Hypoxie-ischémie cérébrale néonatale : rôle de la voie de JNK et implication de l'autophagie dans la mort neuronale

Résumé : Malgré les immenses progrès réalisés depuis plusieurs années en médecine obstétricale ainsi qu'en réanimation néonatale et en recherche expérimentale, l'asphyxie périnatale, une situation de manque d'oxygène autour du moment de la naissance, reste une cause majeure de mortalité et de morbidité neurologique à long terme chez l'enfant (retard mental, paralysie cérébrale, épilepsie, problèmes d'apprentissages) sans toutefois de traitement pharmacologique réel. La nécessité de développer de nouvelles stratégies thérapeutiques pour les complications de l'asphyxie périnatale est donc aujourd'hui encore essentielle. Le but général de ce travail est l'identification de nouvelles cibles thérapeutiques impliquées dans des mécanismes moléculaires pathologiques induits par l'hypoxie-ischémie (HI) dans le cerveau immature. Pour cela, le modèle d'asphyxie périnatale (proche du terme) le plus reconnu chez le rongeur a été développé (modèle de Rice et Vannucci). Il consiste en la ligature permanente d'une artère carotide commune (ischémie) chez le raton de 7 jours combinée à une période d'hypoxie à 8% d'oxygène. Il permet ainsi d'étudier les lésions de type hypoxique-ischémique dans différentes régions cérébrales dont le cortex, l'hippocampe, le striatum et le thalamus. La première partie de ce travail a abordé le rôle de deux voies de MAPK, JNK et p38, après HI néonatale chez le raton à l'aide de peptides inhibiteurs. Tout d'abord, nous avons démontré que D-JNKI1, un peptide inhibiteur de la voie de JNK présentant de fortes propriétés neuroprotectrices dans des modèles d'ischémie cérébrale adulte ainsi que chez le jeune raton, peut intervenir sur différentes voies de mort dont l'activation des calpaïnes (marqueur de la nécrose précoce), l'activation de la caspase-3 (marqueur de l'apoptose) et l'expression de LC3-II (marqueur de macroautophagie). Malgré ces effets positifs le traitement au D-JNKI1 ne modifie pas l'étendue de la lésion cérébrale. L'action limitée de D-JNKI1 peut s'expliquer par une implication modérée des JNKs (faiblement activées et principalement l'isotype JNK3) après HI néonatale sévère. Au contraire, l'inhibition de la voie de nNOS/p38 par le peptide DTAT-GESV permet une augmentation de 20% du volume du tissu sain à court et long terme. Le second projet a étudié les effets de l'HI néonatale sur l'autophagie neuronale. En effet, l'autophagie est un processus catabolique essentiel au bien-être de la cellule. Le type principal d'autophagie (« macroautophagie » , que nous appellerons par la suite « autophagie ») consiste en la séquestration d'éléments à dégrader (protéines ou organelles déficients) dans un compartiment spécialisé, l'autophagosome, qui fusionne avec un lysosome pour former un autolysosome où le contenu est dégradé par les hydrolases lysosomales. Depuis peu, l'excès ou la dérégulation de l'autoptiagie a pu être impliqué dans la mort cellulaire en certaines conditions de stress. Ce travail démontre que l'HI néonatale chez le raton active fortement le flux autophagique, c'est-à-dire augmente la formation des autophagosomes et des autolysosomes, dans les neurones en souffrance. De plus, la relation entre l'autophagie et l'apoptose varie selon la région cérébrale. En effet, alors que dans le cortex les neurones en voie de mort présentent des caractéristiques mixtes apoptotiques et autophagiques, ceux du CA3 sont essentiellement autophagiques et ceux du CA1 sont principalement apoptotiques. L'induction de l'autophagie après HI néonatale semble donc participer à la mort neuronale soit par l'enclenchement de l'apoptose soit comme mécanisme de mort en soi. Afin de comprendre la relation pouvant exister entre autophagie et apoptase un troisième projet a été réalisé sur des cultures primaires de neurones corticaux exposés à un stimulus apoptotique classique, la staurosporine (STS). Nous avons démontré que l'apoptose induite par la STS était précédée et accompagnée par une forte activation du flux autophagique neuronal. L'inhibition de l'autophagie de manière pharmacologique (3-MA) ou plus spécifiquement par ARNs d'interférence dirigés contre deux protéines autophagiques importantes (Atg7 et Atg5) a permis de mettre en évidence des rôles multiples de l'autophagie dans la mort neuronale. En effet, l'autophagie prend non seulement part à une voie de mort parallèle à l'apoptose pouvant être impliquée dans l'activation des calpaïnes, mais est également partiellement responsable de l'induction des voies apoptotiques (activation de la