In vivo and in vitro characterization of the B1 and B2 zinc-binding domains from the acute promyelocytic leukemia protooncoprotein PML.

Acute promyelocytic leukemia (APL) has been ascribed to a chromosomal translocation event which results in a fusion protein comprising the PML protein and retinoic acid receptor alpha. PML is normally a component of a nuclear multiprotein complex which is disrupted in the APL disease state. Here, two newly defined cysteine/histidine-rich protein motifs called the B-box (B1 and B2) from PML have been characterized in terms of their effect on PML nuclear body formation, their dimerization, and their biophysical properties. We have shown that both peptides bind Zn2+, which induces changes in the peptides' structures. We demonstrate that mutants in both B1 and B2 do not form PML nuclear bodies in vivo and have a phenotype that is different from that observed in the APL disease state. Interestingly, these mutations do not affect the ability of wild-type PML to dimerize with mutant proteins in vitro, suggesting that the B1 and B2 domains are involved in an additional interaction central to PML nuclear body formation. This report in conjunction with our previous work demonstrates that the PML RING-Bl/B2 motif plays a fundamental role in formation of a large multiprotein complex, a function that may be common to those unrelated proteins which contain the motif.

Svenska landsmål ock svenskt folkliv.

Founded by J. A. Lundell.

Bulletin.

Each lettered series also has a distinctive title.

Espionage against Poland in the Documents and Analyses of the Polish Special Services (1944 - 1989) - as Illustrated by the Intelligence Activities of the USA

The text analyses the intelligence activity against Poland in the period 1944-1989. The paper also contains a case study, i.e. an analysis of the American intelligence service activity held against Poland. While examining the research thesis, the author used the documents and analyses prepared by the Ministry of Internal Affairs. In order to best illustrate the point, the author presented a number of cases of persons who spied for the USA, which was possible thanks to the analysis of the training materials of the Ministry of Internal Affairs directed to the officers of the Security Service and the Citizens’ Militia. The text tackles the following issues: (1) to what extent did the character of the socio-political system influence the number of persons convicted for espionage against Poland in the period under examination?, (2) what was the level of interest of the foreign intelligence services in Poland before the year 1990?, (3) is it possible to indicate the specificity of the U.S. intelligence activity against Poland? 1) The analysis of data indicates that the period 1946-1956 witnessed a great number of convictions for espionage, which is often associated with the peculiar political situation in Poland of that time. Up to 1953, the countries of the Eastern bloc had reproduced the Stalin’s system, which only ceased due to the death of Stalin himself. Since then, the communist systems gradually transformed into the system of nomenklatura. Irrespective of these changes, Poland still witnessed a wave of repressions, which resulted from the threats continuously looming over the communist authorities – combating the anti-communist underground movement, fighting with the Ukrainian Insurgent Army, the Polish government-in-exile, possible revisionism of borders, social discontent related to the socio-political reforms. Hence, a great number of convictions for espionage at that time could be ascribed to purely political sentences. Moreover, equally significant was the fact that the then judicial practice was preoccupied assessing negatively any contacts and relations with foreigners. This excessive number of convictions could ensue from other criminal-law provisions, which applied with respect to the crimes against the State, including espionage. What is also important is the fact that in the Stalin’s period the judiciary personnel acquired their skills and qualifications through intensive courses in law with the predominant spirit of the theory of evidence and law by Andrey Vyshinsky. Additionally, by the decree of 1944 the Penal Code of the Polish Armed Forces was introduced; the code envisaged the increase in the number of offences classified as penalised with death penalty, whereas the high treason was subject to the military jurisdiction (the civilians were prosecuted in military courts till 1955; the espionage, however, still stood under the military jurisdiction). In 1946, there was introduced the Decree on particularly dangerous crimes in the period of the State’s recovery, which was later called a Small Penal Code. 2) The interest that foreign intelligence services expressed in relation to Poland was similar to the one they had in all countries of Eastern and Central Europe. In the case of Poland, it should be noted that foreign intelligence services recruited Polish citizens who had previously stayed abroad and after WWII returned to their home country. The services also gathered information from Poles staying in immigrant camps (e.g. in FRG). The activity of the American intelligence service on the territory of FRG and West Berlin played a key role. The documents of the Ministry of Internal Affairs pointed to the global range of this activity, e.g. through the recruitment of Polish sailors in the ports of the Netherlands, Japan, etc. In line with the development in the 1970s, espionage, which had so far concentrated on the defence and strategic sectors, became focused on science and technology of the People’s Republic of Poland. The acquisition of collaborators in academic circles was much easier, as PRL opened to academic exchange. Due to the system of visas, the process of candidate selection for intelligence services (e.g. the American) began in embassies. In the 1980s, the activity of the foreign intelligence services concentrated on the specific political situation in Poland, i.e. the growing significance of the “Solidarity” social movement. 3) The specificity of the American intelligence activity against Poland was related to the composition of the residency staff, which was the largest in comparison to other Western countries. The wide range of these activities can be proved by the quantitative data of convictions for espionage in the years 1944-1984 (however, one has to bear in mind the factors mentioned earlier in the text, which led to the misinterpretation of these data). Analysing the data and the documents prepared by the Ministry of Internal Affairs, one should treat them with caution, as, frequently, the Polish counter-intelligence service used to classify the ordinary diplomatic practice and any contacts with foreigners as espionage threats. It is clearly visible in the language of the training materials concerned with “secret service methods of the intelligence activity” as well as in the documents on operational activities of the Security Service in relation to foreigners. The level of interest the USA had in Poland was mirrored in the classification of diplomatic posts, according to which Warsaw occupied the second place (the so-called Group “B”) on the three-point scale. The CIA experienced spectacular defeats during their activity in Poland: supporting the Polish underground anti-communist organisation Freedom and Independence and the so-called Munich-Berg episode (both cases took place in the 1950s). The text focuses only on selected issues related to the espionage activities against Poland. Similarly, the analysis of the problem has been based on selected sources, which has limited the research scope - however, it was not the aim of the author to present the espionage activity against Poland in a comprehensive way. In order to assess the real threat posed by the espionage activity, one should analyse the case of persons convicted for espionage in the period 1944-1989, as the available quantitative data, mentioned in the text, cannot constitute an explicit benchmark for the scale of espionage activity. The inaccuracies in the interpretation of data and variables, which can affect the evaluation of this phenomenon, have been pointed out in the text.

