motilidade total (MT, %) e progressiva (MP, %), ao vigor (1-5) e à integridade funcional (choque hisposmótico com água destilada, %) e estrutural (corante supravital eosina, %) das membranas espermáticas. As avaliações de vigor e funcionalidade de membrana não diferiram (P>0,05) entre os grupos. Entretanto, os grupos T4 (P<0,01) e T5 (P<0,05) foram superiores ao CTR para os parâmetros MT, MP e integridade estrutural de membrana, o que confirma que as LDLs podem ser alternativas eficientes para substituição da gema de ovo em diluidores para criopreservação de sêmen ovino.(AU)

The use of egg yolk makes it difficult to standardize extenders and presents biological hazards. Thus, this study evaluated different concentrations of low-density lipoprotein (LDL) to replace yolk extenders for production of sperm for cryopreservation in ovine. A control extender was used (CTR= 20% yolk) and five treatments, replacing the yolk by different ratios of LDL (T1= 6%; T2= 8%, T3= 12%; T4= 16%; T5= 20%) all based on TRIS-glycerol. For the study, two ejaculates from six Santa Ines breeding were used. Sixty days after cryopreservation, the samples were thawed and evaluated for total motility (MT, %) and progressive motility (MP, %), vigor (1-5) and the functional integrity (hyposmotic shock with distilled water, %) and structural (supravital dye eosin, %) of the sperm membranes. The evaluations of strength and membrane functionality didn't differ (P> 0.05) between groups. However, T4 (P< 0.01) and T5 (P< 0.05) groups were superior to the CTR for the MT, MP, and membrane structural integrity parameters, which confirms that LDLs can be efficient alternatives for yolk replacement in extenders for cryopreservation of ovine semen.(AU)

SNPs rs471462296, rs456245081 and rs438495570 in the 5'UTR region of DGAT1 gene in Nelore / SNPs rs471462296, rs456245081 e rs438495570 na região 5'UTR do gene DGAT1 em Nelore

O objetivo desta pesquisa foi avaliar os SNPs rs471462296, rs456245081 e rs438495570 do gene DGAT1 em bovinos Nelore. Foram analisados 109 bovinos. A extração do DNA genômico foi realizada do sangue dos animais, usando-se o kit Ilustra Blood Genomic Prep Mini Spin® (GE Healthcare, UK). A concentração e o grau de pureza do DNA foram determinados por meio de espectrofotômetro (Nanodrop - Thermo Fisher Scientifc, USA). A genotipagem dos SNPs ocorreu mediante o emprego do ensaio Taqman® (Applied Biosystems, USA). Na análise genômica, não foram encontradas alterações nas frequências alélicas e genotípicas (P≥0,05) para os SNPs testados. Dessa forma, a região 5'UTR analisada apresentou-se monomórfica e a variação de SNPs não foi observada, o que limita seu uso como marcadores moleculares para o gene DGAT1 em Nelore.(AU)

Changes in energetic metabolism of Biomphalaria glabrata (Mollusca, Planorbidae) in response to exogenous calcium / Alterações no metabolismo energético de Biomphalaria glabrata em resposta ao cálcio exógeno

Abstract Calcium is considered an essential element for the metabolism of aquatic snail Biomphalaria glabrata (Say, 1818), intermediate host of Schistosoma mansoni Sambon, 1907 in Brazil, and represents a limiting factor to its distribution and adaptation to the environment. This study investigated the effect of different concentrations of exogenous CaCO3 on the energetic metabolism of B. glabrata for better understanding the physiological interference of chemical elements dissolved in the environment with the physiology of this species. Sixty-day-old snails were distributed into six groups, five exposed to different concentrations of CaCO3 (20, 40, 60, 80 and 100 mg/L) and a control group. The exposure to CaCO3 was assessed over time, with analysis of 15 snails of each group in the following intervals: 1, 14, 21 or 30 days for hemolymph extraction. Concentrations of calcium and glucose in the hemolymph were determined by commercial kits, and organic acids were extracted using an ion exchange column and analyzed by high-performance liquid chromatography. Concentration of calcium in the hemolymph showed no significant difference (p>0.05) from the control group and between the concentrations tested. Concentration of glucose decreased (p<0.05) in the treatments of exposure to 20 and 40 mg/L and increased when exposed to 80 and 100 mg/L CaCO3 compared to control and to other concentrations tested over 30 days. The organic acids pyruvate, oxaloacetate, citrate, succinate, fumarate, beta-hydroxybutyrate and lactate presented increased concentrations, while propionate and acetoacetate, decreased concentrations, when exposed to CaCO3 compared to control. Considering the influence of different periods of exposure to CaCO3, on the 14th day, there were stronger alterations in the metabolism of B. glabrata. In conclusion, exposure to CaCO3 reduced the concentration of glucose, which is metabolized into pyruvate, the final product of glycolysis, and also influenced the energetic metabolism pathways, indicating an aerobic or partially anaerobic functioning.

