Leukocyte-endothelial cell interaction is necessary for photodynamic therapy induced vascular permeabilization.

BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT) affects vascular barrier function and thus increases vessel permeability. This phenomenon may be exploited to facilitate targeted drug delivery and may lead to a new clinical application of photodynamic therapy. Here, we investigate the role of leukocyte recruitment for PDT-induced vascular permeabilization. STUDY DESIGN/MATERIAL AND METHODS: Fluorescein isothiocyanate dextran (FITC-D, 2,000 kDa) was injected intravenously 120 minutes after focal PDT on striated muscle in nude mice bearing dorsal skinfold chambers (Visudyne® 800 µg/kg, fluence rate 300 mW/cm2 , light dose of 200 J/cm2). Leukocyte interaction with endothelial cells was inhibited by antibodies functionally blocking adhesion molecules ("MABS-PDT" group, n = 5); control animals had PDT but no antibody injection (group "PDT", n = 7). By intravital microscopy, we monitored leukocyte rolling and sticking in real-time before, 90 and 180 minutes after PDT. The extravasation of FITC-D from striated muscle vessels into the interstitial space was determined in vivo during 45 minutes to assess treatment-induced alterations of vascular permeability. RESULTS: PDT significantly increased the recruitment of leukocytes and enhanced the leakage of FITC-D. Neutralization of adhesion molecules before PDT suppressed the rolling of leukocytes along the venular endothelium and significantly reduced the extravasation of FITC-D as compared to control animals (156 ± 27 vs. 11 ± 2 (mean ± SEM, number of WBC/30 seconds mm vessel circumference; P < 0.05) at 90 minutes after PDT and 194 ± 21 vs. 14 ± 4 at 180 minutes after PDT). In contrast, leukocyte sticking was not downregulated by the antibody treatment. CONCLUSION: Leukocyte recruitment plays an essential role in the permeability-enhancing effect of PDT.

Microbial biomass and soil fauna during the decomposition of cover crops in no-tillage system

The decomposition of plant residues is a biological process mediated by soil fauna, but few studies have been done evaluating its dynamics in time during the process of disappearance of straw. This study was carried out in Chapecó, in southern Brazil, with the objective of monitoring modifications in soil fauna populations and the C content in the soil microbial biomass (C SMB) during the decomposition of winter cover crop residues in a no-till system. The following treatments were tested: 1) Black oat straw (Avena strigosa Schreb.); 2) Rye straw (Secale cereale L.); 3) Common vetch straw (Vicia sativa L.). The cover crops were grown until full flowering and then cut mechanically with a rolling stalk chopper. The soil fauna and C content in soil microbial biomass (C SMB) were assessed during the period of straw decomposition, from October 2006 to February 2007. To evaluate C SMB by the irradiation-extraction method, soil samples from the 0-10 cm layer were used, collected on eight dates, from before until 100 days after residue chopping. The soil fauna was collected with pitfall traps on seven dates up to 85 days after residue chopping. The phytomass decomposition of common vetch was faster than of black oat and rye residues. The C SMB decreased during the process of straw decomposition, fastest in the treatment with common vetch. In the common vetch treatment, the diversity of the soil fauna was reduced at the end of the decomposition process.

History of the Trappist Abbey of New Melleray, in Dubuque County, Iowa,1892

Written by William Rufus Perkins this book is the history of the Trappist Abbey of New Melleray in Dubuque County, Iowa. New Melleray is a Cistercian (Trappist) monastery located in the rolling farmland south of Dubuque

Long-Term Community Recovery Report, Parkersburg, Iowa, December 2008

At approximately 5pm on May 25, 2008, City of Parkersburger was struck by a powerful EF5 tornado, estimated to be three-quarters of a mile wide. As it moved across the southern half of the City, the tornado cased severe damage, destroying at least twenty businesses and over two hundred fifty homes along with city Hall and Aplington-Parkersburg High School. Numerous other homes, businesses and civic buildings were damaged. Approximately two weeks later, Beaver Creek was among the many Iowa streams to flood. While Parkersburg was not affected significantly by flooding, a number of properties were damaged, including Beaver Meadows Golf Course and a City park with athletic fields.

