A human immune data-informed vaccine concept elicits strong and broad T-cell specificities associated with HIV-1 control in mice and macaques

Background: None of the HIV T-cell vaccine candidates that have reached advanced clinical testing have been able to induce protective T cell immunity. A major reason for these failures may have been suboptimal T cell immunogen designs. Methods: To overcome this problem, we used a novel immunogen design approach that is based on functional T cell response data from more than 1,000 HIV-1 clade B and C infected individuals and which aims to direct the T cell response to the most vulnerable sites of HIV-1. Results: Our approach identified 16 regions in Gag, Pol, Vif and Nef that were relatively conserved and predominantly targeted by individuals with reduced viral loads. These regions formed the basis of the HIVACAT T-cell Immunogen (HTI) sequence which is 529 amino acids in length, includes more than 50 optimally defined CD4+ and CD8+ T-cell epitopes restricted by a wide range of HLA class I and II molecules and covers viral sites where mutations led to a dramatic reduction in viral replicative fitness. In both, C57BL/6 mice and Indian rhesus macaques immunized with an HTI-expressing DNA plasmid (DNA.HTI) induced broad and balanced T-cell responses to several segments within Gag, Pol, and Vif. DNA.HTI induced robust CD4+ and CD8+ T cell responses that were increased by a booster vaccination using modified virus Ankara (MVA.HTI), expanding the DNA.HTI induced response to up to 3.2% IFN-γ T-cells in macaques. HTI-specific T cells showed a central and effector memory phenotype with a significant fraction of the IFN-γ+ CD8+ T cells being Granzyme B+ and able to degranulate (CD107a+). Conclusions: These data demonstrate the immunogenicity of a novel HIV-1 T cell vaccine concept that induced broadly balanced responses to vulnerable sites of HIV-1 while avoiding the induction of responses to potential decoy targets that may divert effective T-cell responses towards variable and less protective viral determinants.

Biotransformation and endocrine disruptive effects of contaminants in ringed seals - implications for monitoring and risk assessment

Marine mammals are exposed to persistent organic pollutants (POPs), which may be biotransformed to metabolites some of which are highly toxic. Both POPs and their metabolites may lead to adverse health effects, which have been studied using various biomarkers. Changes in endocrine homeostasis have been suggested to be sensitive biomarkers for contaminant-related effects. The overall objective of this doctoral thesis was to investigate biotransformation capacity of POPs and their potential endocrine disruptive effects in two contrasting ringed seal populations from the low contaminated Svalbard area and from the highly contaminated Baltic Sea. Biotransformation capacity was studied by determining the relationships between congener-specific patterns and concentrations of polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers (PBDEs) and their hydroxyl (OH)- and/or methylsulfonyl (MeSO₂)-metabolites, and catalytic activities of hepatic xenobiotic-metabolizing phase I and II enzymes. The results suggest that the biotransformation of PCBs, PBDEs and toxaphenes in ringed seals depends on the congener-specific halogen-substitution pattern. Biotransformation products detected in the seals included OH-PCBs, MeSO₂-PCBs and –DDE, pentachlorophenol, 4-OHheptachlorostyrene, and to a minor extent OH-PBDEs. The effects of life history state (moulting and fasting) on contaminant status and potential biomarkers for endocrine disruption, including hormone and vitamin homeostasis, were investigated in the low contaminated ringed seal population from Svalbard. Moulting/fasting status strongly affected thyroid, vitamin A and calcitriol homeostasis, body condition and concentrations of POPs and their OH-metabolites. In contrast, moulting/fasting status was not associated with variations in vitamin E levels. Endocrine disruptive effects on multiple endpoints were investigated in the two contrasting ringed seal populations. The results suggest that thyroid, vitamin A and calcitriol homeostasis may be affected by the exposure of contaminants and/or their metabolites in the Baltic ringed seals. Complex and non-linear relationships were observed between the contaminant levels and the endocrine variables. Positive relationships between circulating free and total thyroid hormone concentration ratios and OH-PCBs suggest that OH-PCBs may mediate the disruption of thyroid hormone transport in plasma. Species differences in thyroid and bone-related effects of contaminants were studied in ringed and grey seals from low contaminated references areas and from the highly contaminated Baltic Sea. The results indicate that these two species living at the same environment approximately at the same trophic level respond in a very different way to contaminant exposure. The results of this thesis suggest that the health status of the Baltic ringed seals has still improved during the last decade. PCB and DDE levels have decreased in these seals and the contaminant-related effects are different today than a decade ago. The health of the Baltic ringed seals is still suggested to be affected by the contaminant exposure. At the present level of the contaminant exposure the Baltic ringed seals seem to be at a zone where their body is able to compensate for the contaminant-mediated endocrine disruption. Based on the results of this thesis, several recommendations that could be applied on monitoring and assessing risk for contaminant effects are provided. Circulating OH-metabolites should be included in monitoring and risk assessment programs due to their high toxic potential. It should be noted that endogenous variables may have complex and highly variable responses to contaminant exposure including non-linear responses. These relationships may be further confounded by life history status. Therefore, it is highly recommended that when using variables related to endocrine homeostasis to investigate/monitor or assess the risk of contaminant effects in seals, the life history status of the animal should be carefully taken into consideration. This applies especially when using thyroid, vitamin A or calcitriolrelated parameters during moulting/fasting period. Extrapolations between species for assessing risk for contaminant effects in phocid seals should be avoided.