Resumo O cálcio é considerado um elemento essencial no metabolismo do molusco aquático Biomphalaria glabrata (Say, 1818), principal hospedeiro intermediário de Schistosoma mansoni Sambon, 1907 no Brasil e, tem sido descrito como um fator limitante na distribuição e adaptação desse molusco no ambiente. O presente trabalho avaliou o efeito de diferentes concentrações de carbonato de cálcio (CaCO3) exógeno ao metabolismo energético de B. glabrata, a fim de subsidiar uma melhor compreensão da interferência de elementos químicos dissolvidos no meio aquático na fisiologia destes moluscos. Foram utilizados moluscos com sessenta dias de vida, distribuídos em seis grupos, cinco expostos a diferentes concentrações de CaCO3 (20, 40, 60, 80 e 100mg/L) e um controle. A exposição ao CaCO3 foi avaliada em função do tempo, sendo retirados 15 moluscos de cada grupo nos seguintes intervalos: 1, 14, 21 ou 30 dias para extração da hemolinfa. As concentrações de cálcio e glicose na hemolinfa foram determinadas usando-se kits comercial e os ácidos orgânicos foram extraídos por meio da coluna de troca iônica e analisados através cromatografia líquida de alta eficiência. Os resultados demonstraram que a concentração de cálcio na hemolinfa não apresentou diferença significativa (p>0,05) em relação ao controle e nas concentrações testadas. A concentração de glicose diminuiu (p<0,05) nas exposições a 20 mg e 40 mg/L e aumentou nas exposições a 80 mg e 100 mg/L de CaCO3 em relação ao controle e demais concentrações testadas ao longo de 30 dias. Os ácidos orgânicos piruvato, oxaloaceato, citrato, succinato, fumarato, β-hidroxibutirato e lactato tiveram suas concentrações aumentadas, enqunato, propionato e acetoacetato tiveram suas concentrações diminuídas na exposição ao CaCO3 comparada ao controle. Quanto a influência dos diferentes períodos de exposição ao CaCO3, aos 14 dias, as alterações no metabolismo de B. glabrata foram mais expressivas. Conclui-se que as exposições ao CaCO3 influenciaram na redução de glicose, sendo esta metabolizada a piruvato, produto final da glicólise e alteraram as vias de metabolismo energético, indicando um funcionamento aeróbio ou parcialmente anaeróbio.

Association of exercise training and angiotensin-converting enzyme 2 activator improves baroreflex sensitivity of spontaneously hypertensive rats

The present study sought to determine cardiovascular effects of aerobic training associated with diminazene aceturate (DIZE), an activator of the angiotensin converting enzyme 2, in spontaneously hypertensive rats (SHRs). Male SHRs (280–350 g) were either subjected to exercise training or not (sedentary group). The trained group was subjected to 8 weeks of aerobic training on a treadmill (five times a week, lasting 60 min at an intensity of 50–60% of maximum aerobic speed). In the last 15 days of the experimental protocol, these groups were redistributed into four groups: i) sedentary SHRs with daily treatment of 1 mg/kg DIZE (S+D1); ii) trained SHRs with daily treatment of 1 mg/kg DIZE (T+D1); iii) sedentary SHRs with daily treatment of vehicle (S+V); and iv) trained SHRs with daily treatment of vehicle (T+V). After treatment, SHRs were anesthetized and subjected to artery and femoral vein cannulation prior to the implantation of ECG electrode. After 24 h, mean arterial pressure (MAP) and heart rate (HR) were recorded; the baroreflex sensitivity and the effect of double autonomic blockade (DAB) were evaluated in non-anesthetized SHRs. DIZE treatment improved baroreflex sensitivity in the T+D1 group as compared with the T+V and S+D1 groups. The intrinsic heart rate (IHR) and MAP were reduced in T+D1 group as compared with T+V and S+D1 groups. Hence, we conclude that the association of exercise training with DIZE treatment improved baroreflex function and cardiovascular regulation.

