A sepse é a principal causa de morte em unidades de terapia intensiva (UTIs) no mundo. A reduzida disponibilidade do aminoácido mais abundante do organismo, a glutamina contribui para o complicado estado catabólico da sepse. No presente estudo investigamos os efeitos da suplementação oral com L-glutamina e L-alanina (GLN+ALA), ambos na norma livre e como dipeptídeo, L-alanil-L-glutamina (DIP), sobre o eixo glutamina-glutationa (GSH), sistema imune, inflamação, proteínas de choque térmico (HSPs) e expressão de genes envolvidos com vias de sinalização proteica em animais endotoxêmicos. Camundongos C57/B6 foram submetidos à endotoxemia (Escherichia coli LPS, 5 mg.kg-1, grupo LPS) e suplementados por 48 horas com L-glutamina (1 g.kg-1) e L-alanina (0,61 g.kg-1, grupo GLN+ALA-LPS) ou 1,49 g.kg-1 de DIP (grupo DIP-LPS). A endotoxemia promoveu depleção da concentração de glutamina no plasma (71%), músculo esquelético (50%) e fígado (49%), quando comparado ao grupo CTRL, sendo restauradas nos grupos DIP-LPS e GLN+ALA-LPS (P<0,05), fato que atenuou a redução da GSH e o estado redox (taxa GSSG/GSH) em eritrócitos circulantes, musculo e fígado (P<0,05). A suplementação em animais endotoxêmicos resultou em uma upregulation dos genes GSR, GPX1 e GCLC no músculo e fígado. A concentração das citocinas plasmáticasTNF-&#945;, IL-6, IL-1&#946; e IL-10 foi atenuada pelas suplementações, bem como a expressão de mRNAs envolvidos com a resposta inflamatória, ativadas pela via do NF-&#954;B(P<0,05). Concomitantemente, verificou-se aumento da capacidade proliferativa de linfócitos T e B circulantes nos grupos GLN+ALA-LPS e DIP-LPS. A expressão de mRNAs e a concentração de HSPs no tecido muscular foi restabelecida pelas suplementações, contudo, a expressão mRNAs relacionados às vias de síntese e degradação proteica foi somente estimulada no tecido hepático(P<0,05). Os resultados do presente estudo demonstram que a suplementação por via oral com GLN+ALA ou DIP podem ser utilizados clinicamente como métodos nutricionais em reverter o quadro de depressão da disponibilidade de glutamina corporal da sepse induzida por LPS, tendo impacto no eixo glutamina-glutationa, sistema imune e inflamatório. / Sepsis is the leading cause of death inintensive care units (ICUs) in the world.The availability ofthe most abundant amino acid in the body, glutamine, is reduced in this situation, fact that contribute to the complicated catabolic state of sepsis. In the present study, we investigated the effects of oral supplementation with L-glutamine and L-alanine (GLN+ALA), both in their free form and as a dipeptide, L-alanyl-L-glutamine (DIP) on glutamine-glutathione axis (GSH), immune and inflammatory system, heat shock proteins (HSPs) expression and gene expressions involved in protein signaling pathways during endotoxemia. C57/B6 mice were subjected to endotoxemia (Escherichia coli LPS, 5 mg.kg-1, LPS group) and supplemented for 48 hours with L-glutamine (1 g.kg-1) plus L-alanine(0.61 g.kg-1, GLN+ALA-LPS group) or 1.49 g.kg-1of DIP (DIP-LPS group). Endotoxemia promoted depletion glutamine concentration in plasma (71%), skeletal muscle (50%) and liver (49%), when compared to the CTRL group, and was restored in the DIP-LPS e GLN+ALA-LPS (P<0.05), fact that attenuate the reduction of GSH and the redox state (GSSG/GSH rate) in circulating erythrocytes, liver and muscle (P<0.05). Supplementations in endotoxemic mice resulted in upregulation of GSR, GCLC and GPX1 genes in muscle and liver. Plasma concentration of TNF-&#945;, IL-6, IL-1&#946; and IL-10 were attenuated by supplementation as well as the expression of mRNAs involved in the inflammatory response, activated by NF&#954;-B pathway (P <0.05). At the same time, high proliferative capacity of circulating T and B lymphocytes GLN+ALA-LPS e DIP-LPS were observed. HSPs (protein and mRNAs) and in muscle were restored by the supplements, however, the mRNAs expression related to the synthesis and degradation of protein pathways was only stimulated in the liver (P <0.05). Our results demonstrate that oral supplementation with GLN+ALA or DIP can be used as clinically nutritional methods to reverse the depression of body glutamine availability during sepsis induced by LPS, impacting on the glutamine-glutathione axis, immune and inflammatory system.