Estudo químico e atividade antibacteriana do caule de Aristolochia esperanzae kuntze (Aristolochiaceae)

From the ethanolic extract of the stem of A. esperanzae ethyl and methyl fatty acid esters, fatty acids, aristolochic I and II acids, and β-cubebin were isolated. In addiction asarinin, populifolic and 2-oxo-populifolic acids, aristolactams AIa and AII, and sitosterol 3-O-β-D-glucopyranoside were also isolated and firstly described in the species. Asarinin and β-cubebin showed antibacterial activity against Bacillus cereus and aristolochic acid I against Staphylococcus aureus and Listeria monocitogenes.

Copper electrodeposition on glassy carbon and highly oriented pyrolytic graphite substrates from perchlorate solutions

In present work, we analyzed the copper electrodeposition onto GCE (System I) and HOPGE (System II) from perchlorate solutions. The current density transients obtained from system I and II were well described through a kinetic mechanism that involves four different contributions: (a) a Langmuir type adsorption process, b) an electron transfer from Cu2+→Cu+, (c) a 3D nucleation limited by a mass transfer reaction and (d) a proton reduction process. It was observed that the values of the nucleation rate, the number of active nucleation sites were increased with the overpotential and they are bigger onto GCE in comparison with HOPGE.

Preparative separation of C19-diterpenoid alkaloids from Aconitum carmichaelii Debx by pH‑zone-refining counter-current chromatography

The technique of pH-zone-refining counter-current chromatography was successfully applied to preparatively separate three C19-diterpenoid alkaloids from the crude extracts of Aconitum carmichaelii for the first time using a two-phase solvent system of petroleum ether-ethyl acetate-methanol-water (5:5:1:9, v/v/v/v). Mesaconitine (I), hypaconitine (II), and deoxyaconitine (III) were obtained from 2.5 g of the crude alkaloids in a one-step separation; the yields were 4.16%, 16.96%, and 5.05%, respectively. The purities of compounds I, II, and III were 93.0%, 95%, and 96%, respectively, as determined by HPLC. The chemical structures of the three compounds were identified by electrospray ionization mass spectrometry (ESI-MS) and NMR.

Freqüência de biovares de ralstonia solanacearum em diferentes cultivares e épocas de cultivo de batata no rio grande do sul

A presença das biovares I e/ou II de Ralstonia solanacearum em uma lavoura de batatas (Solanum tuberosum) tem influência direta no sucesso das medidas adotadas para controlar a murcha bacteriana. As biovares diferem entre si em relação à agressividade, latência e sobrevivência. Assim, um experimento de campo foi conduzido em uma área naturalmente infestada em duas épocas de cultivo com os objetivos de verificar (1) a incidência de biovares I e/ou II, (2) a relação entre biovar e época de plantio e (3) a relação entre biovar e cultivar de batata. Os isolados obtidos de plantas das cultivares Achat, Baronesa, Elvira, Macaca, Monte Bonito e Trapeira foram identificados como biovar I ou II através da PCR, utilizando os oligonucleotídeos iniciadores T3A e T5A. Ambas as biovares foram encontradas na área naturalmente infestada. De 73 isolados de R. solanacearum, 94,5% foram identificados como biovar II e 5,5% como biovar I. A biovar II foi isolada dos cultivos de primavera e de outono, independente da cultivar, mas a I apenas do cultivo de primavera e de plantas assintomáticas das cultivares Achat e Macaca. A maior população da biovar I nestas duas cultivares pode ser uma evidência da possível relação entre biovar e cultivar.

