Effects of chronic restraint stress on the global DNA methylation profile of rat lung cells: Modulation by physical exercise.

The potential of behavioral stress to affect epigenetic mechanisms of non-encephalic tissues is still underestimated. In the present study we evaluated the effects of chronic behavioral stress on the DNA methylation profile of rat lung cells. Furthermore, we evaluated the potential of physical exercise to modulate the changes evoked by behavioral stress in lung cells. Male Wistar rats were divided into four experimental groups: (1) animals submitted to chronic restraint stress (CRS) (ST group) during the period of the 67th-80th postnatal day (PND); (2) animals submitted to physical exercise (EX group) during the 53rd-79th PND; (3) animals submitted to swimming during the 53rd-79th PND and to CRS during the 67th-80th PND (EX-ST group); and (4) animals not submitted to stress or swimming protocols (CTL). Global DNA methylation was quantified using an ELISA-based approach and gene expression was evaluated by real time PCR. A decreased global DNA methylation profile was observed in the ST group, however physical exercise demonstrated protection of lung cells from this stress-related hypomethylation. Increased expression of the Dnmt1 gene was evidenced in the ST group, whereas physical exercise was shown to protect lung cells from this stress-related effect in the EX-ST group. Comparative analysis of the ST and EX groups revealed opposite effects on the expression of Dnmt3a and Dnmt3b; however, a stress-related increase in expression of Dnmt3a and Dnmt3b was not seen in the EX-ST group. Our data showed that behavioral stress induced significant changes in the DNA methylation profile of rat lung cells and that this could be modulated by physical exercise.

Application of a central composite design to evaluate the influence of colouring agents in lipstick formulation.

OBJECTIVES: The aim of this study was to evaluate by central composite design the influence of colouring agents in lipstick colour, expressed by L*, a*, b* parameters (CIELab system) where L* indicates lightness, and a* and b* are the chromaticity coordinates. The a* indicates colour direction from red to green and b* from yellow to blue. METHODS: Lipsticks were formulated as described by (Recent Adv. Prosp. Potent Med. Plants, 2009 and 39). The combined effect of three variables (dye, pigment and opacifier) was evaluated by different formulations in a central composite design. Colour parameters (L*, a*, b*) were analysed by reflectance spectrophotometry. Lipsticks were characterized by visual analyses and melting point. RESULTS: All formulations were integrate and homogeneous. The pigments and dye do not influence in colour transfer neither in melting point of lipsticks. On the other hand, results indicated that variables studied show influence only in parameter b*, whereas for L* and a* values there was no significant difference (P < 0.05). CONCLUSION: It was possible to verify that only the colour parameter b* was influenced by the variation in colouring agent's concentrations in lipstick formulation, leading to the production of the colour ranging between violet and light red. Such results are useful for developing new lipstick formulations to obtain the desired colour in the final product.

Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter.

The human papillomavirus (HPV) L1 major capsid protein, which forms the basis of the currently available vaccines against cervical cancer, self-assembles into virus-like particles (VLPs) when expressed heterologously. We report the development of a biotechnology platform for HPV16 L1 protein expression based on the constitutive PGK1 promoter (PPGK1) from the methylotrophic yeast Pichia pastoris. The L1 gene was cloned under regulation of PPGK1 into pPGKΔ3 expression vector to achieve intracellular expression. In parallel, secretion of the L1 protein was obtained through the use of an alternative vector called pPGKΔ3α, in which a codon optimized α-factor signal sequence was inserted. We devised a work-flow based on the detection of the L1 protein by dot blot, colony blot, and western blot to classify the positive clones. Finally, intracellular HPV VLPs assembly was demonstrated for the first time in yeast cells. This study opens up perspectives for the establishment of an innovative platform for the production of HPV VLPs or other viral antigens for vaccination purposes, based on constitutive expression in P. pastoris.

