Deleterious Actions Of Vegf On The Blood Retinal Barrier And Photoreceptor Survival In The Light-damage Model

Purpose: The retinal balance between pro- and anti-angiogenic factors is critical for angiogenesis control, but is also involved in cell survival. We previously reported upregulation of VEGF and photoreceptor (PR) cell death in the Light-damage (LD) model. Preliminary results showed that anti-VEGF can rescue PR from cell death. Thus, we investigated the role of VEGF on the retina and we herein described the effect of anti-VEGF antibody delivered by lentiviral gene transfer in this model.Methods: To characterize the action of VEGF during the LD, we exposed Balb/c mice subretinally injected with LV-anti-VEGF, or not, to 5'000 lux for 1h. We next evaluated the retinal function, PR survival and protein expression (VEGF, VEGFR1/2, Src, PEDF, p38MAPK, Akt, Peripherin, SWL-opsin) after LD. We analyzed Blood retinal barrier (BRB) integrity on flat-mounted RPE and cryosections stained with β-catenin, ZO-1, N-cadherin and albumin.Results: Results indicate that the VEGF pathway is modulated after LD. LD leads to extravascular albumin leakage and BRB breakdown: β-catenin, ZO-1 and N-cadherin translocate to the cytoplasm of RPE cells showing loss of cell cohesion. This phenomenon is in adequacy with the VEGF time-course expression. Assessment of the retinal function reveals that PR rescue correlates with the level of LV-anti-VEGF expression. Rhodopsin content was higher in the LV-anti-VEGF group than in controls and measures of the ONL thickness indicate that LV-anti-VEGF preserves by 82% the outer nuclear layer from degeneration. Outer segments (OS) appeared well organized with an appropriate length in the LV-anti-VEGF group compared to controls, and the expression of SWL-opsin is maintained in the OS without being mislocalized as in the LV-GFP group. Finally, LV-anti-VEGF treatment prevents BRB breakdown and maintained RPE cell integrity.Conclusions: This study involves VEGF in LD and highlights the prime importance of the BRB integrity for PR survival. Taken together, these results show that anti-VEGF is neuroprotective in this model and maintains functional PR layer in LD-treated mice.

Testing the effects of genetic crossing distance on embryo survival within a metapopulation of brown trout (Salmo trutta)

Predicting progeny performance from parental genetic divergence can potentially enhance the efficiency of supportive breeding programmes and facilitate risk assessment. Yet, experimental testing of the effects of breeding distance on offspring performance remains rare, especially in wild populations of vertebrates. Recent studies have demonstrated that embryos of salmonid fish are sensitive indicators of additive genetic variance for viability traits. We therefore used gametes of wild brown trout (Salmo trutta) from five genetically distinct populations of a river catchment in Switzerland, and used a full factorial design to produce over 2,000 embryos in 100 different crosses with varying genetic distances (FST range 0.005-0.035). Customized egg capsules allowed recording the survival of individual embryos until hatching under natural field conditions. Our breeding design enabled us to evaluate the role of the environment, of genetic and nongenetic parental contributions, and of interactions between these factors, on embryo viability. We found that embryo survival was strongly affected by maternal environmental (i.e. non-genetic) effects and by the microenvironment, i.e. by the location within the gravel. However, embryo survival was not predicted by population divergence, parental allelic dissimilarity, or heterozygosity, neither in the field nor under laboratory conditions. Our findings suggest that the genetic effects of inter-population hybridization within a genetically differentiated meta-population can be minor in comparison to environmental effects.

East 1 St Street Interchange and I-35 Widening, From MP 94.3 to Oralabor Road, Polk County, Iowa, Project # IM-035-4(140)92--13-77, Environmental Assessment and Section 4(f) De Minimis Impact Finding, 2015

From Proposed Action: "The proposed action consists of reconstructing the East 1st Street/I-35 interchange with a Diverging Diamond interchange, widening I-35 from four (4) lanes to six (6) lanes, and widening East 1st Street from four (4) lanes to five (5) lanes from Delaware Avenue to Frisk Drive. The project also proposes to reconstruct the intersections of East 1st Street/Creekview Drive and East 1st Street/Frisk Drive."

West 1 St Street/ IA 57 from Highland Drive to Center Street/Franklin Street, Blackhawk County, Iowa, Project # STP-57-2(28)--2C-07, Environmental Assessment and Section 4(f) De Minimis Impact Finding, 2015

From Proposed Action: "Iowa Northland Regional Council of Governments (INRCOG) and the City of Cedar Falls, in coordination with the Iowa Department of Transportation (Iowa DOT) and the Federal Highway Administration (FHWA), are proposing to upgrade and modernize an approximate 4,900-foot segment of Iowa Highway 57 (IA 57), locally known as West 1st Street, in Cedar Falls, Black Hawk County, Iowa."

