Distribution de la fraction d'excrétion rénale du lithium et de l'acide urique dans l'étude de population Hercules

Problématique¦L'évaluation fonctionnelle du tubule proximal du rein est intéressante pour la compréhension de la physiopathologie des anomalies du transport du sodium à ce niveau du néphron. Ces anomalies participent au développement de l'hypertension artérielle (HTA) et de la sensibilité au sel. Environ 60% à 80% du sodium est réabsorbé proximalement dans le néphron. Chez l'animal, la fonction du tube proximal peut être estimée directement par microponction. Une telle approche directe n'étant pas possible chez l'homme, seul des approches indirectes permettent de recueillir des informations sur la physiologie de ce segment du néphron, comme par exemple par la mesure de la clairance du lithium (endogène ou exogène) ou de l'acide urique. La fraction d'excrétion du lithium (FELi) et la fraction d'excrétion de l'acide urique (FEAU) permettent d'estimer la réabsorption proximale de sodium chez l'homme. Plusieurs études expérimentales et cliniques ont montré l'existence d'une relation linéaire entre la FELi et la fraction excrétée du sodium (FENa) chez l'animal et l'homme.¦Objectif¦Décrire la distribution de la FELi et de la FEAU et analyser les associations de ces deux variables avec l'âge, le sexe, la consommation d'alcool, le tabagisme, la pression artérielle et l'IMC dans l'étude de population Hercules.¦Méthodes¦Sélection au hasard d'un sous-groupe de participants de l'étude CoLaus (N=6188). Nombre de participants à l'étude Hercules: 437, dont 229 femmes et 208 hommes. Les participants ont effectué une récolte d'urine sur 24 heures afin de déterminer la fonction rénale et les FELi et FEAU. Le TFG a été mesuré à l'aide de la clairance de la créatinine. Le test des rangs signés de Wilcoxon pour données appariées ( Wilcoxon matched-pairs signed rank test) et le test de comparaison des médianes ont été utilisés pour la comparaison entre groupes. Le coefficient de corrélation de Spearman a été utilisé pour évaluer la relation entre les variables continues. Les box-plots ont été utilisés pour la description de la distribution du lithium et de l'acide urique selon l'âge, le sexe et la période de la journée. Le logiciel Stata 11.0 a été utilisé pour les analyses statistiques.¦Résultats¦La prévalence de l'HTA dans la population de l'étude Hercules était 33%, la prévalence du diabète était 8%, la prévalence de l'obésité était 15.3%, la prévalence du tabagisme était 23%. La FELi reste stable avec l'âge et est similaire dans les deux sexes. Chez les hommes, la FELi est plus grande la nuit que le jour, c'est-à-dire qu'ils résorbent plus de sodium proximalement le jour que la nuit. Un IMC plus grand est associé à une FELi plus basse dans les deux sexes. Il y a une corrélation négative entre la FEAU et l'IMC (significative) et la consommation d'alcool.¦Conclusion¦Les résultats obtenus avec les données de l'étude Hercules sont similaires à ceux qu'on retrouve dans la littérature sur la FELi et la FEAU , ce qui rassure sur la qualité des données. La FELi varie peu avec l'âge et le sexe, contrairement à la FEAU, qui varie fortement avec l'âge et le sexe. L'HTA, le diabète, l'obésité, le tabagisme et la consommation d'alcool sont associées à une FELi et FEAU plus basses. Les sujets qui présentent ces caractéristiques réabsorbent donc plus de sodium au niveau proximal.

Historical reconstruction of polychlorinated biphenyl (PCB) exposures for workers in a capacitor manufacturing plant

