Competition between SOCS36E and Drk modulates Sevenless receptor tyrosine kinase activity

Modulation of signalling pathways can trigger different cellular responses, including differences in cell fate. This modulation can be achieved by controlling the pathway activity with great precision to ensure robustness and reproducibility of the specification of cell fate. The development of the photoreceptor R7 in the Drosophila melanogasterretina has become a model in which to investigate the control of cell signalling. During R7 specification, a burst of Ras small GTPase (Ras) and mitogen-activated protein kinase (MAPK) controlled by Sevenless receptor tyrosine kinase (Sev) is required. Several cells in each ommatidium express sev. However, the spatiotemporal expression of the boss ligand and the action of negative regulators of the Sev pathway will restrict the R7 fate to a single cell. The Drosophila suppressor of cytokine signalling 36E (SOCS36E) protein contains an SH2 domain and acts as a Sev signalling attenuator. By contrast, downstream of receptor kinase (Drk), the fly homolog of the mammalian Grb2 adaptor protein, which also contains an SH2 domain, acts as a positive activator of the pathway. Here, we apply the Förster resonance energy transfer (FRET) assay to transfected Drosophila S2 cells and demonstrate that Sev binds directly to either the suppressor protein SOCS36E or the adaptor protein Drk. We propose a mechanistic model in which the competition between these two proteins for binding to the same docking site results in either attenuation of the Sev transduction in cells that should not develop R7 photoreceptors or amplification of the Ras-MAPK signal only in the R7 precursor.

Avaliação de compostos funcionais e atividade antioxidante em farinhas de polpa de goiabas

A goiabeira (Psidium guajava) é uma planta nativa brasileira, e seus frutos são ricos em compostos antioxidantes os quais podem causar a oxidação das lipoproteínas, reduzindo o estresse oxidativo. Objetivou-se com este trabalho determinar essas substâncias e a atividade antioxidante das farinhas dos frutos das cultivares Pedro Sato, Paluma e Século XXI. Foram selecionados 35 frutos de cada cultivar, levando-se em consideração o grau de maturação. Os frutos foram lavados, picados, congelados em nitrogênio líquido e liofilizados. Posteriormente, foram trituradas para a obtenção das farinhas. Os parâmetros analisados foram o teor de compostos fenólicos, vitamina C, betacaroteno, fibras alimentares, minerais (ferro, potássio, cobre, magnésio, manganês, zinco e cálcio) e atividade antioxidante, pelos métodos DPPH e betacaroteno/ácido linoleico. Os teores de compostos fenólicos foram maiores para a cultivar Século XXI. O teor de betacaroteno e fibra alimentar não variaram entre as cultivares analisadas. As cultivares Século XXI e Paluma destacaram- se na atividade antioxidante pelos dois métodos. Em relação aos minerais, não houve diferença significativa para o potássio, cobre e zinco. O mineral cálcio não foi detectado pelo método empregado. Os teores de ferro, magnésio e manganês foram maiores para as cultivares Paluma, Pedro Sato e Século XXI, respectivamente.

Connexin43 Inhibition Prevents Human Vein Grafts Intimal Hyperplasia.

Venous bypass grafts often fail following arterial implantation due to excessive smooth muscle cells (VSMC) proliferation and consequent intimal hyperplasia (IH). Intercellular communication mediated by Connexins (Cx) regulates differentiation, growth and proliferation in various cell types. Microarray analysis of vein grafts in a model of bilateral rabbit jugular vein graft revealed Cx43 as an early upregulated gene. Additional experiments conducted using an ex-vivo human saphenous veins perfusion system (EVPS) confirmed that Cx43 was rapidly increased in human veins subjected ex-vivo to arterial hemodynamics. Cx43 knock-down by RNA interference, or adenoviral-mediated overexpression, respectively inhibited or stimulated the proliferation of primary human VSMC in vitro. Furthermore, Cx blockade with carbenoxolone or the specific Cx43 inhibitory peptide 43gap26 prevented the burst in myointimal proliferation and IH formation in human saphenous veins. Our data demonstrated that Cx43 controls proliferation and the formation of IH after arterial engraftment.

