Hypersensitive Cell Death and Papilla Formation in Barley Attacked by the Powdery Mildew Fungus Are Associated with Hydrogen Peroxide but Not with Salicylic Acid Accumulation1

We analyzed the pathogenesis-related generation of H2O2 using the microscopic detection of 3,3-diaminobenzidine polymerization in near-isogenic barley (Hordeum vulgare L.) lines carrying different powdery mildew (Blumeria graminis f.sp. hordei) resistance genes, and in a line expressing chemically activated resistance after treatment with 2,6-dichloroisonicotinic acid (DCINA). Hypersensitive cell death in Mla12 and Mlg genotypes or after chemical activation by DCINA was associated with H2O2 accumulation throughout attacked cells. Formation of cell wall appositions (papillae) mediated in Mlg and mlo5 genotypes and in DCINA-activated plants was paralleled by H2O2 accumulation in effective papillae and in cytosolic vesicles of up to 2 μm in diameter near the papillae. H2O2 was not detected in ineffective papillae of cells that had been successfully penetrated by the fungus. These findings support the hypothesis that H2O2 may play a substantial role in plant defense against the powdery mildew fungus. We did not detect any accumulation of salicylic acid in primary leaves after inoculation of the different barley genotypes, indicating that these defense responses neither relied on nor provoked salicylic acid accumulation in barley.

Evolutionarily conserved Galphabetagamma binding surfaces support a model of the G protein-receptor complex.

The pivotal role of G proteins in sensory, hormonal, inflammatory, and proliferative responses has provoked intense interest in understanding how they interact with their receptors and effectors. Nonetheless, the locations of the receptors and effector binding sites remain poorly characterized, although nearly complete structures of the alphabetagamma heterotrimeric complex are available. Here we apply evolutionary trace (ET) analysis [Lichtarge, O., Bourne, H. R. & Cohen, F. E. (1996) J. Mol. Biol. 257, 342-358] to propose plausible locations for these sites. On each subunit, ET identifies evolutionarily selected surfaces composed of residues that do not vary within functional subgroups and that form spatial clusters. Four clusters correctly identify subunit interfaces, and additional clusters on Galpha point to likely receptor or effector binding sites. Our results implicate the conformationally variable region of Galpha in an effector binding role. Furthermore the range of predicted interactions between the receptor and Galphabetagamma, is sufficiently limited that we can build a low resolution and testable model of the receptor-G protein complex.

Regulated expression of the obese gene product (leptin) in white adipose tissue and 3T3-L1 adipocytes.

A mutation within the obese gene was recently identified as the genetic basis for obesity in the ob/ob mouse. The obese gene product, leptin, is a 16-kDa protein expressed predominantly in adipose tissue. Consistent with leptin's postulated role as an extracellular signaling protein, human embryonic kidney 293 cells transfected with the obese gene secreted leptin with minimal intracellular accumulation. Upon differentiation of 3T3-L1 preadipocytes into adipocytes, the leptin mRNA was expressed concomitant with mRNAs encoding adipocyte marker proteins. A factor(s) present in calf serum markedly activated expression of leptin by fully differentiated 3T3-L1 adipocytes. A 16-hr fast decreased (by approximately 85%) the leptin mRNA level of adipose tissue of lean (ob/+ or +/+) mice but had no effect on the approximately 4-fold higher level in obese (ob/ob) littermates. Since the mutation at the ob locus fails to produce the functional protein, yet its cognate mRNA is overproduced, it appears that leptin is necessary for its own downregulation. Leptin mRNA was also suppressed in adipose tissue of rats during a 16-hr fast and was rapidly induced during a 4-hr refeeding period. Insulin deficiency provoked by streptozotocin also markedly down-regulated leptin mRNA and this suppression was rapidly reversed by insulin. These results suggest that insulin may regulate the expression of leptin.

Evidence for a role of pituitary ATP receptors in the regulation of pituitary function.

Despite a rapidly increasing acceptance for a role of ATP as an extracellular mediator in several biological systems, the present report shows that ATP may mediate physiological responses in pituitary cells. We have now been able to demonstrate a specific action of ATP receptors to mediate the release of luteinizing hormone from gonadotropes and have coupled them with further studies that clearly show that ATP can be exocytotically released from cultured rat pituitary cells. Both ATP and UTP (100 microM) caused a > 14-fold increase in the rate of luteinizing hormone release from superfused cells. Adenosine 5'-[alpha, beta-methylene]triphosphate and 5'-[beta,gamma-methylene triphosphate were ineffective, and 2-methylthio-ATP had only a modest stimulatory effect. Homologous and heterologous desensitization occurred with UTP and ATP, and these did not have additive effects. Thus, nucleotides can be effective stimulators of luteinizing hormone release through a single class of ATP receptor (P2U subtype). The calcium ionophore A23187 provoked release of a substantial amount of ATP from pituitary cells in a concentration- and Ca(2+)-dependent manner, which was desensitized by pretreatment with A23187. This implies a possible paracrine and/or autocrine mechanism by which nucleotides may exert their effects on pituitary cells. In conclusion, we have provided strong evidence for a novel role of extracellular nucleotides as mediators in pituitary--in particular, in gonadotrope--function.

