Rôle des molécules de co-stimulations et de CD93 dans la différenciation en plasmocytes

Le développement des cellules B est constitué d'une première phase qui se déroule dans la moelle en absence d'antigène et d'une deuxième phase qui se déroule dans les organes lymphoïdes secondaires et qui débute uniquement en présence d'antigène. Cette deuxième partie est extrêmement importante et doit être très bien régulée pour lutter efficacement contre les pathogènes, ainsi que pour éviter de nombreuses maladies de type auto-immunes. Ce travail est basé à l'origine sur l'étude de souris mutantes dans lesquelles une protéine des cellules T est modifiée, impliquant une très forte activation des cellules B en absence d'antigène et de manière non spécifique. Ces souris constituent donc un outil de travail très intéressant pour étudier tout d'abord le mécanisme aboutissant à l'activation des cellules B dans ce contexte particulier. De plus comme ces souris contiennent énormément de cellules sécrétant des anticorps, à savoir les plasmocytes, il est facile d'étudier leur phénotype. Cela nous a permis de démontrer qu'un récepteur membranaire, CD93 est exprimé à leur surface. Cette observation a ensuite été confirmée dans des souris normales, de type sauvage. L'utilisation de ce marqueur de surface nous a permis de caractériser plus en détail les étapes du développement des plasmocytes. De plus nous avons tenté de trouver la fonction jouée par cette molécule à la surface de ces cellules, en utilisant des souris dans lesquelles ce récepteur a été supprimé. Si les premières étapes de l'activation des cellules B étaient normales, ces souris n'étaient par contre pas capables de produire des anticorps à long-terme dans le sang. Nous avons pu montrer que la survie des plasmocytes en l'absence de CD93 est moins efficace dans la moelle, probablement du au fait qu'en absence de cette molécule, les plasmocytes ont plus de difficultés à adhérer dans ce que l'on appelle des niches de survie. Nous avons essayé ensuite de déterminer si CD93 peut être utilisé comme cible thérapeutique dans le cadre de maladies auto-immunes ou de lymphomes. Bien que CD93 soit exprimé à la surface des cellules d'intérêt dans les souris souffrant de lupus, il n'a pas été possible de les éliminer avec un anticorps dirigé contre CD93. De plus nous n'avons pas pu mettre en évidence l'expression de CD93 à la surface des plasmocytes humains induits in vitro. SUMMARY : Antigen dependent B cell activation is a key aspect of the adaptive immunity which is involved in the efficient response against pathogens, but also in vaccination and in numerous pathologies. The aim of this project was to investigate two key aspects of the late B cell development, namely the role of costimulatory molecules in the immunological synapse between T and B cells and the characterization of a new plasma cell marker, CD93. This work was initially based on the study of the LatY136F mutant mouse. The latter harbors a point mutation in the LAT adaptor protein which is involved in T cell receptor signaling. As a consequence of this mutation, CD4 T cells in the periphery expand strongly and are polarized in a TH2 manner leading to a normal but exaggerated B cell response. For this reason, these mice provide a useful tool to investigate different aspects of the late B cell development. The first part of the project was focused on the role played by costimulatory molecules in LotY136F CD4 T cell mediated B cell activation. In vitro studies showed that CD80/CD86, IL-4 and LFA-1 were required for LatY136FT cells to activate B cells whereas CD40 and IcosL were not necessary. In vivo we showed that CD80/CD86 was required for initial T cell expansion whereas CD40 and IcosL deficiency led to a less efficient B cell activation. The large amount of plasma cells present in LatY136F mice allows investigating in more details their phenotype and CD93 was found to be expressed on their surface, This observation was confirmed in wild type B cells activated either in vivo or in vitro with T-independent or T-dependent antigens. Moreover we found that CD93 expression can occur either before CD138/Blimp-1 induction or after, showing that two independent pathways can lead to the formation of CD93/CD138 double positive population, which was shown to be the more mature. Indeed, their phenotype correlated with modified transcriptional network, high isotype switched antibody secretion and cell cycle arrest. Analysis of CD93 deficient mice demonstrated that the initial B cell activation after immunization was normal, but also showed that these mice failed to maintain a high antibody secretion level at later time points both after primary and boost immunization. This was shown to be due to a less efficient survival of the long-lived plasma cells in the bone marrow niches, most likely related with a defective adhesion process in absence of CD93. We investigated the possibility to use CD93 as a target to treat plasma cell pathologies, but even if this molecule is expressed on cells of interest in the bone marrow of lupus mice, it was not possible to deplete them using anti-CD93 antibodies. Moreover we were not able to show its expression on the surface of in vitro activated B cells and multiple myeloma cell lines of human origin. In conclusion, our data helped understand both the mechanisms leading to the polyclonal B cell activation occurring in the LatY136F KI mouse and the role played by CD93 on the surface of plasma cells, which could potentially open the way to therapeutic application.