caspase-3 et translocation nucléaire d'AIF). En conclusion, ce travail a montré que l'inhibition de JNK par D-JNKI1 n'est pas un outil neuroprotecteur efficace pour diminuer la mort neuronale provoquée par l'asphyxie périnatalé sévère, et met en lumière deux autres voies thérapeutiques beaucoup plus prometteuses, l'inhibition de nNOS/p38 ou de l'autophagie. ABSTRACT : Despite enormous progress over the last«decades in obstetrical and neonatal medicine and experimental research, perinatal asphyxia, a situation of lack of oxygen around the time of the birth, remains a major cause of mortality and long term neurological morbidity in children (mental retardation, cerebral palsy, epilepsy, learning difficulties) without any effective treatment. It is therefore essential to develop new therapeutic strategies for the complications of perinatal asphyxia. The overall aim of this work was to identify new therapeutic targets involved in pathological molecular mechanisms induced by hypoxia-ischemia (HI) in the immature brain. For this purpose, the most relevant model of perinatal asphyxia (near term) in rodents has been developed (model of Rice and Vannucci). It consists in the permanent ligation of one common carotid artery (ischemia) in the 7-day-old rat combined with a period of hypoxia at 8% oxygen. This model allows the study of the hypoxic-ischemic lesion in different brain regions including the cortex, hippocampus, striatum and thalamus. The first part of this work addressed the role of two MAPK pathways (JNK and p38) after rat neonatal HI using inhibitory peptides. First, we demonstrated that D-JNKI1, a JNK peptide inhibitor presenting strong neuroprotective properties in models of cerebral ischemia in adult and young rats, could affect different cell death mechanisms including the activation of calpain (a marker of necrosis) and caspase-3 (a marker of apoptosis), and the expression of LC3-II (a marker of macroautophagy). Despite these positive effects, D-JNKI1 did not modify the extent of brain damage. The limited action of D-JNKI1 can be explained by the fact that JNKs were only moderately involved (weakly activated and principally the JNK3 isotype) after severe neonatal HI. In contrast, inhibition of nNOS/p38 by the peptide D-TAT-GESV increased the surviving tissue volume by around 20% at short and long term. The second project investigated the effects of neonatal HI on neuronal autophagy. Indeed, autophagy is a catabolic process essential to the well-being of the cell. The principal type of autophagy ("macroautophagy", that we shall henceforth call "autophagy") involves the sequestration of elements to be degraded (deficient proteins or organelles) in a specialized compartment, the autophagosome, which fuses with a lysosome to form an autolysosome where the content is degraded by lysosomal hydrolases. Recently, an excess or deregulation of autophagy has been implicated in cell death in some stress conditions. The present study demonstrated that rat neonatal HI highly enhanced autophagic flux, i.e. increased autophagosome and autolysosome formation, in stressed neurons. Moreover, the relationship between autophagy and apoptosis varies according to the brain region. Indeed, whereas dying neurons in the cortex exhibited mixed features of apoptosis and autophagy, those in CA3 were primarily autophagíc and those in CA1 were mainly apoptotic. The induction of autophagy after neonatal HI seems to participate in neuronal death either by triggering apoptosis or as a death mechanism per se. To understand the relationships that may exist between autophagy and apoptosis, a third project has been conducted using primary cortical neuronal cultures exposed to a classical apoptotic stimulus, staurosporine (STS). We demonstrated that STS-induced apoptosis was preceded and accompanied by a strong activation of neuronal autophagic flux. Inhibition of autophagy pharmacologically (3-MA) or more specifically by RNA interference directed against two important autophagic proteins (Atg7 and AtgS) showed multiple roles of autophagy in neuronal death. Indeed, autophagy was not only involved in a death pathway parallel to apoptosis possibly involved in the activation of calpains, but was also partially responsible for the induction of apoptotic pathways (caspase-3 activation and AIF nuclear translocation). In conclusion, this study showed that JNK inhibition by D-JNKI1 is not an effective neuroprotective tool for decreasing neuronal death following severe perinatal asphyxia, but highlighted two more promising therapeutic approaches, inhibition of the nNOSlp38 pathway or of autophagy.