Irradiação de drogas vegetais: aspectos microbiológicos e químicos

Apesar da industrialização no setor farmacêutico, o emprego de drogas vegetais constitui desafio atual quando considerado alternativa terapêutica para as populações de baixa renda ou aquelas que apresentam tradição no uso dessas drogas. Além disso, tendências modernas valorizam a variedade de espécies com propriedades curativas, em particular as espécies brasileiras, desafiando os pesquisadores a intensificar investigações nessa área e induzindo à população um crescente consumo. Assim, questões relacionadas à qualidade dessas drogas apresentam fundamental importância. Devido à origem, a carga microbiana detectada nas mesmas é normalmente elevada, oferecendo riscos potenciais ao usuário. Desta forma, a avaliação de sua qualidade sanitária constitui etapa obrigatória no que se refere ao aspecto de segurança ao consumidor. Além disso, a eficácia terapêutica pode igualmente ser comprometida por decomposição de componentes, decorrente da ação de microrganismos. Com o objetivo de eliminar os efeitos decorrentes da biocarga presente nas drogas vegetais, agentes descontaminantes, de natureza física ou química, têm sido empregados. A utilização de tais procedimentos de descontaminação, prevista na legislação vigente, requer estudos relacionados à estabilidade dos princípios ativos após exposição ao agente selecionado. Dentre os agentes destaca-se a irradiação gama, amplamente utilizada em função de sua aplicabilidade na ausência de água e de temperatura elevada, além de apresentar alta penetrabilidade e reduzir, com eficácia, a carga microbiana viável. Os objetivos do presente trabalho foram avaliar os efeitos de diferentes doses de radiação ionizante sobre a carga microbiana de quatro espécies de drogas vegetais: alcachofra (Cynara scolymus L.), camomila (Matricaria recutita L.), ginkgo (Ginkgo biloba L.) e guaraná (Paullinia cupana H.B.K.), bem como detectar possíveis alterações provocadas pela radiação sobre os teores de seus princípios ativos. As análises microbiológicas e químicas foram realizadas antes e após irradiação com doses médias de 5,5 kGy, 11,4 kGy e 17,8 kGy. Os resultados obtidos anteriormente à irradiação revelaram elevados níveis de contaminação: média de 4,1 x106 para microrganismos aeróbicos totais e 3,3x105 para fungos. Após descontaminação, a dose média de 11,4 kGy, reduziu a carga de microrganismos aeróbicos totais a níveis menores ou iguais a 102 em todas as drogas, com exceção da camomila proveniente do fornecedor B (3,2x104). Para os fungos, a menor dose aplicada (5,5 kGy) foi suficiente para reduzir a contagem a níveis da ordem de 10. Com relação à determinação dos marcadores nas drogas vegetais, os resultados obtidos não revelaram alterações significativas nos teores de cafeína no guaraná e de glicosídeos flavonoídicos no ginkgo. Para a camomila, as amostras antes a após irradiação, apresentaram o mesmo teor de óleo volátil bem como ausência de diferenças significativas no teor de α-bisabolol. Em contraste, observou-se redução no teor de 7-glicosil apigenina após submissão à radiação ionizante, indicando degradação decorrente do processo. Com relação à alcachofra, permanece ainda desconhecida a influência da radiação devido à ausência de metodologias adequadas para extração e determinação da cinarina.