We developed a semiquantitative job exposure matrix (JEM) for workers exposed to polychlorinated biphenyls (PCBs) at a capacitor manufacturing plant from 1946 to 1977. In a recently updated mortality study, mortality of prostate and stomach cancer increased with increasing levels of cumulative exposure estimated with this JEM (trend p values = 0.003 and 0.04, respectively). Capacitor manufacturing began with winding bales of foil and paper film, which were placed in a metal capacitor box (pre-assembly), and placed in a vacuum chamber for flood-filling (impregnation) with dielectric fluid (PCBs). Capacitors dripping with PCB residues were then transported to sealing stations where ports were soldered shut before degreasing, leak testing, and painting. Using a systematic approach, all 509 unique jobs identified in the work histories were rated by predetermined process- and plant-specific exposure determinants; then categorized based on the jobs' similarities (combination of exposure determinants) into 35 job exposure categories. The job exposure categories were ranked followed by a qualitative PCB exposure rating (baseline, low, medium, and high) for inhalation and dermal intensity. Category differences in other chemical exposures (solvents, etc.) prevented further combining of categories. The mean of all available PCB concentrations (1975 and 1977) for jobs within each intensity rating was regarded as a representative value for that intensity level. Inhalation (in microgram per cubic milligram) and dermal (unitless) exposures were regarded as equally important. Intensity was frequency adjusted for jobs with continuous or intermittent PCB exposures. Era-modifying factors were applied to the earlier time periods (1946-1974) because exposures were considered to have been greater than in later eras (1975-1977). Such interpolations, extrapolations, and modifying factors may introduce non-differential misclassification; however, we do believe our rigorous method minimized misclassification, as shown by the significant exposure-response trends in the epidemiologic analysis.

JARID2 is a direct target of the PAX3-FOXO1 fusion protein and inhibits myogenic differentiation of rhabdomyosarcoma cells.

Rhabdomyosarcomas (RMS) are the most frequent soft-tissue sarcoma in children and characteristically show features of developing skeletal muscle. The alveolar subtype is frequently associated with a PAX3-FOXO1 fusion protein that is known to contribute to the undifferentiated myogenic phenotype of RMS cells. Histone methylation of lysine residues controls developmental processes in both normal and malignant cell contexts. Here we show that JARID2, which encodes a protein known to recruit various complexes with histone-methylating activity to their target genes, is significantly overexpressed in RMS with PAX3-FOXO1 compared with the fusion gene-negative RMS (t-test; P < 0.0001). Multivariate analyses showed that higher JARID2 levels are also associated with metastases at diagnosis, independent of fusion gene status and RMS subtype (n = 120; P = 0.039). JARID2 levels were altered by silencing or overexpressing PAX3-FOXO1 in RMS cell lines with and without the fusion gene, respectively. Consistent with this, we demonstrated that JARID2 is a direct transcriptional target of the PAX3-FOXO1 fusion protein. Silencing JARID2 resulted in reduced cell proliferation coupled with myogenic differentiation, including increased expression of Myogenin (MYOG) and Myosin Light Chain (MYL1) in RMS cell lines representative of both the alveolar and embryonal subtypes. Induced myogenic differentiation was associated with a decrease in JARID2 levels and this phenotype could be rescued by overexpressing JARID2. Furthermore, we that showed JARID2 binds to and alters the methylation status of histone H3 lysine 27 in the promoter regions of MYOG and MYL1 and that the interaction of JARID2 at these promoters is dependent on EED, a core component of the polycomb repressive complex 2 (PRC2). Therefore, JARID2 is a downstream effector of PAX3-FOXO1 that maintains an undifferentiated myogenic phenotype that is characteristic of RMS. JARID2 and other components of PRC2 may represent novel therapeutic targets for treating RMS patients.