Objetivou-se com este estudo compreender as dinâmicas das temperaturas corporais em fêmeas da raça Nelore, e relacionar os aspectos fisiológicos da termorregulação com as respostas celulares pela expressão de proteínas de choque térmico. O projeto foi desenvolvido no Laboratório de Biometeorologia e Etologia, da Faculdade de Zootecnia e Engenharia de Alimentos da Universidade de São Paulo, Pirassununga-SP. Foram utilizadas 20 vacas Nelore pluríparas, cíclicas, mantidas em sistema de pastejo. O período experimental precedeu a estação de monta de 2013 e terminou na inseminação artificial. Para classificar os animais quanto ao gerenciamento de calor foi monitorada a temperatura vaginal durante seis dias por meio de 16 data loggers (n=16). Com o intuito de melhor compreender o gerenciamento de calor, outras respostas fisiológicas, nomeadamente temperatura retal, caudal, ocular, frequência respiratória e taxa de sudação foram colhidas durante os quatro dias finais do monitoramento da temperatura vaginal. Para expressão gênica relativa colheram-se amostras de sangue (n=20). Foram realizados choques térmicos in vitro a 38ºC, 40ºC e 42ºC por duas horas. Após tratamento térmico obteve-se o pellet de leucócitos para posterior extração do RNA pelo método TRIzol. Foram escolhidas 10 vacas com os melhores resultados de concentração de RNA, 5 pertencentes ao grupo de vacas eficientes quanto ao gerenciamento de calor, e 5 não eficientes (n=10). A RT-PCR foi realizada com o kit SuperScript III. A reação polimerase em cadeia em tempo real (qPCR) ocorreu no aparelho StepOnePlus® Applied Biosystem utilizando como marcador fluorescente o SYBR® Green para os genes alvo validados HSPA1A, HSPD1, HSP90AA1, HIF1A e endógenos ACTB, RPL15 e PPIA. Os dados foram analisados por ANOVA, regressão e correlação do SAS 9.2. As vacas foram classificadas em eficientes e não eficientes por meio de coeficientes angulares, advindos da regressão das temperaturas vaginais durante períodos de ganho e perda de calor. As vacas eficientes apresentaram menores temperaturas retais 37,65ºC (P<0,01) e maior taxa de sudação 528,73 g. m-². h-¹ (P<0,06). A nível celular, o aumento programado in vitro da temperatura não aumentou quantidade de transcritos, promovendo manutenção à 38ºC e 40ºC e declínio à 42ºC. A ordem decrescente da abundância de transcritos foi HSPA1A, HSPD1 e HSP90AA1. Pode-se concluir que vacas Nelore com diferentes gerenciamentos de calor apresentam respostas termorregulatórias diferentes. A HSPA1A apresentou a maior abundância de transcritos sendo considerada como marcador molecular para termotolerância, por ser a mais sensível à temperatura e bem conservada. Não foi observado diferença nas expressões gênicas relativas das proteínas de choque térmico entre os animais classificados quanto ao gerenciamento de calor. / The aim of this study was to understand the dynamics of body temperatures in Nellore females, and relate the physiological aspects of thermoregulation with cellular responses by the expression of heat shock proteins. The project was developed in Biometeorology and Ethology Laboratory, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga-SP. Were used 20 Nellore pluriparous cows, cyclical, kept in grazing system. The experimental period preceded the 2013\' breeding season and ended up in artificial insemination. To classify animals in relation to heat management the vaginal temperature was monitored for six days through 16 data loggers (n=16). In order to better understand the heat management, other physiological responses, including rectal, tail and eye temperatures, respiratory rate and sweating rate were taken during the final four days of vaginal temperature monitoring. For gene expression was harvested blood samples (n=20). in vitro heat shocks were performed at 38ºC, 40ºC and 42ºC during two hours. After the heat treatment was obtained the leukocyte pellet for later RNA extraction by TRIzol method. Ten cows were chosen with the best RNA concentration results, five belonging to the group of cows efficient on heat management and five inefficient (n = 10). RT-PCR was performed with SuperScript III kit. The real-time polymerase chain reaction (qPCR) occurred in StepOnePlus® Applied Biosystems instrument using the SYBR® Green as fluorescent marker to the target validated genes HSPA1A, HSPD1, HSP90AA1, HIF1A and the endogenous ACTB, RPL15 and PPIA. Data were analyzed using ANOVA, regression and correlation SAS 9.2. The cows were classified as efficient and inefficient through angular coefficients, derived