Vasa Svenska Lyceums bibliotek

Biblioteket härstammar från i huvudsak två skolor, Wasa trivialskola (grundad 1684) och Wasa gymnasium (grundad 1844). Skolornas bibliotek byggdes upp med donationer. Vid Vasa brand 1852 förstördes största delen av gymnasiets stora boksamling. Av 6.000 böcker kunde bara 1.000 räddas. Biblioteket kunde ändå snabbt byggas upp igen med nya donationer, byten och inköp. Åren 1872-74 sammanslogs trivialskolan och gymnasiet till Vasa Svenska Lyceum. Också boksamlingarna sammanslogs. Biblioteket fungerade som ett lånebibliotek ända till 1974 då lyceet upphörde med sin verksamhet i samband med införandet av grundskolan. Skolan ändrades till Vasa Övningsskola som en del av lärarutbildningen vid Åbo Akademi i Vasa. Det gamla skolbiblioteket blev nu endast en historisk samling. Biblioteket hade 1974, 52.140 volymer (årsberättelsen 1973-1974). I denna siffra ingår även Ranckens samling (Se skild beskrivning). Under årens lopp har material avskrivits. En liten del av den nyare litteraturen finns deponerad på Övningsskolans gymnasiums bibliotek. Tidskrifter och tidningar har förkommit vid flyttningar. Biblioteket har idag ca. 25.000 volymer böcker (640 hm). Dessutom finns ca. 6 hm kartor och planschverk, 26,5 hm avhandlingar från 1900-talet samt ett hundratal främst inhemska dagstidningar från 1800-talet. Av de äldsta och raraste böckerna har ca. 4.000 katalogiserats i Tria. Äldsta böckerna är från medlet av 1500-talet och de nyaste från 1970-talet. Biblioteket är till sitt innehåll akademiskt och humanistiskt med tyngdpunkten på teologi (12 %), historia (23%), filologi och litteratur (30%). Största delen av böckerna är på svenska, tyska och latin. En del finns också på grekiska och hebreiska samt en mindre del på finska, franska, engelska och ryska, framförallt i klassen filologi och litteratur.

Efeito de frações parcialmente purificadas de Saccharomyces cerevisiae na germinação de conídios e formação de apressórios por Colletotrichum sublineolum e Colletotrichum lagenarium

O trabalho teve por objetivo verificar o efeito de preparações ou de frações parcialmente purificadas obtidas de S. cerevisiae, autoclavada por 4 horas seqüencialmente, submetidas à cromatografia de troca iônica (CTI) utilizando tampão Tris-HCl ou bicarbonato de amônio, na germinação de esporos (GE) e formação de apressórios (FA) in vitro por C. lagenarium ou C. sublineolum. Para isto, 40 µL de cada preparação ou fração foram colocados em pocinhos de placa de ELISA, juntamente com 40 µL de uma suspensão de esporos (1 x 10(5) conídios/mL) de C. lagenarium ou de C. sublineolum. Após incubação, determinou-se GE e FA. Água destilada esterilizada foi utilizada como controle. Todas as preparações da levedura autoclavada promoveram estímulo da GE, sem a formação de apressórios por ambos os fitopatógenos. Frações provenientes da CTI, com tampão Tris-HCl, induziram a GE por C. sublineolum e C. lagenarium. Na FA de C. lagenarium houve estímulo pelas frações IV, V e VI, sem diferença, no entanto, na FA de C. sublineolum. Para as frações obtidas por CTI, utilizando tampão bicarbonato de amônio, houve estímulo da GE por C. lagenarium nas frações I e IV e efeito inibitório da germinação pelas frações V, VI e VII. Não houve FA na fração I e as demais frações apresentaram efeito inibitório da FA por C. lagenarium. As frações I e II estimularam a GE e a FA por C. sublineolum e demais frações apresentaram efeito inibitório. Assim, evidencia-se a importância da escolha de tampões no processo de purificação de frações de S. cerevisiae, o que pode resultar em frações que estimulem a germinação de esporos de fitopatógenos fúngicos ou em frações com atividade inibitória da germinação, podendo contribuir futuramente no controle de doenças causadas por esses fungos.