Estudo dos espectros vibracionais de poliacetilenos substituídos

Neste trabalho foram sintetizados a poli-2-etinilpiridina (P2EP), poli-4etinilpiridina (P4EP), o iodeto de poli(2-N-t-butilpiridiniumilacetileno) (P2EPtBu) e a poli-β-etinilnaftaleno (Pβ:EN), os quais são poliacetilenos substituídos. Estes polímeros, juntamente com o cloreto de poli(2-piridínio-2-piridilacetileno) (P2EPH), foram caracterizados por espectroscopia vibracional no infravermelho e Raman. Estes polímeros apresentaram variação na posição das bandas Raman com a energia da radiação excitante - chamada dispersão Raman ou fotosseletividade - da ordem de 10 cm-1, bem inferior ao apresentado pelo poliacetileno (cerca de 60 cm-1). Este deslocamento foi interpretado utilizando-se dois dos modelos existentes para descrever este fenômeno: o Modelo de Modo de Amplitude (AMM) e o Modelo de Coordenada de Conjugação Efetiva (ECCM), os quais fornecem informações sobre a estrutura polimérica e sobre seus níveis eletrônicos. Utilizando-se o AMM foi possível obter informações sobre os níveis eletrônicos excitados de mesma simetria que o estado eletrônico fundamental. Por outro lado, o ECCM, com a ajuda de cálculos DFT, mostrou diferenças na extensão da conjugação e no grau de dimerização entre o P2EP na forma cis e trans e indicou que este polímero apresentava, predominantemente, a estrutura cis, fato este confirmado pelos espectros no infravermelho. A dopagem com I2 provocou efeitos diferentes na estrutura dos polímeros. Os espectros no infravermelho dos polímeros dopados indicaram que o P2EP e o P2EPH apresentaram aumento na quantidade de segmentos cis enquanto o P2EPtBu apresentou diminuição na quantidade desses segmentos. Os espectros Raman dos polímeros dopados confirmaram os dados dos espectros no infravermelho. Esta diferença foi interpretada como sendo devida à diferença no volume do substituinte, pois grupos volumosos favorecem o isômero trans-cisóide onde a distância entre os substituintes é maior. A dopagem também levou a um aumento na condutividade dos polímeros, porém os valores de condutividade obtidos foram bem inferiores que os apresentados pelo poliacetileno dopado (10-5 a 10-7 contra 102 S cm-1, tipicamente).

2012: una odisea urbana. La nueva sede de la Confederación de Empresarios de Albacete (FEDA)

La nueva sede de la Confederación de Empresarios de Albacete (FEDA), 2007-2011, es un edificio del equipo de arquitectos COR & Asociados, formado por Jesús Olivares y Miguel Rodenas, titulados por la Universidad de Alicante. Situado en un retal de la ciudad de Albacete (España), con 3800 m2 y un coste de poco menos de 1000 €/m2, este equipamiento público, de manifiesta calidad como pieza arquitectónica, suscita a los autores del presente artículo una reflexión en profundidad sobre el alcance de su impacto en el medio indeciso en que se inserta y sobre su capacidad para urbanizar un entorno falto de carácter.

Isobaric vapor–liquid–liquid–solid equilibrium of the water + NaCl + 1-butanol system at 101.3 kPa

A mixture of water + NaCl + 1-butanol at 101.3 kPa is studied in order to determine the influence of salt on its experimental vapor–liquid–liquid–solid equilibrium. A detailed analysis of the evolution with temperature of the different equilibrium regions is carried out. The study is conducted at a constant pressure of 101.3 kPa in a recirculating still that has been modified by our research group. The changes in the 1-butanol/water composition ratio in the vapor phase that are provoked by the salt are studied as a function of equilibrium region. In addition, the mutual solubility of 1-butanol and water is assessed in the liquid–liquid and solid–liquid regions.

John Warren's copy of votes from a meeting of the Boston Medical Society, 1784 May 3

This document lists the eleven votes cast at a meeting of the Boston Medical Society on May 3, 1784. It was authorized as a "true coppy" by Thomas Kast, the Secretary of the Society. The following members of the Society were present at the meeting, all of them doctors: James Pecker, James Lloyd, Joseph Gardner, Samuel Danforth, Isaac Rand, Jr., Charles Jarvis, Thomas Kast, Benjamin Curtis, Thomas Welsh, Nathaniel Walker Appleton, and doctors whose last names were Adams, Townsend, Eustis, Homans, and Whitwell. The document indicates that a meeting had been held the previous evening, as well (May 2, 1784), at which the topics on which votes were taken had been discussed. The votes, eleven in total, were all related to the doctors' concerns about John Warren and his involvement with the emerging medical school (now Harvard Medical School), that school's relation to almshouses, the medical care of the poor, and other related matters. The tone and content of these votes reveals anger on the part of the members of the Boston Medical Society towards Warren. This anger appears to have stemmed from the perceived threat of Warren to their own practices, exacerbated by a vote of the Harvard Corporation on April 19, 1784. This vote authorized Warren to apply to the Overseers of the Poor for the town of Boston, requesting that students in the newly-established Harvard medical program, where Warren was Professor of Anatomy and Surgery, be allowed to visit the hospital of the almshouse with their professors for the purpose of clinical instruction. Although Warren believed that the students would learn far more from these visits, in regards to surgical experience, than they could possibly learn in Cambridge, the proposal provoked great distrust from the members of the Boston Medical Society, who accused Warren of an "attempt to direct the public medical business from its usual channels" for his own financial and professional gain.