Tornabenea ribeirensis (Apiaceaeb a new species from São Nicolau, Cape Verde Islands (West Africa)

Many species of Apiaceae are found in the Macaronesien Region. Several have been introduccd by human activities, but a number of taxa is endemic to the different archipelagos or even rrstrictcd to a single island. The following enumeration is based mainly on HANSEN & SUNDING ( 1993). In the Arores 28 different taxa of Apiaceae occur; among them four endemic species [AUIMI; hrrrrii WATSON, A. trifoliatum (WATSON) TKEL., Clrtrc~,y~l~~ll~r~~~ cl:oricrm TREL.. SOErich trwrictr GUTIINICK ex SEUB.]. In Madeira the Apiaceae are very diverse and consist ol’ 29 species and subspecies. From the archipelago two monotypic genera, rC/c/trtio. velitru~rr t/ccipicvr.s (SCHRAD. & J. C. WENDL.) Ho~+hl. md kJorli:ia edu[is LOWE and ~hrcc cndcmic species [Oemmrlre diwricore (R. BR.) MABB.. I/nperrr/orio lotvei COSS. and Burrirr~r hre~$~lirrnr LOWE] are described. The Canary Islands have the highest numbcr of plant-species and a high level of endemism. 5-l taxa of Apiaceae are recorded including three endemic genera (Rtrrheopsis A. HANSEX & KUNKEL, Todm-oa PARL. and Tiqyrmm PARL.) and further I5 endemic taxa. The Apiaceae are represented in the Cape Verde Islands by I2 species. Most of the taxa have been introduced by human activities (LOBIN & ZIZKA 1957) like Amvhm grm’eo- 1efr.s L., Apirm grmvolerrs L, Foerricrrhrr urlgore MILL.. Corimrtlru~t~ srrtirvrrrr L. or Petrosilerrm crisprrm (FRILL.) A.W.HILL. These species are cultivated and some of them later became \\esdy. Other species like Ciclosper- UWL /e/~fo/~/l~ll~rrtr (PER%) SPRAGUE (= Apimr leproplr~llrr~rr) are weeds of cultivated grounds or wasted lands. All these species are today widespread in temperate. subtropical or tropical regions all over the world. The only native species are to be found in the endemic genus To~wI~~I~~~ PARL.

Cedar Rapids-Iowa City Goose Hunting Zone, 2005

Regulations for hunting in Iowa.

Engineered aprotinin for improved stability of fibrin biomaterials.

Fibrin has been long used clinically for hemostasis and sealing, yet extension of use in other applications has been limited due to its relatively rapid resorption in vivo, even with addition of aprotinin or other protease inhibitors. We report an engineered aprotinin variant that can be immobilized within fibrin and thus provide extended longevity. When recombinantly fused to a transglutaminase substrate domain from α(2)-plasmin inhibitor (α(2)PI(1-8)), the resulting variant, aprotinin-α(2)PI(1-8), was covalently crosslinked into fibrin matrices during normal thrombin/factor XIIIa-mediated polymerization. Challenge with physiological plasmin concentrations revealed that aprotinin-α(2)PI(1-8)-containing matrices retained 78% of their mass after 3 wk, whereas matrices containing wild type (WT) aprotinin degraded completely within 1 wk. Plasmin challenge of commercial sealants Omrixil and Tisseel, supplemented with aprotinin-α(2)PI(1-8) or WT aprotinin, showed extended longevity as well. When seeded with human dermal fibroblasts, aprotinin-α(2)PI(1-8)-supplemented matrices supported cell growth for at least 33% longer than those containing WT aprotinin. Subcutaneously implanted matrices containing aprotinin-α(2)PI(1-8) were detectable in mice for more than twice as long as those containing WT aprotinin. We conclude that our engineered recombinant aprotinin variant can confer extended longevity to fibrin matrices more effectively than WT aprotinin in vitro and in vivo.

