Melanopsin and Clock Genes: Regulation by Light and Endothelin in the Zebrafish ZEM-2S Cell Line

It is well known that clocks are present in brain regions other than the suprachiasmatic nucleus and in many peripheral tissues. In the teleost, Danio rerio, peripheral oscillators can be directly synchronized by light. Danio rerio ZEM-2S embryonic cells respond to light with differential growth: cells kept in constant light exhibited a strong inhibition of proliferation, whereas in cells kept in light:dark (LD) cycles (14L:10D and 10L:14D) or in constant darkness (DD), the doubling times were not statistically different. We demonstrated by RT-PCR followed by PCR that ZEM-2S cells express two melanopsins, Opn4x and Opn4m, and the six Cry genes. The presence of the protein OPN4x was demonstrated by immunocytochemistry. The pattern of temporal expression of the genes Opn4x, Per1, Cry1b, and Clock was studied in ZEM-2S cells kept for five days in 12L:12D or DD. In 12L:12D, the clock genes Per 1 and Cry1b exhibited robust circadian expression, while Opn4x and Clock expression seemed to vary in an ultradian pattern. Both Per1 and Cry1b genes had higher expression during the L phase; Clock gene had an increase in expression coincident with the D phase, and during the subjective night. In DD, the temporal variation of Per1 and Cry1b genes was greatly attenuated but not extinguished, and the higher expressions were shifted to the transition times between subjective day and night, demonstrating that Per and Cry1b were synchronized by the LD cycle. Clock and Opn4x kept the ultradian oscillation, but the rhythm was not statistically significant. As endothelins (ET) have been reported to be a potent stimulator of Per genes in rodents, we investigated the effect of endothelin on ZEM-2S cells, which express ETA receptors. Cells were kept in 12D:12L for five days, and then treated with 10-11 to 10-8M ET-1 for 24h. ET-1 exhibited a biphasic effect on Opn4x expression. At 10-11M, the hormone exerted a highly significant stimulation of Opn4x expression during the L phase and introduced a circadian oscillatory pattern. At 10-10M, a significant increase was seen at ZT21 and ZT0 (i.e., at the end of the D phase and beginning of the L phase), whereas 10-9 and 10-8M ET-1 inhibited the expression of Opn4x at most ZTs. Clock expression was unaffected by 10-8M ET-1; however, in the presence of lower concentrations, the expression was enhanced at some ZTs, strengthening the ultradian oscillation. ET-1 at 10-11 and 10-10M had no effect on Per1 circadian expression; however, 10-9 and 10-8M ET-1 reduced the amplitude of Per1 expression in the beginning of the L phase. ET-1 effects were less evident on Cry 1b. For both genes, the reduction in expression was not sufficient to abolish the circadian oscillatory pattern. Based on these results and data in the literature, a link between ET-1 stimulation of ETA receptors may be established by E4BP4 binding to the promoters and consequent inhibition of gene expression.

The Immune-Pineal Axis Stress as a Modulator of Pineal Gland Function

The temporal organization of mammals presents a daily adjustment to the environmental light/dark cycle. The environmental light detected by the retina adjusts the central clock in the suprachiasmatic nuclei, which innervate the pineal gland through a polysynaptic pathway. During the night, this gland produces and releases the nocturnal hormone melatonin, which circulates throughout the whole body and adjusts several bodily functions according to the existence and duration of darkness. We have previously shown that during the time frame of an inflammatory response, pro-inflammatory cytokines, such as tumor necrosis factor-a, inhibit while anti-inflammatory mediators, such as glucocorticoids, enhance the synthesis of melatonin, interfering in the daily adjustment of the light/dark cycle. Therefore, injury disconnects the organism from environmental cycling, while recovery restores the light/dark information to the whole organism. Here, we extend these observations by evaluating the effect of a mild restraint stress, which did not induce macroscopic gastric lesions. After 2 h of restraint, there was an increase in circulating corticosterone, indicating activation of the hypothalamus-pituitary-adrenal (HPA) axis. In parallel, an increase in melatonin production was observed. Taking into account the data obtained with models of inflammation and stress, we reinforce the hypothesis that the activity of the pineal gland is modulated by the state of the immune system and the HPA axis, implicating the darkness hormone melatonin as a modulator of defense responses.