Fanconi anemia and genome stability : the role of FANCM

RÉSUMÉ: Le génome de toute cellule est susceptible d'être attaqué par des agents endogènes et exogènes. Afin de préserver l'intégrité génomique, les cellules ont développé des multitudes de mécanismes. La réplication de l'ADN, une étape importante durant le cycle cellulaire, constitue un stress et présente un danger important pour l'intégrité du génome. L'anémie de Fanconi est une maladie héréditaire rare dont les protéines impliquées semblent jouer un rôle crucial dans la réponse au stress réplicatif. La maladie est associée à une instabilité chromosomique ainsi qu'à une forte probabilité de développer des cancers. Les cellules des patients souffrant de l'anémie de Fanconi sont sensibles à des agents interférant avec la réplication de l'ADN, et plus particulièrement àdes agents qui fient les deux brins d'ADN d'une manière covalente. L'anémie de Fanconi est une maladie génétiquement hétérogène. Treize protéines ont pu être identifiées. Elles semblent figurer dans une même voie de signalisation qui est aussi connue sous le nom de « FA/BRCA pathway », car un des gènes est identique au gène BRCA2 (breast cancer susceptibility gene 2). Huit protéines forment un complexe nucléaire dont l'intégrité est nécessaire à la monoubiquitination de deux autres protéines, FANCD2 et FANCI, en réponse à un stress réplicatif. A ce jour, la fonction moléculaire des protéines du « FA/BRCA pathway »reste encore mal décrite. Au début de mon travail de thèse, nous avons donc décidé de purifier les protéines du complexe nucléaire et d'étudier leurs propriétés biochimiques. Nous avons tout d'abord étudié les cinq protéines connues à l'époque qui sont FANCA, FANCC, FANCE, FANCF et FANCG. Par la suite, nous avons étendu notre étude à des protéines découvertes plus récemment, FANCL, FANCM et FAAP24, en concentrant finalement notre travail sur la caractérisation de FANCM. FANCM, contrairement aux autres protéines du complexe, est constituée de deux domaines conservés suggérant un rôle important dans le métabolisme de l'ADN. Il s'agit d'un domaine « DEAH box hélicase »situé dans la partie N-terminale et d'un domaine « ERCC4 nuclease »situé dans la partie C-terminale de la protéine. Dans cette étude, nous avons purifié avec succès la protéine FANCM entière à partir d'un système hétérologue. Nous montrons que FANCM s'attache de manière spécifique à des jonctions de Holliday et des fourches de réplication. De plus, nous démontrons que FANCM peut déplacer le point de jonction de ces structures via son domaine hélicase de manière dépendante de l'ATP. FANCM est aussi capable de dissocier de grands intermédiaires de la recombinaison, via la migration de jonctions de Holliday à travers une région d'homologie de 2.6 kb. Tous ces résultats suggèrent que FANCM peut s'attacher spécifiquement à des fourches de réplication et à des jonctions de Holliday in vitro et que son domaine hélicase est associé à une activité migratoire efficace. Nous pensons que FANCM peut avoir un rôle direct sur les intermédiaires de réplication. Ceci est en accord avec l'idée que les protéines de l'anémie de Fanconi coordonnent la réparation de l'ADN au niveau des fourches de réplication arrêtées. Nos résultats donnent une première indication quant au rôle de FANCM dans la cellule et peuvent contribuer à élucider la fonction de cette voie de signalisation peu comprise jusqu'à présent. SUMMARY: The genome of every cell is subject to a constant offence by endogenous and exogenous agents. Not surprisingly; cells have evolved a multitude of mechanisms which aim at preserving genomic integrity. A key step during the life cycle of a cell, DNA replication itself, constitutes a special danger to the integrity of the genome. The proteins defective in the rare hereditary disease Fanconi anemia (FA) are suspected to play a crucial role in the cellular response to DNA replication stress. The disease is associated with chromosomal instability and pronounced cancer susceptibility. Cells from Fanconi anemia patients are sensitive to a variety of agents which interfere with DNA replication, DNA interstrand cross-linking agents being particularly threatening to their survival. Fanconi anemia is a genetically heterogeneous disease with 13 different proteins identified, which seem to work together in a common pathway. Since one of the FA genes is identical to the breast cancer susceptibility gene BRCA2, it is also referred to as the FA/BRCA pathway. Eight proteins form a nuclear complex, whose integriry is required for the monoubiquitination of two other FA proteins, FANCD2 and FANCI, in response to DNA replication stress. Despite intensive research, the function of the FA/BRCA pathway at a molecular level has remained largely elusive so far. At the beginning of my thesis, we therefore decided to purify the proteins of the FA core complex and to investigate their biochemical properties. We started with the five proteins which were known at that time, FANCA, FANCC, FANCE, FANCF, and FACG. Later on, we extended our studies to the newly discovered proteins FANCL, FANCM, and FAAP24, and eventually focused our work on the characterisation of FANCM. In contrast to the other core complex proteins, FANCM contains two conserved domains, which point to a role in DNA metabolism: an N-terminal DEAH box helicase domain and a C-terminal ERCC4 nuclease domain. In this study, we have successfully purified full-length FANCM from a recombinant source. We show that purified FANCM binds to branched DNA molecules, such as Holliday junctions and replication forks, with high specificity and affinity. In addition, we demonstrate that FANCM can translocate the junction point of branched DNA molecules due to its helicase domain in an ATPase-dependent manner. FANCM can even dissociate large recombination intermediates, via branch migration of Holliday junctions through a 2.6 kb region of homology. Taken together, our data suggest that FANCM can specifically bind to replication forks and Holliday junctions in vitro, and that its DEAH box helicase domain is associated with a potent branch migration activity. We propose that FANCM might have a direct role in the processing of DNA replication intermediates. This is consistent with the current view that FA proteins coordinate DNA repair at stalled replication forks. Our findings provide a first hint as to the context in which FANCM might play a role in the cell. We are optimistic that they might be key to further elucidate the function of a pathway which is far from being understood.