Insect feeding preferences on Piperaceae species observed in São Paulo city, Brazil

Piperaceae species have been placed among the basal angiosperm and are adapted to a variety of habitats including moist forests, secondary vegetation and dry high lands. The major anatomical/morphology features are of small trees, vines, and shrubs for Piper species, while the epiphytic and succulent characteristics are predominant forms among Peperomia species. Their secondary chemistry can be mostly represented by amides, phenylpropanoids/lignoids, and chromenes in addition to a phletoria of biosynthetically mixed-origin secondary compounds. Although several amides and lignans are known as insecticides, several phytophagous insects, among which some considered pests of economic importance, have been observed feeding vigorously on Piperaceae species. Herein we describe the feeding preferences of fourteen phytophagous species of Coleoptera, Lepidoptera and Hemiptera over approximately fifty Piperaceae species observed in São Paulo, SP, Brazil, in a long-term basis.

Mini-STRs em genética forense

A análise do ADN baseada nos STRs (Short Tandem Repeats) tem provado ser uma ferramenta extremamente útil em estudos forenses. A aplicação dos multiplex de STRs resulta em perfis genéticos completos, para a maior parte das amostras, desde que contenham uma quantidade apreciável de ADN. Contudo, em amostras com ADN degradado, a análise tornase mais complicada, uma vez que os STRs com maior peso molecular (acima dos 250 pb), podem não ser amplificados nestas condições. Como a metodologia de Mini-STRs não estava, ainda, implementada nos Laboratórios Forenses portugueses, o objectivo principal deste estudo consistiu na aplicação de um multiplex, designado por Mini-SGM, que contém os marcadores TH01, FGA, D18S51, D16S359, D2S1338 e a Amelogenina, para implementação na prática forense, tendo os resultados sido comparados com a metodologia de rotina. O trabalho experimental teve início com um estudo populacional, que compreendeu uma amostra de 110 indivíduos de origem portuguesa, não relacionados e uma amostra de 52 indivíduos, não relacionados, oriundos de Cabo Verde, encontrando-se, ambas as populações, em Equilíbrio de Hardy-Weinberg, excepto para o locus D18S51, na população portuguesa e o locus D2S1338 na população de origem africana. Em relação aos parâmetros estatísticos forenses, este estudo demonstrou que o poder de discriminação varia entre 0,903 e 0,959, para a população portuguesa, e entre 0,908 e 0,958, para a população de Cabo Verde. Após o estudo populacional, foram analisados 39 casos forenses, devidamente anonimizados, da casuística do Serviço de Genética e Biologia Forense da Delegação de Lisboa do INML, com diferentes tipos de vestígios biológicos, como sangue, ossos, pulmão, coração, tecidos embrionários, tecidos inclusos em parafina, fígado, rim e dentes, de modo a ilustrar a utilidade desta nova metodologia nos casos forenses. No estudo de amostras degradadas, o uso da metodologia de rotina pode,

Seasonal and spatial species richness variation of dung beetle (Coleoptera, Scarabaeidae s. str.) in the Atlantic Forest of southeastern Brazil

