Modulation of plasma matrix metalloproteinase 9 and its inhibitors by vitamin D.

PhDMatrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are upregulated in a variety of diseases. Hypothesis: As TIMP-1 levels are elevated in liver fibrosis, might a similar process occur in essential hypertension driven left ventricular hypertrophy and furthermore may TIMP-1 be a marker of vascular disease? If TIMP-1 levels are a potential marker of cardiovascular disease could their levels be modulated by vitamin D? Methods: Plasma TIMP-1 levels and aldosterone were measured a) in patients with essential hypertension who had never been on treatment or had been off treatment for 1 month and b) healthy controls. All participants underwent echocardiography. To assess whether TIMP-1 was a marker of vascular disease insulin, sCRP, fibrinogen, homocysteine, PAI-1 were measured in Bangladeshis pre supplementation with vitamin D. TIMP-1, MMP2, 9 and 25 hydroxyvitamin D 25(OH)vitD were also measured pre and post supplementation. Subsequent studies included measurements of MMP2, 9 and TIMP-1and 4 in submariners pre and post patrol and MMP9 and 25(OH)vitD in patients who re-stenosed post angioplasty. TIMP-4 was validated using a radioimmunoassay, 25(OH)vitD measured using a triple quad MS and other assays using ELISAs. Results: Plasma TIMP-1 was higher in hypertensive patients than in the controls (p<0.0001) and was correlated with left ventricular hypertrophy and with aldosterone. In the Bangladeshi study,. TIMP-1 was not correlated with other markers of vascular disease. TIMP-1 was correlated with systolic blood pressure (p<0.007) There was an inverse correlation of 25(OH)vitD with MMP9 (P<0.001) and TIMP-1 (p<0.05) and sCRP (p<0.05). The inverse relationship between MMP9 and 25(OH)vitD was also repeated in the submariner and restenosis studies. Conclusions: Plasma TIMP-1 may be an important determinant in essential hypertension and 25(OH)vit D may have a positive effect in reducing the inflammatory response as measured by MMP9. The increased 25(OH)vitD may also act by reducing aldosterone levels and thus suppressing TIMP-1 level

A proposed roadmap for the control of infections in wildlife using Chlamydia vaccine development in koalas Phascolarctos cinereus as a template

Vaccination strategies provide a crucial tool for managements of disease risks in wildlife, but have been utilized mostly for domestic species. However, a significant body of work has now been published describing the successful development of an anti-chlamydial vaccine for the koala Phascolarctos cinereus, Goldfuss, 1817. As such, vaccinations against these infections in the koala, can provide important insights into the use of vaccines for wildlife. Chlamydia infections in the koala have been intensively studied for over 30 years. Infections cause severe disease states, such as kerato-conjunctivitis (blindness) and reproductive tract disease (infertility), and/or mortality; and are contributing significantly to population declines. We aim to use the plethora of data available from koala chlamydial studies as a template to propose a roadmap for the development of vaccines for other wildlife species, especially in this era of antibiotic resistance. As such we have outlined the important steps that have led to significant milestones resulting in the successful development of a vaccine against an infectious disease in a non-domestic species. We hope to thus provide, not only a timely review on Chlamydia vaccines in koalas, but also an important conservation and management roadmap to help guide future researchers that are considering the development of a vaccine for a wild species.publishedVersio

The Sustainable Beef Profit Partnership Approach to the Adoption of New Beef Industry Technologies

Technology adoption in the Australian beef industry has been low and slow compared to the intensive livestock and cropping industries. The principles of accelerated adoption provide an innovative solution to this problem. In the Beef CRC, Sustainable Beef Profit Partnership (BPP) members will meet regularly to measure their current performance, set targets for future productivity increases, and use a profitability framework to assess the potential impact of new technology. Capacity building and partnership outcomes will also be assessed. The BPP teams will be supported with appropriate tools and resources. The information generated will be used to underpin the achievement of Beef CRC commercialisation outputs and profitability outcomes.Accelerated adoption, continuous improvement and innovation, beef industry, profit, Livestock Production/Industries,

Keratinocyte EGF signalling dominates in atopic dermatitis lesions: A comparative RNAseq analysis

Atopic dermatitis (AD) remains a highly heterogenous disorder with a multifactorial aetiology. Whilst keratinocytes are known to play a fundamental role in AD, their contribution to the overall immune landscape in moderate‐to‐severe AD is still poorly understood. In order to design new therapeutics, further investigation is needed into common disease pathways at the molecular level. We used publicly available whole‐tissue RNAseq data (4 studies) and single‐cell RNAseq keratinocyte data to identify genes/pathways that are involved in keratinocyte responses in AD and after dupilumab treatment. Transcripts present in both keratinocytes (single‐cell) and whole‐tissue, referred to as the keratinocyte‐enriched lesional skin (KELS) genes, were analysed using functional/pathway analysis. Following statistical testing, 2049 genes (16.8%) were differentially expressed in KELS. Enrichment analyses predicted increases in not only type‐1/type‐2 immune signalling and chemoattraction, but also in EGF‐dominated growth factor signalling. We identified complex crosstalk between keratinocytes and immune cells involving a dominant EGF family signature which converges on keratinocytes with potential immunomodulatory and chemotaxis‐promoting consequences. Although keratinocytes express the IL4R, we observed no change in EGF signalling in KELS after three‐month treatment with dupilumab, indicating that this pathway is not modulated by dupilumab immunotherapy. EGF family signalling is significantly dysregulated in AD lesions but is not associated with keratinocyte proliferation. EGF signalling pathways in AD require further study