The central histaminergic neurons in vitro, and their neuroprotective role in excitotoxic cell death in the postnatal rat hippocampus.

Histamine acts as a neurotransmitter in the central nervous system. Brain histamine in synthesized in neurons located to the posterior hypothalamus, from where these neurons send their projections to different parts of the brain. Released histamine participates in the regulation of several physiological functions such as arousal, attention and body homeostasis. Disturbances in the histaminergic system have been detected in diseases such as epilepsy, sleep disorders, anxiety, depression, Alzheimer’s disease, and schizophrenia. The purpose of this thesis was to develop optimal culture conditions for the histaminergic neurons, to study their detailed morphology, and to find out their significance in the kainic acid (KA)-induced neuronal death in the immature rat hippocampus. The morphology of the histaminergic neurons in vitro was comparable with the earlier findings. Histamine-containing vesicles were found in the axon but also in the cell body and dendrites suggesting a possibility for the somatodendritic release. Moreover, histamine was shown to be colocalized with the vesicular monoamine transporter 2 (VMAT2) suggesting that VMAT2 transports histamine to the subcellular storage vesicles. Furthermore, histamine was localized with γ-aminobutyric acid (GABA) in distinct storage vesicles and with neuropeptide galanin partly in the same storage vesicles suggesting different corelease mechanisms for GABA and galanin with histamine. In the organotypic hippocampal slice cultures, KA-induced neuronal death was first detected 12 h after the treatment being restricted mainly to the CA3 subregion. Moreover, cell death was irreversible, since the 48 h recovery period did not save the cells, but instead increased the damage. Finally, neuronal death was suggested to be necrotic, since intracellular apoptotic pathways were not activated, and the morphological changes detected with the electron microscopy were characteristic for necrosis. In the coculture system of the hippocampal and posterior hypothalamic slices, histaminergic neurons significantly decreased epileptiform burst activity and neuronal death in the hippocampal slices, this effect being mediated by histamine 1 (H1) and 3 (H3) receptors. In conclusion, the histaminergic neurons were maintained succesfully in the in vitro conditions exhibiting comparable morphological characteristics as detected earlier in vivo. Moreover, they developed functional innervations within the hippocampal slices in the coculture system. Finally, the KA-induced regionspecific, irreversible and necrotic hippocampal pyramidal cell damage was significantly decreased by the histaminergic neurons through H1 and H3 receptors.

Abnormal Returns of Dividend Announcements during a Boom and a Recession. Empirical Evidence from U.S. from the years of 2000 - 2002 and 2005 - 2007, including Finnish Extra Dividends

This study examines the short time price effect of dividend announcements during a boom and a recession. The data being used here is gathered from the years of 2000 - 2002 when it was a recession after the techno bubble burst and from the years 2005 - 2007 when investors experienced large capital gains all around the world. The data consists of dividend increases and intact observations. The aim is to find out differences in abnormal returns between a boom and a recession. Second, the study examines differences between different dividend yield brackets. Third, Finnish extra dividends, mainly being delivered to shareholders in 2004 are included to the empirical test. Generally stated, the aim is to find out do investors respect dividends more during a recession than a boom and can this be proved by using dividend yield brackets. The empirical results from U.S shows that the abnormal returns of dividend increase announcements during the recession in the beginning of this decade were larger than during the boom. Thus, investors seem to respect dividend increases more when stock prices are falling. Substantial abnormal returns of dividend increases during the time period of 2005 - 2007 could not be found. The results from the overall samples state that the abnormal returns during the recession were positively slightly higher than during the boom. No clear and strong evidence was found between different dividend yield brackets. In Finland, there were substantial abnormal returns on the announcement day of the extra dividends. Thus, indicating that investors saw the extra dividends as a good thing for shareholders' value. This paper is mostly in line with the theory that investors respect dividends more during bad times than good times.