Loss of insulin synthesis and beta cell survival induced by oxidized LDL require activation of reticulum endoplasmic stress: a mechanism countered by HDL

Elevated circulating concentrations in modified LDL-cholesterol particles (e.g. oxidised LDL) and low levels in HDL increase not only the risk for diabetic patients to develop cardiovascular diseases but also may contribute to development and progression of diabetes by directly having adverse effects on β-cells. Chronic exposure of β-cells to 2 mM human oxidised LDL-cholesterol (oxLDL) increases the rate of apoptosis, reduce insulin biosynthesis and the secretory capacity of the cells in response to nutrients. In line with the protective role, HDL efficiently antagonised the harmful effects of ox- LDL, suggesting that low levels of HDL would be inefficient to protect β-cells against oxLDL attack in patients. Activation of endoplasmic reticulum (ER) stress is pointed out to contribute to β-cell dysfunction elicited by environmental stressors. In this study we investigated whether activation of ER stress is required for oxLDL to mediate detrimental effects on β-cells and we tested the potential antagonist properties of HDL: The mouse MIN6 insulin-secreting cells were cultured with 2 mM of LDL-cholesterol preparation (native or in vitro oxidized) in the presence or absence of 1 mM of HDL-cholesterol or the ER stress inhibitor 4-phenylbutyrate (4-PBA): Prolonged exposure of MIN6 cells to 2 mM oxLDL-cholesterol for 48 hours led to an increase in expression of ER stress markers such as ATF4, CHOP and p58 and stimulated the splicing of XBP-1 whereas, induction of these markers was not observable in the cells cultured with native LDL. Treatment of the cells with the 4-PBA chemical chaperone molecule efficiently blocked activation of the ER stress markers induced by oxLDL. The latter mediates β-cell dysfunction and apoptosis by diminishing the expression of islet brain 1 (IB1) and Bcl2. The levels of these two proteins were preserved in the cells that were co-treated with oxLDL and the 4-PBA. Consistent with this result we found that blockade of ER stress activation alleviated the loss of insulin synthesis and abolished apoptosis evoked by oxLDL. However incubation of the cells with 4-PBA did not prevent impairment of insulin secretion elicited by oxLDL, indicating that ER stress is not responsible for the oxLDL-mediated defect of insulin secretion. Co-incubation of the cells with HDL mimicked the effects of 4-PBA on the expression of IB1 and Blc2 and thereby counteracted oxLDL attacks on insulin synthesis and cell survivals. We found that HDL efficiently inhibited activation of the ER stress mediated by oxLDL: These data highlight the contribution of the ER stress in the defects of insulin synthesis and cell survivals induced by oxLDL and emphasize the potent role of HDL to counter activation of the oxLDL-mediated ER-stress activation:

A role for the transcriptional repressor ICER in the molecular pathogenesis of type 2 diabetes