Perspectives interculturelles et interlinguistiques sur le discours académique – Crosscultural and Crosslinguistic Perspectives on Academic Discourse. Volume 1

The Department of French Studies of the University of Turku (Finland) organized an International Bilingual Conference on Crosscultural and Crosslinguistic Perspectives on Academic Discourse from 2022 May 2005. The event hosted specialists on Academic Discourse from Belgium, Finland, France, Germany, Italy, Norway, Spain, and the USA. This book is the first volume in our series of publications on Academic Discourse (AD hereafter). The following pages are composed of selected papers from the conference and focus on different aspects and analytical frameworks of Academic Discourse. One of the motivations behind organizing the conference was to examine and expand research on AD in different languages. Another one was to question to what extent academic genres are culturebound and language specific or primarily field or domain specific. The research carried out on AD has been mainly concerned with the use of English in different academic settings for a long time now – mainly written contexts – and at the expense of other languages. Alternatively the academic genre conventions of English and English speaking world have served as a basis for comparison with other languages and cultures. We consider this first volume to be a strong contribution to the spreading out of researches based on other languages than English in AD, namely Finnish, French, Italian, Norwegian and Romanian in this book. All the following articles have a strong link with the French language: either French is constitutive of the AD corpora under examination or the article was written in French. The structure of the book suggests and provides evidence that the concept of AD is understood and tackled to varying degrees by different scholars. Our first volume opens up the discussion on what AD is and backs dissemination, overlapping and expansion of current research questions and methodologies. The book is divided into three parts and contains four articles in English and six articles in French. The papers in part one and part two cover what we call the prototypical genre of written AD, i.e. the research article. Part one follows up on issues linked to the 13 Research Article (RA hereafter). Kjersti Fløttum asks wether a typical RA exists and concentrates on authors’ voices in RA (self and other dimensions), whereas Didriksen and Gjesdal’s article focuses on individual variation of the author’s voice in RA. The last article in this section is by Nadine Rentel and deals with evaluation in the writing of RA. Part two concentrates on the teaching and learning of AD within foreign language learning, another more or less canonical genre of AD. Two aspects of writing are covered in the first two articles: foreign students’ representations on rhetorical traditions (Hidden) and a contrastive assessment of written exercices in French and Finnish in Higher Education (Suzanne). The last contribution in this section on AD moves away from traditional written forms and looks at how argumentation is constructed in students’ oral presentations (Dervin and Fauveau). The last part of the book continues the extension by featuring four articles written in French exploring institutional and scientific discourses. Institutional discourses under scrutiny include the European Bologna Process (Galatanu) and Romanian reform texts (Moilanen). As for scientific discourses, the next paper in this section deconstructs an ideological discourse on the didactics of French as a foreign language (Pescheux). Finally, the last paper in part three reflects on varied forms of AD at university (Defays). We hope that this book will add some fuel to continue discussing diverse forms of and approches to AD – in different languages and voices! No need to say that with the current upsurge in academic mobility, reflecting on crosscultural and crosslinguistic AD has just but started.