Casual but Smart: The Court’s new clothes in the Area of Freedom Security and Justice (AFSJ) after the Lisbon Treaty. Research Papers in Law, 2/2008

Introduction. Unintended as it was, the European Court of Justice (ECJ, the Court, the Court of the EU) has played an extremely important role in the construction of the Area of Freedom Security and Justice (AFSJ). The AFSJ was set up by the Treaty of Amsterdam in 1997 and only entered into force in May 1999. The fact that this is a new field of EU competence, poses afresh all the fundamental questions – both political and legal – triggered by European integration, namely in terms of: a) distribution of powers between the Union and its Member States, b) attribution of competences between the various EU Institutions, c) direct effect and supremacy of EU rules, d) scope of competence of the ECJ, and e) extent of the protection given to fundamental rights. The above questions have prompted judicial solutions which take into account both the extremely sensible fields of law upon which the AFSJ is anchored, and the EU’s highly inconvenient three-pillar institutional framework.1 The ECJ is the body whose institutional role is to benefit most from this upcoming ‘depilarisation’, possibly more than that of the European Parliament. This structure is on the verge of being abandoned, provided the Treaty of Lisbon enters into force.2 However spectacular this formal boost of the Court’s competence, the changes in real terms are not going to be that dramatic. This apparent contradiction is explained, to a large extent, by the fact that the Court has in many ways ‘provoked’, or even ‘anticipated’, the depilarisation of its own jurisdictional role, already under the existing three-pillar structure. Simply put, under the new – post Treaty of Lisbon – regime, the Court will have full jurisdiction over all AFSJ matters, as those are going to be fully integrated in what is now the first pillar. Some limitations will continue to apply, however, while a special AFSJ procedure will be institutionalised. Indeed, if we look into the new Treaty we may identify general modifications to the Court’s structure and jurisdiction affecting the AFSJ (section 2), modifications in the field of the AFSJ stemming from the abolition of the pillar structure (section 3) and, finally, some rules specifically applicable to the AFSJ (section 4).

The EU's Group-to-Group Dialogue with the Southern Mediterranean and ASEAN – How much have they achieved? A Comparative Analysis. EU Centre Singapore Working Paper No, 18, December 2013

Four decades of the EU's group-to-group dialogues with the Southern Mediterranean grouping of countries and with ASEAN have produced different dynamics and outcomes, despite the EU’s common strategy to use economic soft power to achieve their goals for the partnerships. Diverging conditions in the two regions created inconsistency in the EU's application of the common approach. The EU's neighbourhood security concerns forced it to relax its political stand with their Southern Mediterranean partners. For ASEAN, geographical distance dilutes the EU’s security concerns it that region and has afforded the EU to be more ideological and assertive on democracy and human rights practices. These issues have provoked disagreements in EU-ASEAN dialogues, but both sides have also tried to remain pragmatic in order to achieve some progress in the partnership. In contrast, the protracted the Arab-Israeli conflict continues to hamper the Euro-Mediterranean dialogue, resulting in little progress. Social upheavals in the Southern Mediterranean also brought their partnership to a standstill. The EU's cooperation with former authoritarian regimes like Libya and Syria have only caused damage to its credibility in the Southern Mediterranean, and future Euro-Mediterranean dialogues are likely to be affected by it.

In a state of necessity. How has Orban changed Hungary. OSW Point of View Number 41, April 2014

Viktor Orban’s sweeping victory in the 2010 election ensured his party, Fidesz, a constitutional two-thirds majority in parliament. The party took over the rule of the country from the discredited political left when Hungary was plunged in political and economic crisis. Claiming that the circumstances were unusual and that it had a strong electoral mandate, Fidesz introduced radical changes in the country and thus challenged the previous economic and political order. These changes have led to an unprecedented concentration of power and provoked a discussion on the limits of democracy and the rule of law in the European Union. The state’s economic role has strengthened. The Orban government has been unable to overcome economic stagnation but it has managed to stabilise Hungary’s budget situation, which needs to deal with the high debt. Hungary’s relations with most partners in the EU and NATO have cooled due to controversial moves made by its government. As regards foreign policy and economic co-operation, Orban has granted high priority to the ‘Eastern opening’, where Russia has assumed the leading role.