The origin of the Land grant act of 1862 (the so-called Morrill act) and some account of its author, Jonathan B. Turner,

Appendix A, Letter from Senator Morrill.--Appendix B, Extract from Forquer's letter.--Appendix C, The Turner pamphlet.

Eating ethically: Emmanuel Levinas and Simone Weil (Exploring the ethical realms of anorexia and so-called states of grace)

Emmanuel Levinas's work on the ethical responsibility of the face-to-face relation offers an illuminating context or clearing within which we might better appreciate the work of Simone Weil. Levinas's subjectivity of the hostage, the one who is responsible for the other before being responsible for the self, provides us with a way of re-encountering the categories of gravity and grace invoked in Weil's original account. In this paper I explore the terrain between these thinkers by raising the question of eating as, in part, an ethical act. Weil's conception of grace refers to the state of decreation in which the utter humility of the self moves toward a kind of disintegration and weightlessness. this weightlessness, which Weil contrasts to the gravity of terrestrial weight, might be thought of in terms of the subject's fundamental responsibility for the other, especially in terms of the injunction Thou shalt neither kill nor take the food of thy neighbour. Taking the place of the other, taking the food from the mouth of the other, is the ethical dilemma facing the subject as hostage and an elaboration of this situation may provide us with steps toward a radical questioning of anorexia as - at least in part - an ethical rather than purely medical condition.

A gestão dos custos da qualidade: Modelo A.B.Q.C. aplicado à Mercauto

Mestrado em Controlo e Gestão dos Negócios

Antitumor activity of hierridin B, a cyanobacterial secondary metabolite found in both filamentous and unicellular marine strains

Cyanobacteria are widely recognized as a valuable source of bioactive metabolites. The majority of such compounds have been isolated from so-called complex cyanobacteria, such as filamentous or colonial forms, which usually display a larger number of biosynthetic gene clusters in their genomes, when compared to free-living unicellular forms. Nevertheless, picocyanobacteria are also known to have potential to produce bioactive natural products. Here, we report the isolation of hierridin B from the marine picocyanobacterium Cyanobium sp. LEGE 06113. This compound had previously been isolated from the filamentous epiphytic cyanobacterium Phormidium ectocarpi SAG 60.90, and had been shown to possess antiplasmodial activity. A phylogenetic analysis of the 16S rRNA gene from both strains confirmed that these cyanobacteria derive from different evolutionary lineages. We further investigated the biological activity of hierridin B, and tested its cytotoxicity towards a panel of human cancer cell lines; it showed selective cytotoxicity towards HT-29 colon adenocarcinoma cells.

Antitumor activity of hierridin B, a cyanobacterial secondary metabolite found in both filamentous and unicellular marine strains

Cyanobacteria are widely recognized as a valuable source of bioactive metabolites. The majority of such compounds have been isolated from so-called complex cyanobacteria, such as filamentous or colonial forms, which usually display a larger number of biosynthetic gene clusters in their genomes, when compared to free-living unicellular forms. Nevertheless, picocyanobacteria are also known to have potential to produce bioactive natural products. Here, we report the isolation of hierridin B from the marine picocyanobacterium Cyanobium sp. LEGE 06113. This compound had previously been isolated from the filamentous epiphytic cyanobacterium Phormidium ectocarpi SAG 60.90, and had been shown to possess antiplasmodial activity. A phylogenetic analysis of the 16S rRNA gene from both strains confirmed that these cyanobacteria derive from different evolutionary lineages. We further investigated the biological activity of hierridin B, and tested its cytotoxicity towards a panel of human cancer cell lines; it showed selective cytotoxicity towards HT-29 colon adenocarcinoma cells.