Effect of calcium addition and pH on yield and texture of Minas cured cheese / Efeito da adição de cálcio e do pH sobre o rendimento e a textura de queijo-de-Minas curado

Milk calcium concentration is a factor related to cheese texture, an important rheological property of cheese quality as perceived by consumers. This study aimed to evaluate the effect of different pH conditions (5.8 and 6.6) and calcium addition (0, 150, 300 ppm of CaCl2), on yield and nutrient retention of the clots obtained and on the texture of Minascured cheese. Clots were analyzed for wet and dry yield, percentage content and retention of protein, fat and calcium. The texture of the cheese was evaluated by instrumental and sensorial tests. No differences were observed on the wet and dry yields, or on the protein content, fat and calcium retention of clots produced in the different experimental conditions. The instrumental evaluation showed that calcium addition significantly influenced the texture of cheeses, regardless of the pH of milk clotting. The sensory panel did not find a difference in the hardness ofcheeses produced at the same pH of milk clotting in function of CaCl2 addition. There was no difference in the texture of Minascured cheese due to the calcium addition to milk for dairy product consumers, which brings a new perspective on manufacture for cheese markers.

Avaliou-se o efeito de diferentes condições de pH ­5,8 e 6,6 ­e da adição de cálcio ­0, 150, 300ppm de CaCl2 - sobre o rendimento, a retenção de nutrientes nos coágulos produzidos e a textura de queijo-de-Minas curado. Foram analisados nos coágulos os rendimentos úmido e seco, o percentual e a retenção de proteína, de gordura e de cálcio. A textura dos queijos foi avaliada por testes instrumental e sensorial. Não foram observadas diferenças nos rendimentos úmido e seco, no conteúdo percentual e na retenção de proteína, gordura e cálcio nos coágulos produzidos em diferentes condições experimentais. A análise instrumental mostrou que a adição ou não de cálcio influenciou a textura dos queijos, independentemente do pH de coagulação do leite. O painel sensorial não diferiu quanto à dureza dos queijos produzidos em um mesmo valor de pH de coagulação do leite mediante a adição de CaCl2. Para os consumidores, não houve diferença na textura do queijo Minas curado em razão da adição de cálcio ao leite. Isto traz nova perspectiva na fabricação para os produtores de queijo.

Differences in Evolution of Children with Non-severe Acute Lower Respiratory Tract Infection With and Without Radiographically Diagnosed Pneumonia.

Objective: To identify differences in the evolution of children with non-severe acute lower respiratory tract infection between those with and without radiographically diagnosed pneumonia. Design: Prospective cohort study. Setting: A public university pediatric hospital in Salvador, Northeast Brazil. Patients: Children aged 2-59 months. Methods: By active surveillance, the pneumonia cases were prospectively identified in a 2-year period. Each case was followed-up for changes in various clinical symptoms and signs. Demographic, clinical and radiographic data were recorded in standardized forms. Exclusion was due to antibiotic use in the previous 48 hours, signs of severe disease, refusal to give informed consent, underlying chronic illness, hospitalization in the previous 7 days or amoxicillin allergy. Chest X-ray (CXR) was later read by at least 2 independent pediatric radiologists. Main Outcome Measures: Radiographic diagnosed pneumonia based on agreed detection of pulmonary infiltrate or pleural effusion in 2 assessments. Results: A total of 382 patients receiving amoxicillin were studied, of whom, 372 (97.4%) had concordant radiographic diagnosis which was pneumonia (52%), normal CXR (41%) and others (7%). By multivariate analysis, age (OR=1.03; 95% CI: 1.02- 1.05), disease ≥ 5days (OR = 1.04; 95% CI: 1.001-1.08), reduced pulmonary expansion (OR = 3.3; 95% CI: 1.4-8.0), absence of wheezing (OR = 0.5; 95% CI: 0.3-0.9), crackles on admission (OR = 2.0; 95% CI: 1.2-3.5), inability to drink on day 1 (OR = 4.2; 95% CI: 1.05-17.3), consolidation percussion sign (OR = 7.0; 95% CI: 1.5-32.3), tachypnea (OR = 2.0; 95% CI: 1.09-3.6) and fever (OR = 3.6; 95% CI: 1.4-9.4) on day 2 were independently associated with pneumonia. The highest positive predictive value was at the 2nd day of evolution for tachypnea (71.0%) and fever (81.1%). Conclusion: Persistence of fever or tachypnea up to the second day of amoxicillin treatment is predictive of radiographically diagnosed pneumonia among children with non-severe lower respiratory tract diseases.