XERODERMA PIGMENTOSUM: LIVING IN THE DARK BUT WITH HOPE IN THERAPY

Xeroderma pigmentosum patients suffer from extreme photosensitivity caused by a genetic defect in DNA repair pathways. This condition obliges them to live in darkness and avoid sunshine. Although the molecular basis of the defect has been known for more than 40 years now, the treatment possibilities are very limited, and to date all have been focused on the skin. Herein, we summarize the effects of sunlight and the molecular mechanisms implicated in the defects that lead to this syndrome, as well as the strategies that have been tested to alleviate skin manifestations, including cancer. Preclinical attempts to correct genetic defects by means of different gene therapy approaches are also described. All these efforts are now bringing hope and some light into the life of patients and their families.

PANA V : Evaluation of a Literacy Project

PANA V Evaluation of a Literacy ProjectSUMMARY AND CONCLUSIONSThis evaluation set out to explore the impact of the literacy work carried out through PANA V. It focussed on clarifying effects such as empowerment and poverty reduction in relation to the civil society. Two specific objectives were to evaluate the methodological approach and the didactic materials and to evaluate the sustainability of the project.Although the focus of the evaluation has been PANA V, the project has been evaluated in its context, as one in a series of five projects located in Rwanda ten years after the war and genocide. The conclusion will consider future plans in this field.The evaluator has striven to create a holistic picture of the effects of the project, although the given time for the evaluation was short. Only three weeks were spent in the field study and only ten days in the actual field. Although there were some organisational and logistic problems, as is common when carrying out a study like this in a poor country, many literacy sites were visited and quite many participators were interviewed. The overall impression from the study is overwhelmingly positive. So many people commit themselves in this task of teaching Rwandans reading, writing and numeracy. Despite harsh conditions learners strive to learn and group leaders devote themselves to the task. Many leaders on different levels try their very best to manage their difficult and demanding task. The main objective was to explore the impact of the project on poverty reduction, particularly on empowerment and strategies for everyday life. Women were to be regarded particularly. From the results it is clear that the project has a strong, positive impact both on poverty reduction and empowerment of marginalised groups. Among those who have benefited from the alphabetisation are mainly women. Unfortunately, when it comes to leaders in PANA, who may also be said to have benefited from the project, only a small minority is women. This is something that is recommended that it be reconsidered inside the organisation. As a majority of the targeted learners are women, and as the economic and social situation of women in Rwanda is generally weak, this is a question that I recommend the Pentesostal church and ADEPR to look particularly into. With many women being single breadwinners of their households, it is important that also women get access to positions that may bring benefits of different kind.It is also clear that the project has positive effects for the civil society. In the present situation in Rwanda, during the process of reconciliation and rapid progress, basic education for the poor majority is a democratic issue. In a country with a plethora of internet-cafés in the capital and a small minority that use cars and mobile-telephones to communicate nation-wide, it is of outmost importance that the majority acquires basic education, of which literacy is a central part. To strengthen the civil society in Rwanda literacy is important. One central issue is then that Rwanda develops toward becoming a country where literacy is used for the benefit of the citizens and it is a democratic issue that all citizens get an opportunity to participate. Crucial for this is that strong efforts are put into primary schools nation-wide. Literacy projects for adults, like PANA, may only complement these efforts, but they constitute important and necessary complements. Other relevant ways to promote literacy are campaigns in Radio and TV and through cultural events such as festivals, music and theatre. News papers, magazines and books are natural parts of such campaigns as well as adult education. As stated under the results not much can be said about the didactics in this evaluation. On the whole the methodology and the materials fill their function well and receive a high reputation. As people learn to read and write under very simple conditions, obviously the approach is appropriate. A few suggestions may be given from the study:•Focus groups leaders’ attention on clearness, that they show very clearly what is to be read. Good structuring is probably of great importance for many learners.•Make clear what is tested in the tests and consider the possibility to use a holistic test that would be more congruent with the methodology. The possibility to use only one grade, pass, would enable a more practical test, such as reading a short, relevant text, writing something relevant and solving practical mathematic problems. Avoid tests that demand school knowledge.•Avoid using methaphors such as “fight against illiteracy” and connections between illiteracy/literacy and darkness/light. It is not true that illiteracy causes bad things and that literacy only brings good. •Be prepared that it may be more difficult in the future to achieve the goals as it may be the case that the early learners where the ones who achieved easily. The goal of “literacy in six month” in PANA will probably hold only for some learners but also those who do not manage in six months need literacy skills.A third objective was to secure sustainability. As for sustainability of the project in itself, and of the literacy process, the main conclusion is that there is a good potential. The commitment and devotedness among many involved in PANA proves good. One weakness is individual leaders in ADEPR who do not see this as an important task for the Pentecostal church in Rwanda. Other weaknesses are the unwillingness to mention explicitly the wish, for example among group leaders, to get some kind of incentive and the fear of loosing believers by cooperation with other organisations. A higher degree of transparency in this issue would probably solve some irritations and tensions.As for the sustainability of the literacy skills much may be done to improve. The acquired skills seem to be comparably relevant. The level achieved, and the level tested, may be defined as basic literacy skills, consisting of basic reading, writing and numeracy skills. However, these skills are very restricted and there is a high risk that the skills will decline, which means that there is a high risk that people will forget how to read and write because of lack of exercising. From these conclusions a few suggestions for future development will be given.