Biomass production and essential oil yield from leaves, fine stems and resprouts using pruning the crown of Aniba canelilla (H.B.K.) (Lauraceae) in the Central Amazon

Aniba canelilla (H.B.K.) Mez. is a tree species from Amazon that produces essential oil. The oil extraction from its leaves and stems can be an alternative way to avoid the tree cutting for production of essential oil. The aim of this study was to analyse factors that may influence the essential oil production and the biomass of resprouts after pruning the leaves and stems of A. canelilla trees. The tree crowns were pruned in the wet season and after nine months the leaves and stems of the remaining crown and the resprouts were collected, in the dry season. The results showed that the essential oil yield and chemical composition differed among the stems, leaves and resprouts. The stems' essential oil production differed between the seasons and had a higher production in the resprouting stems than the old stems of the remaining crown. The production of essential oil and leaf biomass of resprouts were differently related to the canopy openness, indicating that light increases the production of the essential oil and decreases the biomass of resprouting leaves. This study revealed that plant organs differ in their essential oil production and that the canopy openness must be taken into account when pruning the A. canelilla tree crown in order to achieve higher oil productivity.

A concentração e a quantidade de micronutrientes e de alumínio no cafeeiro, Coffea arabica, L., variedade mundo novo (B.Rodr.) Choussy, aos dez anos de idade

O presente trabalho relata os dados obtidos sôbre a concentração e a quantidade de micronutrientes, boro (B), cloro (Cl), cobre (Cu), ferro (Fe), manganês (Mn), molibdênio (Mo) e zinco (Zn), no tronco, ramos, fôlhas e frutos do cafeeiro, Coffea arabica, L., variedade mundo novo (B.Rodr.) Choussy, aos dez anos de idade e crescendo em solo latosólico da Escola Superior de Agricultura "Luiz de Queiroz", em Piracicaba. Além dos micronutrientes, são apresentados dados sobre a concentração e a quantidade de alumínio, nas citadas partes do cafeeiro.

The role of the Estrogen-Responsive B box protein in skin morphogenesis, wound repair and skin disease