AbstractType 2 diabetes (T2D) is a metabolic disease which affects more than 200 millions people worldwide. The progression of this affection reaches nowadays epidemic proportions, owing to the constant augmentation in the frequency of overweight, obesity and sedentary. The pathogenesis of T2D is characterized by reduction in the action of insulin on its target tissues - an alteration referred as insulin resistance - and pancreatic β-cell dysfunction. This latter deterioration is defined by impairment in insulin biosynthesis and secretion, and a loss of β-cell mass by apoptosis. Environmental factors related to T2D, such as chronic elevation in glucose and free fatty acids levels, inflammatory cytokines and pro-atherogenic oxidized low- density lipoproteins (LDL), contribute to the loss of pancreatic β-cell function.In this study, we have demonstrated that the transcription factor Inducible Cyclic AMP Early Repressor (ICER) participates to the progression of both β-cell dysfunction and insulin resistance. The expression of this factor is driven by an alternative promoter and ICER protein represents therefore a truncated product of the Cyclic AMP Response Element Modulator (CREM) family which lacks transactivation domain. Consequently, the transcription factor ICER acts as a passive repressor which reduces expression of genes controlled by the cyclic AMP and Cyclic AMP Response Element Binding protein (CREB) pathway.In insulin-secreting cells, the accumulation of reactive oxygen species caused by environmental factors and notably oxidized LDL - a process known as oxidative stress - induces the transcription factor ICER. This transcriptional repressor hampers the secretory capacity of β-cells by silencing key genes of the exocytotic machinery. In addition, the factor ICER reduces the expression of the scaffold protein Islet Brain 1 (IB 1 ), thereby favouring the activation of the c-Jun N-terminal Kinase (JNK) pathway. This triggering alters in turn insulin biosynthesis and survival capacities of pancreatic β-cells.In the adipose tissue of mice and human subjects suffering from obesity, the transcription factor ICER contributes to the alteration in insulin action. The loss in ICER protein in these tissues induces a constant activation of the CREB pathway and the subsequent expression of the Activating Transcription Factor 3 (ATF3). In turn, this repressor reduces the transcript levels of the glucose transporter GLUT4 and the insulin-sensitizer peptide adiponectin, thereby contributing to the diminution in insulin action.In conclusion, these data shed light on the important role of the transcriptional repressor ICER in the pathogenesis of T2D, which contributes to both alteration in β-cell function and aggravation of insulin resistance. Consequently, a better understanding of the molecular mechanisms responsible for the alterations in ICER levels is required and could lead to develop new therapeutic strategies for the treatment of T2D.RésuméLe diabète de type 2 (DT2) est une maladie métabolique qui affecte plus de 200 millions de personnes dans le monde. La progression de cette affection atteint aujourd'hui des proportions épidémiques imputables à l'augmentation rapide dans les fréquences du surpoids, de l'obésité et de la sédentarité. La pathogenèse du DT2 se caractérise par une diminution de l'action de l'insuline sur ses tissus cibles - un processus nommé insulino-résistance - ainsi qu'une dysfonction des cellules β pancréatiques sécrétrices d'insuline. Cette dernière détérioration se définit par une réduction de la capacité de synthèse et de sécrétion de l'insuline et mène finalement à une perte de la masse de cellules β par apoptose. Des facteurs environnementaux fréquemment associés au DT2, tels l'élévation chronique des taux plasmatiques de glucose et d'acides gras libres, les cytokines pro-inflammatoires et les lipoprotéines de faible densité (LDL) oxydées, contribuent à la perte de fonction des cellules β pancréatiques.Dans cette étude, nous avons démontré que le facteur de transcription « Inducible Cyclic AMP Early Repressor » (ICER) participe à la progression de la dysfonction des cellules β pancréatiques et au développement de Pinsulino-résistance. Son expression étant gouvernée par un promoteur alternatif, la protéine d'ICER représente un produit tronqué de la famille des «Cyclic AMP Response Element Modulator » (CREM), sans domaine de transactivation. Par conséquent, le facteur ICER agit comme un répresseur passif qui réduit l'expression des gènes contrôlés par la voie de l'AMP cyclique et des « Cyclic AMP Response Element Binding protein » (CREB).Dans les cellules sécrétrices d'insuline, l'accumulation de radicaux d'oxygène libres, soutenue par les facteurs environnementaux et notamment les LDL oxydées - un processus appelé stress oxydatif- induit de manière ininterrompue le facteur de transcription ICER. Ainsi activé, ce répresseur transcriptionnel altère la capacité sécrétoire des cellules β en bloquant l'expression de gènes clés de la machinerie d'exocytose. En outre, le facteur ICER favorise l'activation de la cascade de signalisation « c-Jun N- terminal Kinase » (JNK) en réduisant l'expression de la protéine « Islet Brain 1 » (IB1), altérant ainsi les fonctions de biosynthèse de l'insuline et de survie des cellules β pancréatiques.Dans le tissu adipeux des souris et des sujets humains souffrant d'obésité, le facteur de transcription ICER contribue à l'altération de la réponse à l'insuline. La disparition de la protéine ICER dans ces tissus entraîne une activation persistante de la voie de signalisation des CREB et une induction du facteur de transcription « Activating Transcription Factor 3 » (ATF3). A son tour, le répresseur ATF3 inhibe l'expression du transporteur de glucose GLUT4 et du peptide adipocytaire insulino-sensibilisateur adiponectine, contribuant ainsi à la diminution de l'action de l'insuline en conditions d'obésité.En conclusion, à la lumière de ces résultats, le répresseur transcriptionnel ICER apparaît comme un facteur important dans la pathogenèse du DT2, en participant à la perte de fonction des cellules β pancréatiques et à l'aggravation de l'insulino-résistance. Par conséquent, l'étude des mécanismes moléculaires responsables de l'altération des niveaux du facteur ICER pourrait permettre le développement de nouvelles stratégies de traitement du DT2.Résumé didactiqueL'énergie nécessaire au bon fonctionnement de l'organisme est fournie par l'alimentation, notamment sous forme de sucres (glucides). Ceux-ci sont dégradés en glucose, lequel sera distribué aux différents organes par la circulation sanguine. Après un repas, le niveau de glucose sanguin, nommé glycémie, s'élève et favorise la sécrétion d'une hormone appelée insuline par les cellules β du pancréas. L'insuline permet, à son tour, aux organes, tels le foie, les muscles et le tissu adipeux de capter et d'utiliser le glucose ; la glycémie retrouve ainsi son niveau basai.Le diabète de type 2 (DT2) est une maladie métabolique qui affecte plus de 200 millions de personnes dans le monde. Le développement de cette affection est causée par deux processus pathologiques. D'une part, les quantités d'insuline secrétée par les cellules β pancréatiques, ainsi que la survie de ces cellules sont réduites, un phénomène connu sous le nom de dysfonction des cellules β. D'autre part, la sensibilité des tissus à l'insuline se trouve diminuée. Cette dernière altération, l'insulino-résistance, empêche le transport et l'utilisation du glucose par les tissus et mène à une accumulation de ce sucre dans le sang. Cette stagnation de glucose dans le compartiment sanguin est appelée hyperglycémie et favorise l'apparition des complications secondaires du diabète, telles que les maladies cardiovasculaires, l'insuffisance rénale, la cécité et la perte de sensibilité des extrémités.Dans cette étude, nous avons démontré que le facteur ICER qui contrôle spécifiquement l'expression de certains gènes, contribue non seulement à la dysfonction des cellules β, mais aussi au développement de l'insulino-résistance. En effet, dans les cellules β pancréatiques en conditions diabétiques, l'activation du facteur ICER altère la capacité de synthèse et de sécrétion d'insuline et réduit la survie ces cellules.Dans le tissu adipeux des souris et des sujets humains souffrant d'obésité, le facteur ICER contribue à la perte de sensibilité à l'insuline. La disparition d'ICER altère l'expression de la protéine qui capte le glucose, le transoprteur GLUT4, et l'hormone adipocytaire favorisant la sensibilité à l'insuline, nommée adiponectine. Ainsi, la perte d'ICER participe à la réduction de la captation de glucose par le tissue adipeux et au développement de l'insulino-résistance au cours de l'obésité.En conclusion, à la lumière de ces résultats, le facteur ICER apparaît comme un contributeur important à la progression du DT2, en soutenant la dysfonction des cellules β pancréatiques et l'aggravation de l'insulino-résistance. Par conséquent, l'étude des mécanismes responsables de la dérégulation du facteur ICER pourrait permettre le développement de nouvelles stratégies de traitement du DT2.