Control of fungal diseases and chilling injury in post-harvest tahiti lime / Controle de doenças fúngicas e de danos por frio em pós-colheita de lima ácida Tahiti

The Tahiti lime appears very susceptible to attack by post-harvest diseases, primarily by the fungi Penicillium and Phomopsis, and also because of its high sensitivity to storage at low temperatures. In order to reduce such damage, the present study aimed to verify the efficiency of heat treatment and disinfection of pathogens in the prevention of post-harvest chilling injury of this cultivar and to compare this treatment with other products using the conventional fungicides. The heat treatments were studied with hot-water temperatures ranging between 48 and 56° C. Water at room temperature was used as a control treatment. After treatment, the fruits were kept under cold temperature at 10° C and RH 90% for about 45 days. For comparison, three other treatments were carried out simultaneously, one using imazalil, one with baking soda, and a third with sodium carbonate, these three products being applied by baths in cold water. Two groups of fruit were evaluated, one treated by immersion considering pathogens coming from the field and another by inoculation with spores of the previously isolated pathogens. For the evaluation of physical and chemical parameters of fruits, determinations were made of the skin color, texture, weight loss, size, juice yield, soluble solids, total acidity and vitamin C content. The determination of the sensitivity of the fruit to cold was made by their exposure at temperatures inducing cold damage. The design was a randomized block design with nine treatments, analyzed by the Statgraphics statistical package. Heat treatments, especially at 52° C, were shown to be more promising in the control of pathogenic fungi and cold damage, surpassing the conventional fungicides. No changes were found in the intrinsic and extrinsic parameters in relation to the application of the different treatments.

A lima ácida Tahiti apresenta-se muito susceptível ao ataque de doenças pós-colheita e, também, por sua elevada sensibilidade, ao armazenamento em baixas temperaturas. Com a finalidade de reduzir estes danos, este trabalho objetivou verificar a eficiência do tratamento térmico na desinfestação de patógenos e na prevenção de danos por frio na pós-colheita deste cultivar e comparar este tratamento com outros utilizando os fungicidas convencionais. Os tratamentos térmicos estudados foram com água quente variando as temperaturas entre 48 a 56° C. A testemunha consistiu de tratamento com água em temperatura ambiente. Depois de tratados, os frutos foram divididos em dois lotes e mantidos em câmara fria com temperatura de 10° C e UR de 90%, por aproximadamente 45 dias. Para comparação foram feitos três outros tratamentos simultaneamente: um utilizando imazalil, outro com bicarbonato de sódio e o terceiro com carbonato de sódio, sendo as aplicações destes três produtos feitas em banhos com água em temperatura ambiente. Foram avaliados 2 grupos de frutos, um tratado por imersão considerando os patógenos oriundos do campo, e outro por inoculação com esporos dos fungos previamente isolados. Os parâmetros físico-químicos dos frutos avaliados foram a coloração da casca, a textura, a perda de massa, o tamanho, o rendimento de suco, de sólidos solúveis, o °brix, o ratio, a acidez total e a vitamina C. A determinação da sensibilidade dos frutos ao frio foi feita pela exposição deles a temperaturas indutoras de dano por frio. O delineamento experimental utilizado foi blocos ao acaso, com nove tratamentos, analisado pelo pacote estatístico Statgraphics. Verificram-se que os tratamentos térmicos, principalmente a 52° C, (...)

Expression of the bovine papillomavirus type 1, 2 and 4 L1 genes in the yeast Pichia pastoris.

Papillomaviruses are known to cause benign or malignant lesions in various animals. In cattle, bovine papillomavirus (BPV) is the etiologic agent of papillomatosis and neoplasia of the upper gastrointestinal tract and urinary bladder. Currently, there are no standard diagnostic tests or prophylactic vaccines. Protection against papillomavirus infection is conferred by neutralizing antibodies directed towards the major structural protein L1. These antibodies can be efficiently induced by immunization with virus-like particles that are formed spontaneously after L1 gene expression in recombinant systems. The yeast Pichia pastoris is known to provide an efficient system for expression of proteins due to reduced cost and high levels of protein production. We evaluated P. pastoris for expression of the L1 gene from BPV1, BPV2 and BPV4. After methanol induction, the recombinants were able to produce L1 proteins of the three different BPV types. To increase heterologous L1 protein levels, a codon optimization strategy was used for production under bioreactor conditions. The BPV1 L1 protein was identified by monoclonal antibody anti-6xHis. This is the first report of BPV L1 expression in yeast.

The arboreal component of a dry forest in Northeastern Brazil.