Abnormal Return of Dividend Announcements during a Boom and a Recession. Empirical evidence from the years of 2002 - 2002 and 2005 - 2007, including the Finnish extra dividends

Tutkimuksen tavoitteena on tutkia epänormaalien tuottojen esiintymistä nousu- ja laskusuhdanteen aikana osingonilmoituspäivän ympärillä. Osinkoilmoitukset ovat kerätty Yhdysvaltojen markkinalta (NYSE) ajanjaksoilta 2000 - 2002, jolloin pörssit laskivat teknokuplan jälkeen ja 2005 - 2007, jolloin sijoittajat kokivat suuria kurssivoittoja. Osinkoilmoitushavainnot koostuvat yhtiöistä, jotka nostivat tai pitivät osinko per osake paikallaan. Tavoitteena on tutkia eroja epänormaaleissa tuotoissa näiden kahden ajanjakson välillä. Toiseksi, tavoitteena on tutkia miten epänormaalit tuotot poikkeavat toisistaan eri osinkotuottoluokissa. Kolmanneksi, tavoitteena on tutkia esiintyikö markkinoilla epänormaaleja tuottoja kun suomalaiset yritykset ilmoittivat ylimääräisistä osingoista, pääasiassa vuonna 2004. Yksinkertaisesti ja lyhyesti sanottuna tavoitteena on tutkia arvostavatko sijoittajat osinkoja enemmän laskukauden vai nousukauden aikana. Rahoitusteorian mukaan sijoittajien tulisi arvostaa laskukauden aikana enemmän yhtiöitä, jotka pystyvät maksamaan huonosta taloustilanteesta huolimatta hyvää osinkoa. Empiiriset testit Yhdysvalloista osoittavat, että osingon nostamisesta johtuvat epänormaalit tuotot olivat suuremmat laskusuhdanteen aikana kuin noususuhdanteen aikana. Tämä on linjassa teorian kanssa. Osingon-nostot aiheuttivat nousukauden aikana vähäisiä epänormaaleja tuottoja. Selviä eroja eri osingontuottoluokkien välillä ei pystytty havaitsemaan. Tulokset yhdistetystä aineistosta osoittavat, että sijoittajat kokivat vähäisiä positiivisia epänormaaleja tuottoja laskukauden aikana. Nousukautena tuotot olivat lähellä nollaa. Suomen markkinoilla havaittiin selvä epänormaalituotto osingonilmoituspäivänä. Tulokset ovat pääpiirteittäin linjassa teorian kanssa. Sijoittajat arvostavat osinkoja hieman enemmän lasku- kuin noususuhdanteen aikana.

Coma post-anoxique après arrêt cardiaque et hypothermie, peut-on encore prédire le pronostic en 2014 ?