Russia's position on the events in Kyrgyzstan (April-June 2010). OSW Commentary No. 38, 2010-07-27

Russia was the first state in the world to de facto recognise the regime change in Kyrgyzstan that took place on 7 April 2010. This recognition, along with a previous campaign by the Russian media against the then President Kurmanbek Bakiyev, has given rise to suspicion that the events of April were provoked by Russia. However, it seems no more than reasonable to say that Russia provided some inspiration and lobbying in that direction. Russia offered support to the new Kyrgyz government almost immediately, albeit conditionally. Russia’s relations with Roza Otunbayeva’s government have been changing in nature; they are currently much cooler than they had been immediately after the coup. There are many indications that this change was a reaction to the extension of the lease agreement for the American military base in the Manas airport. At the same time, Moscow remains in contact with the political rivals to the current regime, which suggests that the Kremlin is preparing for different developments, and does not regard the current crisis as having been fully resolved. Despite the interim government’s plea for help, Russia refused to undertake military intervention in southern Kyrgyzstan, which plunged into ethnic unrest in June. This shows that Russia is wary of being dragged into a long-standing and bloody conflict in the region, which could entail considerable expenses and jeopardise Russia’s authority. It should be expected that after the October parliamentary elections in Kyrgyzstan, Russia will return to its plans to establish a second military base in this country (in addition to the Kant base) to reinforce its dominant position in the region. This is the first time that Russia has had a real chance to play a stabilising role in the CIS area. How Russia copes with this challenge may decide its position in post-Soviet Central Asia – and in a wider context, its relations with NATO, the USA and China.

The prospects for the EU-Ukraine free trade agreement. OSW Commentary No. 94, 2012-10-15

The European Union and Ukraine initialled the Deep and Comprehensive Free Trade Area Agreement (DCFTA) on 19 July 2012. The scope of the agreement which the EU and Ukraine reached following their negotiations is much more extensive than that of a typical free trade agreement. It envisages not only the lifting of tariff and extra-tariff barriers but also, more importantly, Kyiv adopting EU legal solutions and standards in this area. Whether the agreement will be signed and implemented is still an open question and depends on the existing political conditions. On the one hand, the repression imposed by the government in Kyiv on its political opponents (including the detention of the former prime minister, Yulia Tymoshenko) has provoked criticism from the EU, which refuses to sign the agreement if the government in Kyiv continues to violate democratic principles. The manner in which Ukraine’s parliamentary elections are conducted this October will be the key test. On the other hand, Russia is increasingly active in its efforts to involve Ukraine in the integration projects it has initiated (the Customs Union and the Eurasian Economic Community). It should be noted that Moscow has effective instruments to exert its will, such as the dependence of the Ukrainian economy on supplies of Russian oil and gas and on exports to the Russian market. Besides, Moscow also has political instruments at its disposal. It is impossible to participate in integration projects both with the EU and with Russia. Therefore, Kyiv will have to make a strategic decision and choose the direction of its economic integration. Unless Ukraine takes concrete action to implement its agreements with the EU, primarily including the free trade agreement, its economic dependence on Russia will grow, and it will be more likely to join the Russian integration projects.

Determinants of Farm Labour Use: A Comparison between Ireland and Italy. Factor Markets Working Documents No. 60, August 2013

This paper examines the effect of the decoupling of farm direct payments upon the off-farm labour supply decisions of farmers in both Ireland and Italy, using panel data from the Farm Business Survey (REA) and FADN database covering the period from 2002 to 2009 to model these decisions. Drawing from the conceptual agricultural household model, the authors hypothesise that the decoupling of direct payments led to an increase in off-farm labour activity despite some competing factors. This hypothesis rests largely upon the argument that the effects of changes in relative wages have dominated other factors. At a micro level, the decoupling-induced decline in the farm wage relative to the non-farm wage ought to have provoked a greater incentive for off-farm labour supply. The main known competing argument is that decoupling introduced a new source of non-labour income i.e. a wealth effect. This may in turn have suppressed or eliminated the likelihood of increased off-farm labour supply for some farmers. For the purposes of comparative analysis, the Italian model utilises the data from the REA database instead of the FADN as the latter has a less than satisfactory coverage of labour issues. Both models are developed at a national level. The paper draws from the literature on female labour supply and uses a sample selection corrected ordinary least squares model to examine both the decisions of off-farm work participation and the decisions regarding the amount of time spent working off-farm. The preliminary results indicate that decoupling has not had a significant impact on off-farm labour supply in the case of Ireland but there appears to be a significantly negative relationship in the Italian case. It still remains the case in both countries that the wealth of the farmer is negatively correlated with the likelihood of off-farm employment.