The dry forests of northeastern Brazil are found near the coastal zone and on low, isolated mountains inland amid semi-arid vegetation. The floristic composition of these dry montane forests, as well as their relationship to humid forests (Atlantic forest sensu stricto) and to the deciduous thorn woodlands (Caatinga sensu stricto) of the Brazilian northeast are not yet well known. This paper sought to determine if the arboreal plants in a dry forest growing on a low mountain in the semi-arid inland region (Serra Negra, 8 degrees 35 - 8 degrees 38 S and 38 degrees 02 - 38 degrees 04 W) between the municipalities of Floresta and Inajá, state of Pernambuco have the same floristic composition and structure as that seen in other regional forests. In fifty 10 x 20 m plots all live and standing dead trees with trunk measuring > 5 cm diameter at breast height were measured. Floristic similarities between the forest studied and other regional forests were assessed using multivariate analysis. The results demonstrate that the dry forest studied can be classified into two groups that represent two major vegetational transitions: (1) a humid forest/dry forest transition; and (2) a deciduous thorn-woodland/ dry forest transition.

The arboreal component of a dry forest in Northeastern Brazil

As florestas secas do Nordeste do Brasil são encontradas próximo à zona costeira, nos planaltos e em montanhas baixas e isoladas no interior do semi-árido nordestino. A composição dessas florestas secas e sua relação com as florestas úmidas (Floresta Atlântica sensu stricto) e a vegetação arbustivo-arbórea caducifólia espinhosa (Caatinga sensu stricto) do Nordeste do Brasil ainda não estão resolvidas. Este trabalho procura determinar se a flora arbórea da floresta seca que cresce em uma montanha baixa da região semi-árida localizada entre os municípios de Floresta e Inajá, Pernambuco (Serra Negra, 8° 35’ - 8° 38’ S e 38° 02’ - 38° 04’ W) apresenta a mesma composição florística e estrutura observada em outras florestas da região. Foram amostrados todos os indivíduos vivos ou mortos, ainda de pé, com diâmetro do tronco à altura do peito > 5 cm, presentes em 50 parcelas (10 x 20 m). A similaridade florística entre Serra Negra e outras florestas da região foram avaliadas usando análises multivariadas. Os resultados demonstram que as florestas secas englobam dois conjuntos florísticos que representam duas transições: (1) transição florestas úmidas/ florestas secas; e (2) transição vegetação arbustivo-arbórea caducifólia espinhosa/floresta seca, onde está localizada Serra Negra.

Anogenital warts in children: sexual abuse or unintentional contamination?

Anogenital warts (AGW) were recently recognized in children, and their significance as an index of childhood sexual abuse is controversial. We report our transdisciplinary approach (including a pediatric surgeon, psychologist, social worker, ethics expert, and occasionally law enforcement agents) and its results in a group of 17 children with AGW treated at the public pediatric referral hospital in Rio de Janeiro, Brazil, during a 3-year period (1996-1999). All children were treated by electrocauterization of the warts, tested for other STDs, and submitted to perineal examination under anesthesia. Families received psycho-social counseling as necessary and cases were referred to child protection and law enforcement agents when indicated according to Brazilian legislation. We identified a high incidence of sexual abuse (8 children, 5/7 > 5 years old), with 3 patients inconclusive as to sexual abuse and 7 cases of perinatal transmission (5/8 < 4 years old). We conclude that AGW are indeed a strong sign of suspicion for sexual abuse in children, especially but not exclusively > 5 years of age. However, strong support and a transdisciplinary approach to the children and their families is necessary to identify it.

Avaliação cardiorrespiratória em indivíduos sedentários durante programa de condicionamento físico / Cardiorespiratory evaluation in sedentary subjects during a physical conditioning program

Foram estudados 20 pacientes assintomaticos, sedentarios, do sexo masculino, submetidos a um programa de condicionamento fisico por 4 meses. A prescricao de exercicios foi feita segundo o proposto por Balke em 1974. Foram analisados: frequencia cardiaca de repouso, frequencia cardiaca maxima, pressao arterial em repouso, consumo de oxigenio, tempo de esforco e carga maxima atingida. Ficaram evidenciadas as adaptacoes organicas consequentes ao condicionamento fisico, caracterizadas por reducao significativa de frequencia cardiaca de repouso e de esforco para uma determinada carga, queda de valores tensionais, elevacao do consumo maximo de oxigenio e aumento da capacidade de desenvolver trabalho, determinado pela aptidao do individuo em alcancar maiores valores de carga (watts), antes de atingir a fadiga muscular