Levantamento etnobotânico de plantas medicinais em área de caatinga no município de São José de Espinharas, Paraíba, Brasil / Ethnobotanical survey of medicinal plants in a caatinga area in São José de Espinharas Municipality, Paraíba State, Brazil

Este trabalho teve como objetivo realizar levantamento das plantas medicinais utilizadas pela comunidade de São José de Espinharas, a fim de registrar e preservar o conhecimento popular. A metodologia usada foi de entrevistas semi-estruturadas, com observações participantes, coleta e identificação do material botânico e preparação de exsicatas, resultando em lista de 82 espécies de 38 famílias diferentes. São fornecidas as informações nomenclatura popular e botânica, uso terapêutico, parte utilizada, forma de uso, modo de preparo e doenças tratadas. As raízes (30 por cento) foram ás partes mais utilizadas e a forma de preparo foi lambedor (32 por cento). O conhecimento sobre os usos e modos de preparo provém em geral, dos familiares (85 por cento). Com esses resultados, verifica-se a interação da população local com a flora e utilização relacionada a aspectos sociais, econômicos, culturais e às mudanças ambientais.

The aim of this study was to perform a survey of medicinal plants used by the community of São José de Espinharas, Paraíba State, Brazil, in order to record and preserve the folk knowledge. The adopted methodology was based on semi-structured interviews, with participating observations, botanical material collection and identification, and voucher preparation, resulting in a list of 82 species of 38 different families. The following information was provided: folk and botanical nomenclature, therapeutic application, used plant part, forms of use, method of preparation and treated diseases. Roots (30 percent) constituted the part most frequently used and syrup (32 percent) was the predominant method of preparation. The knowledge of uses and methods of preparation are generally handed down in the family (85 percent). Based on those results, there is an interaction of the local population with the flora and its use is related to social, economical and cultural aspects and environmental changes.

Induction of cell migration and activation in mice by the freshwater sponge Drulia uruguayensis Bonetto & Ezcurra de Drago, 1968 (Porifera: Metaniidae)

Freshwater sponges are abundant in the Amazon region and they have been known to cause dermatitis (acute inflammation) since the beginning of the 20th century. To determine whether additional constituents, besides their body spicules, cause dermatological reactions in humans, an experimental study was developed and carried out using mice and Drulia uruguayensis prepared in three different forms: intact sponges (IS), macerated sponges (MS) or isolated spicules - megascleres (ISM). The cells most commonly involved in inflammatory reactions (mast cells, eosinophils and neutrophils), as well as intraepithelial lymphocytes and degranulated mast cells, were counted so that they could be used as parameters to determine which of the sponge preparations induced the greatest reaction. The effects of the sponge on the skin were then determined by histological analysis. The results obtained showed that IS caused the greatest inflammatory reaction (p = 0.000005), activating mainly mast cells (p = 0.0018). The histopathological analysis revealed a slight loss of continuity of the epidermis when ISM or IS were applied. These findings allow us to conclude that a structurally intact sponge can cause a greater inflammatory reaction in the first contact because of its ability to perforate the skin and allow inflammatory agents to enter. Other proteins present in dried sponge bodies could induce allergic but not toxic responses (in contact with the entire sponge, a large number of pharmacologically inert proteins may be introduced, with a potential allergen).

Detecção fo genoma de HPV em pacientes com carcinoma espino-celular da laringe: comparação entre os ensaios de PCR convencional e em tempo real / Detection of HPV genome in patients with squamous cell carcinoma of the larynx: comparison between conventional and real time PCR assays

A interação do genoma do HPV com o genoma da célula hospedeira ou de proteínas virais com proteínas celulares necessárias ao controle do ciclo celular desencadeia a morte celular ou age como fator de iniciação e progressão de processos malignos. Diversostrabalhos têm sido desenvolvidos no sentido de associar a infecção de HPV aos cânceres de cabeça e pescoço. O objetivo do presente estudo foi comparar a detecção do genoma de HPV em pacientes com CEC da laringe, considerando os ensaios de PCR convencional e em tempo real. O grupo amostral foi constituído de 15 pacientes do Registro de Câncer de Base Populacional da ACCG. Das peças cirúrgicas, foi obtido DNA total, usado na investigação molecular, para detecção do genoma de HPV, utilizando os primers genéricos GP05/06, que detectam todos os tipos de HPV. O genoma de HPV foi amplificado em 30 dos casos por PCR convencional e no ensaio de PCR em tempo real apresentaram amplificação em 46,7 das amostras, evidenciando uma diferença em 40 . Os estudosde associação entre o HPV e os cânceres da laringe são importantes no sentido de ampliar o conhecimento acerca dos mecanismos de infecção, iniciação e promoção tumoral potencializada por estes vírus.

