Unexpected role of surface transglutaminase type II in celiac disease

Background & Aims: In celiac disease (CD), transglutaminase type II (TG2) has 2 fundamental roles: (1) as the autoantigen recognized by highly specific autoantibodies and (2) the modifier of pathogenic gliadin T-cell epitopes. It follows that inhibition of TG2 might represent an attractive strategy to curb the toxic action of gliadin. Here we studied the validity of this strategy using the organ culture approach. Methods: Duodenal biopsy specimens from 30 treated patients with CD, 33 untreated patients with CD, and 24 controls were cultured with or without gliadin peptides p31-43, pα-9, and deamidated pα-9 for 20 minutes, 3 hours, and 24 hours. In 31 patients with CD and 16 controls, TG2 inhibitor R283 or anti-TG CUB 7402 or anti-surface TG2 (6B9) mAbs were used in cultures. T84 cells were also cultured with or without peptides with or without TG inhibitors. Mucosal modifications after culture were assessed by immunofluorescence, in situ detection of TG activity, confocal microscopy, and fluorescence-activated cell sorter analysis. Results: The enzymatic inhibition of TG2 only controlled gliadin-specific T-cell activation. The binding of surface TG2 contained gliadin-specific T-cell activation and p31-43-induced actin rearrangement, epithelial phosphorylation, and apoptosis, both in organ cultures and T84 cells. Conclusions: These data indicate a novel and unexpected biological role for surface TG2 in the pathogenesis of CD suggesting a third role for TG2 in CD. These results have a specific impact for celiac disease, with wider implications indicating a novel biologic function of TG2 with possible repercussions in other diseases. © 2005 by the American Gastroenterological Association.

Kinetic studies of the metathesis of 1-hexene using Re2O7/γ-Al2O3 and the synthesis and metathesis of oxazolines

The kinetics of the metathesis of 1-hexene using Re2O7/-Al_2O_3 as the catalyst were investigated under a variety of conditions. The experiments were carried out under high vacuum conditions. The product solutions were characterised by gas liquid chromatography and mass spectroscopy. The initial kinetics of the metathesis of 1-hexene showed that the reaction was first order in the weight of the catalyst and second order in the concentration of 1-hexene. A kinetic scheme which correlated the experimental data with the metallocarbene chain mechanism postulated by Herisson and Chauvin and the kinetics of the reaction was explained using a model based on the Langmuir-Hinshelwood theory. The low conversion of 1-hexene to its products is due to termination reactions which most likely occur by the decomposition of the metallocyclobutane intermediate to produce a cyclopropane derivative and an inactive centre. The optimum temperature for the metathesis of 1-hexene over Re_2O_7/-Al2O3 is 45oC and above this temperature, the rate of metathesis decreases rapidly. Co-catalysts alter the active sites for metathesis so that the catalyst is more selective to the metathesis of 1-hexene. However, the regeneration of metathesis activity is much worse for promoted catalysts than for the unpromoted. The synthesis and metathesis of 4,4-dimethyl-2-allowbreak (9-decenyl)-1,3-oxazoline and 4,4-dimethyl-2-allowbreak (3-pentenyl)-1,3-oxazoline was attempted and the products were analysed by thin layer chromatography, infra-red, 13C and 1H nmr and mass spectroscopy. Obtaining the oxazolines in a good yield with high purity was difficult and consequently metathesis of the impure products did not occur.

Subunit vaccines distearoylphosphatidylcholine-based liposomes entrapping antigen offer a neutral alternative to dimethyldioctadecylammonium-based cationic liposomes as an adjuvant delivery system

The adjuvanticity of liposomes can be directed through formulation to develop a safe yet potent vaccine candidate. With the addition of the cationic lipid dimethyldioctadecylammonium bromide (DDA) to stable neutral distearoylphosphatidylcholine (DSPC):cholesterol (Chol) liposomes, vesicle size reduces while protein entrapment increases. The addition of the immunomodulator, trehalose 6,6-dibehenate (TDB) to either the neutral or cationic liposomes did not affect the physiochemical characteristics of these liposome vesicles. However, the protective immune response, as indicated by the amount of IFN-? production, increases considerably when TDB is present. High levels of IFN-? were observed for cationic liposomes; however, there was a marked reduction in IFN-? release over time. Conversely, for neutral liposomes containing TDB, although the initial amount of IFN-? was slightly lower than the cationic equivalent, the overall protective immune responses of these neutral liposomes were effectively maintained over time, generating good levels of protection. To that end, although the addition of DSPC and Chol reduced the protective immunity of DDA:TDB liposomes, relatively high protection was observed for the neutral counterpart, DSPC:Chol:TDB, which may offer an effective neutral alternative to the DDA:TDB cationic system, especially for the delivery of either zwitterionic (neutral) or cationic molecules or antigens.