The knowledge on Atlantic Forest scarab beetle fauna is quite limited. This biome is strongly degraded and these insects can be used as bioindicators since they are sensitive to forest destruction and show distinct organizational patterns in forest fragments or in areas that have been deteriorated by human activity. Thus, a study of the Scarabaeidae (sensu stricto) dung beetles fauna that inhabit Serra do Japi, São Paulo, Brazil (23º12'-23º22' S and 46º53'-47º03'W) was carried out; the monthly species richness was analyzed in six areas during one year and the vegetation's structural physiognomy was described. The areas included a conserved and a degraded valley, a northward and a southward hillside, a hilltop, and an area of secondary forest growing under eucalyptus trees. The specimens were collected using four pitfall traps baited with human feces, which remained at each spot during 48 hours. Between September, 1997 and August, 1998, 3524 individuals of 39 species were collected; the most abundant were: Canthidium trinodosum, Eurysternus cyanescens, Uroxys kratochvili, Scybalocanthon nigriceps, Uroxys lata, Canthonella sp., Dichotomius assifer, Deltochilum furcatum, Canthidium sp.2, Canthon latipes, Deltochilum rubripenne, Eurysternus sp., and Dichotomius sp.1. The number of individuals and species was greater in the hot, rainy season, when there was a correlation between the number of species and the mean annual temperature [r²= 0.69; p<0.01]. The lower winter richness was most pronounced in the conserved valley, while richness remained relatively constant in the degraded valley; abundance was much higher in the degraded valley. The cluster analysis showed that the valleys and hillsides are the most similar in relation to species composition and abundance, yet different from the secondary forest with eucalypts and the hilltop.

Evaluation of Remediation Recommendations: Stakeholder Workshop 3 Ribeira Seca - Cape Verde

In Cape Verde, the low soil cover and inadequate practices on rain fed agricultural lands constitute major problems related to desertification. To the fragility of the land associates severe water erosion, causing tons of land to be washed away from the fields every year during the rainy season. Therefore, the aim in the scope of combating desertification is to provide a certain degree of permanent soil cover to serve as shield for the impact of rain. During the selection workshop several technologies, all related to vegetative cover either as strips or surface cover were discussed. Only two technologies were selected: vegetation strip with pigeon pea and afforestation with fruit trees.  Technology 1: Pigeon pea (cajanus cajan) barriers/strips. It consists in planting seeds of pigeon pea, a leguminous perennial shrub that has dual purpose of protecting the soil and feed people. It is planted in association with maize crop. After the maize is harvested, the soil remains with some degree of cover. Though the objective was to plant as strip barriers, six meters apart, most farmers planted it as surface cover.  Technology 2: Afforestation with fruit trees. It consists in the plantation of different fruit tree species in humid areas to provide both soil cover and feed for farmers. Since fruit trees require several years to provide effective cover, and though it was implemented in some areas, it was not evaluated during the project’s period.

Evaluation of Remediation Recommendations: Stakeholder Workshop 3 Ribeira Seca - Cape Verde

In Cape Verde, the low soil cover and inadequate practices on rain fed agricultural lands constitute major problems related to desertification. To the fragility of the land associates severe water erosion, causing tons of land to be washed away from the fields every year during the rainy season. Therefore, the aim in the scope of combating desertification is to provide a certain degree of permanent soil cover to serve as shield for the impact of rain. During the selection workshop several technologies, all related to vegetative cover either as strips or surface cover were discussed. Only two technologies were selected: vegetation strip with pigeon pea and afforestation with fruit trees.  Technology 1: Pigeon pea (cajanus cajan) barriers/strips. It consists in planting seeds of pigeon pea, a leguminous perennial shrub that has dual purpose of protecting the soil and feed people. It is planted in association with maize crop. After the maize is harvested, the soil remains with some degree of cover. Though the objective was to plant as strip barriers, six meters apart, most farmers planted it as surface cover.  Technology 2: Afforestation with fruit trees. It consists in the plantation of different fruit tree species in humid areas to provide both soil cover and feed for farmers. Since fruit trees require several years to provide effective cover, and though it was implemented in some areas, it was not evaluated during the project’s period.

Erosão dos solos em Cabo Verde - Estudos dos Processos e Quantificação de Três Bacias na Ilha de Santiago