Iowa Rail Route Alternatives Analysis, June 1998

The development of new rail systems in the first part of the 21st century is the result of a wide range of trends that are making it increasingly difficult to maintain regional mobility using the two dominant intercity travel modes, auto and air. These trends include the changing character of the economic structure of industry. The character of the North American industrial structure is moving rapidly from a manufacturing base to a service based economy. This is increasing the need for business travel while the increase in disposable income due to higher salaries has promoted increased social and tourist travel. Another trend is the change in the regulatory environment. The trend towards deregulation has dramatically reduced the willingness of the airlines to operate from smaller airports and the level of service has fallen due to the creation of hub and spoke systems. While new air technology such as regional jets may mitigate this trend to some degree in medium-size airports, smaller airports will continue to lose out. Finally, increasing environmental concerns have reduced the ability of the automobile to meet intercity travel needs because of increased suburban congestion and limited highway capacity in big cities. Against this background the rail mode offers new options due to first, the existing rail rights-of-way offering direct access into major cities that, in most cases, have significant capacity available and, second, a revolution in vehicle technology that makes new rail rolling stock faster and less expensive to purchase and operate. This study is designed to evaluate the potential for rail service making an important contribution to maintaining regional mobility over the next 30 to 50 years in Iowa. The study evaluates the potential for rail service on three key routes across Iowa and assesses the impact of new train technology in reducing costs and improving rail service. The study also considers the potential for developing the system on an incremental basis. The service analysis and recommendations do not involve current Amtrak intercity service. That service is presumed to continue on its current route and schedule. The study builds from data and analyses that have been generated for the Midwest Rail Initiative (MWRI) Study. For example, the zone system and operating and capital unit cost assumptions are derived from the MWRI study. The MWRI represents a cooperative effort between nine Midwest states, Amtrak and the Federal Railroad Administration (FRA) contracting with Transportation Economics & Management Systems, Inc. to evaluate the potential for a regional rail system. The 1 The map represents the system including the decision on the Iowa route derived from the current study. Iowa Rail Route Alternatives Analysis TEMS 1-2 system is to offer modern, frequent, higher speed train service to the region, with Chicago as the connecting hub. Exhibit 1-1 illustrates the size of the system, and how the Iowa route fits in to the whole.

Multiple sulfatase deficiency with neonatal manifestation.

Multiple Sulfatase Deficiency (MSD; OMIM 272200) is a rare autosomal recessive inborn error of metabolism caused by mutations in the sulfatase modifying factor 1 gene, encoding the formyglycine-generating enzyme (FGE), and resulting in tissue accumulation of sulfatides, sulphated glycosaminoglycans, sphingolipids and steroid sulfates. Less than 50 cases have been published so far. We report a new case of MSD presenting in the newborn period with hypotonia, apnoea, cyanosis and rolling eyes, hepato-splenomegaly and deafness. This patient was compound heterozygous for two so far undescribed SUMF1 mutations (c.191C¿>¿A; p.S64X and c.818A¿>¿G; p.D273G).

Heparin attenuates TNF-alpha induced inflammatory response through a CD11b dependent mechanism

Background In addition to its anticoagulant properties, heparin has anti-inflammatory effects, the molecular and mechanistic bases of which are incompletely defined. AIMS The current studies were designed to test the hypothesis that heparin abrogates the expression or function of leucocyte-endothelial adherence molecules which are fundamental to the acute inflammatory response. Methods The effects of heparin on tumour necrosis factor alpha (TNF-¿) induced leucocyte rolling, adhesion, and migration as well as vascular permeability were assessed in rat mesenteric venules using intravital microscopy. Expression of adhesion molecules was quantitated using a double radiolabelled monoclonal antibody (mAb) binding technique in vivo (P-selectin, intercellular cell adhesion molecule type 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1)) or flow cytometry (CD11a, CD11b, and L-selectin). Ex vivo binding of heparin to neutrophils was assessed by flow cytometry. RESULTS TNF-alpha induced a significant increase in leucocyte rolling, adhesion, and migration, and vascular permeability, coincident with a significant increase in expression of P-selectin, ICAM-1, and VCAM-1. Ex vivo assessment of blood neutrophils showed significant upregulation of CD11a and CD11b and significant downregulation of L-selectin within five hours of TNF-¿ administration. Heparin pretreatment significantly attenuated leucocyte rolling, adhesion, and migration but did not affect expression of cell adhesion molecules or vascular permeability elicited by TNF-¿ administration. Binding of heparin was significantly increased on blood neutrophils obtained five hours after TNF-¿ administration. Preincubation with an anti-CD11b mAb but not with an anti-CD11a or anti-L-selectin antibody significantly diminished heparin binding ex vivo.

Disrupting the EMMPRIN (CD147)-cyclophilin A interaction reduces infarct size and preserves systolic function after myocardial ischemia and reperfusion.