Health consultation : Review of on-site air monitoring data during the removal action at Le Mars coal gas site, Le Mars coal gas plant Le Mars, Plymouth County, Iowa EPA facility ID: IA0001032556 (2005)

The Iowa Department of Public Health (IDPH), Hazardous Waste Site Health Assessment Program was asked by the US Environmental Protection Agency (EPA) to review a round of air sampling data. The air data was collected and analyzed during a removal action at the Le Mars Coal Gas Site in Le Mars, Iowa. EPA asked IDPH to determine from the air data if additional monitoring is necessary throughout the removal action to protect nearby residents from exposure.

Environmental control of the Pom1-dependent cell-size regulation pathway in fission yeast

Cells couple their growth and division rate in response to nutrient availability to maintain a constant size. This co-ordination happens either at the G1-S or the G2-M transition of the cell cycle. In the rod-shaped fission yeast, size regulation happens at the G2-M transition prior to mitotic commitment. Recent studies have focused on the role of the DYRK-family protein kinase Pom1, which forms gradients emanating from cell poles and inhibits the mitotic activator kinase Cdr2, present at the cell middle. Pom1 was proposed to inhibit Cdr2 until cells reached a critical size before division. However when and where Pom1 inhibits Cdr2 is not clear as medial Pom1 levels do not change during cell elongation. Here I show that Pom1 gradients are susceptible to environmental changes in glucose. Specifically, upon glucose limitation, Pom1 re-localizes from the poles to the cell sides where it delays mitosis through regulating Cdr2. This re-localization occurs due to microtubule de- stabilization and lateral catastrophes leading to transient deposition of the Pom1 gradient nucleator Tea4 along the cell cortex. As Tea4 localization to cell sides is sufficient to recruit Pom1, this explains the mechanism of Pom1 re-localization. Microtubule destabilization and consequently Tea4 and Pom1 spread depends on the activity of the cAMP-dependent Protein Kinase A (PKA/Pka1), as pka1 mutant cells have stable microtubules and retain polar Tea4 and Pom1 under limited glucose. PKA signaling negatively regulates the microtubule rescue factor CLASP/Cls1, thus reducing its ability to stabilize microtubules. Thus PKA signaling tunes CLASP activity to promote microtubule de-stabilization and Pom1 re-localization upon glucose limitation. I show that the side-localized Pom1 delays mitosis and balances the role of the mitosis promoting, mitogen-associated protein kinase (MAPK) protein Sty1. Thus Pom1 re-localization may serve to buffer cell size upon glucose limitation. -- Afin de maintenir une taille constante, les cellules régulent leur croissance ainsi que leur taux de division selon les nutriments disponibles dans le milieu. Dans la levure fissipare, cette régulation de la taille précède l'engagement mitotique et se fait à la transition entre les phases G2 à M du cycle cellulaire. Des études récentes se sont focalisées sur le rôle de la protéine Pom1, membre de la famille des DYRK kinase. Celle-ci forme un gradient provenant des pôles de la cellule et inhibe l'activateur mitotique Cdr2 présent au centre de la cellule. Le model propose que Pom1 inhibe Cdr2 jusqu'à atteindre une taille critique avant la division. Cependant quand et à quel endroit dans la cellulle Pom1 inhibe Cdr2 n'était pas clair car les niveaux médians de Pom1 ne changent pas au cours de la l'élongation des cellules. Dans cette étude, je montre que les gradients de Pom1 sont sensibles aux changements environnementaux du taux de glucose. Plus spécifiquement, en conditions limitantes de glucose, Pom1 se relocalise des pôles de la cellule pour se distribuer sur les côtés de celle-ci. Par conséquent, un délai d'entrée en mitose est observé dû à l'inhibition Cdr2 par Pom1. Cette délocalisation est due à la déstabilisation des microtubules qui va conduire à une déposition transitoire de Tea4, le nucléateur du gradient de Pom1, tout au long du cortex de la cellule. Comme la localisation de Tea4 sur les côtés de la cellule est suffisante pour recruter la protéine Pom1, ceci explique le mécanisme de relocalisation de celle-ci. La déstabilisation des microtubules et par conséquent la diffusion de Tea4 et Pom1 dépendent de l'activité de la protéine kinase A dépendante de l'AMP cyclique (PKA/Pka1). En absence de pka1, la stabilité des microtubules n'est pas affectée ce qui permet la rétention de Tea4 et Pom1 aux pôles de la cellule même en conditions limitantes de glucose. La signalisation via PKA régule négativement le facteur de sauvetage des microtubules CLASP/Cls1 et permet donc de réduire sa fonction de déstabilisation des microtubules. Ainsi la signalisation via PKA affine l'activité des CLASP pour promouvoir la déstabilisation des microtubules et la relocalisation de Pom1 en conditions limitantes de glucose. Je montre que