Fatores da administração do sistema penitenciário do Rio de Janeiro que os impedem de cumprirem com sua missão de recuperação do detento?

Since some years ago, the penitentiany systen of Rio de Janeiro is going through the nuest sendus administrative crisis,leading more and more to chaotic situations, over imagined before by homan being. Nowadays all the factors and components of the existing models are still medievaIs, and even distant fron the human needs. The daily life ofthe condemned is a sway betuen lack of hygiene, disrespect to homan rigts, lack of modem corrective practices and also, lack of psychological support. How, then can me demand from the state ( Govemment) the rehabilitation of the imprisoned and their preparation to face society if they are treated manny times as victins of this same society? This society impose a life style enjoyed only by a privileged social class which forget about then when sent to the darkness. Many of then, join the penitentary systen because of minor crimes, and when they serve their tem, um for funately they go back to prison accused for move violent crimes. The penitentian models haven't developed the same way the society where they worked at has. There, the present brazilian penitentiary model has showed obsolete and inefficient alone its principal mission ofrehabilitation and re - education ofthe imprisoned. Our main objetive will be the construction and analysis of the penitential administrative model, as being able to fullfill the necesity of the penitentiary administrator (Manager). Analy zing on a specific reasoning which focus showld not be that of the theories and isolated projects of the topic, we will develop a progran far form religions, judiciary on political technics by building na administrtive penitentiary model strictly professional where we coul a have a deep analysis of the topic. We will try to approach the aspects of organization existing to day inorder to understand them and criate, a model wich will adjust betten to the necesitives of this thesis.

Apocalypse now e o coração das trevas: o embate entre indivíduo e sociedade

O filme Apocalypse Now (1979), de Francis Ford Coppola, inspirado no romance O Coração das Trevas (1902), de Joseph Conrad, discute o conflito entre indivíduo e sociedade no contexto da Guerra do Vietnã, nos anos 60 e 70 do século XX. Este trabalho pretende mostrar, embasado pela Semiótica francesa, as estratégias utilizadas pelo diretor para explicitar essa oposição na linguagem cinematográfica e preservar a fidelidade temática em relação às questões abordadas pelo livro, passado durante o Imperialismo britânico na África, no século XIX.

Effects of photoperiod on reproduction of Siamese fighting fish Betta splendens

This study investigated the effect of different photoperiods (24 h of light (L):0 h of darkness (D); 20L:4D; 16L:8D; 12L:12D; and 8L:16D) on the reproduction and growth of Betta splendens. The results showed that spawning frequency was significantly higher in couples reared under 16L:8D and 12L:12D, in comparison with other treatments. The highest number of eggs per spawn was obtained under 16L:8D (544.76±375.23) and 12L:12D (471.13±261.52), and the lowest values were detected for 24L:0D (128.55±58.14) and 20L:4D (187.87±103.84). Fertility and fecundity also showed significantly higher average values in 16L:8D and 12L:12D when compared with 24L:0D and 20L:4D treatments. Egg volume and perivitelline space were significantly higher in 24L:0D treatments that showed the lowest numbers of eggs per spawn, while the vitelline volume did not show significant differences. Other variables such as breeders weight gain and condition factor (K) were not statistically different. Moreover, the final length varies according to photoperiod and gender. These results demonstrated a key role for the photoperiod upon B. splendens reproduction. The best reproductive performance is achieved under the photoperiods that best approached those that occur in spring and summer (16L:8D and 12L:12D), coinciding with their best seasons for reproduction.