from regression of vaginal temperatures for the gain and heat loss periods. Efficient cows had lower rectal temperature 37.65ºC (P<0.01), higher sweating rate 528.73 g. m ². h-¹ (P<0.06). At cellular level, the increase of in vitro programmed temperature has not increased the transcripts amount, promoting the maintenance at 38ºC and 40ºC and decline at 42ºC. The decreasing order of transcripts amount was HSPA1A, HSPD1 and HSP90AA1. It can be concluded that Nellore cows with different heat managements has different thermoregulatory responses. The HSPA1A showed the highest transcripts abundance being considered as a molecular marker for thermotolerance, for being more sensitive to temperature and better conserved. There was no difference in gene expression for heat shock proteins between animals classified by heat management.

O objetivo deste trabalho foi quantificar as concentrações de heat shock protein (HSP) 27 e 70 no músculo Longissimus thoracis bovino e avaliar o perfil proteico das amostras no decorrer do processo de maturação, bem como, a relação com as características de qualidade da carne. Foram utilizados 191 animais ½ Simental Sul Africano x ½ Nelore (½ S x ½ N), castrados e terminados em confinamento. Para qualidade da carne mensurou-se os valores de pH, cor (L*, a* e b*), perda de água por cocção (PAC) e força de cisalhamento (FC) em dois tempos de maturação (um e 14 dias). Para a quantificação de HSP 27 e 70 foram selecionadas 40 amostras em função dos valores de FC, e separadas em dois grupos (mais e menos macia) com 20 amostras cada, dentro de cada tempo de maturação (48h e 14 dias). Para realização da análise proteômica foram selecionados três animais de cada grupo e tempo de maturação. A concentração de HSP 27 apresentou diferença significativa (P &lt; 0,05) entre os tempos de maturação dentro dos dois grupos estudados. Já para a HSP 70 verificou-se diferença significativa (P &lt; 0,05) apenas para as amostras do grupo mais macia. A análise de correlação mostrou que as HSP 27 e 70 não interferiram nas características de qualidade da carne. A análise proteômica identificou seis proteínas diferencialmente abundantes (P &lt; 0,05) e o enriquecimento funcional demonstrou que essas proteínas estão relacionadas com processos biológicos e componentes celulares. A desmina e a glicerol-3-fosfato desidrogenase demonstraram correlação com a FC e coloração da carne, respectivamente. Os resultados indicam que as HSP 27 e 70 não podem ser apontadas como biomarcadores eficientes para qualidade de carne em animais ½ S x ½ N. Já a glicerol-3-fosfato desidrogenase demonstrou forte potencial em predizer a estabilidade da cor da carne. Desmina pode ser apontada como um importante biomarcador candidato associado à maciez da carne em animais ½ S x ½ N. / The aim of this work was to quantify the concentrations of heat shock protein (HSP) 27 and 70 in Longissimus thoracis muscle and evaluate the protein profile of the samples during the aging process and its relationship with meat quality characteristics. A total of 191 steers ½ Simmental South African x ½ Nellore (½ S x ½ N) feedlot-finished, were used in this work. For meat quality, pH values, color values (L *, a * and b *), cooking loss (CL) and shear force (SF) were measured in two aging times (one and 14 days). For quantification of HSP 27 and 70, 40 samples were selected according to the SF values and separated into two groups (more and less tender) with 20 samples each, according to the aging time (one and 14 days). To perform the proteomic analysis were selected three animals from each group and aging time. The concentration of HSP 27 showed a significant difference (P &lt; 0.05) between aging times within both groups. As for the HSP 70, there was a significant difference (P &lt; 0.05) for the tenderness samples. Correlation analysis showed that the HSP 27 and 70 didn\'t affect the meat quality traits. The proteomic analysis identified six proteins differentially expressed (P &lt; 0.05) and functional enrichment analysis demonstrated that these proteins are associated with biological processes and cellular components. Desmin and glycerol-3-phosphate dehydrogenase showed correlation with meat traits of SF and color, respectively. The results indicate that HSP 27 and 70 could not be identified as effective biomarkers for meat quality in ½ S x ½ N. However, glycerol-3-phosphate dehydrogenase showed a strong potential in predicting meat color stability. Desmin can be shown as a candidate biomarker associated with meat tenderness in ½ S x ½ N.