la localisation sur les côtés retarde l'entrée en mitose et compense l'action de la protéine Sty1, connue pour être une MAPK qui induit l'entrée en mitose. Ainsi, la relocalisation de Pom1 pourrait servir à tamponner la taille de la cellule en condition limitantes de glucose. -- Various cell types in the environment such as bacterial, plant or animal cells have a distinct cellular size. Maintaining a constant cell size is important for fitness in unicellular organisms and for diverse functions in multicellular organisms. Cells regulate their size by coordinating their growth rate to their division rate. This coupling is important otherwise cells would get progressively smaller or larger after each successive cell cycle. In their natural environment cells may face fluctuations in the available nutrient supply. Thus cells have to coordinate their division rate to the variable growth rates shown under different nutrient conditions. During my PhD, I worked with a single-celled rod shaped yeast called the fission yeast. These cells are longer when the nutrient supply is abundant and shorter when the nutrient supply is scarce. A protein that senses changes in the external carbon source (glucose) is called Protein Kinase A (PKA). The rod shape of fission yeast cells is maintained thanks to a structural backbone called the cytoskeleton. One of the components of this backbone is called microtubules, which are small tube like structures spanning the length of the cell. They transport a protein called Tea4, which in turn is important for the proper localization of another protein Pom1 to the cell ends. Pom1 helps to maintain proper shape and size of these rod shaped yeast cells. My thesis work showed that upon reduction in the external nutrient (glucose) levels, microtubules become less stable and show an alteration in their organization. A significant percentage of the microtubules contact the side of the cell instead of touching only the cell tip. This leads to the spreading of the protein Pom1 away from the tips all around the cell periphery. This helps fission yeast cells to maintain the proper size required under these conditions of limited glucose supply. I further showed that the protein PKA regulates microtubule stability and organization and thus Pom1 spreading and maintenance of proper cell size. Thus my work led to the discovery of a novel pathway by which fission yeast cells maintain their size under limited supply of glucose. -- Divers types cellulaires dans l'environnement tels que les bactéries, les plantes ou les cellules animales ont une taille précise. Le maintien d'une taille cellulaire constante est importante pour le fitness des organismes unicellulaire ainsi que pour multiples fonctions dans les organismes multicellulaires. Les cellules régulent leur taille en coordonnant le taux de croissance avec le taux de division. Ce couplage est essentiel sinon les cellules deviendraient progressivement plus petites ou plus grandes après chaque cycle cellulaire. Dans leur habitat naturels les cellules peuvent faire face a des fluctuations dans le taux de nutriment disponible. Les cellules doivent donc coordonner leur taux de division aux taux variables de croissances perçus dans les différentes conditions nutritionnels. Pendant ma thèse, j'ai travaillée sur une levure unicellulaire, en forme de bâtonnet, nommé levure fissipare ou levure de fission. La taille de ces cellules est plus grande quand le taux de nutriments est grand et plus courte quand celui-ci est plus faible. Une protéine qui perçoit les changements dans le taux externe de la source de carbone (glucose) est nommée PKA pour protéine kinase A. La forme en bâtonnet de la cellule est due aux caractères structuraux du cytosquelette. Une composante importante de ce cytosquelette sont les microtubules, dont la structures ressemble à des petit tubes qui vont d'un bout à l'autre de la cellule. Ces microtubules transportent une protéine importante nommée Tea4 qui à leur tour importante pour la bonne localisation d'une autre protéine Pom1 aux extrémités de la cellule. La protéine Pom1 aide à maintenir la taille appropriée des levures fissipares. Mon travail de thèse a montré qu'en présence de taux faible de nutriments (glucose) les microtubules deviennent de moins en moins stables et montrent une désorganisation globale. Un pourcentage significatif des microtubules touche les côtés de la cellule aux lieu d'atteindre uniquement les extrémités. Ceci a pour conséquence une diffusion de Pom1 tout au long du cortex de la cellule. Ceci aide les levures fissipares à maintenir la taille appropriée pendant ce stress nutritionnel. De plus, je montre que PKA régule la stabilité et l'organisation des microtubules et par conséquent la diffusion de Pom1 et le maintien d'une taille constante. En conclusion, mon travail a conduit à la découverte d'un nouveau mécanisme par lequel la levure fissipare maintient sa taille dans des conditions limitantes en glucose.