L’archipel du Cap Vert constitué de 10 îles volcaniques appartient à la zone sahélienne qui s’étend de l’atlantique jusqu’à la mer rouge. Depuis, plusieurs décennies le Cap Vert est affecté par la désertification causée en grande partie par la récession climatique et l’érosion des sols. Ces facteurs, associés à la forte pression anthropique sur les ressources, à l’orographie accidentée et à des pluies tropicales parfois diluviennes, provoquent de sérieuses pertes du patrimoine foncier. Cependant, depuis son Indépendance en 1975, le Gouvernement a mené un vaste programme d’arborisation, de restauration des terres et d’aménagement des cours d’eau. Pourtant, très peu de recherches ont été menées pour évaluer les actions de protection et de conservation des sols et des eaux. Par conséquent, il n’existe quasiment pas de données sur la problématique de la dégradation des terres ni de bilans. Dans le cadre de ce travail, nous avons étudié les différents facteurs qui contrôlent l’érosion hydrique des sols. Nous avons plus particulièrement cherché à différencier les effets des activités humaines, notamment agricoles, de ceux des facteurs climatiques comme les précipitations et la génération des écoulements. Nous avons également établi les premiers bilans d’exportations de matières en suspension et en solution dans le contexte de l’archipel du Cap Vert. L’étude a été menée à l’échelle de trois bassins versants de l’ile de Santiago Cap-Vert. Ces trois bassins versant (Longueira, Grande et Godim) sont localisés dans la partie centrale de l’île de Santiago et représentatifs des divers modes d’occupation du sol et des différents climats de l’île. Il existe un gradient climatique entre les trois bassins versants. En effet, Longueira qui présente une superficie de 4,18 km2, une pente moyenne de 47 %, se localise dans une zone humide couverte à 69 % par une forêt et une surface agricole de 17 %. Grande avec une superficie de 1,87 km2, se localise en zone sub humide pour une pente moyenne de 50 %, il est essentiellement agricole. Godim, avec une superficie de 2,0 km2, se localise en zone semi aride, il est particulièrement agricole et sa pente moyenne est de 32 %. Pour ces trois bassins versants, les écoulements de crue à l’exutoire ont été mesurés et échantillonnés de 2004 à 2009. Le bassin versant de Longueira a fait l’objet d’un suivi plus poussé, notamment en termes de fréquence d’échantillonnage et de suivi des écoulements hors crue. Sur chaque échantillon nous avons procédé à la détermination de la concentration des matières en suspension ainsi qu’à l’analyse des éléments majeurs. Les résultats obtenus montrent que l’érosion mécanique dans les 3 bassins versants est caractérisée par une forte variabilité spatiale et temporelle. Sur la période 2005-2009, le bilan moyen annuel pour les bassins versants de Longueira, Grande et Godim est de : 4266, 157 et 10,1 t.km2.an-1 respectivement. La saison humide 2006 a été la plus érosive pour l’ensemble des trois bassins versants et particulièrement dans Longueira avec 2 crues exceptionnelles qui ont généré une concentration moyenne de matières en suspension supérieure à 100 g/l. En revanche, les saisons 2005 et 2008 ont été dans l’ensemble peu érosives car les concentrations moyennes ne dépassèrent pas 20 g/l. Par ailleurs, il n’y a pas eu de lames d’eau écoulées pour les saisons 2005 et 2007 pour le bassin de Godim. Sur le bassin de Longueira, l’étude des phénomènes d’hystérésis permet de caractériser chaque crue et de montrer que l’évolution temporelle des exportations de matières en suspension au cours de la saison est fortement influencée par les activités agricoles. En effet, la première crue provoque l’exportation massive des sédiments disponibles et localisés dans le lit du cours d’eau. En conséquence, la seconde est moins exportatrice de sédiments. Un mois après les premières pluies, les activités de sarclage diminuent la densité du couvert végétal et destructurent la partie superficielle des sols, ce qui provoque à nouveau une très forte exportation de sédiments lors de la troisième crue. Les résultats de l’érosion chimique sur le bassin de Longueira indiquent que le taux d’érosion chimique moyen s’élève à 45 t.km2.an-1 avec une forte variabilité temporelle. En effet, les saisons les plus humides de 2006 et 2007 sont les plus exportatrices de matières en solution, alors que 2005 a eu une faible exportation. L’utilisation du modèle de mélanges EMMA (End-Members Mixing Analysis) montre que les écoulements hypodermique et profond, qui alimentent le cours d’eau en éléments dissous, sont les principaux facteurs de l’érosion chimique. On montre ainsi que les écoulements hors crue sont à l’origine de plus de 90% des flux d’érosion chimique. L’écoulement superficiel, qui contribue à environ 70 % du débit du cours d’eau en crue, constitue un facteur de premier plan de l’érosion mécanique des sols.