Antibacterial activity of indole alkaloids from Aspidosperma ramiflorum

We evaluated the antibacterial activities of the crude methanol extract, fractions (I-V) obtained after acid-base extraction and pure compounds from the stem bark of Aspidosperma ramiflorum. The minimum inhibitory concentration (MIC) was determined by the microdilution technique in Mueller-Hinton broth. Inoculates were prepared in this medium from 24-h broth cultures of bacteria (10(7) CFU/mL). Microtiter plates were incubated at 37°C and the MICs were recorded after 24 h of incubation. Two susceptibility endpoints were recorded for each isolate. The crude methanol extract presented moderate activity against the Gram-positive bacteria B. subtilis (MIC = 250 µg/mL) and S. aureus (MIC = 500 µg/mL), and was inactive against the Gram-negative bacteria E. coli and P. aeruginosa (MIC > 1000 µg/mL). Fractions I and II were inactive against standard strains at concentrations of <=1000 µg/mL and fraction III displayed moderate antibacterial activity against B. subtilis (MIC = 500 µg/mL) and S. aureus (MIC = 250 µg/mL). Fraction IV showed high activity against B. subtilis and S. aureus (MIC = 15.6 µg/mL) and moderate activity against E. coli and P. aeruginosa (MIC = 250 µg/mL). Fraction V presented high activity against B. subtilis (MIC = 15.6 µg/mL) and S. aureus (MIC = 31.3 µg/mL) and was inactive against Gram-negative bacteria (MIC > 1000 µg/mL). Fractions III, IV and V were then submitted to bioassay-guided fractionation by silica gel column chromatography, yielding individual purified ramiflorines A and B. Both ramiflorines showed significant activity against S. aureus (MIC = 25 µg/mL) and E. faecalis (MIC = 50 µg/mL), with EC50 of 8 and 2.5 µg/mL for ramiflorines A and B, respectively, against S. aureus. These results are promising, showing that these compounds are biologically active against Gram-positive bacteria.

Sex in fungi: lessons of gene regulation

Fungi have been very useful for gene regulation studies. Mating implicates in a series of events influenced by many types of environmental input that are interpreted into regulatory pathways, including signal transduction. Although various aspects of mating and signal transduction in the yeast Saccharomyces cerevisiae have long been characterized, recent findings in filamentous fungi indicate that pheromones and pheromone receptors may be essential for mating partner recognition and also for nucleus recognition in sorting before meiosis. A brief overview on mating-type genes of ascomycete fungi and recent contributions to the understanding of their role in the regulation of multicellularity and sexual dimorphism is presented in this review

Origem anômala da artéria pulmonar direita em aorta ascendente (hemitruncus) / Anomalous origin of right pulmonary artery from ascending aorta (hemitruncus)

A origem da artéria pulmonar direita em aorta ascendente é uma anomalia congênita rara, com poucos casos descritos na literatura. Descrevemos dois casos desta malforamçäo, um associado à interrupçäo de arco aórtico tipo B. Em ambos, o diagnóstico foi realizado mediante ecocardigrafia bidimensional, com confirmaçäo angiográfica em um. A origem da artéria pulmonar direita era próxima à valva aórtica, anomalia patogeneticamente diferente da origem perto da artéria inominada.

Persistência do canal arterial e endarterite da artéria pulmonar / Patent ductus arteriosus and pulmonary artery endarteritis

Descrevemos uma paciente de sete anos com diagnóstico ecocardiográfico e confirmação cirúrgica de endarterite de artéria pulmonar, citada como complicação da persistência do canal arterial e ilustramos a necessidade de ressecção cirúrgica completa do processo infeccioso para evitar outras complicaçöes, como embolia pulmonar.

Defeito parcial do septo atrioventricular associado a doença de Ebstein da valva atrioventricular direita / Partial Atrioventricular Septal Defect Associated with Right Atrioventricular Valve Ebstein's Disease

A associaçäo de doença de Ebstein e defeito septal atrioventricular é extremamente rara, havendo 13 casos na literatura e nenhum em nosso meio. Descrevemos uma paciente de 15 anos com diagnóstico eco e angiográfico e confirmaçäo cirúrgica. A näo identificaçäo de uma das lesöes poderia comprometer o planejamento e resultado cirúrgico

Infarto do ventrículo direito. Apresentação de um caso / Infarction of the right ventricle. Report of a case

Os autores apresentam um caso de infarto do miocardio comprometendo o ventriculo direito e comentam os aspectos clinicos, o comportamento dos exames subsidiarios, assim como a terapeutica