Objective-Inflammation and proteolysis crucially contribute to myocardial ischemia and reperfusion injury. The extracellular matrix metalloproteinase inducer EMMPRIN (CD147) and its ligand cyclophilin A (CyPA) may be involved in both processes. The aim of the study was to characterize the role of the CD147 and CyPA interplay in myocardial ischemia/reperfusion (I/R) injury.Methods and Results-Immunohistochemistry showed enhanced expression of CD147 and CyPA in myocardial sections from human autopsies of patients who had died from acute myocardial infarction and from mice at 24 hours after I/R. At 24 hours and 7 days after I/R, the infarct size was reduced in CD147(+/-) mice vs CD147(+/+) mice (C57Bl/6), in mice (C57Bl/6) treated with monoclonal antibody anti-CD147 vs control monoclonal antibody, and in CyPA(-/-) mice vs CyPA(+/+) mice (129S6/SvEv), all of which are associated with reduced monocyte and neutrophil recruitment at 24 hours and with a preserved systolic function at 7 days. The combination of CyPA(-/-) mice with anti-CD147 treatment did not yield further protection compared with either inhibition strategy alone. In vitro, treatment with CyPA induced monocyte chemotaxis in a CD147-and phosphatidylinositol 3-kinase-dependent manner and induced monocyte rolling and adhesion to endothelium (human umbilical vein endothelial cells) under flow in a CD147-dependent manner.Conclusion-CD147 and its ligand CyPA are inflammatory mediators after myocardial ischemia and reperfusion and represent potential targets to prevent myocardial I/R injury.

Iowa Byways: The Official Guide to Iowa Byways, 2013

Browse through this guide and you’ll find the distinct flavor of what is available along each byway. Discover recreational, historic, cultural and scenic attractions using the maps and lists provided in the guide. You’ll find numbered attractions for each byway in or near the town listed. For a comprehensive list of byway features, visit www.iowabyways.org. Friendly local contacts are provided to help you along the way. Iowa Transportation Maps clearly tracking all the Iowa byways with red dotted lines are available at Iowa’s official welcome centers. Traveling Iowa’s byways you will experience small town America, while enjoying diverse landscapes and unique landforms that have been shaped over thousands of years. Iowa’s cultural heritage also plays a major role across all 11 byways, boasting hundreds of historic sites, national landmarks and interpretive centers, each telling Iowa’s stories from the first Native Americans through European immigrants to modern times. Glaciers once covered much of Iowa, shaping the broad flat plains of the prairie. These massive sheets of ice missed the northeast corner of the state, leaving the land along the Driftless Area Byway rugged and hilly with rock outcroppings, springs and cold water trout streams. Rivers coursed their way through the land, carving deep furrows in some places and leaving gently rolling hills in others. In western Iowa, wind has shaped fine sand into the impressive Loess Hills, a rare land form found in only one other place on earth. Iowa’s two national scenic byways and nine state byways offer unique varieties of scenic features, and more for you to see and do. View three states from atop a Mississippi River bluff, stop at a modern art museum and then tour a working farm. Explore a historic mill, visit a national aquarium, take a boat ride in a cave, purchase locally crafted pottery and wares from local artisans or trace the footsteps of Lewis and Clark. Experience the actual wagon ruts of the Mormon Trail, ride your bike 13 stories high, canoe a water trail, star gaze under Iowa’s darkest sky, and marvel at mounds built by prehistoric cultures. Agriculture wraps Iowa’s byways with an abundance of farmland vistas and fills Iowa lands with ever-changing crops and activities for you to “harvest.” You’ll see croplands on the vast flat plains and farmsteads sprinkled across rolling hills reminiscent of a Grant Wood painting. Along the way, you might wander in a corn maze, rest at a bed and breakfast, study farming in museums, discover the Iowa barn quilt collection or visit a working Amish farm. When you are ready to step outside your vehicle, you’ll find much more to do and see. Prairie, forests, rivers and public lands are abundant along Iowa’s byways; providing opportunities for you to stop and play in the outdoors with hiking, biking, kayaking and trout fishing. Classic hometowns with pride for their unique lore and offerings are found all along the byways. They invite you to taste local food, enjoy their architecture, and immerse yourself in the rich history and culture that defines them. Why not plan your next journey off the beaten path? No matter how you choose to make the most of every moment, we know that time spent along Iowa’s byways is sure to grow your love for Iowa’s diverse, beautiful vistas and authentic communities. Happy driving!

