Identification of novel innate immune mechanisms regulating inflammation in the gastrointestinal tract

The Gastro-Intestinal (GI) tract is a unique region in the body. Our innate immune system retains a fine homeostatic balance between avoiding inappropriate inflammatory responses against the myriad commensal microbes residing in the gut while also remaining active enough to prevent invasive pathogenic attack. The intestinal epithelium represents the frontline of this interface. It has long been known to act as a physical barrier preventing the lumenal bacteria of the gastro-intestinal tract from activating an inflammatory immune response in the immune cells of the underlying mucosa. However, in recent years, an appreciation has grown surrounding the role played by the intestinal epithelium in regulating innate immune responses, both in the prevention of infection and in maintaining a homeostatic environment through modulation of innate immune signalling systems. The aim of this thesis was to identify novel innate immune mechanisms regulating inflammation in the GI tract. To achieve this aim, we chose several aspects of regulatory mechanisms utilised in this region by the innate immune system. We identified several commensal strains of bacteria expressing proteins containing signalling domains used by Pattern Recognition Receptors (PRRs) of the innate immune system. Three such bacterial proteins were studied for their potentially subversive roles in host innate immune signalling as a means of regulating homeostasis in the GI tract. We also examined differential responses to PRR activation depending on their sub-cellular localisation. This was investigated based on reports that apical Toll-Like Receptor (TLR) 9 activation resulted in abrogation of inflammatory responses mediated by other TLRs in Intestinal Epithelial Cells (IECs) such as basolateral TLR4 activation. Using the well-studied invasive intra-cellular pathogen Listeria monocytogenes as a model for infection, we also used a PRR siRNA library screening technique to identify novel PRRs used by IECs in both inhibition and activation of inflammatory responses. Many of the PRRs identified in this screen were previously believed not to be expressed in IECs. Furthermore, the same study has led to the identification of the previously uncharacterised TLR10 as a functional inflammatory receptor of IECs. Further analysis revealed a similar role in macrophages where it was shown to respond to intracellular and motile pathogens such as Gram-positive L.monocytogenes and Gram negative Salmonella typhimurium. TLR10 expression in IECs was predominantly intracellular. This is likely in order to avoid inappropriate inflammatory activation through the recognition of commensal microbial antigens on the apical cell surface of IECs. Moreover, these results have revealed a more complex network of innate immune signalling mechanisms involved in both activating and inhibiting inflammatory responses in IECs than was previously believed. This contribution to our understanding of innate immune regulation in this region has several direct and indirect benefits. The identification of several novel PRRs involved in activating and inhibiting inflammation in the GI tract may be used as novel therapeutic targets in the treatment of disease; both for inducing tolerance and reducing inflammation, or indeed, as targets for adjuvant activation in the development of oral vaccines against pathogenic attack.

Hydrogen sulfide is a novel mediator of lipopolysaccharide-induced inflammation in the mouse

Hydrogen sulfide (H2S) is synthesized in the body from L-Cysteine by several enzymes including cystathionine-gamma-lyase (CSE). To date, there is little information about the potential role of H2S in inflammation. We have now investigated the part played by H2S in endotoxin-induced inflammation in the mouse. E. coli lipopolysaccharide (LPS) administration produced a dose (10 and 20 mg/kg ip)- and time (6 and 24 h)-dependent increase in plasma H2S concentration. LPS (10 mg/kg ip, 6 h) increased plasma H2S concentration from 34.1 +/- 0.7 mu M to 40.9 +/- 0.6 mu M (n=6, P

SEROTONINERGIC AND PEPTIDERGIC NERVE ELEMENTS IN THE PROTOSCOLEX OF ECHINOCOCCUS-GRANULOSUS (CESTODA, CYCLOPHYLLIDEA)