Influence of soil pH on inorganic phosphorus sorption and desorption in a humid brazilian Ultisol

Liming is a common practice to raise soil pH and increase phosphorus (P) bioavailability in tropical regions. However, reports on the effect of liming on P sorption and bioavailability are controversial. The process of phosphorus desorption is more important than P sorption for defining P bioavailability. However few studies on the relationship between soil pH and P desorption are available, and even fewer in the tropical soils. The effects of soil pH on P sorption and desorption in an Ultisol from Bahia, Brazil, were investigated in this study. Phosphorus sorption decreased by up to 21 and 34 % with pH increases from 4.7 to 5.9 and 7.0, respectively. Decreasing Langmuir K parameter and decreasing partition coefficients (Kd) with increasing pH supported this trend. Phosphorus desorption was positively affected by increased soil pH by both the total amount of P desorbed and the ratio of desorbed P to initially sorbed P. A decreased K parameter and increased Kd value, particularly at the highest pH value and highest P-addition level, endorsed this phenomenon. Liming the soil had the double effect of reducing P sorption (up to 4.5 kg ha-1 of remaining P in solution) and enhancing P desorption (up to 2.7 kg ha-1 of additionally released P into solution).

Hydrologic Simulations of the Maquoketa River Watershed Using SWAT Working Paper 09-WP 49,June 2009

This paper describes the application of the Soil and Water Assessment Tool (SWAT) model to the Maquoketa River watershed, located in northeast Iowa. The inputs to the model were obtained from the Environmental Protection Agency’s geographic information/database system called Better Assessment Science Integrating Point and Nonpoint Sources (BASINS). Climatic data from six weather stations located in and around the watershed, and measured streamflow data from a U.S. Geological Survey gage station at the watershed outlet were used in the sensitivity analysis of SWAT model parameters as well as its calibration and validation for watershed hydrology and streamflow. A sensitivity analysis was performed using an influence coefficient method to evaluate surface runoff and base flow variations in response to changes in model input hydrologic parameters. The curve number, evaporation compensation factor, and soil available water capacity were found to be the most sensitive parameters among eight selected parameters when applying SWAT to the Maquoketa River watershed. Model calibration, facilitated by the sensitivity analysis, was performed for the period 1988 through 1993, and validation was performed for 1982 through 1987. The model performance was evaluated by well-established statistical methods and was found to explain at least 86% and 69% of the variability in the measured stream flow data for the calibration and validation periods, respectively. This initial hydrologic modeling analysis will facilitate future applications of SWAT to the Maquoketa River watershed for various watershed analysis, including water quality.

Expression of E-Selectin and Vascular Cell Adhesion Molecule-1 in So-Called 'Negative' Appendices: First Results to Support the Pathological Diagnosis in Borderline Cases.

Background: Since the rate of histologically 'negative' appendices still ranges between 15 and 20%, appendicitis in 'borderline' cases remains a challenging disease. As previously described, cell adhesion molecule expression correlates with different stages of appendicitis. Therefore, it was of interest to determine whether the 'negative' appendix correlated with the absence of E-selectin or vascular cell adhesion molecule-1 (VCAM-1). Methods: Nineteen grossly normal appendices from a series of 120 appendectomy specimens from patients with suspected appendicitis were analysed in frozen sections for the expression of E-selectin and VCAM-1. As control, 5 normal appendices were stained. Results: This study showed a coexpression of E-selectin and VCAM-1 in endothelial cells in early and recurrent appendicitis. In patients with symptoms for less than 6 h, only E-selectin was detected. Cases with fibrosis and luminal obliteration were only positive for VCAM-1. In cases of early appendicitis with symptoms of less than 6 h duration, a discordance between histological and immunohistochemical results was found. Conclusions: This report indicates that E-selectin and VCAM-1 expression could be useful parameters in the diagnosis of appendicitis in borderline cases.

Biological control of soil-borne pathogens by fluorescent pseudomonads.