Estudi sobre l'eficàcia dels mètodes de reescalfament in situ en el tractament de la hipotèrmia accidental en muntanya

En la última dècada, els esports de muntanya, en qualsevol de les seves modalitats, s’han vist incrementats en nombre de practicants. Aquest fet ha comportat un important augment d’incidència de la hipotèrmia accidental. La hipotèrmia accidental és defineix com un estat patològic caracteritzat per la disminució de la temperatura corporal central per sota dels 35ºC, com a resultat de la incapacitat de l’organisme per generar el calor necessari per garantir el manteniment de les funcions fisiològiques. A través d’aquest estudi es vol demostrar que l’actuació in situ, a través de certes mesures de reescalfament, de la víctima que pateix hipotèrmia, milloren la supervivència i les complicacions secundàries d’aquesta patologia, a més a més d’avaluar l’eficàcia d’aquestes tècniques. És a dir, volem demostrar l’efectivitat de l’aplicació terapèutica extrahospitalaria de les tècniques de reescalfament. Realitzarem un estudi quantitatiu observacional analític de caràcter prospectiu a la zona dels Alps francesos d’una mostra de 237 casos d’hipotèrmia en persones practicants d’esports de muntanya que reuneixin uns criteris d’inclusió i exclusió. Els professionals de rescat aplicaran les mesures de reescalfament prèviament pautades, per les quals hauran estat formats, i enregistraran en el full de recollida de dades tota la informació sobre les variables d’estudi. És un estudi on la intervenció principal es porta a terma al medi natural i que per tant les condicions climatològiques i medioambientals poden actuar com a gran limitador de l’estudi, cal afegir també com a limitació el fet que el participant no vulgui formar part de l’estudi o que el professional de rescat que ha de portar a terme el pla terapèutic no ho faci adequadament.

Combined drug action of 2-phenylimidazo[2,1-b]benzothiazole derivatives on cancer cells according to their oncogenic molecular signatures

The development of targeted molecular therapies has provided remarkable advances into the treatment of human cancers. However, in most tumors the selective pressure triggered by anticancer agents encourages cancer cells to acquire resistance mechanisms. The generation of new rationally designed targeting agents acting on the oncogenic path(s) at multiple levels is a promising approach for molecular therapies. 2-phenylimidazo[2,1-b]benzothiazole derivatives have been highlighted for their properties of targeting oncogenic Met receptor tyrosine kinase (RTK) signaling. In this study, we evaluated the mechanism of action of one of the most active imidazo[2,1-b]benzothiazol-2-ylphenyl moiety-based agents, Triflorcas, on a panel of cancer cells with distinct features. We show that Triflorcas impairs in vitro and in vivo tumorigenesis of cancer cells carrying Met mutations. Moreover, Triflorcas hampers survival and anchorage-independent growth of cancer cells characterized by 'RTK swapping' by interfering with PDGFRβ phosphorylation. A restrained effect of Triflorcas on metabolic genes correlates with the absence of major side effects in vivo. Mechanistically, in addition to targeting Met, Triflorcas alters phosphorylation levels of the PI3K-Akt pathway, mediating oncogenic dependency to Met, in addition to Retinoblastoma and nucleophosmin/B23, resulting in altered cell cycle progression and mitotic failure. Our findings show how the unusual binding plasticity of the Met active site towards structurally different inhibitors can be exploited to generate drugs able to target Met oncogenic dependency at distinct levels. Moreover, the disease-oriented NCI Anticancer Drug Screen revealed that Triflorcas elicits a unique profile of growth inhibitory-responses on cancer cell lines, indicating a novel mechanism of drug action. The anti-tumor activity elicited by 2-phenylimidazo[2,1-b]benzothiazole derivatives through combined inhibition of distinct effectors in cancer cells reveal them to be promising anticancer agents for further investigation.