The localisation and distribution of 5-hydroxytryptamine (5-HT, or serotonin) and neuropeptides in the nervous system of the protoscolex of the hydatid organism Echinococcus granulosus were determined by an indirect immunofluorescence technique. Nerve-cell bodies immunoreactive for 5-HT occurred in the lateral ganglia and in association with the lateral longitudinal nerve cords. 5-HT immunostaining was also evident in the central nerve ring, in the rostellar nerves and in the nerve plexus innervating the suckers. Of the antisera used to screen the protoscolex for neuropeptide immunoreactivity (IR), immunostaining was obtained with those raised against pancreatic polypeptide (PP), peptide YY (PYY), substance P (SP), peptide histidine isoleucine (PI-II) and vasoactive intestinal peptide (VIP). The most extensive pattern of IR occurred with antisera to PP and PYY. Immunoreactive nerve elements were evident in the lateral ganglia, central nerve ring, rostellar nerves, rostellar ganglia, sucker plexus and longitudinal nerve cords. The distribution of SP-, PHI- and VIP-IRs was more restricted: SP-IR occurred in the lateral ganglia and sucker nerves, whilst PHI- and VIP-immunoreactive nerve elements were associated with the lateral longitudinal nerve cords. Protoscoleces cultured in vitro for 29 days were also examined and neuroanatomical changes noted. A greater development of the longitudinal nerve cords and their cross-connectives in the body of the worm was evident, and a group of nerve cells were seen to develop at the posterior end of the main lateral nerve cords.

Biotransformation and accumulation of arsenic in soil amended with seaweed

For many coastal regions of the world, it has been common practice to apply seaweed to the land as a soil improver and fertilizer. Seaweed is rich in arsenosugars and has a tissue concentration of arsenic up to 100 micro/g g(-1). These arsenic species are relatively nontoxic to humans; however, in the environment they may accumulate in the soil and decompose to more toxic arsenic species. The aim of this study was to determine the fate and biotransformation of these arsenosugars in soil using HPLC-ICP-MS analysis. Data from coastal soils currently manured with seaweeds were used to investigate if arsenic was accumulating in these soils. Long-term application of seaweed increased arsenic concentrations in these soils up to 10-fold (0.35 mg of As kg(-1) for nonagronomic peat, 4.3 mg of As kg(-1) for seaweed-amended peat). The biotransformation of arsenic was studied in microcosm experiments in which a sandy (machair) soil, traditionally manured with seaweed, was amended with Laminaria digitata and Fucus vesiculosus. In both seaweed species, the arsenic occurs in the form of arsenosugars (85%). The application of 50 g of seaweed to 1 kg of soil leads to an increase of arsenic in the soils, and the dominating species found in the soil pore water were dimethylarsinic acid (DMA(V)) and the inorganic species arsenate (As(V)) and arsenite (As(III)) after the initial appearance of arsenosugars. A proposed decomposition pathway of arsenosugars is discussed in which the arsenosugars are transformed to DMA(V) and further to inorganic arsenic without appreciable amounts of methylarsonic acid (MA(V)). Commercially available seaweed-based fertilizers contain arsenic concentration between 10 and 50 mg kg(-1). The arsenic species in these fertilizers depends on the manufacturing procedure. Some contain mainly arsenosugars while others contain mainly DMA(V) and inorganic arsenic. With the application rates suggested by the manufacturers, the application of these fertilizers is 2 orders of magnitude lower than the maximum permissible sewage sludge load for arsenic (varies from 0.025 kg ha(-1) yr(-1) in Styria, Austria, to 0.7 kg ha(-1) yr(-1) in the U.K.), while a direct seaweed application would exceed the maximum arsenic load by at least a factor of 2.

Physical Exercise Program for Elderly Subjects in the “Club Nueva Vida”, in Tunja City

A descriptive, case series intervention study was performed to identify changes in physical qualities, aerobic power, flexibility, strength and body composition amongst 20 elderly subjects belonging to the “Club Nueva Vida” in Tunja City, during the second period of 2005. A PRE and POST assessment of their physical qualities was applied after a 16 week physical exercise program.Results showed an increase in the average of the static muscle streneth from 0.27 to 0.30 Newtons. Standard deviation diminished in 0.1. The dynamic strength measure by the push up test improved in 10% a 35% improved in 10% a 35% improved their. Cardiovascular capability until to high levels of 80% there were not significant changes in the body weight between measures taken before and after the program application. The results obtained with the sit and reach test did not show statistically significant differences. Hower in other flexibility tests the results changed from bad ranges to middle and good ones. The enrolled population was stable during the development of the exercise program. This study concludes that elderly adults acquire changes in their physical quality features after exercise intervention. Physical exercise programs in elderly adults must involve strength activities for upper limbs, trunk and aerobic power activities. The “Beliefs Health Model” is a adherence to reach compliance and maintenance in physical exercise.