Dual affinity method for plasmid DNA purification in aqueous two-phase systems

The DNA binding fusion protein, LacI-His6-GFP, together with the conjugate PEG-IDA-Cu(II) (10 kDa) was evaluated as a dual affinity system for the pUC19 plasmid extraction from an alkaline bacterial cell lysate in poly(ethylene glycol) (PEG)/dextran (DEX) aqueous two-phase systems (ATPS). In a PEG 600-DEX 40 ATPS containing 0.273 nmol of LacI fusion protein and 0.14% (w/w) of the functionalised PEG-IDA-Cu(II), more than 72% of the plasmid DNA partitioned to the PEG phase, without RNA or genomic DNA contamination as evaluated by agarose gel electrophoresis. In a second extraction stage, the elution of pDNA from the LacI binding complex proved difficult using either dextran or phosphate buffer as second phase, though more than 75% of the overall protein was removed in both systems. A maximum recovery of approximately 27% of the pCU19 plasmid was achieved using the PEG-dextran system as a second extraction system, with 80-90% of pDNA partitioning to the bottom phase. This represents about 7.4 microg of pDNA extracted per 1 mL of pUC19 desalted lysate.

Zirconia and alumina based catalysts for steam reforming of naphthalene

The present paper deals with experimentation of ZrO2 and Al2O3-supported catalysts for conversion of naphthalene, chosen as tar model compound of pyrolysis or gasification syngas. In particular, the reforming capacity of active metals and promoters such as Co, Ni, Fe, Cr, Ce and Pt was tested in a fixed bed reactor at temperature from 400 to 900 °C. As regards ZrO2-supported catalysts, the best results were achieved by the Ni/Fe/Pt catalyst with 96% naphthalene conversion, 78% and 280% as CO and H2 production yield at 800 °C. Regarding Al2O3-supported catalysts, they were more active on average than the zirconia ones, achieving a very good performance even at 500 °C (90–100% naphthalene conversion, 30–40% CO yield and 300–350% H2 yield at 550 °C). Influence of different amounts of alumina, montmorillonite and carbon on carrier composition as well as pellets’ size were also studied. Both zirconia and alumina catalysts showed deactivation at higher temperatures due to coke deposition, resulting in a strong H2 production drop. Regeneration of catalysts by O2 and steam as well as activation by H2 were also studied. The activated catalyst was able to convert more than 99% naphthalene at 450 °C with a CO and H2 production yield of 26% and 420%, respectively.

A comparative study of cationic liposome and niosome-based adjuvant systems for protein subunit vaccines:characterisation, environmental scanning electron microscopy and immunisation studies in mice

Vesicular adjuvant systems composing dimethyldioctadecylammonium (DDA) can promote both cell-mediated and humoral immune responses to the tuberculosis vaccine fusion protein in mice. However, these DDA preparations were found to be physically unstable, forming aggregates under ambient storage conditions. Therefore there is a need to improve the stability of such systems without undermining their potent adjuvanticity. To this end, the effect of incorporating non-ionic surfactants, such as 1-monopalmitoyl glycerol (MP), in addition to cholesterol (Chol) and trehalose 6,6′-dibehenate (TDB), on the stability and efficacy of these vaccine delivery systems was investigated. Differential scanning calorimetry revealed a reduction in the phase transition temperature (T c) of DDA-based vesicles by ∼12°C when MP and cholesterol (1:1 molar ratio) were incorporated into the DDA system. Transmission electron microscopy (TEM) revealed the addition of MP to DDA vesicles resulted in the formation of multi-lamellar vesicles. Environmental scanning electron microscopy (ESEM) of MP-Chol-DDA-TDB (16:16:4:0.5 μmol) indicated that incorporation of antigen led to increased stability of the vesicles, perhaps as a result of the antigen embedding within the vesicle bilayers. At 4°C DDA liposomes showed significant vesicle aggregation after 28 days, although addition of MP-Chol or TDB was shown to inhibit this instability. Alternatively, at 25°C only the MP-based systems retained their original size. The presence of MP within the vesicle formulation was also shown to promote a sustained release of antigen in-vitro. The adjuvant activity of various systems was tested in mice against three subunit antigens, including mycobacterial fusion protein Ag85b-ESAT-6, and two malarial antigens (Merozoite surface protein 1, MSP1, and the glutamate rich protein, GLURP). The MP- and DDA-based systems induced antibody responses at comparable levels whereas the DDA-based systems induced more powerful cell-mediated immune responses. © 2006 The Authors.