Alkaline phosphatase activity as a tool for assessing nutritional conditions in the seagrass Posidonia oceanica.(L.) Delile

The effects of experimental phosphorus enrichments on alkaline phosphatase activity (APA) of the seagrass Posidonia oceanica (L.) Delile were tested. Short-term additions (as phosphate, 12 hours, in the laboratory) decreased APA by 18-2896, depending on the plant part considered (roots, young leaves or old leaves). The values of APA after this treatment were well correlated with interna1 phosphorus pools (as P concentration in plant tissues). Long-term additions (as phosphate, added to the sediment, 1 month in situ) decreased APA by 40-75%, also depending on the plant part. We conclude that alkaline phosphatase activity is a good indicator of P deficiency in this seagrass. We used this indicator to assess the P-nutritional status in a series of meadows in the NW Mediterranean, finding a high geographical variability, but correlations between APA and basic features of the meadows (carbonate content of the sediment, organic content of the sediment, shoot density, etc.) were not significant. Consequently, phosphorus deficiency does not seem to be directly related to these descriptors.

Spatial distribution and biomass of aquatic rooted macrophytes and their relevance in the metabolism of a Mediterranean coastal lagoon

This work aims to characterise the current autotrophic compartment of the Albufera des Grau coastal lagoon (Menorca, Balearic Islands) and to assess the relationship between the submerged macrophytes and the limnological parameters of the lagoon. During the study period the submerged vegetation was dominated by the macrophyte Ruppia cirrhosa, which formed dense extensive meadows covering 79% of the surface. Another macrophyte species, Potamogeton pectinatus, was also observed but only forming small stands near the rushing streams. Macroalgae were only occasionally observed. Macrophyte biomass showed a clear seasonal trend, with maximum values in July. The biomass of R. cirrhosa achieved 1760 g DW m-2, the highest biomass ever reported for this species in the literature. The seasonal production-decomposition cycle of the macrophyte meadows appears to drive the nutrient dynamics and carbon fluxes in the lagoon. Despite the significant biomass accumulation and the absence of a washout of nutrients and organic matter to the sea, the lagoon did not experience a dystrophic collapse. These results indicate that internal metabolism is more important than exchange processes in the lagoon.

Initiation de l'adhésion leucocytaire : rôle des fucosyltransférases et des ligands des sélectines