Arresting developments in the cardiac myocyte cell cycle: Role of cyclin-dependent kinase inhibitors

Like most other cells in the body, foetal and neonatal cardiac myocytes are able to divide and proliferate. However, the ability of these cells to undergo cell division decreases progressively during development such that adult myocytes are unable to divide. A major problem arising from this inability of adult cardiac myocytes to proliferate is that the mature heart is unable to regenerate new myocardial tissue following severe injury, e.g. infarction, which can lead to compromised cardiac pump function and even death. Studies in proliferating cells have identified a group of genes and proteins that controls cell division. These proteins include cyclins, cyclin-dependent kinases (CDKs) and CDK inhibitors (CDKIs), which interact with each other to form complexes that are essential for controlling normal cell cycle progression. A variety of other proteins, e.g. the retinoblastoma protein (pRb) and members of the E2F family of transcription factors, also can interact with, and modulate the activities of, these complexes. Despite the major role that these proteins play in other cell types, little was known until recently about their existence and activities in immature (proliferating) or mature (non-proliferating) cardiac myocytes. The reason(s) why cardiac myocytes lose their ability to divide during development remains unknown, but if strategies were developed to understand the mechanisms underlying cardiac myocyte growth, it could open up new avenues for the treatment of cardiovascular disease. In this article, we shall review the function of the cell cycle machinery and outline some of our recent findings pertaining to the involvement of the cell cycle in modulating cardiac myocyte growth and hypertrophy.

Manipulation in Newspaper Articles : A Political Discourse Analysis of Lexical Choice and Manipulation in Japanese Newspaper Crisis Reporting in the case of North Korea

This paper analyzes some forms of linguistic manipulation in Japanese in newspapers when reporting on North Korea and its nuclear tests. The focus lies on lexical ambiguity in headlines and journalist’s voices in the body of the articles, that results in manipulation of the minds of the readers. The study is based on a corpus of nine articles from two of Japan’s largest newspapers Yomiuri Online and Asahi Shimbun Digital. The linguistic phenomenon that contribute to create manipulation are divided into Short Term Memory impact or Long Term Memory impact and examples will be discussed under each of the categories.The main results of the study are that headlines in Japanese newspapers do not make use of an ambiguous, double grounded structure. However, the articles are filled with explicit and implied attitudes as well as attributed material from people of a high social status, which suggests that manipulation of the long term memory is a tool used in Japanese media.

Chemical and physical speciation of arsenic in a small pond receiving gold mine waste effluent

The chemical and physical speciation of arsenic in a small pond that receives wastewater from a gold mine operation in western Victoria, Australia was studied using differential pulse polarography. By using different sample pretreatments, distinction between the physical states (dissolved or particulate As), between the oxidation states (As(III) or As(V)), and between the degrees of lability (labile or strongly bound) was achieved. The results are interpreted in terms of the physicochemical properties with reference to the use of the pond as a settlement dam for gold mining effluent. The speciation of arsenic was found to vary markedly with the physicochemical properties of the water. A model for the behavior of arsenic in the pond is proposed.

International examples of large-scale systems-theory and practice IV: B2B e-commerce implementation in the Australian context

Two of the most commonly used classifications in IS research are the factor approach and the process approach. While many studies of IS implementation are undertaken using a factor approach, little research so far examines the process of e-commerce implementation, especially in relation to the implementation of e-commerce in business-to-business (B2B) relationships. A holistic understanding of implementation which combines both the factor and process approaches using a case study method, is suggested as particularly suitable because of its ability to capture the reality of e-commerce implementation in an organisation's natural environment and in much greater detail than is possible using one of these approaches alone. In this paper, therefore, we endeavour to contribute to what we perceive as a gap in the body of theory surrounding the implementation process in the business-to-business e-commerce literature. We describe the findings of multiple case studies involving ten major Australian e-commerce initiators. In addition to confirming our earlier finding of the importance of non-technical factors for the success of the implementation process we also present, through our case studies, the various management and business issues associated with the success or failure of B2B e-commerce implementation.