Particular bacterial strains in certain natural environments prevent infectious diseases of plant roots. How these bacteria achieve this protection from pathogenic fungi has been analysed in detail in biocontrol strains of fluorescent pseudomonads. During root colonization, these bacteria produce antifungal antibiotics, elicit induced systemic resistance in the host plant or interfere specifically with fungal pathogenicity factors. Before engaging in these activities, biocontrol bacteria go through several regulatory processes at the transcriptional and post-transcriptional levels.

Nitrogen application timing and soil inorganic nitrogen dynamics under no-till oat/maize sequential cropping

The timing of N application to maize is a key factor to be considered in no-till oat/maize sequential cropping. This study aimed to evaluate the influence of pre-planting, planting and sidedress N application on oat residue decomposition, on soil N immobilisation and remineralisation and on N uptake by maize plants in no-till oat/maize sequential cropping. Undisturbed soil cores of 10 and 20 cm diameter were collected from the 0-15 cm layer of a no-till Red Latossol, when the oat cover crop was in the milk-grain stage. Two greenhouse experiments were conducted simultaneously. Experiment A, established in the 10 cm diameter cores and without plant cultivation, was used to asses N dynamics in soil and oat residues. Experiment B, established in the 20 cm diameter cores and with maize cultivation, was used to assess plant growth and N uptake. An amount of 6.0 Mg ha-1 dry matter of oat residues was spread on the surface of the cores. A rate of 90 kg N ha-1 applied as ammonium sulphate in both experiments was split in pre-planting, planting and sidedress applications as follows: (a) 00-00-00 (control), (b) 90-00-00 (pre-planting application, 20 days before planting), (c) 00-90-00 (planting application), (d) 00-30-60 (split in a planting and a sidedress application 31 days after emergence), (e) 00-00-00* (control, without oat residue) and (f) 90-00-00* (pre-planting application, without oat residue). The N concentration and N content in oat residues were not affected during decomposition by N fertilisation. Most of the fertiliser NH4+-N was converted into NO3--N within 20 days after application. A significant decrease in NO3--N contents in the 0-4 cm layer was observed in all treatments between 40 and 60 days after the oat residue placement on the soil surface, suggesting the occurrence of N immobilisation in this period. Considering that most of the inorganic N was converted into NO3- and that no immobilisation of the pre planting fertiliser N occurred at the time of its application, it was possible to conclude that pre-planting applied N was prone to losses by leaching. On the other hand, with split N applications, maize plants showed N deficiency symptoms before sidedress application. Two indications for fertiliser-N management in no-till oat/maize sequential cropping could be suggested: (a) in case of split application, the sidedress should be earlier than 30 days after emergence, and (b) if integral application is preferred to save field operations, this should be done at planting.

Qualidade do solo avaliada pelo "Soil Quality Kit Test" em dois experimentos de longa duração no Rio Grande do Sul

A avaliação da qualidade do solo (QS) é importante estratégia no planejamento agrícola, possibilitando a identificação e o aprimoramento de sistemas de manejo com características de alta produtividade e de preservação ambiental. O presente estudo foi realizado em dois experimentos de longa duração (10 e 15 anos) conduzidos no Sul do Brasil e teve por objetivo avaliar o efeito de sistemas de manejo na QS, utilizando um kit de análise expedita de qualidade de solo (KQS), desenvolvido pelo Instituto de Qualidade do Solo-USDA-ARS. A eficiência desse kit foi avaliada pela comparação com os métodos tradicionais utilizados na ciência do solo. Nas duas áreas experimentais investigou-se um total de 12 tratamentos, os quais englobaram sistemas de preparo com diferentes intensidades de revolvimento do solo (preparo convencional, preparo reduzido e plantio direto) e sistemas de culturas com ampla faixa de adição de resíduos vegetais ao solo, além da aplicação de doses anuais de N-uréia, variando de 0 a 144 kg ha-1. Em cada base experimental uma área sob campo natural foi avaliada, servindo como referência da condição do solo na ausência de interferência antrópica. Como indicadores de QS, foram avaliados infiltração de água, respiração do solo, densidade do solo, teor de nitrato+nitrito (N-NO3- + N-NO2-), estabilidade de agregados em água e pH. De maneira geral, os coeficientes de correlação entre os métodos do KQS e os métodos tradicionais foram elevados, sendo o mais alto para o indicador pH (r = 0,98) e o menor para o indicador infiltração de água no solo (r = 0,42). Os tratamentos selecionados foram teoricamente ordenados em ordem crescente de QS, a qual foi reproduzida de forma eficiente pelo índice de estoque de carbono (IEC), calculado pela razão entre o estoque de C orgânico do solo, na camada de 0-5 cm, de cada tratamento e o estoque de C orgânico no solo sob campo natural. Os indicadores estabilidade de agregados, N-NO3- + N-NO2- e respiração do solo foram os mais eficientes em discriminar a QS. O KQS foi eficiente em avaliar a QS dos tratamentos nas duas áreas experimentais. Os níveis mais elevados de QS foram alcançados nos tratamentos com plantio direto e consórcio de gramíneas e leguminosas tropicais, devido à cobertura do solo e às elevadas adições de C e N via resíduos culturais.