L'encéphalopathie post-anoxique après arrêt cardiaque (AC) est une cause féquente d'admission pour coma en réanimation. Depuis les recommandations de 2003, l'hypothermie thérapeutique (HT) est devenue un standard de traitement après AC et est à l'origine de l'amélioration du pronostic au cours de cette derniere décennie. Les élements prédicteurs de pronostic validés par l'Académie Américaine de Neurologie avant l'ère de l'HT sont devenus moins précis. En effet, l'HT et la sédation retardent la reprise de la réponse motrice et peuvent altérer la valeur prédictive des réflexes du tronc cérébral. Une nouvelle approche est nécessaire pour établir un pronostic après AC et HT. L'enregistrement (pendant l'HTou peu après) d'une activité électroencéphalographique réactive et/ou continue est un bon prédicteur de récupération neurologique favorable après AC. Au contraire, la présence d'un tracé non réactif ou discontinu de type burst-suppression, avec une réponse N20 absente bilatérale aux potentiels évoqués somatosensoriels, sont presqu'à 100 % prédictifs d'un coma irréversible déjà à 48 heures après AC. L'HT modifie aussi la valeur prédictive de l'énolase neuronale spécifique (NSE), principal biomarqueur sérique de la lésion cérébrale post-anoxique. Un réveil avec bonne récupération neurologique a été récemment observé par plusieurs groupes chez des patients présentant des valeurs de NSE>33 μg/L à 48-72 heures : ce seuil ne doit pas être utilisé seul pour guider le traitement. L'imagerie par résonance magnétique de diffusion peut aider à prédire les séquelles neurologiques à long terme. Un réveil chez les patients en coma post-anoxique est de plus en plus observé, malgré l'absence précoce de signes moteurs et une élévation franche des biomarqueurs neuronaux. En 2014, une nouvelle approche multimodale du pronostic est donc nécessaire, pour optimiser la prédiction d'une évolution clinique favorable après AC. Hypoxic-ischemic encephalopathy after cardiac arrest (CA) is a frequent cause of intensive care unit (ICU) admission. Incorporated in all recent guidelines, therapeutic hypothermia (TH) has become a standard of care and has contributed to improve prognosis after CA during the past decade. The accuracy of prognostic predictors validated in 2006 by the American Academy of Neurology before the era of TH is less accurate. Indeed, TH and sedation may delay the recovery of motor response and alter the predictive value of brainstem reflexes. A new approach is needed to accurately establish prognosis after CA and TH. A reactive and/or continuous electroencephalogram background (during TH or shortly thereafter) strongly predicts good outcome. On the contrary, unreactive/spontaneous burst-suppression electroencephalogram pattern, together with absent N20 on somatosensory evoked potentials, is almost 100% predictive of irreversible coma. TH also affects the predictive value of neuronspecific enolase (NSE), the main serum biomarker of postanoxic injury. A good outcome can occur despite NSE levels >33 μg/L, so this cutoff value should not be used alone to guide treatment. Diffusion magnetic resonance imagery may help predict long-term neurological sequelae. Awakening from postanoxic coma is increasingly observed, despite the absence of early motor signs and pathological elevation of NSE. In 2014, a multimodal approach to prognosis is recommended to optimize the prediction of outcome after CA.

Standardized EEG interpretation accurately predicts prognosis after cardiac arrest.

OBJECTIVE: To identify reliable predictors of outcome in comatose patients after cardiac arrest using a single routine EEG and standardized interpretation according to the terminology proposed by the American Clinical Neurophysiology Society. METHODS: In this cohort study, 4 EEG specialists, blinded to outcome, evaluated prospectively recorded EEGs in the Target Temperature Management trial (TTM trial) that randomized patients to 33°C vs 36°C. Routine EEG was performed in patients still comatose after rewarming. EEGs were classified into highly malignant (suppression, suppression with periodic discharges, burst-suppression), malignant (periodic or rhythmic patterns, pathological or nonreactive background), and benign EEG (absence of malignant features). Poor outcome was defined as best Cerebral Performance Category score 3-5 until 180 days. RESULTS: Eight TTM sites randomized 202 patients. EEGs were recorded in 103 patients at a median 77 hours after cardiac arrest; 37% had a highly malignant EEG and all had a poor outcome (specificity 100%, sensitivity 50%). Any malignant EEG feature had a low specificity to predict poor prognosis (48%) but if 2 malignant EEG features were present specificity increased to 96% (p < 0.001). Specificity and sensitivity were not significantly affected by targeted temperature or sedation. A benign EEG was found in 1% of the patients with a poor outcome. CONCLUSIONS: Highly malignant EEG after rewarming reliably predicted poor outcome in half of patients without false predictions. An isolated finding of a single malignant feature did not predict poor outcome whereas a benign EEG was highly predictive of a good outcome.