Characterization of cationic liposomes based on dimethyldioctadecylammonium and synthetic cord factor from M. tuberculosis (trehalose 6,6′- dibehenate) - A novel adjuvant inducing both strong CMI and antibody responses:a novel adjuvant inducing both strong CMI and antibody responses

Incorporation of the glycolipid trehalose 6,6′-dibehenate (TDB) into cationic liposomes composed of the quaternary ammonium compound dimethyldioctadecylammonium (DDA) produce an adjuvant system which induces a powerful cell-mediated immune response and a strong antibody response, desirable for a high number of disease targets. We have used differential scanning calorimetry (DSC) to investigate the effect of TDB on the gel-fluid phase transition of DDA liposomes and to demonstrate that TDB is incorporated into DDA liposome bilayers. Transmission Electron Microscopy (TEM) and cryo-TEM confirmed that liposomes were formed when a lipid film of DDA containing small amounts of TDB was hydrated in an aqueous buffer solution at physiological pH. Furthermore, time development of particle size and zeta potential of DDA liposomes incorporating TDB during storage at 4°C and 25°C, indicates that TDB effectively stabilizes the DDA liposomes. Immunization of mice with the mycobacterial fusion protein Ag85B-ESAT-6 in DDA-TDB liposomes induced a strong, specific Th1 type immune response characterized by substantial production of the interferon-γ cytokine and high levels of IgG2b isotype antibodies. The lymphocyte subset releasing the interferon-γ was identified as CD4 T cells.

Mathematical Model and Simulation of a Pneumatic Apparatus for In-Drilling Alignment of an Inertial Navigation Unit during Horizontal Well Drilling

Conventional methods in horizontal drilling processes incorporate magnetic surveying techniques for determining the position and orientation of the bottom-hole assembly (BHA). Such means result in an increased weight of the drilling assembly, higher cost due to the use of non-magnetic collars necessary for the shielding of the magnetometers, and significant errors in the position of the drilling bit. A fiber-optic gyroscope (FOG) based inertial navigation system (INS) has been proposed as an alternative to magnetometer -based downhole surveying. The utilizing of a tactical-grade FOG based surveying system in the harsh downhole environment has been shown to be theoretically feasible, yielding a significant BHA position error reduction (less than 100m over a 2-h experiment). To limit the growing errors of the INS, an in-drilling alignment (IDA) method for the INS has been proposed. This article aims at describing a simple, pneumatics-based design of the IDA apparatus and its implementation downhole. A mathematical model of the setup is developed and tested with Bloodshed Dev-C++. The simulations demonstrate a simple, low cost and feasible IDA apparatus.

The Church of Our Blessed Redeemer Who Walked Upon the Waters

The Church of Our Blessed Redeemer Who Walked Upon the Waters is a collection of short stories about Elwyn Parker, a devout pianist who becomes a worldly car salesman. "Thirty Fingers," "My Father's Business," and "Apostate" introduce Elwyn, a saint at church and a trouble-making evangelist at school, who nevertheless finds himself in a love affair with an older woman, Sister Morrisohn. In "Captivity," Elwyn, a college freshman, experiences worldliness, then grows to resent and ultimately reject Sister Morrisohn. In "The Leap," Elwyn is back at the piano, but unemployed and unhappily married. He finds comfort only in his decade-old affair. In "The Lord of Travel," Elwyn, a car salesman, appears bereft of his former morals until he hoodwinks Ida, who reminds him of the now deceased Sister Morrisohn. Elwyn repairs Ida's car, redeeming himself in the process.