Combined drug action of 2-phenylimidazo[2,1-b]benzothiazole derivatives on cancer cells according to their oncogenic molecular signatures

The development of targeted molecular therapies has provided remarkable advances into the treatment of human cancers. However, in most tumors the selective pressure triggered by anticancer agents encourages cancer cells to acquire resistance mechanisms. The generation of new rationally designed targeting agents acting on the oncogenic path(s) at multiple levels is a promising approach for molecular therapies. 2-phenylimidazo[2,1-b]benzothiazole derivatives have been highlighted for their properties of targeting oncogenic Met receptor tyrosine kinase (RTK) signaling. In this study, we evaluated the mechanism of action of one of the most active imidazo[2,1-b]benzothiazol-2-ylphenyl moiety-based agents, Triflorcas, on a panel of cancer cells with distinct features. We show that Triflorcas impairs in vitro and in vivo tumorigenesis of cancer cells carrying Met mutations. Moreover, Triflorcas hampers survival and anchorage-independent growth of cancer cells characterized by 'RTK swapping' by interfering with PDGFRβ phosphorylation. A restrained effect of Triflorcas on metabolic genes correlates with the absence of major side effects in vivo. Mechanistically, in addition to targeting Met, Triflorcas alters phosphorylation levels of the PI3K-Akt pathway, mediating oncogenic dependency to Met, in addition to Retinoblastoma and nucleophosmin/B23, resulting in altered cell cycle progression and mitotic failure. Our findings show how the unusual binding plasticity of the Met active site towards structurally different inhibitors can be exploited to generate drugs able to target Met oncogenic dependency at distinct levels. Moreover, the disease-oriented NCI Anticancer Drug Screen revealed that Triflorcas elicits a unique profile of growth inhibitory-responses on cancer cell lines, indicating a novel mechanism of drug action. The anti-tumor activity elicited by 2-phenylimidazo[2,1-b]benzothiazole derivatives through combined inhibition of distinct effectors in cancer cells reveal them to be promising anticancer agents for further investigation.

Environmental policy instruments concerning industrial end-use energy efficiency

Energy efficiency and saving energy are the main question marks when thinking of reducing carbon dioxide emissions or cutting costs. The objective of thesis is to evaluate policy instruments concerning end-use energy efficiency of heavy industry in European Union. These policy instruments may be divided in various ways, but in this thesis the division is to administrative, financial, informative and voluntary instruments. Administrative instruments introduced in this thesis are Directive on Integrated Pollution Prevention and Control, Directive on Energy End-use Efficiency and Energy Services, and Climate and Energy Package. Financial means include energy and emission taxation, EU Emission Trading Scheme and diverse support systems. Informative instruments consist of horizontal BAT Reference Document for Energy Efficiency, as well as substantial EU documents including Green Paper on Energy Efficiency, Action Plan for Energy Efficiency and An Energy Policy for Europe. And finally, voluntary instruments include environmental managements systems like ISO 14001 and EMAS, energy auditing and benchmarking. The efficiency of different policy instruments vary quite a lot. Informative instruments lack the commitment from industry and are thus almost ineffective, contrary to EU Emission Trading Scheme, which is said to be the solution to climate problems. The efficiency of administrative means can be placed between those mentioned and voluntary instruments are still quite fresh to be examined fruitfully. However, each instrument has their potential and challenges. Cases from corporate world strengthen the results from theoretical part. Cases were written mainly on the basis of interviews. The interviewees praised the energy efficiency contract of Finnish industry, but the EU ETS takes the leading role of policy instruments. However, for industry the reductions do not come easily.

Characterizing Propionibacterium freudenreichii ssp. shermanii JS and Lactobacillus rhamnosus LC705 as a new probiotic combination: basic properties of JS and pilot in vivo assessment of the combination