Metric Plan Reading Course, HR-324, 1997

This Plan Reading Course was developed by the Department of Civil and Construction Engineering of Iowa State University under contract with the Iowa Highway Research Board, Project HR-324. It is intended to be an instructional tool for Iowa DOT, county and municipal employees within the state of Iowa. Under this contract, a previous Plan Reading Course, prepared for the Iowa State Highway Commission in 1965, has been completely revised using a new format, new plans, updated specifications, and new material. This course is a self-taught course consisting of two parts; Highway Plans, and Bridge and Culvert plans. Each part consists of a self-instruction book, a set of plans, a question booklet, and an answer booklet. This is the self-instruction book for the Bridge and Culvert Plans part of the course. The example structures included in this part of the course are a prestressed concrete beam bridge and a reinforced concrete box culvert.

Seleção de variáveis em modelos matemáticos dos parâmetros de cultivo do camarão marinho Litopenaeus vannamei

O objetivo deste trabalho foi selecionar as variáveis de manejo do camarão marinho Litopenaeus vannamei que mais influenciaram nas variáveis-respostas ao cultivo (produção, produtividade, peso final e taxa de sobrevivência), em modelos matemáticos. O banco de dados foi composto por 83 cultivos, realizados no período de 2003 a 2005, obtidos de uma fazenda comercial localizada no litoral sul de Pernambuco. Para estimar os parâmetros dos modelos, utilizou-se a técnica dos mínimos quadrados. A seleção das variáveis foi realizada com o processo "backward elimination" associado ao método de transformação de Box e Cox. A adequação das equações e os pressupostos de normalidade e homocedasticidade, para os erros, foram analisadas com base na análise de variância e análise de resíduo. É possível relacionar essas variáveis e estabelecer predições com as equações.

Highway Applications for Rammed Aggregate Piers in Iowa Soils, TR-443, 2003

This report describes a study to evaluate Geopier® soil reinforcement technology in transportation construction. Three projects requiring settlement control were chosen for evaluation—an embankment foundation, a box culvert, and a bridge approach fill. For each project, construction observations, in situ soil testing, laboratory material characterization, and performance monitoring were carried out. For the embankment foundation project, Geopier elements were installed within and around an abutment footprint for the new I-35 overpass at the US Highway 5/Interstate 35 interchange in Des Moines, Iowa. Although the main focus of this investigation was to evaluate embankment foundation reinforcement using Geopier elements, a stone column reinforced soil provided an opportunity to compare systems. In situ testing included cone penetration tests (CPTs), pressuremeter tests (PMTs), Ko stepped blade tests, and borehole shear tests (BSTs), as well as laboratory material testing. Comparative stiffness and densities of Geopier elements and stone columns were evaluated based on full-scale modulus load tests and standard penetration tests. Vibrating wire settlement cells and total stress cells were installed to monitor settlement and stress concentration on the reinforcing elements and matrix soil. Settlement plates were also monitored by conventional optical survey methods. Results show that the Geopier system and the stone columns performed their intended functions. The second project involved settlement monitoring of a 4.2 m wide x 3.6 m high x 50 m long box culvert constructed beneath a bridge on Iowa Highway 191 south of Neola, Iowa. Geopier elements were installed to reduce total and differential settlement while ensuring the stability of the existing bridge pier foundations. Benefits of the box culvert and embankment fill included (1) ease of future roadway expansion and (2) continual service of the roadway throughout construction. Site investigations consisted of in situ testing including CPTs, PMTs, BSTs, and dilatometer tests. Consolidated drained triaxial compression tests, unconsolidated undrained triaxial compression test, oedometer tests, and Atterberg limit tests were conducted to define strength and consolidation parameters and soil index properties for classification. Vibrating wire settlement cells, total stress cells, and piezometers were installed for continuous monitoring during and after box culvert construction and fill placement. This project was successful at controlling settlement of the box culvert and preventing downdrag of the bridge foundations, but could have been enhanced by reducing the length of Geopier elements at the ends of the box culvert. This would have increased localized settlement while reducing overall differential settlement. The third project involved settlement monitoring of bridge approach fill sections reinforced with Geopier elements. Thirty Geopier elements, spaced 1.8 m apart in six rows of varying length, were installed on both sides of a new bridge on US Highway 18/218 near Charles City, Iowa. Based on the results of this project, it was determined that future applications of Geopier soil reinforcement should consider extending the elements deeper into the embankment foundation fill, not just the fill itself.