Epigean ant communities in Atlantic Forest remnants of São Paulo: a comparative study using the guild concept

Epigean ant communities in Atlantic Forest remnants of São Paulo: a comparative study using the guild concept. The guilds constitute a valuable ecological tool, because they allow conducting comparisons among environments under different conditions. The ants can be used as ecological indicators, mainly for the monitoring of degraded forest areas. The aim of this research was to study guild organization among the epigeous Formicidae living in Atlantic forest remnants of the State of São Paulo, Brazil. Ant collections were performed in three distinct Atlantic forest biome areas: arboreal littoral vegetation ("restinga") (Cananéia), semideciduous seasonal forest (Piracicaba) and dense ombrophylousforest (Pariquera-Açu). After identification, the ants were grouped into guilds, based on the ecological attributes of behavior and habit, according to the literature. Nine guilds were found; the semideciduous seasonal forest ecosystem presented eight of them, followed by the arboreal sandbank (7) and dense ombrophylous forest (6). The guilds found were: litter omnivorous and scavengers, granivorous species, specialist predators living in litter and soil, litter generalist predators, subterranean mealybug-dependent species, army ants, dominant or subdominants arboreal, that occasionally forage on the ground, soil or litter dominant and fungus-growers, using feces and insect body fragments. The guilds found can be used in the monitoring of the mirmecofauna in the Atlantic Forest biome, supplying insights for further ecological studies.

Draft environmental report on Cape Verde

Faced with recurrent drought and famine during five centuries of human occupation, the small and densely populated Cape Verde Islands have a history of severe environmental problems. The arid climate and steep, rocky terrain provide scant resources for traditional subsistance farming under the best conditions, and in years of low rainfall the failure of rainfed crops causes massive food shortages. Agricultural use of steep slopes where rainfall is highest has led to soil erosion, as has removal of the island's vegetation for fuel and livestock. Pressure on the vegetation is particularly severe in dry years. International aid can provide relief from famine, and the introduction of modern agricultural and conservation techniques can improve the land and increase yield, but it is unlikely that Cape Verde can ever be entirely self -sufficient in food. Ultimately, the solution of Cape Verde's economic and environmental problems will probably require the development of productive urban jobs so the population can shift away from the intensive and destructive use of land for subsistance farming. In the meantime, the people of Cape Verde can best be served by instituting fundamental measures to conserve and restore the land so that it can be used to its fullest potential. The primary environmental problems in Cape Verde today are: 1. Soil degradation. Encouraged by brief but heavy rains and steep slopes, soil erosion is made worse by lack of vegetation. Soils are also low in organic matter due to the practice of completely removing crop plants and natural vegetation for food, fuel or livestock feed. 2. Water shortage. Brief and erratic rainfall in combination with rapid runoff makes surface water scarce and difficult to use. Groundwater supplies can be better developed but capabilities are poorly known and the complex nature of the geological substrate makes estimation difficult. Water is the critical limiting factor to the agricultural capability of the islands. 3. Fuel shortage. Demand for fuel is intense and has resulted in the virtual elimination of native vegetation. Fuelwood supplies are becoming more and more scarce and costly. Development of managed fuelwood plantations and alternate energy sources is required. 4. Inappropriate land use. Much of the land now used for raising crops or livestock is too steep or too arid for these purposes, causing erosion and destruction of vegetation. Improving yield in more appropriate areas and encouraging less damaging uses of the remaining marginal lands can help to alleviate this problem.

Infestation and natural parasitism of aphids in single and mixed pastures of black oats and ryegrass

Some species of aphids are major pests on cereal crops and grass pastures. Usually these pests are not adequately controlled in pasture lands that become sources of aphid infestations to cereal crops. The dynamics of aphids and the incidence of natural enemies are less known in pasture systems than in cereal fields. The objective of this work was to assess the aphid infestation and natural aphid parasitism in different pasture composition. Three hypotheses were tested: 1- the aphid species composition in pastures may vary according to the cereal species in the field; 2- the mixture of two plant species can modify the amount and diversity of aphids; 3- the plant species composition of pasture fields influences the parasitism of aphids. Empirical data were obtained from three Poaceae fields: black oats (Avena strigosa L.), ryegrass (Lolium multiflorum L.), and a mixed field of black oats and ryegrass. The most abundant aphid species was Rhopalosiphum padi followed by Sitobion avenae. Plant species composition increases the amount and the parasitism rates of aphids. The mixture of heavily infested black oats with a poorly infested ryegrass resulted in reduction of aphid infestation in comparison with heavily-infested single plant species field. This is possible because the conditions are favorable for the development of parasitoid populations. Aphidius colemani was the main parasitoid found in these areas.