Characterizing Propionibacterium freudenreichii ssp. shermanii JS and Lactobacillus rhamnosus LC705 as a new probiotic combination: basic properties of JS and pilot in vivo assessment of the combination Each candidate probiotic strain has to have the documentation for the proper identification with current molecular tools, for the biological properties, for the safety aspects and for the health benefits in human trials if the intention is to apply the strain as health promoting culture in the commercial applications. No generalization based on species properties of an existing probiotic are valid for any novel strain, as strain specific differences appear e.g. in the resistance to GI tract conditions and in health promoting benefits (Madsen, 2006). The strain evaluation based on individual strain specific probiotic characteristics is therefore the first key action for the selection of the new probiotic candidate. The ultimate goal in the selection of the probiotic strain is to provide adequate amounts of active, living cells for the application and to guarantee that the cells are physiologically strong enough to survive and be biologically active in the adverse environmental conditions in the product and in GI tract of the host. The in vivo intervention studies are expensive and time consuming; therefore it is not rational to test all the possible candidates in vivo. Thus, the proper in vitro studies are helping to eliminate strains which are unlikely to perform well in vivo. The aims of this study were to characterize the strains of Propionibacterium freudenreichii ssp. shermanii JS and Lactobacillus rhamnosus LC705, both used for decades as cheese starter cultures, for their technological and possible probiotic functionality applied in a combined culture. The in vitro studies of Propionibacterium freudenreichii ssp. shermanii JS focused on the monitoring of the viability rates during the acid and bile treatments and on the safety aspects such as antibiotic susceptibility and adhesion. The studies with the combination of the strains JS and LC705 administered in fruit juices monitored the survival of the strains JS and LC705 during the GI transit and their effect on gut wellbeing properties measured as relief of constipation. In addition, safety parameters such as side effects and some peripheral immune parameters were assessed. Separately, the combination of P. freudenreichii ssp. shermanii JS and Lactobacillus rhamnosus LC705 was evaluated from the technological point of view as a bioprotective culture in fermented foods and wheat bread applications. In this study, the role ofP. freudenreichii ssp. shermanii JS as a candidate probiotic culture alone and in a combination with L. rhamnosus LC705 was demonstrated. Both strains were transiently recovered in high numbers in fecal samples of healthy adults during the consumption period. The good survival through the GI transit was proven for both strains with a recovery rate from 70 to 80% for the JS strain and from 40 to 60% for the LC705 strain from the daily dose of 10 log10 CFU. The good survival was shown from the consumption of fruit juices which do not provide similar matrix protection for the cells as milk based products. The strain JS did not pose

Eucalyptus ESTs involved in mechanisms against plant pathogens and environmental stresses

Eucalypt plantation has high economical importance in Brazil; however, it has been attacked by various pathogens under different environmental stress conditions. Disease resistance and survival under unfavorable environmental conditions have revealed that the eucalypt has developed highly efficient defense systems. Here we show the results of the Eucalyptus ESTs Genome Project (FORESTs). Using the expressed sequence tags (ESTs) obtained by the Project, contigs of similar sequences from each cDNA library induced and not induced by stress agents were formed, and cDNA sequences similar to other already known molecules, such as plant-signaling molecules, phytoalexins, lignin biosynthesis pathways, PR-proteins and putative genes corresponding to enzymes involved in the detoxification of reactive oxygen species, were identified. We also present general considerations about the mechanisms of Eucalyptus defense against biotic and abiotic stresses. These data are of extreme importance for future eucalypt breeding programs aimed at developing plants with enhanced resistance against pathogens and environmental stresses.

Superoxide dismutase 3-mediated cell survival and proliferation

This dissertation studies the signaling events mediated by the extracellular superoxide dismutase (SOD3). SOD3 is an antioxidant enzyme which converts the harmful superoxide into hydrogen peroxide. Overproduction of these reactive oxygen species (ROS) in the cellular environment as a result of tissue injury or impaired antioxidant defense system has detrimental effects on tissue integrity and function. However, especially hydrogen peroxide is also an important signaling agent. Ischemic injury in muscle causes acute oxidative stress and inflammation. We investigated the ability of SOD3 to attenuate ischemia induced inflammation and to promote recovery of skeletal muscle tissue. We found that SOD3 can downregulate the expression of several inflammatory cytokines and cell adhesion molecules thus preventing the accumulation of oxidant-producing inflammatory cells. Secondly, SOD3 was able to promote long-term activation of the mitogenic Erk pathway, but increased only briefly the activity of pro-survival Akt pathway at an early stage of ischemic inflammation, thus reducing apoptosis. SOD3 is a prominent antioxidant in the thyroid gland where oxidative stress is constantly present. We investigated the role of SOD3 in normal thyroid follicular cells and the changes in its expression in various hyperproliferative disorders. We first showed that SOD3 is TSH-responsive which indicated its participation in thyroid function. Its principal function seems to be in follicular cell proliferation since knockdown cells were deficient in proliferation. Additionally, it was overexpressed in goiter tissue. However, SOD3 was consistently downregulated in thyroid cancer cell lines and tissues. In conclusion, SOD3 is involved in tissue maintenance, cell proliferation and inflammatory cell migration. Its mechanisms of action are the activation of known proliferation/survival pathways, inhibition of apoptosis and regulation of adhesion molecule expression.