RESUME La dissémination extramédullaire des cellules blastiques est une complication majeure des leucémies myéloïdes (LMA) ou lymphoïdes aiguës (LLA). La migration des cellules blastiques dépend de mécanismes semblables à ceux qui régulent la migration des leucocytes dans un site d'inflammation. Parmi ceux-ci, les oligosaccharides fucosylés décorant les ligands des sélectines jouent un rôle clé en interagissant avec les sélectines. PSGL-1 (P-Selectin Glycoprotein Ligand-1) est une protéine de 240 kD, exprimée à la surface des leucocytes, permettant de soutenir le roulement leucocytaire sur les sélectines, le long de la paroi vasculaire. L'interaction de PSGL-1 avec les sélectines nécessite des modifications post-traductionnelles de type sialylation, sulfatation , N et 0-glycosylation. Parmi les enzymes impliqués, les α1,3-fucosyltransférases jouent un rôle important dans la biosynthèse d'oligosaccharides fucosylés, ligands des sélectines (sLex, Lex, VIM-2, CLA). Comme l'expression des α1,3-fucosyltransférases par les cellules blastiques leucémiques n'a pas été étudiée précédemment, nous l'avons recherchée dans 120 cas de leucémies aiguës. Les ARNm des FucT-IV et -VII ont été détectés, par RT-PCR, dans tous les cas testés. L'ARNm de la FucT-IX n'a été observé que dans 40% des leucémies aiguës (48/120). L'ARNm de la FucT-IX est détecté dans 65% des LMA (47/72) et, moins fréquemment, dans 26% des LLA (11/42). A noter que les cas de LLA exprimant la FucT-IX correspondent essentiellement à des LLA secondaires à la transformation d'une leucémie myéloïde chronique ou des LLA de la lignée B de type leucémie/lymphome de Burkitt. L'expression de PSGL-1 et des oligosaccharides fucosylés par les blastes varie significativement parmi les LMA et les LLA : Lex, VIM-2 et sLex étant exprimés plus fréquemment par les myéloblastes que par les lymphoblastes. Le rôle des FucT-IV, -VII et -IX dans la synthèse des Lex, VIM-2, CLA et sLex a été examiné en exprimant l'ADNc de chaque FucT dans des cellules CHO. L'immunophénotypisation des transfectants indique que la FucT-VII synthétise sLex et CLA, mais pas Lex et VIM-2. Lex et VIM-2 sont générés par la FucT-IV. La FucT-IX ne participe qu'à la synthèse de Lex, sa capacité de synthèse de VIM-2 dans les cellules CHO est très faible. Le rôle de la FucT-IX dans la régulation du roulement cellulaire dépendant des sélectines a été testé dans des conditions de flux. Les vitesses de roulement des cellules CHO co-exprimant la FucT-LX, la core-2 01,6-N-acetylglucosaminyltransferase et PSGL-1 sont très élevées sur la P-sélectine (médiane : 497.95 µm/s, n=96) alors qu'elles sont beaucoup plus lentes sur la E-sélectine (médiane 7 µm/s, n=64). Les recrutements sur la E-sélectine des cellules CHO-C2F9PSGL¬1 et des CHO-C2F7PSGL-1 sont similaires (moyenne ± SEM : 127.44 ± 4.38 vs. 151.16 ± 3.16 cellules/min/mm2, n=5). Celui des cellules CHO-C2F4PSGL-1 est par contre plus faible (54.20 ± 2.13 cellules/min/mm2, n=5). Ces résultats indiquent que la FucT-IX est impliquée dans la biosynthèse de Lex, VIM-2 et CLA et qu'elle régule l'interaction des cellules CHO avec la E-sélectine. Contrairement aux FucT-IV et -VII, la FucT-IX ne joue qu'un rôle mineur dans la régulation du roulement cellulaire sur la L- et la P-sélectine. L'expression fréquente de la FucT-IX par les myéloblastes suggère qu'elle pourrait participer avec les FucT-IV et -VII à la régulation de la migration cellulaire dépendant de la E-sélectine. Finalement, ce travail de thèse a été étendu à l'identification des protéines cytoplasmiques qui interagissent avec le domaine cytoplasmique de PSGL-1 et qui pourraient être impliquées dans la transmission de signaux intracellulaires. Les ligands intracellulaires de PSGL-1 seront identifiés par la technique du double hybride qui nous a déjà permis de confirmer que syk et la N-moésine se lient au domaine cytoplasmique de PSGL-1. Des ligands supplémentaires seront identifiés employant une librairie provenant des cellules souches hématopoïétiques comme proie. ABSTRACT Blast cell dissemination is a major complication of acute myeloblastic (AML) and lymphoblastic leukemia (ALL). Blast cell migration is dependent on mechanisms that are similar to those which regulate leukocyte migration into inflammatory lesions. Among them, fticosylated oligosaccharides that decorate selectin ligands play a key role by interacting with selectins. PSGL-1 (P-Selectin Glycoprotein Ligand-1) is a 240 kD glycoprotein constitutively expressed on leucocytes and which supports leukocyte rolling on selectins. PSGL-1 interaction with selectins is dependent on post-translational modifications such as sialylation, sulfation, N- and 0-glycosylation. Among the involved enzymes, the α1,3-fucosyltransferases (FucT) play a major role in generating cell surface glycoconjugates carrying fucosylated oligosaccharides which interact with selectins (sLex, Lex, VIM-2, CLA). Since no information is available on the expression of α1,3-fucosyltransferases by leukemic blast cells, we examined it in 120 