Dual Action of BPC194: A Membrane Active Peptide Killing Bacterial Cells

Membrane active peptides can perturb the lipid bilayer in several ways, such as poration and fusion of the target cell membrane, and thereby efficiently kill bacterial cells. We probe here the mechanistic basis of membrane poration and fusion caused by membrane-active, antimicrobial peptides. We show that the cyclic antimicrobial peptide, BPC194, inhibits growth of Gram-negative bacteria and ruptures the outer and inner membrane at the onset of killing, suggesting that not just poration is taking place at the cell envelope. To simplify the system and to better understand the mechanism of action, we performed Förster resonance energy transfer and cryogenic transmission electron microscopy studies in model membranes and show that the BPC194 causes fusion of vesicles. The fusogenic action is accompanied by leakage as probed by dual-color fluorescence burst analysis at a single liposome level. Atomistic molecular dynamics simulations reveal how the peptides are able to simultaneously perturb the membrane towards porated and fused states. We show that the cyclic antimicrobial peptides trigger both fusion and pore formation and that such large membrane perturbations have a similar mechanistic basis

Candidate counterparts to the soft gamma-ray flare in the direction od LSI+61303

A short duration burst reminiscent of a soft gamma-ray repeater/anomalous X-ray pulsar behaviour was detected in the direction of LS I +61 303 by the Swift satellite. While the association with this well known gamma-ray binary is likely, a different origin cannot be excluded. Aims. We explore the error box of this unexpected flaring event and establish the radio, near-infrared and X-ray sources in our search for any peculiar alternative counterpart. Methods. We carried out a combined analysis of archive Very Large Array radio data of LS I +61 303 sensitive to both compact and extended emission. We also reanalysed previous near infrared observations with the 3.5 m telescope of the Centro Astronómico Hispano Alemán and X-ray observations with the Chandra satellite. Results. Our deep radio maps of the LS I +61 303 environment represent a significant advancement on previous work and 16 compact radio sources in the LS I +61 303 vicinity are detected. For some detections, we also identify near infrared and X-ray counterparts. Extended emission features in the field are also detected and confirmed. The possible connection of some of these sources with the observed flaring event is considered. Based on these data, we are unable to claim a clear association between the Swift-BAT flare and any of the sources reported here. However, this study represents the most sophisticated attempt to determine possible alternative counterparts other than LS I +61 303.

Human disease and drug pharmacology, complex as real life

In the past decades drug discovery practice has escaped from the complexity of the formerly used phenotypic screening in animals to focus on assessing drug effects on isolated protein targets in the search for drugs that exclusively and potently hit one selected target, thought to be critical for a given disease, while not affecting at all any other target to avoid the occurrence of side-effects. However, reality does not conform to these expectations, and, conversely, this approach has been concurrent with increased attrition figures in late-stage clinical trials, precisely due to lack of efficacy and safety. In this context, a network biology perspective of human disease and treatment has burst into the drug discovery scenario to bring it back to the consideration of the complexity of living organisms and particularly of the (patho)physiological environment where protein targets are (mal)functioning and where drugs have to exert their restoring action. Under this perspective, it has been found that usually there is not one but several disease-causing genes and, therefore, not one but several relevant protein targets to be hit, which do not work on isolation but in a highly interconnected manner, and that most known drugs are inherently promiscuous. In this light, the rationale behind the currently prevailing single-target-based drug discovery approach might even seem a Utopia, while, conversely, the notion that the complexity of human disease must be tackled with complex polypharmacological therapeutic interventions constitutes a difficult-torefuse argument that is spurring the development of multitarget therapies.

Ocorrência das biovares 1 e 2 de Ralstonia solanacearum em lavouras de batata no estado do Rio Grande do Sul

Com a finalidade de verificar a ocorrência e a distribuição de biovares de Ralstonia solanacearum no Estado do Rio Grande do Sul, plantas com sintomas de murcha-bacteriana foram coletadas, na primavera de 1999, em 25 lavouras de batata (Solanum tuberosum) em dez municípios das quatro áreas de produção: Serra do Nordeste, Planalto Superior, Depressão Central e Grandes Lagoas. A determinação da espécie foi feita com teste sorológico através de ELISA e da biovar com base no metabolismo oxidativo de açúcares e álcoois. Noventa e quatro porcento dos 490 isolados foram identificados como biovar 2. A ocorrência da biovar 1 foi registrada na Serra do Nordeste e Depressão Central. A ocorrência das biovares não mostrou relação com a temperatura média local ou cultivar de batata.