Insights into the Design and Characteristics of the Sedimentation Process at Multi-Barrel Culverts in Iowa, TR-596, 2010

The present study is an integral part of a broader study focused on the design and implementation of self-cleaning culverts, i.e., configurations that prevent the formation of sediment deposits after culvert construction or cleaning. Sediment deposition at culverts is influenced by many factors, including the size and characteristics of material of which the channel is composed, the hydraulic characteristics generated under different hydrology events, the culvert geometry design, channel transition design, and the vegetation around the channel. The multitude of combinations produced by this set of variables makes the investigation of practical situations a complex undertaking. In addition to the considerations above, the field and analytical observations have revealed flow complexities affecting the flow and sediment transport through culverts that further increase the dimensions of the investigation. The flow complexities investigated in this study entail: flow non-uniformity in the areas of transition to and from the culvert, flow unsteadiness due to the flood wave propagation through the channel, and the asynchronous correlation between the flow and sediment hydrographs resulting from storm events. To date, the literature contains no systematic studies on sediment transport through multi-box culverts or investigations on the adverse effects of sediment deposition at culverts. Moreover, there is limited knowledge about the non-uniform, unsteady sediment transport in channels of variable geometry. Furthermore, there are few readily useable (inexpensive and practical) numerical models that can reliably simulate flow and sediment transport in such complex situations. Given the current state of knowledge, the main goal of the present study is to investigate the above flow complexities in order to provide the needed insights for a series of ongoing culvert studies. The research was phased so that field observations were conducted first to understand the culvert behavior in Iowa landscape. Modeling through complementary hydraulic model and numerical experiments was subsequently carried out to gain the practical knowledge for the development of the self-cleaning culvert designs.

Valproate treatment of human cord blood CD4-positive effector T cells confers on them the molecular profile (microRNA signature and FOXP3 expression) of natural regulatory CD4-positive cells through inhibition of histone deacetylase.

Regulatory T cells (Tregs) play a key role in immune system homeostasis and tolerance to antigens, thereby preventing autoimmunity, and may be partly responsible for the lack of an appropriate immune response against tumor cells. Although not sufficient, a high expression of forkhead box P3 (FOXP3) is necessary for their suppressive function. Recent reports have shown that histones deacetylase inhibitors increased FOXP3 expression in T cells. We therefore decided to investigate in non-Tregs CD4-positive cells, the mechanisms by which an aspecific opening of the chromatin could lead to an increased FOXP3 expression. We focused on binding of potentially activating transcription factors to the promoter region of FOXP3 and on modifications in the five miRs constituting the Tregs signature. Valproate treatment induced binding of Ets-1 and Ets-2 to the FOXP3 promoter and acted positively on its expression, by increasing the acetylation of histone H4 lysines. Valproate treatment also induced the acquisition of the miRs Tregs signature. To elucidate whether the changes in the miRs expression could be due to the increased FOXP3 expression, we transduced these non-Tregs with a FOXP3 lentiviral expression vector, and found no changes in miRs expression. Therefore, the modification in their miRs expression profile is not due to an increased expression of FOXP3 but directly results from histones deacetylase inhibition. Rather, the increased FOXP3 expression results from the additive effects of Ets factors binding and the change in expression level of miR-21 and miR-31. We conclude that valproate treatment of human non-Tregs confers on them a molecular profile similar to that of their regulatory counterpart.