cases of acute leukemia. FucT-IV and -VII mRNAs were detected, by RT-PCR, in all tested cases. In contrast, the presence of FucT-IX mRNA was shown in only 40% of patients with acute leukemia (48/120). FucT-IX mRNA was detected in 65% of AML (47/72) and, less frequently, in 26% of ALL (11/42). Importantly, all ALL cases expressing FucT-IX were either secondary leukemia resulting from the transformation of chronic myelocytic leukemia in acute lymphoblastic leukemia or mature B-ALL (FAB L3 subtype or Burkitt lymphoma/leukemia according to WHO classification). FucT-IX was not detected in precursor B or T-ALL. The expression of PSGL-1 and fucosylated epitopes was significantly different among AML and ALL, Lex, VIM-2 and sLex being more frequently expressed by myeloblasts than by lymphoblasts. The role of FucT-IV, -VII and -IX in the biosynthesis of Lex, VIM-2, CLA and sLex was examined by expressing the cDNA of each α1,3-FucT in CHO cells. Immunophenotypic analysis of CHO transfectants indicated that FucT-VII synthesizes sLex and CLA but not Lex or VIM-2. Lex and CLA were generated by both FucT-IV and -IX. FucT-IV and FucT-IX differed in their ability to synthesize VIM-2, FucT-IX being less efficient than FucT-IV. The role of FucT-IX in regulating selectin-dependent rolling was assessed under hydrodynamic flow conditions. P-selectin-dependent interactions were transient and occurred at high velocities (median: 497.95 1,µm/s, n=96). In contrast, much slower rolling velocities were observed on E-selectin (median: 7 µm/s, n=64). The recruitment of CHO-C2F9PSGL-1 and CHO-C2F7PSGL-1 cells was similar on E-selectin (mean ± SEM: 127.44 ± 4.38, n=5 vs 151.16 ± 3.16 cells/min/mm2, n=5). In the other hand, CHO-C2F4PSGL-1 cells were less efficiently recruited on E-selectin (54.20 ± 2.13 cells/min/mm2, n=5). This results indicate that FucT-IX is involved in the biosynthesis of Lex, VIM-2 and CLA and that it confers E-selectin binding activity to CHO cells. By contrast to FucT-IV and -VII, FucT-IX had a minor role in regulating P- and L-selectin-dependent rolling on CHO transfectants. The frequent expression of FucT-IX in myeloblasts suggests that it may participate with FucT-IV and -VII in regulating E-selectin-dependent cell migration into tissues. Finally, this thesis work was extended to the identification of the cytoplasmic proteins interacting with cytoplasmic domain of PSGL-1 that may be involved in transducing intracellular signals. We planned to identify these intracellular ligands of PSGL-1 by using the double hybrid technique and already confirmed that syk and N-moesin bind to the cytoplasmic domain of PSGL-1. Additional PSGL-1 ligands will be sought by the same technique using a CD34+ stem cell library as pray. RESUME DESTINE A UN LARGE PUBLIC : L'adhésion et la migration leucocytaire sont nécessaires à de nombreux processus cellulaires comme la régulation de l'hématopoïèse, mais aussi dans la pathogenèse de l'artériosclérose, des maladies inflammatoires et de la métastatisation des cellules cancéreuses. Les molécules impliquées constituent depuis peu des cibles pour la thérapie du cancer. La migration leucocytaire vers un site d'inflammation dépend de mécanismes complexes, se déroulant en plusieurs étapes, nécessitant l'interaction séquentielle de molécules d'adhésion leucocytaires et endothéliales. Ainsi, chronologiquement, suite à un stimulus inflammatoire, les leucocytes « roulent » sur les cellules endothéliales, sont activées, s'arrêtent et traversent la paroi endothéliale (diapédèse) pour migrer dans les tissus environnants inflammés selon un gradient chimiotactique. La première étape de roulement met en jeu deux molécules principales : PSGL-1 (P-Sélectine Glycoprotéine Ligand-1) du coté des leucocytes et les sélectines du coté de l'endothélium de la paroi vasculaire. L'interaction entre ces deux molécules nécessite des décorations de ces protéines par des sucres, des résidus sulfates et des acides sialiques. Le sucre essentiel à la liaison demeure le fucose qui est attaché aux protéines grâce à des enzymes de la famille des fucosyltransferases. Actuellement, neuf fucosyltransférases humaines ont été identifiées et désignées sous FucT-I à IX. La FucT-IX, dernière fucosyltransférase clonée, a un faible degré d'homologie avec les autres fucosyltransférases mais sa séquence est extrêmement conservée entre les espèces. Ceci traduit son importance par une forte résistance à la pression évolutive. L'examen de son expression au sein de 120 cas de leucémies aiguës a mis en évidence son comportement atypique. En effet, alors que les autres FucTs sont toujours présentes, la FucT¬IX ne s'exprime que dans un cas sur deux en moyenne avec une préférence plus importante pour les leucémies myéloïdes. Ainsi, une étude plus approfondie de cet enzyme à mis en évidence sa capacité à induire une interaction cellulaire plus spécifique de la E-sélectine. Elle décore non seulement des protéines de surface, mais aussi certainement les glycolipides constituant la membrane cellulaire.