Association of nuclear matrix proteins with granular and threaded nuclear bodies in cell lines undergoing apoptosis.

The granules which appear in the nucleolar area in apoptotic HL-60 cells after camptothecin administration (Zweyer et al., Exp. Cell Res. 221,27-40, 1995) were detected also in several other cell lines induced to undergo apoptosis by different stimuli, such as MOLT-4 treated with staurosporine, K-562 incubated with actinomycin D, P-815 exposed to temperature causing heat shock, Jurkat cells treated with EGTA, U-937 growing in the presence of cycloheximide and tumor necrosis factor-alpha, and HeLa cells treated with etoposide. Using immunoelectron microscopy techniques, we demonstrate that, besides the already described nuclear matrix proteins p125 and p160, these granules contain other nucleoskeletal polypeptides such as proliferating cell nuclear antigen, a component of ribonucleoprotein particles, a 105-kDa constituent of nuclear spliceosomes, and the 240-kDa nuclear mitotic apparatus-associated protein referred to as NuMA. Moreover, we also found in the granules SAF-A/hn-RNP-U and SATB1 proteins, two polypeptides that have been reported to bind scaffold-associated regions DNA sequences in vitro, thus mediating the formation of looped DNA structures in vivo. Fibrillarin and coilin are not present in these granules or the PML protein. Thus, the granules seen during the apoptotic process apparently are different from coiled bodies or other types of nuclear bodies. Furthermore, these granules do not contain chromatin components such as histones and DNA. Last, Western blotting analysis revealed that nuclear matrix proteins present in the granules are not proteolytically degraded except for the NuMA polypeptide. We propose that these granules might represent aggregates of nuclear matrix proteins forming during the apoptotic process. Moreover, since the granules are present in several cell lines undergoing apoptosis, they could be considered a previously unrecognized morphological hallmark of the apoptotic process.

Vasopressin-inducible ubiquitin-specific protease 10 increases ENaC cell surface expression by deubiquitylating and stabilizing sorting nexin 3

Adjustment of Na+ balance in extracellular fluids is achieved by regulated Na+ transport involving the amiloride-sensitive epithelial Na+ channel (ENaC) in the distal nephron. In this context, ENaC is controlled by a number of hormones, including vasopressin, which promotes rapid translocation of water and Na+ channels to the plasma membrane and long-term effects on transcription of vasopressin-induced and -reduced transcripts. We have identified a mRNA encoding the deubiquitylating enzyme ubiquitin-specific protease 10 (Usp10), whose expression is increased by vasopressin at both the mRNA and the protein level. Coexpression of Usp10 in ENaC-transfected HEK-293 cells causes a more than fivefold increase in amiloride-sensitive Na+ currents, as measured by whole cell patch clamping. This is accompanied by a three- to fourfold increase in surface expression of alpha- and gamma-ENaC, as shown by cell surface biotinylation experiments. Although ENaC is well known to be regulated by its direct ubiquitylation, Usp10 does not affect the ubiquitylation level of ENaC, suggesting an indirect effect. A two-hybrid screen identified sorting nexin 3 (SNX3) as a novel substrate of Usp10. We show that it is a ubiquitylated protein that is degraded by the proteasome; interaction with Usp10 leads to its deubiquitylation and stabilization. When coexpressed with ENaC, SNX3 increases the channel's cell surface expression, similarly to Usp10. In mCCD(cl1) cells, vasopressin increases SNX3 protein but not mRNA, supporting the idea that the vasopressin-induced Usp10 deubiquitylates and stabilizes endogenous SNX3 and consequently promotes cell surface expression of ENaC