Dans le contexte du changement climatique, l’augmentation de la fréquence et de l’intensité des canicules a un impact négatif sur la physiologie des plantes, du fait de la perturbation structurale des protéines et des membranes par les fortes températures. Au cours de cette thèse, nous avons développé et caractérisé un système expérimental original dans lequel des plantules d’Arabidopsis thaliana sont arrêtées dans leur développement du fait d’une carence minérale. Ces plantules présentent une grande plasticité métabolique, notamment au niveau énergétique, qui leur permet de survivre pendant des semaines en état stationnaire. Nous avons ensuite réalisé une étude intégrative des processus qui permettent à ces plantules de survivre à un stress thermique normalement létal (43°C, 2 h), grâce à l’application d’un prétraitement à une température non létale (38°C, 2 h). Le pré-traitement à 38°C permet la protection du métabolisme énergétique ainsi que la reprise de la dynamique des organites lors de la phase de récupération. Au niveau transcriptionnel, les plantules pré-acclimatées surexpriment notamment des protéines chaperones et des gènes impliqués dans la photosynthèse et la régulation de l’expression des génomes mitochondriaux et plastidiaux. Au niveau protéique, l’accumulation d’HSPs et d’autres protéines de stress favorisent la récupération des plantules, tandis qu’en l’absence d’acclimatation, le choc thermique provoque la diminution des protéines ribosomales et l’accumulation de protéines impliquées dans la dégradation des protéines. Cette étude illustre l’intérêt de l’analyse multi-échelle pour décrypter les mécanismes de réponse des plantes aux stress. / In the context of climate change, the increased frequency and intensity of heat waves will likely have a negative impact on plant physiology, due to the structural destabilization of proteins and membranes caused by high temperatures. As part of this thesis, we developed and characterized an original experimental setup in which Arabidopsis thaliana seedlings are arrested in their development because of mineral starvation. These seedlings exhibit a high metabolic plasticity, especially for energy metabolism, which allows them to survive in a steady state for weeks. Then, we performed an integrative analysis of the processes that allow these seedlings to survive an otherwise lethal heat stress (43°C, 2 h), thanks to a priming treatment at a nonlethal temperature (38°C, 2 h). Priming protects the energy metabolism and permits the recovery of organelle dynamics after stress. At the transcriptional level, primed seedlings overexpress many chaperone proteins and genes involved in photosynthesis, and in the regulation of the expression of mitochondrial and plastidial genomes. At the protein level, the accumulation of HSPs and other stress proteins favour seedling recovery, whereas in the absence of acclimation, heat shock provokes the decrease of ribosomal proteins and the accumulation of proteins implicated in protein degradation. This study highlights the relevance of multi-scale analysis to decipher mechanisms of stress response in plants.