Regulation of P-selectin glycoprotein ligand 1 (PSGL-1) interactions with selectins : role of the decameric repeats and of the cytoplasmic domain

SUMMARY BACKGROUND: P-selectin glycoprotein ligand 1 (PSGL-1) is a major selectin ligand, mediating leukocyte rolling along inflamed vascular wall. It is a mucin-like homodimer composed of a N-terminal domain which binds selectins, followed by 14-16 decameric repeats (DR), a transmembrane domain and a cytoplasmic tail, which may be involved in regulating leukocyte rolling and in generating intracellular signals, through its binding to moesin and Syk. P- and L-selectin binding is dependent on core-2 O-glycosylation and tyrosine sulfation of PSGL-1 N-terminus. However, a minor part of E-selectin-mediated rolling is dependent on N-terminal O-glycans; additional binding sites may thus be involved. In this project, we studied whether (1) PSGL-1 DR and (2) PSGL-1 cytoplasmic residues which bind moesin, were also involved in the regulation of selectin-dependent rolling. METHODS: Several mutated cDNAs were obtained: (1) PSGL-1 DR were either deleted, or substituted by platelet GPlba macroglycopeptide, (2) Ser-336, -348, Lys-337 and Arg-338 were mutated to alanine; moreover, truncation mutants retaining only 6 or 2 cytoplasmic residues were also generated. Transfected CHO expressing mutant PSGL-1 were tested for their ability to bind soluble selectin chimeras and to support selectin-dependent rolling under flow conditions. RESULTS: (1) Deletion of the DR had a dramatic effect on P- and L-selectin-dependent cell recruitment and rolling stability, which could only partially be compensated for, by GPlba substitution. In addition, we observed that DR create a binding site for E-selectin and thus support PSGL-1-dependent rolling. (2) Flow assays revealed that the moesin-binding site, in particular Ser-336, plays a crucial role in regulating the recruitment, velocity and rolling stability of PSGL-1-expressing cells on P- and L-selectin. CONCLUSIONS: Data presented here highlight the structure -function relationship of PSGL-1 DR. Moreover, they reveal a crucial role for the moesin-binding residues in regulating P-and L-selectin-dependent rolling. RÉSUMÉ CONTEXTE: PSGL-1 (P-selectin glycoprotein ligand 1) est un ligand majeur des sélectines permettant le roulement des leucocytes le long de la paroi vasculaire enflammée. C'est un homodimère de type mucine, composé d'un domaine N-terminal liant les sélectines, suivi de 14-16 répétitions décamèriques (RD), d'un domaine transmembranaire et d'une queue cytoplasmique qui pourrait être impliquée dans la régulation du roulement leucocytaire et la génération de signaux intracellulaires, via sa liaison à la moésine et à Syk. La liaison à la Pet à la L-sélectine dépend de la présentation par le N-terminus de PSGL-1 de O-glycans sur des structures core-2 et de tyrosines sulfatées. Cependant, une fraction mineure du roulement médié par la E-sélectine dépend des O-glycans N-terminaux; des sites de liaisons supplémentaires pourraient donc être impliqués. Dans ce projet, nous avons étudié si (1) les RD de PSGL-1 ainsi que (2) les résidus cytoplasmiques liant la moésine, étaient impliqués dans la régulation du roulement dépendant des sélectines. MÉTHODES: Plusieurs ADN codant des formes mutées de PSGL-1 ont été obtenus: (1) Les RD de PSGL-1 ont été soit ôtées, soit remplacées par le macroglycopeptide de la GPlba plaquettaire, (2) les Ser-336, -348, la Lys-337 et l'Arg-338 ont été mutées en alanine; par ailleurs, des mutants tronqués ne retenant plus que 6 ou 2 résidus cytoplasmiques ont également été générés. Des CHO transfectées exprimant PSGL-1 muté ont été testées pour leur capacité à lier des sélectines chimériques solubles et à soutenir un roulement dépendant des sélectines dans des conditions de flux. RÉSULTATS: (1) La perte des RD a eu un effet dramatique sur le recrutement cellulaire et la stabilité de roulement dépendant des P- et L-sélectine, qui n'a pu être que partiellement compensé par la substitution par la GPlba. De plus, nous avons observé que les RD forment un site de liaison pour la E-sélectine et soutiennent ainsi le roulement dépendant de PSGL-1. (2) Les tests de flux ont révélé que le site de liaison à la moésine, notamment la Ser-336, joue un rôle crucial dans la régulation du recrutement, de la vitesse et de la stabilité du roulement des cellules exprimant PSGL-1 sur les P- et L-sélectine. CONCLUSIONS; Les données présentées ici ont permis d'éclaircir la relation structure -fonction des RD de PSGL-1. Par ailleurs, elles révèlent un rôle crucial pour les résidus liant la moésine dans le roulement dépendant des P- et L-sélectine. RÉSUMÉ DESTINÉ À UN LARGE PUBLIC Pour accomplir ses fonctions, le sang circule sur un réseau de 96'000 kilomètres; ainsi, il approvisionne les cellules de l'organisme en énergie, il transporte diverses substances, il assure la défense contre les pathogènes et il participe à la régulation de la température corporelle. Le sang contient plusieurs types de cellules: la grande majorité sont les globules rouges, auxquels il faut ajouter les plaquettes (dont le rôle est de colmater les lésions vasculaires) et les globules blancs (leucocytes) qui, bien que présents en très faible quantité (moins de 0.01 %), jouent un rôle crucial en cas d'infection ou d'inflammation. Une attaque par un pathogène provoque plusieurs changements (rougeur, chaleur, gonflement, douleur), qui sont des manifestations de l'inflammation. Pour atteindre l'agent infectieux, des globules blancs spécialisés (les granulocytes) doivent quitter la circulation sanguine. Afin de faciliter leur capture, les vaisseaux sanguins vont exprimer des protéines telles que les sélectines, qui sont reconnues par une protéine leucocytaire appelée PSGL-1 (P-selectin glycoprotein ligand 7). L'interaction des sélectines avec PSGL-1 soutient le roulement du globule blanc le long de la paroi vasculaire, à une vitesse très inférieure à celle du flux sanguin. Ce roulement conduit à l'activation du globule blanc par des molécules de l'inflammation, permettant son adhésion ferme, puis son arrêt. Finalement, le granulocyte va migrer à travers la paroi du vaisseau pour atteindre et éliminer les causes de l'inflammation. L'adhésion est un processus intéressant à caractériser, car outre l'inflammation, il est également impliqué dans l'artériosclérose, l'infarctus, la métastatisation et la thrombose. Dans ce travail, nous nous sommes intéressés à définir les rôles des différents domaines de PSGL-1 dans la régulation de son interaction avec les sélectines. En effet, en plus de son extrémité extracellulaire de haute affinité pour les sélectines, PSGL-1 est composé de plusieurs séquences répétées hautement glycosylées et d'une courte région intracellulaire, dont les fonctions n'avaient pas été étudiées auparavant. En créant des formes mutées de PSGL-1, nous avons pu montrer qu'un roulement efficace des leucocytes nécessite la présence des régions répétitives et du domaine intracellulaire au complet.