Identification of the minimal promoter region of the mouse NKX5-3, a transcription factor implicated in eye development.

Early ocular development is controlled by a complex network of transcription factors, cell cycle regulators, and diffusible signalling molecules. Together, these molecules regulate cell proliferation and apoptosis, and specify retinal fate. NKX5-3 is a homeobox transcription factor implicated in eye development. The analysis of the 5'-flanking region of the mouse Nkx5-3 gene revealed a predicted TATA-less promoter sequence between -416 and -166 of the translation start site. To functionally characterise Nkx5-3 promoter activity, serial deletions of the promoter sequence were introduced in pGL-3 basic vector and promoter activity of these 5'- and 3'-deleted constructions was tested in HeLa and CHO cells. Transactivation assays identified a region between -350 and -296 exhibiting promoter-like activity. Combined analysis by deletions and point mutations showed that this sequence, containing multiple Sp1 binding sites was necessary to promote transcriptional activity. Binding of Sp1 to this region was confirmed by electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation, using an antibody specific for Sp1. Altogether, these results demonstrated that the immediate upstream region of Nkx5-3 gene possessed a strong intrinsic promoter activity in vitro, suggesting a potential role in Nkx5-3 transcription in vivo.

TLR3 and Rig-like receptor on myeloid dendritic cells and Rig-like receptor on human NK cells are both mandatory for production of IFN-gamma in response to double-stranded RNA.

Cross-talk between NK cells and dendritic cells (DCs) is critical for the potent therapeutic response to dsRNA, but the receptors involved remained controversial. We show in this paper that two dsRNAs, polyadenylic-polyuridylic acid and polyinosinic-polycytidylic acid [poly(I:C)], similarly engaged human TLR3, whereas only poly(I:C) triggered human RIG-I and MDA5. Both dsRNA enhanced NK cell activation within PBMCs but only poly(I:C) induced IFN-gamma. Although myeloid DCs (mDCs) were required for NK cell activation, induction of cytolytic potential and IFN-gamma production did not require contact with mDCs but was dependent on type I IFN and IL-12, respectively. Poly(I:C) but not polyadenylic-polyuridylic acid synergized with mDC-derived IL-12 for IFN-gamma production by acting directly on NK cells. Finally, the requirement of both TLR3 and Rig-like receptor (RLR) on mDCs and RLRs but not TLR3 on NK cells for IFN-gamma production was demonstrated using TLR3- and Cardif-deficient mice and human RIG-I-specific activator. Thus, we report the requirement of cotriggering TLR3 and RLR on mDCs and RLRs on NK cells for a pathogen product to induce potent innate cell activation.

The homologous regulators ANR of Pseudomonas aeruginosa and FNR of Escherichia coli have overlapping but distinct specificities for anaerobically inducible promoters.

The anaerobic transcriptional regulator ANR induces the arginine deiminase and denitrification pathways in Pseudomonas aeruginosa during oxygen limitation. The homologous activator FNR of Escherichia coli, when introduced into an anr mutant of P. aeruginosa, could functionally replace ANR for anaerobic growth on nitrate but not for anaerobic induction of arginine deiminase. In an FNR-positive E. coli strain, the ANR-dependent promoter of the arcDABC operon, which encodes the enzymes of the arginine deiminase pathway, was not expressed. To analyse systematically these distinct induction patterns, a lacZ promoter-probe, broad-host-range plasmid containing various -40 regions (the ANR/FNR recognition sequences) and -10 promoter sequences was constructed. These constructs were tested in P. aeruginosa and in E. coli expressing either ANR or FNR. In conjunction with the consensus -10 hexamer of E. coli sigma 70 RNA polymerase (TATAAT), the consensus FNR site (TTGAT ..... ATCAA) was recognized efficiently by ANR and FNR in both hosts. By contrast, when promoters contained the Arc box (TTGAC .... ATCAG), which is found in the arcDABC promoter, or a symmetrical mutant FNR site (CTGAT .... ATCAG), ANR was a more effective activator than was FNR. Conversely, an extended 22 bp, fully symmetrical FNR site allowed better activation with FNR than with ANR. Combination of the arc promoter -10 sequence (CCTAAT) with the Arc box or the consensus FNR site resulted in good ANR-dependent expression in P. aeruginosa but gave practically no expression in E. coli, suggesting that RNA polymerase of P. aeruginosa differs from the E. coli enzyme in -10 recognition specificity. In conclusion, ANR and FNR are able to activate the RNA polymerases of P. aeruginosa and E. coli when the -40 and -10 promoter elements ae identical or close to the E. coli consensus sequences.