SUMMARY: EBBP is a poorly characterized member of the RBCC/TRIM family (RING finger B-box coiled-coilltripartite motif). It is ubiquitously expressed, but particularly high levels are found in keratinocytes. There is evidence that EBBP is involved in inflammatory processes, since it can interact with pro-interleukin-1 ß (prolL-1 ß) in human macrophages and keratinocytes, and its downregulation results in reduced secretion of IL-1 ß. IL-1ß activation and secretion requires the proteolytic cleavage of prolL-1ß by caspase-1, which in turn is actìvated by a protein complex called the inflammasome. As it has been demonstrated that EBBP can bind two different proteins of the inflammasome (NALP-1 and caspase 1), we assumed that EBBP plays a role in the regulation of inflammation and that the inflammasome, which has as yet only been described in ínflammatory cells, may also exist in keratinocytes. Indeed, I could show in my thesis that the inflammasome components are expressed in human keratinócytes at the RNA and protein level and also in vivo in human epidermis. After irradiation with a physiological dose of UVB, keratinocytes activated prolL-1ß and secreted prolL-1 a, IL-1 ß, prolL-18 and inflammasome proteins, although all these proteins lack a classical signal peptide. The secretion was dependent on caspase-1 activity, but not on de novo protein synthesis. Knock-down of NALP1 and -3, caspase-1 and -5, EBBP and Asc strongly reduced the secretion of IL-1 ß, demonstrating that also in keratinocytes inflammasome proteins are directly involved in maturation of this cytokine. These results demonstrate for the first time the presence of an active inflammasome in non-professional immune cells. Moreover, they show that UV irradiation is a stimulus for inflammasome activation in keratinocytes. For the analysis of the ín vivo functions of EBBP, transgenic mice overexpressing EBBP in the epidermis were generated. To examine the influence of EBBP overexpression on inflammatory processes, we subjected the mice to different challenges, which induce inflammation. Wound-healing, UVB irradiation and delayed hypersensitivity were tested, but we did not observe any phenotype in the K14-EBBP mice. Besides, a conditional ebbp knockout mouse has been obtained, which will allow to determine the effects of EBBP gene deletion in different tissues and organs. RESUME: EBBP est un membre encore mal connu de la famille des RBCC/TRIM (RING finger B-box coiled-coil/tripartite motif). Il est exprimé de manière ubiquitaire, et en particulier dans les kératinocytes. EBBP étant capable d'interagir avec la prointerleukine-1 ß (prolL-1 ß) dans les macrophages et les kératinocytes humains et de réguler la sécrétion de l'IL-1 ß, il est très probable que cette protéine est impliquée dans l'inflammation. L'activation et la sécrétion de l'IL-1 ß requièrent le clivage protéolytique de son précurseur prolL-1ß par la caspase-1, qui est elle-même activée par un complexe protéique appelé l'inflammasome. Comme il a été démontré qu'EBBP peut lier deux protéines de l'inflammasome (NALP-1 et caspase-1), nous avons émis l'hypothèse qu'EBBP joue un rôle dans la régulation de l'inflammation et que l'inflammasome, jusqu'ici décrit exclusivement dans des cellules inflammatoires, existe dans les kératinocytes. En effet, j'ai pu montrer dans ma thèse que les composants de l'inflammasome sont exprimés dans les kératinocytes humains ainsi que in vivo dans l'épiderme humain. Après irradiation avec une dose, physiologique d'UVB, les kératinocytes activent la prolL-1 ß et sécrètent la prolL-1a, l'IL-1 ß, la prolL-18 et des protéines de l'inflammasome, bien que toutes ces protéines soient dépourvues de peptide signal. La sécrétion dépend de la caspase-1 mais pas de la synthèse protéique de novo. Le knock-down de NALP-1 et -3, des caspase-1 et -5, d'EBBP et d'Asc réduit de manière marquée la sécrétion d'IL-1 ß, démontrant que dans les kératinocytes également, les protéines de l'inflammasome sont impliquées directement dans la maturation de cette cytokine. Ces résultats démontrent pour la première fois la présence d'un inflammasome actif dans des cellules immunitaires non professionnelles. De plus, ils montrent que l'irradiation aux UV est un stimulus pour l'activation de l'inflammasome dans les kératinocytes. Pour l'analyse des fonctions d'EBBP in vivo, nous avons généré des souris transgéniques qui surexpriment EBBP dans l'épiderme. En vue d'examiner l'influence de la surexpression d'EBBP sur le processus inflammatoire, nous avons soumis ces souris à differents modèles d'inflammation. Nous avons testé cicatrisation, UVB et hypersensibilité retardée, mais n'avons pas observé de phénotype chez les souris transgéniques. En parallèle, nous avons également généré des souris knock-out pour ebbp qui devraient nous permettre de déterminer les effets de la suppression d'EBBP dans différents tissus et organes.

Transcriptional activation of the utrophin promoter B by a constitutively active Ets-transcription factor.

Duchenne muscular dystrophy is an X-linked genetic disease caused by the absence of functional dystrophin. Pharmacological upregulation of utrophin, the autosomal homologue of dystrophin, offers a potential therapeutic approach to treat Duchenne patients. Full-length utrophin mRNA is transcribed from two alternative promoters, called A and B. In contrast to the utrophin promoter A, little is known about the factors regulating the activity of the utrophin promoter B. Computer analysis of this second promoter revealed the presence of several conserved binding motives for Ets-transcription factors. Using electrotransfer of cDNA into mouse muscles, we demonstrate that a genetically modified beta-subunit of the Ets-transcription factor GA-binding protein potently activates a utrophin promoter B reporter construct in innervated muscle fibers in vivo. These results make the GA-binding protein and the signaling cascade regulating its activity in muscle cells, potential targets for the pharmacological modulation of utrophin expression in Duchenne patients.

Evaluation of the Molluscicidal Properties of Euphorbia splendens var. hislopii (N.E.B.) Latex: Experimental Test in an Endemic Area in the State of Minas Gerais, Brazil

Following the positive results obtained regarding the molluscicidal properties of the latex of Euphorbia splendens that were corroborated in laboratory and field tests under restricted conditions, a field study was conducted in experimental streams located in an endemic area. After recording the average annual fluctuations of vectors in three streams, a solution of E. splendens latex at 12 ppm was applied in stream A, a solution of niclosamide at 3 ppm that was applied in stream B and a third stream (C) remained untreated for negative control. Applications of E. splendens and niclosamide resulted in a mortality of 100% among the snails collected in the streams A and B. No dead snails were found in the negative control stream. A monthly follow-up survey conducted during three consecutive months confirmed the return of vectors to both experimental streams treated with latex and niclosamide. This fact has called for a need to repeat application in order to reach the snails that remained buried in the mud substrate or escaped to the water edge, as well as, newly hatched snails that did not respond to the concentration of these molluscicides. Adults snails collected a month following treatment led us to believe that they had migrate from untreated areas of the streams to those previously treated