Inledning: Det högkänsliga draget eller den högkänsliga personen (HSP) är en del av personlighetspsykologin och myntades först av Elaine Aron. Ämnet och draget är relativt outforskat och det finns fortfarande områden att belysa. Syftet med arbetet är att undersöka ett av dessa områden och det är psykoterapeuters erfarenhet av det högkänsliga draget i psykoterapi.  Frågeställning: Vad har psykodynamiskt orienterade psykoterapeuter för erfarenheter av det högkänsliga draget i en psykoterapi?  Metod: Fem legitimerade psykoterapeuter med erfarenhet av området valdes ut genom ett bekvämlighetsurval samt snöbollsmetod. För att besvara studiens fråga användes en kvalitativ metod med en induktiv ansats, samt semistrukturerade frågor och öppna fördjupningsfrågor. Intervjumaterialet strukturerades enligt en tematisk analys.  Resultat: De intervjuande psykoterapeuterna beskrev högkänsliga draget som ett resultat av individens uppväxtmiljö och omständigheter i livet. Det är uttrycket för negativa livsomständigheter, vilka går att arbeta med i en terapi. En integrering av kropp och själ blir i detta sammanhang ett mål för att uppnå en psykisk och fysisk hälsa hos personer med det högkänsliga draget. På grund av studiens storlek bör inga generella slutsatser dras rörande det högkänsliga draget, utan bör enbart vara begränsad till denna studie.  Diskussion: Psykoterapeuters erfarenheter kring det högkänsliga draget visade sig dela mycket av redan existerande föreställningar. Troligen har det sin grund i att definitionen av draget redan finns i befintlig litteratur och haft en viss mediaexponering, men kan även berott på att författaren haft en förförståelse om draget som kan haft en påverkan på respondenterna i studien. Att kroppen och sexualitet kom upp som ett ämne skapade nya frågor som tyvärr inte fick tillräcklig med plats i denna studie. Ämnet som sådant är intressant att studera vidare. Då studien enbart berör fem psykoterapeuters erfarenheter om det högkänsliga draget, bör inte några generella slutsatser dras rörande draget. Resultaten som presenteras bör endast begränsas till denna studie. / Introduction: The Highly Sensitive Trait or Highly Sensitive Person (HSP) is part of personality psychology and was first coined by Elaine Aron. The subjects and the trait is relatively unexplored and there are still areas to highlight. The purpose of this work is to investigate one of these areas and it is psychotherapists experience of patients with the highly sensitive trait.  Question: What are psychodynamic oriented psychotherapists’ experience of the highly sensitive trait in a psychotherapeutic context?  Method: Five licensed psychotherapists with experience in the area were selected through a convenience selection and snowball method. To answer the study’s question. A qualitative method with an inductive approach was used, with semi-structured questions and open in-depth questions. The interview material was structured according to a thematic analysis.  Results: The interviewed psychotherapists described the highly sensitive trait as a result of the individuals upbringing environment and circumstances in life. It is the expression of negative life circumstances, which can be worked with in a therapy. In this context, the integration of the body and mind becomes a goal for achieving mental and physical health in people with the highly sensitive trait. Due to the size of the study no general conclusions should be drawn regarding the highly sensitive trait, but should be limited to this study only.  Discussion: Psychotherapists ́ experience of the highly sensitive trait turned out to share much of already existing perceptions. This was probably due to the fact that the definition of the trait already exists in literature and has had a certain media exposure. But it may also be due to the authors pre-existing understanding of the trait that may have had an impact on the respondents in the study. The fact that the body and sexuality came up as a topic created new questions that unfortunately did not got enough space in this study. The subject is however interesting to study further. As the study is only based on the experience of five psychotherapists about the highly sensitive trait, no general conclusions should be drawn about the trait. The results should only be limited to this study.

Helenaskolan är ett projekt som utmanar hur skolor utformas idag och eftersträvar inkludering av de personer som vanligtvis inte prioriteras i den byggda skolmiljön. Skolans form har inspirerats av generella behov inom den byggda miljön hos barn med HSP och barn inom autismspektrat.  Många av dessa anpassningar grundar sig i möjligheten att själv anpassa storleken på rummet och på den sociala gruppen efter vad som fungerar bäst för ens egen inlärning. Framförallt fick basrummet ett stort fokus, då det är en utav de läromiljöer en elev idag ägnar mest inlärningstid i. Att skapa miljöer som behandlar kontrasten mellan lugn - aktivitet, natur - byggd miljö, socialt - icke socialt, litet - stort etc. var nödvändigt för att kunna skapa dessa anpassningsbara rum. Denna gradvisa förändring i storlek och krav på socialt engagemang blir tydlig om man ser till flödet utifrån och in, där de yttre funktionerna är större, mer socialt utmanande rum och där de inre är mindre, mer intima rum. / Helena School is a project that challenges the nowadays usual way of making schools for extrovert, active children and instead focuses on the pupil’s possibilities to choose the size of both the physical and social rooms of learning. The design of the spaces was very much inspired by analyses of the needs of children with HSP and children in the autism spectrum.  The intention was that every pupil should have the possibility to adapt the size of the room and of the social group to what suits one’s learning best. The homeroom is one of the most used rooms, and therefore it was given much thought. To create environments that address the contrast between stillness - activity, nature - built environment, social - non-social, small - large etc. was necessary to be able to create these adaptable rooms. The change in size and demand of social engagement is obvious when looking to the flow from the outside of the building to the inner parts of it. The outer functions are larger and more socially challenging as opposed to the inner rooms that are both smaller and more intimate.