The Structural Basis for inorganic Pyrophosphatase Catalysis and Regulation

Inorganic pyrophosphatases (PPases) are essential enzymes for every living cell. PPases provide the necessary thermodynamic pull for many biosynthetic reactions by hydrolyzing pyrophosphate. There are two types of PPases: integral membrane-bound and soluble enzymes. The latter type is divided into two non-homologous protein families, I and II. Family I PPases are present in all kingdoms of life, whereas family II PPases are only found in prokaryotes, including archae. Family I PPases, particularly that from <i>Saccharomyces cerevisiaei>, are among the most extensively characterized phosphoryl transfer enzymes. In the present study, we have solved the structures of wild-type and seven active site variants of <i>S. cerevisiaei> PPase bound to its natural metal cofactor, magnesium ion. These structures have facilitated derivation of the complete enzyme reaction scheme for PPase, fulfilling structures of all the reaction intermediates. The main focus in this study was on a novel subfamily of family II PPases (CBSPPase) containing a large insert formed by two CBS domains and a DRTGG domain within the catalytic domain. The CBS domain (named after cystathionine beta-synthase in which it was initially identified) usually occurs as tandem pairs with two or four copies in many proteins in all kingdoms of life. The structure formed by a pair of CBS domains is also known as a Bateman domain. CBS domains function as regulatory units, with adenylate ligands as the main effectors. The DRTGG domain (designated based on its most conserved residues) occurs less frequently and only in prokaryotes. Often, the domain co-exists with CBS domains, but its function remains unknown. The key objective of the current study was to explore the structural rearrangements in the CBS domains induced by regulatory adenylate ligands and their functional consequences. Two CBS-PPases were investigated, one from <i>Clostridium perfringensi> (<i>cpi>CBS-PPase) containing both CBS and DRTGG domains in its regulatory region and the other from <i> Moorella thermoaceticai> <i>(mti> CBS-PPase) lacking the DRTGG domain. We additionally constructed a separate regulatory region of <i>cpi>CBS-PPase (<i>cpi>CBS). Both full-length enzymes and cpCBS formed homodimers. Two structures of the regulatory region of cpCBS-PPase complexed with the inhibitor, AMP, and activator, diadenosine tetraphosphate, were solved. The structures were significantly different, providing information on the structural pathway from bound adenylates to the interface between the regulatory and catalytic parts. To our knowledge, these are the first reported structures of a regulated CBS enzyme, which reveal large conformational changes upon regulator binding. The activator-bound structure was more open, consistent with the different thermostabilities of the activator- and inhibitor-bound forms of <i>cpi>CBS-PPase. The results of the functional studies on wild-type and variant CBS-PPases provide support for inferences made on the basis of structural analyses. Moreover, these findings indicate that CBS-PPase activity is highly sensitive to adenine nucleotide distribution between AMP, ADP and ATP, and hence to the energy level of the cell. CBS-PPase activity is markedly inhibited at low energy levels, allowing PPi energy to be used for cell survival instead of being converted into heat.

Caracterização genética de Calophyllum brasiliense Camb. em duas populações de mata ciliar

Calophyllum brasiliense Camb. é uma espécie predominante de áreas com alta saturação hídrica, sendo comum nos ambientes cilares do sul de Minas Gerais. Com o intuito de se estudar a variabilidade genética dessa espécie, 20 indivíduos reprodutivos de C. brasiliense foram amostrados de duas populações localizadas a montante do Reservatório Hidroelétrico do Funil, localizado no Município de Lavras,MG. A análise de eletroforese isoenzimática permitiu a obtenção de 17 alelos, distribuídos entre oito locos, sendo estes representados em cinco sistemas enzimáticos: alfa-esterase, beta-esterase, fosfatase ácida, glutamato oxalacetato, malato desidrogenase e transaminase. Os índices de diversidade revelaram um baixo número de alelos por loco em ambas as populações (1,75 e 1,50). A porcentagem de locos polimórficos (P) foi de 37,5% e 50% nas populações I e II, respectivamente. A heterozigosidade média observada foi de 0,119 e 0,111 e a esperada, de 0,131 e 0,112. O número de migrantes (Nm) encontrado entre as populações foi de 2,70. O tamanho efetivo estimado foi de 18 indivíduos para a população I e 19 para a II.

Distribuição de biomassa e nutrientes na aréa de Mimosa caesalpiniaefolia Benth

O trabalho foi realizado na Estação Experimental de Itambé, PE, para avaliar a distribuição da biomassa e nutrientes em povoamentos de Mimosa caesalpiniaefolia Benth. Foram estudados dois povoamentos, em solo e topografia similares com oito e 11 anos de idade, nos espaçamentos de 3,0 x 3,0 m e 4,5 x 4,5 m, respectivamente. A biomassa foi determinada para cada componente aéreo. As amostras de folhas, galhos, cascas e fuste foram coletadas e analisadas quimicamente. A biomassa total encontrada foi de 66,22 e 80,78 t/ha, nos povoamentos I e II, respectivamente. O material lenhoso (galhos e lenho) representou 96,52 e 97,98% da biomassa total, nos povoamentos I e II. A distribuição de biomassa nos povoamentos I e II foi, respectivamente; de galhos (44,99 e 53,40%), fuste (51,53 e 44,58%), casca (2,39 e 1,40%) e folhas (1,10 e 0,62%). A ordem de concentração de nutrientes nos dois povoamentos, em todos os componentes da parte aérea, de modo geral, obedeceu à seguinte ordem decrescente: nitrogênio > cálcio > potássio > magnésio > enxofre > fósforo.