Leaf senescence of common bean plants as affected by soil phosphorus supply

Responses of leaf senescence to P supply could constitute adaptive mechanisms for plant growth under P-limiting conditions. The aim of this study was to evaluate the effects of soil P supply on leaf senescence of common bean (Phaseolus vulgaris L.). Eight P levels, ranging from 5 to 640 mg kg-1 P, were applied to pots containing four bean plants of cultivar Carioca in 10 kg of an Oxic Haplustult soil. Attached leaves were counted weekly, abscised leaves were collected every other day, and seeds were harvested at maturity. The number of live leaves increased until 48 days after emergence (DAE) and decreased afterwards, irrespective of applied P levels. At lower applied P levels, the initial increase and the final decrease of leaf number was weak, whereas at higher applied P levels the leaf number increased intensively at the beginning of the growth cycle and decreased strongly after 48 DAE. Dry matter and P accumulated in senesced leaves increased as soil P levels increased until 61 DAE, but differences between P treatments narrowed thereafter. The greatest amounts of dry mass and P deposited by senesced leaves were observed at 48-54 DAE for high P levels, at 62-68 DAE for intermediate P levels and at 69-76 DAE for low P levels. These results indicate that soil P supply did not affect the stage of maximal leaf number and the beginning of leaf senescence of common bean plants, but the stage of greatest deposition of senesced leaves occurred earlier in the growth cycle as the soil P supply was raised.

Maize response to nitrogen fertilization timing in two tillage systems in a soil with high organic matter content

No-tillage systems, associated to black oat as preceding cover crop, have been increasingly adopted. This has motivated anticipated maize nitrogen fertilization, transferring it from the side-dress system at the stage when plants have five to six expanded leaves to when the preceding cover crop is eliminated or to maize sowing. This study was conducted to evaluate the effects of soil tillage system and timing of N fertilization on maize grain yield and agronomic efficiency of N applied to a soil with high organic matter content. A three-year field experiment was conducted in Lages, state of Santa Catarina, from 1999 onwards. Treatments were set up in a split plot arrangement. Two soil tillage systems were tested in the main plots: conventional tillage (CT) and no-tillage (NT). Six N management systems were assessed in the split-plots: S1 - control, without N application; S2 - all N (100 kg ha-1) applied at oat desiccation; S3 - all N applied at maize sowing; S4 - all N side-dressed when maize had five expanded leaves (V5 growth stage); S5 - 1/3 of N rate applied at maize sowing and 2/3 at V5; S6 - 2/3 of nitrogen rate applied at maize sowing and 1/3 at V5. Maize response to the time and form of splitting N was not affected by the soil tillage system. Grain yield ranged from 6.0 to 11.8 t ha-1. The anticipation of N application (S2 and S3) decreased grain yield in two of three years. In the rainiest early spring season (2000/2001) of the experiment, S4 promoted an yield advantage of 2.2 t ha-1 over S2 and S3. Application of total N rate before or at sowing decreased the number of kernels produced per ear in 2000/2001 and 2001/2002 and the number of ears produced per area in 2001/2002, resulting in reduced grain yield. The agronomic efficiency of applied N (kg grain increase/kg of N applied) ranged from 13.9 to 38.8 and was always higher in the S4 than in the S2 and S3 N systems. Short-term N immobilization did not reduce grain yield when no N was applied before or at maize sowing in a soil with high organic matter content, regardless of the soil tillage system.

Classification of acid sulphate soils of Finland according to Soil Taxonomy and the FAO/Unesco legend

Selostus: Suomen happamien sulfaattimaiden kansainvälinen luokittelu

A software to calculate soil hydraulic conductivity in internal drainage experiments (SHC, Version 2.00)

A software for the calculation of unsaturated soil hydraulic conductivity K(theta) is presented for commonly used methods found in the literature, based on field experiments in which a soil profile is submitted to water infiltration followed by internal drainage. The software is available at: dourado@esalq.usp.br.