Most quantum algorithms that are efficient as opposed to their equivalent classical algorithms are solving variants of the Hidden Subgroup Problem (HSP), therefore HSP is a central problem in the field of quantum computing. In this thesis, we offer some interesting results about the subgroup and coset structure of certain groups, including the dihedral group. We describe classical algorithms to solve the HSP over various abelian groups and the dihedral group. We also discuss some existing quantum algorithms to solve the HSP and give our own novel algorithms and ideas to approach the HSP for the dihedral groups.

Exposure of cells to environmental or chemical stressors will initiate the heat shock response, which is mediated by heat shock proteins. Heat shock proteins are molecular chaperones which are classified by size into six main families: HSP100, HSP90, HSP70, HSP60, HSP40 and the small heat shock proteins (sHsps). The sHsp family members bind to denatured proteins and maintain them in a folding competent state such that they may be refolded by other molecular chaperones. <br /><br /> The present study examined the expression and function of two amphibian sHsps, namely, <em>Rana catesbeiana</em> HSP30 and <em>Xenopus laevis</em> HSP27. Initially, an antisense riboprobe was produced to study the mRNA accumulation of <em>Rana hsp30</em> in cultured tongue fibroblast (FT) cells. Results showed that <em>Rana hsp30</em> mRNA was optimally induced when maintained at 35&deg;C for 2 h. An antibody to the recombinant <em>Rana</em> HSP30 protein was also produced in order to study HSP30 protein accumulation in <em>Rana</em> FT cells. Analysis showed that <em>Rana</em> HSP30 was heat-inducible and accumulated maximally at 4 h when maintained at 35&deg;C and then allowed to recover at 22&deg;C for 2 h. Immunocytochemical analysis indicated that <em>Rana</em> HSP30 protein was present primarily in the nucleus, with diffuse localization in the cytoplasm. Additional immunocytochemical analysis showed that <em>Rana</em> HSP30 remained in the nucleus following heat stress and extended periods of recovery. <br /><br /> The molecular chaperone function of <em>Rana</em> HSP30 was also studied. Recombinant <em>Rana</em> HSP30 was found to inhibit the heat induced aggregation of various target proteins including citrate synthase, luciferase and malate dehydrogenase. Also, no major difference was detected between <em>Rana</em> HSP30 and <em>Xenopus</em> HSP30C in the inhibition of heat-induced aggregation of target proteins. <br /><br /> This study also examined the expression and function of <em>Xenopus laevis</em> HSP27. Analysis of the putative amino acid sequence of the <em>Xenopus hsp27</em> cDNA revealed that it had an identity of 71% with chicken, 65% with zebrafish, 63% with human and 53% with topminnow. Most of the identity was located within the &alpha;-crystallin domain of the protein. Interestingly, <em>Xenopus</em> HSP27 shared only a 19% identity with 2 other <em>Xenopus</em> sHsps, HSP30C and HSP30D. <br /><br /> Western blot analysis using an anti-<em>Xenopus</em> HSP27 antibody revealed that HSP27 was not detectable in cultured kidney epithelial cells. However, examination of early <em>Xenopus</em> embryos revealed that HSP27 was first detected in tadpole embryos (stage 44). Heat-inducible HSP27 was also first detected at this stage. The accumulation pattern of <em>Xenopus</em> HSP27 protein was distinct from <em>Xenopus</em> HSP30, which was heat-inducible at midtailbud stage 26, approximately two and a half days earlier in development. <br /><br /> Analysis of recombinant HSP27 by native pore exclusion limit electrophoresis showed that it formed high molecular weight, multimeric complexes. The molecular chaperone function of HSP27 was assessed by means of thermal aggregation assays employing citrate synthase, luciferase and malate dehydrogenase. <em>Xenopus</em> HSP27 inhibited the heat-induced aggregation of all of these target proteins. This study has revealed that <em>Xenopus</em> HSP27 is a member of the HSP27 subfamily of small heat shock proteins in <em>Xenopus</em> and distinct from the HSP30 family. The accumulation of HSP27 under constitutive and stress-inducible conditions is developmentally regulated. Finally, this sHsp appears to function as a molecular chaperone.