Influência da luz e substrato na germinação e desenvolvimento inicial de duas espécies de Calliandra Benth: (Mimosoideae - Leguminosae) endêmicas da Chapada Diamantina, Bahia

O gênero Calliandra Benth. (Leguminosae) possui 134 espécies exclusivamente neotropicais, das quais 42 ocorrem na Chapada Diamantina, sendo 32 endêmicas da área. O objetivo deste trabalho foi avaliar o efeito da luz e substrato na germinação e desenvolvimento inicial de C. viscidula e C. hygrophila. As sementes foram submetidas a três tratamentos em laboratório: luz, escuro e luz/escuro (12 h/12 h). Em viveiro foram utilizados três substratos: (I) areia; (II) terra vegetal + areia; (III) vermiculita + terra vegetal + areia, mantidos a 100%; e 70% de luminosidade. As avaliações da germinação foram diárias durante 21 dias, utilizando-se os parâmetros: germinabilidade, índice de velocidade de germinação (índice de velocidade de emergência para viveiro), tempo médio de germinação e frequência relativa. Os parâmetros analisados no desenvolvimento inicial foram: comprimentos da parte aérea e raiz, número de folhas, massa seca, relação parte aérea/sistema radicular e razão do peso da folha. C. viscidula e C. hygrophila foram classificadas como fotoblásticas neutras e apresentaram melhor desenvolvimento a 70% de luminosidade nos substratos III e I e II , respectivamente.

Regeneração natural em um remanescente de caatinga com diferentes históricos de uso no agreste pernambucano

O objetivo deste trabalho foi comparar a regeneração natural de duas áreas de vegetação de caatinga com diferentes históricos de uso no agreste pernambucano. O estudo foi realizado em duas áreas: Área I de mata nativa, sem evidências históricas de eliminação total da vegetação para fins de cultivos agrícolas; e Área II, anteriormente ocupada com cultivo da palma forrageira, abandonada há cerca de 30 anos e que se encontrava em estágio de sucessão secundária. Para coleta dos dados da vegetação, foram utilizadas 24 parcelas de dimensões de 5 x 5 m, em 12 parcelas em cada área, sendo considerados indivíduos em regeneração natural as plantas que possuíram circunferência a 1,30 m do solo (CAP) < 6,0 cm e altura >1,0 m. No levantamento estrutural dos indivíduos regenerantes das duas áreas foram identificados 581 plantas pertencentes a 14 famílias botânicas, 26 gêneros e 30 espécies. As densidades totais obtidas neste estudo foram de 11.200 ind.ha-1 e 8.116 ind.ha-1, nas Áreas I e II, respectivamente. As espécies que obtiveram maiores densidades de regeneração natural na Área I foram: Croton argyrophyllus, Senegalia bahiensis, Croton blanchetianus e Coutareahexandra; e na Área II: Senegalia bahiensis, Poincianella pyramidalis, Zanthoxyllum sp, Croton blanchetianus e Croton argyrophyllus.

Avaliação do microclima de instalações para gado de leite com diferentes recursos de climatização

Este trabalho foi realizado no verão de 2002 e teve como objetivo avaliar a influência de alguns recursos de climatização no microclima interno das instalações. Foram utilizadas três instalações: i) controle; ii) com aspersão associada à ventilação, e iii) com tela de sombreamento. Os dados climáticos de cada instalação foram registrados e, posteriormente, calculados os índices de temperatura e umidade, do globo negro e umidade e a entalpia. Foram selecionados e analisados 26 dias com entalpia elevada. Os índices de temperatura e umidade (ITU) e de temperatura do globo e umidade (ITGU), nos horários mais quentes do dia, diferenciaram-se para a instalação climatizada (ii), que apresentou melhor desempenho em relação às demais. A entalpia menor foi observada no tratamento com tela (iii).