Division of Soil Conservation Soil and Water Quality Review, 2009

Special Points of Interest: • The Division of Soil Conservation celebrated its 70th anniversary July 1, 2009. The Iowa Soil Conservation: Laws were enacted in 1939 creating the state soil conservation agency and governing committee and providing for the creation of Iowa’s 100 soil and water conservation districts. • The Mines & Minerals Bureau, through the federal Abandoned Mine Land (AML) Program, worked with various watershed groups to again secure an additional $1 million dollars in funding for the construction on projects in Marion, Mahaska and Monroe Counties. • Iowa hosted the Mississippi River/Gulf of Mexico Hypoxia Task Force tour and meeting in September 2009.

Podocalyxin is a novel polysialylated neural adhesion protein with multiple roles in neural development and synapse formation

Neural development and plasticity are regulated by neural adhesion proteins, including the polysialylated form of NCAM (PSA-NCAM). Podocalyxin (PC) is a renal PSA-containing protein that has been reported to function as an anti-adhesin in kidney podocytes. Here we show that PC is widely expressed in neurons during neural development. Neural PC interacts with the ERM protein family, and with NHERF1/2 and RhoA/G. Experiments in vitro and phenotypic analyses of podxl-deficient mice indicate that PC is involved in neurite growth, branching and axonal fasciculation, and that PC loss-of-function reduces the number of synapses in the CNS and in the neuromuscular system. We also show that whereas some of the brain PC functions require PSA, others depend on PC per se. Our results show that PC, the second highly sialylated neural adhesion protein, plays multiple roles in neural development.

Regenerating cortical connections in a dish: the entorhino-hippocampal organotypic slice co-culture as tool for pharmacological screening of molecules promoting axon regeneration

We present a method for using long-term organotypic slice co-cultures of the entorhino-hippocampal formation to analyze the axon-regenerative properties of a determined compound. The culture method is based on the membrane interphase method, which is easy to perform and is generally reproducible. The degree of axonal regeneration after treatment in lesioned cultures can be seen directly using green fluorescent protein (GFP) transgenic mice or by axon tracing and histological methods. Possible changes in cell morphology after pharmacological treatment can be determined easily by focal in vitro electroporation. The well-preserved cytoarchitectonics in the co-culture facilitate the analysis of identified cells or regenerating axons. The protocol takes up to a month.

Temporal changes in the expression and distribution of adhesion molecules during liver development and regeneration

We have compared by immunocytochemistry and immunoblotting the expression and distribution of adhesion molecules participating in cell-matrix and cell-cell interactions during embryonic development and regeneration of rat liver. Fibronectin and the fibronectin receptor, integrin alpha 5 beta 1, were distributed pericellularly and expressed at a steady level during development from the 16th day of gestation and in neonate and adult liver. AGp110, a nonintegrin fibronectin receptor was first detected on the 17th day of gestation in a similar, nonpolarized distribution on parenchymal cell surfaces. At that stage of development haemopoiesis is at a peak in rat liver and fibronectin and receptors alpha 5 beta 1 and AGp110 were prominent on the surface of blood cell precursors. During the last 2 d of gestation (20th and 21st day) hepatocytes assembled around lumina. AGp110 was initially depolarized on the surface of these acinar cells but then confined to the lumen and to newly-formed bile canaliculi. At birth, a marked increase occurred in the canalicular expression of AGp110 and in the branching of the canalicular network. Simultaneously, there was enhanced expression of ZO-1, a protein component of tight junctions. On the second day postpartum, presence of AGp110 and of protein constituents of desmosomes and intermediate junctions, DGI and E-cadherin, respectively, was notably enhanced in cellular fractions insoluble in nonionic detergents, presumably signifying linkage of AGp110 with the cytoskeleton and assembly of desmosomal and intermediate junctions. During liver regeneration after partial hepatectomy, AGp110 remained confined to apical surfaces, indicating a preservation of basic polarity in parenchymal cells. A decrease in the extent and continuity of the canalicular network occurred in proliferating parenchyma, starting 24 h after resection in areas close to the terminal afferent blood supply of portal veins and spreading to the rest of the liver within the next 24 h. Distinct acinar structures, similar to the ones in prenatal liver, appeared at 72 h after hepatectomy. Restoration of the normal branching of the biliary tree commenced at 72 h. At 7 d postoperatively acinar formation declined and one-cell-thick hepatic plates, as in normal liver, were observed.