Systemic scleroderma (SSc) is a systemic connective tissue disease affecting skin and internal organs. The pathogenesis of SSc is characterized by inflammation, vasculopathy and fibrosis. No agent has been proven effective in the treatment of SSc. There is a lack of suitable biomarkers for monitoring the disease activity or the response to the treatment of SSc. Therefore our aim was to analyse the extracellular levels of S100A4, Hsp90 (Heat shock protein 90) and IL-35 (interleukin-35) in SSc. S100A4 and Hsp90 have been initially studied in tumours; in some of them considered as suitable prognostic markers and candidates for future therapies. We have recently described the profibrotic role of S100A4 and Hsp90 in the pathogenesis of SSc. Our results showed that inactivation of S100A4 and Hsp90 effectively prevented the development of experimental skin fibrosis. This was consequently confirmed by the analysis of S100A4 and Hsp90 in the peripheral blood of patients with SSc, where significant associations with disease activity and organ involvement were detected. IL-35 may become another potential biomarker of SSc. We detected increased expression of IL-35 in the affected skin, dermal fibroblasts and in serum of patients with SSc. Moreover, the main profibrotic mediator transforming growth factor...

High temperature stress (HTS) adversely affects reproduction in most plant species studied to date. HTS during flowering may result in an almost total inhibition of seed production in crop plants. Increasing our knowledge of the effects of HTS on seed production will aid the breeding of more thermotolerant crop plants and improve our understanding of the effects of stress on plants. An investigation of the effects of both drought and high temperature stress on the yields of barley, canola, flax, durum and spring wheat in five locations in Saskatchewan over a 25-year period was performed using multivariate analysis. Higher temperatures during June and July, when the plants were flowering, were correlated with reductions in yields of all the crops studied (except barley in June). A positive correlation between yields and precipitation during May and the winter preceding the growing season was observed.<p>In growth chambers, <i>Brassica napus</i> silique and seed production were inhibited during a ramping HTS treatment. This was due to a decrease in pollen germinability rather than a reduction in the number of flowers produced. HTS also caused reductions in megagametophyte fertility and disrupted embryo and/or seed development.<p>Transgenic plants were developed to overcome the effects of HTS on seed production. Two DNA constructs, one with the <i>Arabidopsis thaliana LEAFY</i> (<i>AtLFY</i>) promoter controlling <i>A. thaliana HEAT SHOCK PROTEIN 101</i> (<i>AtHSP101</i>) ORF expression and another with the <i>AtHSP101</i> promoter controlling <i>AtLFY</i> ORF expression, were inserted into <i>B. napus</i>. Other DNA constructs were made, using the constitutively expressed Cauliflower Mosaic Virus <i>35S</i> or the synthetic <i>EntCup4</i> promoters to control expression of the <i>AtHSP101</i> or <i>A. thaliana HEAT SHOCK TRANSCRIPTION FACTOR 3</i> (<i>AtHSF3</i>) ORFs. These constructs were inserted into both <i>B. napus</i> and <i>A. thaliana</i>. Transgenic plants were tested using a ramping temperature regime but were found not to have increased flower thermotolerance. During the manufacture of the DNA constructs it was determined that, in <i>A. thaliana</i>, 573 bp of <i>AtHSP101</i> had been copied between Terminal Inverted Repeats of a <i>Mu-Like Element</i> (<i>MULE</i>). This fragment was named <i>HSP101B</i>. In some transgenic <i>B. napus</i> and <i>A. thaliana</i> lines, containing 2046 bp of the <i>HSP101B</i> upstream regulatory region controlling <i>B</i>-glucuronidase (GUS) expression, cold-inducible GUS expression was observed. Methylation may have a role in control of endogenous <i>HSP101B</i> transcription.