Interactions of juvenile Lumbricus terrestris with adults and their burrow systems in a two-dimensional microcosm

The objective of this work was to evaluate interactions of Lumbricus terrestris juveniles with adults and with inherited burrow systems. An experiment was set up using a two dimensional Evans' boxes microcosm. Adult L. terrestris were added to 16 boxes (one individual per box) and kept in darkness at 17ºC along with eight unoccupied boxes for two months. The adult L. terrestris were removed from eight randomly selected boxes, and L. terrestris juveniles were added (one juvenile per box), composing three treatments with eight replicates: 1, with an adult in an inherited burrow (ABJ); 2, alone in an inherited burrow (BJ); and 3, alone in a previously uninhabited box (J). The proportion of juveniles occupying adult burrows observed was significantly different in treatments ABJ (48%) and BJ (75%). The mean mass of juveniles at experimental termination differed significantly among treatments and was greater in treatment J (4.04±0.39 g) in comparison to the BJ (3.09±0.93 g) and ABJ treatments (2.13±0.64 g). Results suggest a negative influence of both the presence of an adult and its burrow system on juvenile growth. Intraspecific competition partially explained this, but further investigation is required to examine how an inherited environment (i.e. burrow) could negatively affect the growth of juveniles.

Arabidopsis jasmonate signaling pathway.

Jasmonates control defense gene expression and male fertility in the model plant Arabidopsis thaliana. In both cases, the involvement of the jasmonate pathway is complex, involving large-scale transcriptional reprogramming. Additionally, jasmonate signaling is hard-wired into the auxin, ethylene, and salicylate signal networks, all of which are under intense investigation in Arabidopsis. In male fertility, jasmonic acid (JA) is the essential signal intervening both at the level of anther elongation and in pollen dehiscense. A number of genes potentially involved in jasmonate-dependent anther elongation have recently been discovered. In the case of defense, at least two jasmonates, JA and its precursor 12-oxo-phytodienoic acid (OPDA), are necessary for the fine-tuning of defense gene expression in response to various microbial pathogens and arthropod herbivores. However, only OPDA is required for full resistance to some insects and fungi. Other jasmonates probably affect yet more physiological responses. A series of breakthroughs have identified the SKP/CULLIN/F-BOX (SCF), CORONATINE INSENSITIVE (COI1) complex, acting together with the CONSTITUTIVE PHOTOMORPHOGENIC 9 (COP9) signalosome, as central regulatory components of jasmonate signaling in Arabidopsis. The studies, mostly involving mutational approaches, have paved the way for suppressor screens that are expected to further extend our knowledge of jasmonate signaling. When these and other new mutants affecting jasmonate signaling are characterized, new nodes will be added to the Arabidopsis Jasmonate Signaling Pathway Connections Map, and the lists of target genes regulated by jasmonates in Arabidopsis will be expanded.

A minimal i-motif stabilized by minor groove G:T:G:T tetrads

The repetitive DNA sequences found at telomeres and centromeres play a crucial role in the structure and function of eukaryotic chromosomes. This role may be related to the tendency observed in many repetitive DNAs to adopt non-canonical structures. Although there is an increasing recognition of the importance of DNA quadruplexes in chromosome biology, the co-existence of different quadruplex-forming elements in the same DNA structure is still a matter of debate. Here we report the structural study of the oligonucleotide d(TCGTTTCGT) and its cyclic analog d. Both sequences form dimeric quadruplex structures consisting of a minimal i-motif capped, at both ends, by a slipped minor groove-aligned G:T:G:T tetrad. These mini i-motifs, which do not exhibit the characteristic CD spectra of other i-motif structures, can be observed at neutral pH, although they are more stable under acidic conditions. This finding is particularly relevant since these oligonucleotide sequences do not contain contiguous cytosines. Importantly, these structures resemble the loop moiety adopted by an 11-nucleotide fragment of the conserved centromeric protein B (CENP-B) box motif, which is the binding site for the CENP-B.