Da educação infantil ao ensino fundamental: o que contam as crianças sobre essa travessia na cultura de escola

From inquiries concerning the child as an individual with rights, this work takes as its object of study the perception of 5-7 years old children on their journey from kindergarten to elementary school, in a school culture. The objective of the research is, therefore, to investigate what the children tell in narratives drawn into a conversation circle about their experiences of school life in kindergarten and the first grade of elementary school. The participants were 18 children from a public school in the city of Natal (RN). Five rounds of conversation were held in which the children told a little alien, who was unaware of the school culture, what they knew about school and what they did at it. The research is linked to the project "Children's Narratives. What the children tell about childhood schools?"(Passeggi et all, 2011) and adopts epistemological principles and research methods of (auto)biographical education, taking as a working hypothesis the child's ability to reflect on their experiences and understand from their point of view, what happens to them. Analyses were organized based on the concept of school culture (Barroso, 2012). In the narratives of children, the three dimensions of school culture: the functionalist (purpose and rules), structural (structure and pedagogical organization) and the interactional (relations with others, with the spaces and with knowledge) are considered intertwined in their school perceptions and signal experienced tensions in a process of "conversion" from child to student. Children seem to realize the uniqueness of each level of education. They recognize as a characteristic of early childhood education the recreational activities, and as injunctions of the first year of elementary school the "study", "learning to read and write" to "be smart" to "change." The schooling will thus, constitute, in their eyes, as a time and a place where the children's culture gives way to school culture, and in this journey they experience that the desire to play and the duty/want to study cross the three dimensions of school. At the end of the journey, the status of children as cultural beings with rights is confirmed, whose narratives about school and about their experiences of "conversion" in a student, reveal much about the power of reflection on themselves, the school and the society in which they live, legitimizing their place in educational research and in child care policies.

Alcuni applicazioni dell'azione di un gruppo su un insieme

la tesi tratta alcuni risultati e applicazioni relative alla teoria delle azioni di gruppi su insiemi finiti, come l'equazione delle classi e il teorema di Cauchy. Infine illustra l'uso di tali risultati nella rappresentazione tramite permutazioni.

2003 Habitat Maps derived from Grey Nurse Shark (GNS) Project of Wolf Rock, Double Island Point, Queensland, Australia

Surveying habitats critical to the survival of grey nurse sharks in South-East Queensland has mapped critical habitats, gathered species inventories and developed protocols for ecological monitoring of critical habitats in southern Queensland. This information has assisted stakeholders with habitat definition and effective management. In 2002 members of UniDive applied successfully for World Wide Fund for Nature, Threatened Species Network funds to map the critical Grey Nurse Shark Habitats in south east Queensland. UniDive members used the funding to survey, from the boats of local dive operators, Wolf Rock at Double Island Point, Gotham, Cherub's Cave, Henderson's Rock and China Wall at North Moreton and Flat Rock at Point Look Out during 2002 and 2003. These sites are situated along the south east Queensland coast and are known to be key Grey Nurse Shark aggregation sites. During the project UniDive members were trained in mapping and survey techniques that include identification of fish, invertebrates and substrate types. Training was conducted by experts from the University of Queensland (Centre of Marine Studies, Biophysical Remote Sensing) and the Queensland Parks and Wildlife Service who are also UniDive members. The monitoring methods (see methods) are based upon results of the UniDive Coastcare project from 2002, the international established Reef Check program and research conducted by Biophysical Remote Sensing and the Centre of Marine Studies. Habitats were mapped using a combination of towed GPS photo transects, aerial photography, bathymetry surveys and expert knowledge. This data provides georeferenced information regarding the major features of each of Sites mapped including Wolf Rock

El papel de los carteles de la División de Gráfica y Cine de Parques y Recreo Público en la redefinición colonial del Puerto Rico de mediados del siglo XX