An integrated encyclopedia of DNA elements in the human genome.

The human genome encodes the blueprint of life, but the function of the vast majority of its nearly three billion bases is unknown. The Encyclopedia of DNA Elements (ENCODE) project has systematically mapped regions of transcription, transcription factor association, chromatin structure and histone modification. These data enabled us to assign biochemical functions for 80% of the genome, in particular outside of the well-studied protein-coding regions. Many discovered candidate regulatory elements are physically associated with one another and with expressed genes, providing new insights into the mechanisms of gene regulation. The newly identified elements also show a statistical correspondence to sequence variants linked to human disease, and can thereby guide interpretation of this variation. Overall, the project provides new insights into the organization and regulation of our genes and genome, and is an expansive resource of functional annotations for biomedical research.

Iowa Natural Resources, Agency Performance Plan, FY2013

The work of the Department of Natural Resources impacts the lives of all Iowans. Iowans deserve a clean environment and quality natural areas for public use and enjoyment. This report reflects the progress made during fiscal year 2013 (FY13) toward our goals and provides information regarding the condition of our state’s natural resources and the effectiveness of our programs. In FY13, we continued to improve collaboration with other executive branch agencies. The DNR and DOT work very closely on the issuance of permits needed for road and bridge constructions, but recently we have also been working together to meet the administrative needs of the agencies. The DNR is working closely with the DOT to adopt an Electronic Records Management System used by the DOT. This system will improve accessibility to public documents and reduce the amount of paper files retained in storage. The DNR also continues to improve collaboration with other agencies, such as the Iowa Economic Development Authority as we work closely with them on business development in the state. The DNR strives to continually improve our customer service and how we can meet Iowan’s needs. As an example, the online reservation system for campground reservations has grown over the past eight years so that now 88% of the camping reservations are made online. The DNR continues to improve our online presence and accessibility. In FY13 the Iowa Legislature approved paying off the State’s bond debt used to construct Honey Creek Resort State Park. By removing this debt, the DNR will be able to focus more on the future of the Resort, rather than the past debt. Finally, in August of 2012, the DNR was faced with a tragic accident, where a seasonal parks employee died after rolling a mower into a lake. This incident has caused us to establish a Safety Program at the DNR and to review all of our departmental safety trainings, programs, and equipment. By focusing on our employee’s safety and well being, it is another way that we can demonstrate that at the DNR, our employees are our greatest asset.