Copper transporters in the human placenta

Copper is an essential micronutrient that is particularly important during pregnancy for normal fetal development. This study determined the mechanisms by which copper is transported from the maternal circulation to the fetal circulation via the placenta and how this transport is regulated by maternal hormone levels.

The role of copper transport proteins in the small intestines

Physiological copper homeostasis involves striking a balance between absorption and secretion. ATP7A was identified at the trans-Golgi network but relocalized to vesicles under copper exposure in the intestine. This suggests that ATP7A may be a rate limiting step in intestinal uptake of copper.

Control of biodegradation of a Mg alloy in simulated body fluid

Mg alloy AZ31 is an attractive candidate for coronary artery stents, as it possesses excellent biocompatibility in human body and good mechanical properties. However, AZ31 magnesium alloys generally have poor corrosion resistance in the body environment. This paper reports on the early stages of an investigation into the corrosion mechanism and the morphology of corrosion of AZ31 in simulated body fluid (SBF). The investigation will also consider ways of improving corrosion resistance of this alloy in SBF through the use of ionic liquids. The results to date have shown that AZ31 suffers severe localized pitting corrosion in SBF. The pits mainly develop adjacent to the Al-Mn intermetallic second phase in the α matrix. Energy Dispersive X-Ray Spectroscopy results revealed the presence of Mg, O, Ca, and P in the layer of corrosion product. Treatment of the AZ31 alloy prior to corrosion testing in SBF with the ionic liquid trimethyl (butyl) phosphonium diphenyl phosphate (P1444DPP) produced some increase in the corrosion resistance of the alloy.

A potential anti-corrosive ionic liquid coating for MG alloy AZ31 in simulated body fluids

Magnesium alloys are attractive materials for biomedical applications, due to their excellent biocompatibility. However, these alloys show fast corrosion rates in the body that limits their clinical applications. Low-toxic ionic liquid (IL) trimethyl(butyl)phosphonium diphenyl phosphate P1444dpp has been investigated to provide corrosion protection for magnesium alloy AZ31 in simulated body fluids (SBFs). This work reports a preliminary exploration of the influence of different treatment temperatures on the corrosion protection properties of IL films for the magnesium alloy AZ31 in SBFs. Results show that the IL treatment at room temperature did not bring significant improvement in the corrosion performance of the AZ31 in SBF. However, when the treatment temperature was increased to 75°C, the IL treatment resulted in a substantial reduction of the corrosion, in particular the reduction of localized pitting corrosion. The influence of ionic liquid treatment on the corrosion performance of the magnesium alloys AZ31 in SBFs has been investigated by electrochemical impedance spectroscopy (EIS) tests and immersion tests.

Predicting the chemical composition of the body and the carcass of 3/4Boer x 1/4Saanen kids using body components

The determination of the chemical composition of body and carcass is important in nutritional and growth regulation studies. The purpose of this study was to develop equations to predict the chemical composition of body and carcass using chemical composition of body components. Twenty 3/4Boer x 1/4Saanen crossbred male kids, weighing from 20 to 35 kg, were used in this study. The empty body chemical composition was measured by grinding all body components and sampling for chemical analyses. The body components used to estimate body and carcass composition were: neck, fore leg, ribs, loin, hind leg, 9-11 th rib section, non-carcass components (head plus feet, organs plus blood, and hide), visceral fat, and kidney fat. The chemical composition of organs plus blood and 9-11 th rib section had the highest precision to estimate percentage of fat, protein, and water in the body (r(2) of 0.94, 0.82, and 0.90, respectively). For carcass composition, the chemical composition of ribs was the best component to predict all carcass chemical components; however, the equations to estimate the percentages of protein and ash showed a low precision (r(2) = 0.48, 0.44, respectively). The 9-11 th rib section was accurate and precise to estimate carcass fat percentage. We concluded the chemical composition of the body of 3/4Boer x 1/4Saanen crossbred male kids was highly correlated with the composition of body parts, specifically organs plus blood and 9-11 th rib section. Further studies should focus on evaluating these body parts for different breeds and genders under different production scenarios. (C) 2007 Elsevier B.V. All rights reserved.