Calli induction in leaf explants of coffee elite genotypes

Visando a alcançar alta eficiência na indução de calos a partir de explantes foliares de plantas matrizes de C. arabica com alta heterozigose, por meio da embriogênese somática indireta, foram instalados três experimentos. O primeiro experimento foi conduzido em esquema fatorial 2x5, constituído de dois meios de cultura (BOXTEL & BERTHOULY, 1996 e TEIXEIRA et al., 2004) e cinco genótipos de C. arabica. No segundo experimento, foi avaliado o potencial de produção de calos embriogênicos em 10 genótipos, sendo cada genótipo considerado como um tratamento e, no terceiro experimento, foram avaliadas as variações nas concentrações de 2.4-D (2,5 e 20µM) e 2-iP (2,5 e 20µM) nos meios primário e secundário de TEIXEIRA et al. (2004). As culturas foram mantidas a 25oC, sob obscuridade. Para os genótipos 2.2 e 7.2, verificou-se a superioridade do meio de cultura Teixeira et al. (2004) em relação ao meio BOXTEL & BERTHOULY (1996). No genótipo 4.2, observou-se o comportamento inverso, ou seja, a superioridade do meio BOXTEL & BERTHOULY (1996). Os genótipos 3.0 e 5.0 apresentaram o mesmo comportamento em ambos os meios de cultura estudados, evidenciando que a produção de embriões somáticos é fortemente dependente do genótipo. A indução de calos depende da relação de 2-iP e 2.4-D. A combinação de 20.0µM of 2.4-D e 20.0µM of 2-iP promoveu a maior porcentagem de indução de calos embriogênicos.

Fatores que afetam a germinação e a emergência de Xanthium strumarim L.: dormência, qualidade da luz e profundidade de semeadura

Foram conduzidos três ensaios com o objetivo de estudar os efeitos de diferentes métodos de quebra de dormência das sementes, da qualidade da luz e da profundidade de semeadura na germinação e emergência do Xanthium strumarim L. (carrapichão). No primeiro ensaio, sob condições de câmara de crescimento, os tratamentos constaram de 5 métodos de quebra de dormência: mecânico (lixa), químico (H2SO4 conc. por 10 min.), pré-tratamento em água (imersão por 2 horas), físico (choque térmico: água à 80oC por 2 min.) e testemunha, que proporcionaram 100%, 93%, 93%, 87% e 80% de germinação, respectivamente, sem haver diferença estatística entre os mesmos. O pré-tratamento em água apresentou menor índice de velocidade de germinação (IVG) e os demais não diferiram significativamente entre si. No segundo ensaio, também sob condições de câmara de crescimento, os tratamentos de qualidade da luz constaram de 6 filtros de luz, que resultaram nas seguintes porcentagens de germinação: azul - com refletividade máxima a 450 nm (13%), verde-500 nm (33%), vermelho-700 nm (13%), vermelho/distante-760 nm (7%), ausência de luz (26%) e transparente - sem absortividade na região de 380 a 760 nm (testemunha, 73%), indicando sensibilidade das sementes à qualidade de luz. No terceiro ensaio, sob condições de casa-de-vegetação, os tratamentos foram diferentes profundidades de semeadura (0 a 20 cm, em intervalos de 2 cm), sendo que o máximo de emergência ocorreu no intervalo de 0 a 8 cm, no qual foi observado 67% de emergência do carrapichão, com maior índice de velocidade emergência (IVE), sendo que após os 16 cm, praticamente não houve emergência.

Efeitos de substratos e temperaturas na germinação de sementes de tamareira-anã (Phoenix roebelenii O'Brien)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influência da temperatura e da luminosidade no desenvolvimento de Guignardia citricarpa, agente causal da mancha preta dos frutos cítricos