El cartel serigráfico del proyecto de carteles del Taller de Cinema y Gráfica de Parques y Recreo Público se utilizó como medio para educar e insertar a las grandes masas dentro de las reformas coloniales ocurridas como consecuencia de la entrada de Estados Unidos a la Segunda Guerra Mundial y readiestrar a la población puertorriqueña, entonces compuesta por campesinos, para servir como mano de obra barata y dócil al proyecto colonial y a los beneficiarios del proyecto de industrialización de la posguerra. Se ha dicho en innumerables ocasiones que el camino al infierno está repleto de buenas intenciones; este es el relato de un viaje de ida. Los carteles de la División de Cinema y Gráfica de Parques y Recreo Público mantienen desde el origen varios elementos inquietantes de lo que se podría denominar como la política de reconstrucción de una colonia, de un territorio objeto de la sujeción/subyugación por parte de otro país, y presentarlo como un objetivo plausible, de consideración y cumplimiento con los derechos humanos fundamentales...

The effects of agglomeration economies on the productivity of manufacturing local units in Ireland

The purpose of this study is to examine the effects of agglomeration economies on the productivity of manufacturing local units in Ireland. Four types of agglomeration economies are considered in this study. These are internal economies of scale, localization economies, related variety and urbanization economies. This study makes a number of contributions to the literature. Firstly, this is the first study to conduct an investigation of the effects of agglomeration economies on the productivity of manufacturing local units operating in Ireland. Secondly, this study distinguishes between indigenous and foreign-owned local units which is important given the dual nature of the Irish economy (Krugman, 1997). Thirdly, in addition to considering the effects of agglomeration economies, this study examines the impact of spurious agglomeration on the productivity of foreign-owned local units. Using data from the Census of Industrial Local Units and a series of IV GMM estimators to control for endogeneity, the results of the analysis conducted in Chapter 6 reveal that there are differences in the effects of agglomeration economies on the productivity of indigenous and foreign-owned local units. In Chapter 7 the Census of Industrial Local Units is supplemented by additional data sources and more in-depth measures are generated to capture the features of each of the external agglomeration economies considered in this analysis. There is some evidence to suggest that the availability of local inputs has a negative and significant impact on productivity. The NACE based measures of related variety reveal that the availability of local inputs and knowledge spillovers for related sectors have a negative and significant impact on productivity. There is clear evidence to suggest that urbanization economies are important for increasing the productivity of indigenous local units. The findings reveal that a 1% increase in population density in the NUTS 3 region leads to an increase in the productivity of indigenous local units of approximately 0.07% to 0.08%. The results also reveal that there is a significant difference in the effects of agglomeration economies on the productivity of low-tech and medium/high-tech indigenous local units. The more in-depth measures of agglomeration economies used in Chapter 7 are also used in Chapter 8. A series of IV GMM regressions are estimated in order to identify the impact of agglomeration economies and spurious agglomeration on the productivity of foreign-owned local units operating in Ireland. There is some evidence found to suggest that the availability of a pool of skilled labour has a positive and significant on productivity of foreign-owned local units. There is also evidence to suggest that localization knowledge spillovers have a negative impact on the productivity of foreign-owned local units. There is strong evidence to suggest that the availability of local inputs has a negative impact on the productivity. The negative impact is not confined to the NACE 4-digit sector but also extends into related sectors as determined by Porter’s (2003) cluster classification. The cluster based skills measure of related variety has a positive and significant impact on the productivity of foreign-owned local units. Similar to Chapter 7, there is clear evidence to suggest that urbanization economies are important for increasing the productivity of foreign-owned local units. Both the summary measure and each of the more in-depth measures of agglomeration economies have a positive and significant impact on productivity. Spurious agglomeration has a positive and significant impact on the productivity of foreign-owned local units. The results indicate that the more foreign-owned local units of the same nationality in the country the greater the levels of productivity for the local unit. From a policy perspective, urbanization economies are clearly important for increasing the productivity of both indigenous and foreign-owned local units. Furthermore, the availability of a pool of skilled labour appears to be important for increasing the productivity of foreign-owned local units. Another policy implication that arises from these results relates to the differences observed between indigenous local units and foreign-owned local units and also between low-tech and medium/high-tech indigenous local units. These findings indicate that ‘one-size-fits-all’ type policies are not appropriate for increasing the productivity of local units operating in Ireland. Policies should be tailored to the needs of either indigenous or foreign-owned local units and also to specific sectors. This positive finding for own country spurious agglomeration is important from a policy perspective and is one that IDA Ireland should take on board.