Determinação do mecanismo de destruição de células mediado por meso-tetramesitylporfirina, octaetilporfirina, octaetilporfirina de vanadil e luz visível

A Terapia Fotodinâmica é um tratamento clínico empregado no combate a vários tipos de tumores. Nesta técnica, o paciente recebe uma certa dosagem de uma substância fotossensibilizadora que, por sua vez, acumula-se preferencialmente no tecido tumoral. Em seguida irradia-se luz visível, neste tecido, com a finalidade de se excitar o fotossensibilizador. Esta substância, excitada, pode transferir elétrons e/ou energia para o oxigênio, no estado fundamental, gerando espécies reativas de oxigênio como o radical superóxido e oxigênio singlete. O primeiro é gerado no mecanismo tipo I, pela transferência de elétrons, e o segundo no mecanismo tipo II, pela transferência de energia. Estas espécies reativas geram um estresse oxidativo, no tecido tumoral, levando as células cancerígenas à morte. Neste trabalho, foram avaliadas as propriedades fotossensibilizadoras de três porfirinas e constatou-se que as mesmas são capazes de destruir células, na presença de luz e oxigênio. Este evento foi mais pronunciado, na presença de água deuterada, e inibido por supressores de oxigênio singlete, indicando predominância do mecanismo tipo II.

Efeito da complexação de metais aos antiinflamatórios na ação contra agentes oxidativos e radicais livres: ação do cetoprofeno

Os radicais livres são espécies altamente reativas geradas nos organismos vivos com a finalidade de proteção. Porém, em algumas circunstâncias, estes são responsáveis pela ocorrência ou o agravo de danos teciduais. Muitos antiinflamatórios apresentam ação direta sobre radicais livres e espécies reativas não radicalares, o que contribui para suas ações contra a inflamação. O cetoprofeno é um antiinflamatório não esteroidal que gera radicais livres ao sofrer fotoirradiação e tem com isso um efeito hemolítico importante. A complexação de metais a diferentes fármacos tem sido utilizada como estratégia para melhorar a ação farmacológica de diferentes moléculas e reduzir seus efeitos colaterais. Neste trabalho são apresentados resultados do estudo de ação do cetoprofeno e seus complexos de cério e cobre sobre radicais livres e sobre eritrócitos. Observou-se que o cério intensifica as propriedades scavenger do cetoprofeno sobre radicais livres enquanto o cobre intensifica as ações sobre oxidantes não radicalares. O cobre ainda reduziu o efeito hemolítico apresentado pelo cetoprofeno e mantido pelo seu derivado de cério.

Chemical characteristics and Kraft pulping of tension wood from Eucalyptus globulus labill

Tension (TW) and opposite wood (OW) of Eucalyptus globulus trees were analyzed for its chemical characteristics and Kraft pulp production. Lignin content was 16% lower and contained 32% more syringyl units in TW than in OW. The increase in syringyl units favoured the formation of β-O-4 bonds that was also higher in TW than in OW (84% vs. 64%, respectively). The effect of these wood features was evaluated in the production of Kraft pulps from both types of wood. At kappa number 16, Kraft pulps obtained from TW demanded less active alkali in delignification and presented slightly higher or similar pulp yield than pulps made with OW. Fiber length, coarseness and intrinsic viscosity were also higher in tension than in opposite pulps. When pulps where refined to 30°SR, TW pulps needed 18% more revolutions in the PFI mill to achieve the same beating degree than OW pulps. Strength properties (tensile, tear and burst indexes) were slightly higher or similar in tension as compared with opposite wood pulps. After an OD0(EO)D1 bleaching sequence, both pulps achieved up to 89% ISO brightness. Bleached pulps from TW presented higher viscosity and low amount of hexenuronic acids than pulps from OW. Results showed that TW presented high xylans and low lignin content that caused a decrease in alkali consumption, increase pulp strength properties and similar bleaching performance as compared with pulps from OW.