O presente trabalho teve como objetivo avaliar sob condições de laboratório, o efeito de temperatura (15, 18, 20, 21, 24, 25, 27, 30, 33 e 35 ± 1 ºC) nas condições escuro, luz contínua e fotoperíodo 12/12 h, na produção de pseudotécios de isolados de Guignardia citricarpa, provenientes de regiões cítricolas dos Estados de São Paulo e Rio de Janeiro. Discos de folhas de limoeiro 'Cravo' (Citrus limonia), de 12 mm de diâmetro, foram autoclavados e depositadas (parte abaxial) na superfície do meio de cultura constituído por ágar-água 2%. Foram colocados quatro discos de folhas por placa onde, de forma conjunta e intercalar aos mesmos, depositaram-se dois discos obtidos de colônias de Phyllosticta citricarpa, com 21 dias de incubação. Foi, também, estudado o efeito da temperatura e do tempo de incubação (2, 8 e 16 h) na germinação dos ascósporos. Após 21 dias de incubação, a ótima temperatura ajustada pela função beta generalizada, para produção de pseudotécios deu-se a 26 e 22,5 a 27,5 °C, sob condição de escuro e de luz, respectivamente. Observou-se também produção de pseudotécios a 27 ºC em fotoperíodo 12/12 h. em estudo complementar foi verificado que, aos 19 dias, a 27 ºC, cerca de 90% dos pseudotécios haviam alcançado a maturidade, com abundante produção de ascósporos. A maior porcentagem de ascósporos germinados foi constatada na temperatura de 24 ºC, após 16 h de incubação. Dentre as vantagens alcançadas, incluem-se a possibilidade (i) da produção massal de ascósporos em curto período de tempo, e (ii) da padronização do inóculo, tanto qualitativa, quanto quantitativa.

Efeito da temperatura e da luz na germinação de sementes de Acacia polyphylla DC.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Armazenamento e requerimento fotoblástico de sementes de Acacia polyphylla DC

Acacia polyphyla (Mimosaceae) é uma espécie arbórea nativa do Brasil, importante para a recuperação de áreas degradadas. Sementes dessa espécie foram armazenadas em condições naturais e artificiais, com os objetivos de avaliar a longevidade das sementes no solo, conservar a sua qualidade fisiológica pelo período correspondente a duas colheitas e verificar o requerimento fotoblástico das sementes armazenadas. em condições naturais, as sementes foram enterradas em clareira, sob dossel ralo e sob dossel denso. As sementes deterioraram rapidamente, revelando-se incapazes de compor o banco de sementes do solo. em condições artificiais, as sementes foram acondicionadas em diferentes embalagens e armazenadas por dois anos em ambiente não controlado e em câmara fria. Periodicamente, as sementes foram colocadas para germinar a 25ºC, na ausência de luz e sob fotoperíodo de oito horas sob luzes branca e de sombreamento. Durante todo o período de armazenamento, a germinação no escuro foi inferior à constatada sob luzes branca e de sombreamento. A qualidade fisiológica foi conservada por dois anos, quando as sementes foram acondicionadas em embalagem impermeável e armazenadas em câmara fria. O comportamento germinativo das sementes armazenadas por dois anos foi comparado com o de sementes recém-colhidas, em temperaturas constante e alternada, não sendo constatado efeito da idade e do regime de temperatura no requerimento fotoblástico das sementes.

Germinação de sementes de pau-ferro submetidas a diferentes condições de armazenamento, escarificação química, temperatura e luz

Caesalpinia leiostachya (Benth.) Ducke (pau-ferro) é uma planta arbórea nativa do Brasil, cujas sementes possuem dormência causada pela impermeabilidade do tegumento à água. Neste trabalho foram conduzidos dois experimentos, nos quais foram utilizados diferentes períodos de escarificação em ácido sulfúrico concentrado para superar a dormência das sementes. No primeiro experimento, sementes coletadas em agosto de 1997 foram armazenadas por oito meses em ambiente não controlado no interior do próprio fruto, e em câmara seca após serem extraídas dos frutos; a seguir, elas foram imersas em ácido sulfúrico por 0, 10, 20, 40, 60 e 80min e colocadas para germinar nas temperaturas constante de 25ºC e alternada de 20-30°C, sob fotoperíodo de 8h. No segundo experimento, sementes extraídas de frutos recém-coletados em agosto de 1998 foram imersas em ácido sulfúrico por 0, 10, 20, 30, 40 e 60min, seguido do teste de germinação conduzido nas mesmas temperaturas do experimento anterior, na ausência e presença de luz. Foram avaliados a porcentagem final e o índice de velocidade de germinação das sementes. Os resultados mostraram que (a) a manutenção das sementes no interior dos frutos é uma alternativa viável para o armazenamento durante o período adotado; (b) as sementes recém-coletadas são indiferentes à luz, nas duas temperaturas testadas; (c) as sementes recém-coletadas e as armazenadas germinam em maior velocidade a 25ºC; (d) em sementes armazenadas, a imersão em ácido sulfúrico por 10min é suficiente para superar a dormência; (e) em sementes recém-coletadas, a imersão em ácido sulfúrico por 20 a 30min favorece a porcentagem e a velocidade de germinação.

Comparação entre cultivares de carambola para produção de produtos minimamente processados

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)