Trafficking of Estrella lausannensis in human macrophages

Estrella lausannensis is a new member of the Chlamydiales order. Like other Chlamydia-related bacteria, it is able to replicate in amoebae and in fish cell lines. A preliminary study investigating the pathogenic potential of Chlamydia-related bacteria found a correlation between antibody response to E. lausannensis and pneumonia in children. To further investigate the pathogenic potential of E. lausannensis, we determined its ability to grow in human macrophages and its intracellular trafficking. The replication in macrophages resulted in viable E. lausannensis; however, it caused a significant cytopathic effect. The intracellular trafficking of E. lausannensis was analyzed by determining the interaction of the Estrella-containing inclusions with various endocytic markers as well as host organelles. The E. lausannensis inclusion escaped the endocytic pathway rapidly avoiding maturation into phagolysosomes by preventing both EEA-1 and LAMP-1 accumulation. Compared to Waddlia chondrophila, another Chlamydia-related bacteria, the recruitment of mitochondria and endoplasmic reticulum was minimal for E. lausannensis inclusions. Estrella lausannensis appears to use a distinct source of nutrients and energy compared to other members of the Chlamydiales order. In conclusion, we hypothesize that E. lausannensis has a restricted growth in human macrophages, due to its reduced capacity to control programmed cell deat

Enhanced diffusion of Uranium and Thorium linked to crystal plasticity in zircon

The effects of crystal-plasticity on the U-Th-Pb system in zircon is studied by quantitative microstructural and microchemical analysis of a large zircon grain collected from pyroxenite of the Lewisian Complex, Scotland. Electron backscatter diffraction (EBSD) mapping reveals a c.18° variation in crystallographic orientation that comprises both a gradual change in orientation and a series of discrete low-angle (<4°) boundaries. These microstructural data are consistent with crystal-plastic deformation of zircon associated with the formation and migration of dislocations. A heterogeneous pattern of dark cathodoluminescence, with the darkest domains coinciding with low-angle boundaries, mimics the deformation microstructure identified by EBSD. Geochemical data collected using the Sensitive High Resolution Ion MicroProbe (SHRIMP) shows a positive correlation between concentrations of the elements U, Th and Pb (ranging from 20–60 ppm, 30–110 ppm, and 14–36 ppm, respectively) and Th/U ratio (1.13 – 1.8) with the deformation microstructure. The highest measured concentrations and Th/U coincide with low-angle boundaries. This enrichment is interpreted to reflect enhanced bulk diffusion of U and Th due to the formation and migration of high-diffusivity dislocations. (207)Pb/(206)Pb ages for individual analyses show no significant variation across the grain, and define a concordant, combined mean age of 2451 ± 14 Ma. This indicates that the grain was deformed shortly after initial crystallization, most probably during retrograde Inverian metamorphism at amphibolite facies conditions. The elevated Th over U and consistent (207)Pb/(206)Pb ages indicates that deformation most likely occurred in the presence of a late-stage magmatic fluid that drove an increase in the Th/U during deformation. The relative enrichment of Th over U implies that Th/U ratio may not always be a robust indicator of crystallization environment. This study provides the first evidence of deformation-related modification of the U-Th system in zircon and has fundamental implications for the application and interpretation of zircon trace element data

Spatial constraints within the chlamydial host cell inclusion predict interrupted development and persistence

Background. The chlamydial developmental cycle involves the alternation between the metabolically inert elementary body (EB) and the replicating reticulate body (RB). The triggers that mediate the interchange between these particle types are unknown and yet this is crucial for understanding basic Chlamydia biology. Presentation of the hypothesis. We have proposed a hypothesis to explain key chlamydial developmental events whereby RBs are replicating strictly whilst in contact with the host cell membrane-derived inclusion via type three secretion (T3S) injectisomes. As the inclusion expands, the contact between each RB and the inclusion membrane decreases, eventually reaching a threshold, beyond which T3S is inactivated upon detachment and this is the signal for RB-to-EB differentiation. Testing the hypothesis. We explore this hypothesis through the development of a detailed mathematical model. The model uses knowledge and data of the biological system wherever available and simulates the chlamydial developmental cycle under the assumptions of the hypothesis in order to predict various outcomes and implications under a number of scenarios. Implications of the hypothesis. We show that the concept of in vitro persistent infection is not only consistent with the hypothesis but in fact an implication of it. We show that increasing the RB radius, and/or the maximum length of T3S needles mediating contact between RBs and the inclusion membrane, and/or the number of inclusions per infected cell, will contribute to the development of persistent infection. The RB radius is the most important determinant of whether persistent infection would ensue, and subsequently, the magnitude of the EB yield. We determine relationships between the length of the T3S needle and the RB radius within an inclusion, and between the RB radius and the number of inclusions per host cell to predict whether persistent infection or normal development would occur within a host cell. These results are all testable experimentally and could lead to significantly greater understanding of one of the most crucial steps in chlamydial development

Two distinct forms of Chlamydia psittaci associated with disease and infertility in Phascolarctos cinereus (Koala)

While several diseases associated with Chlamydia psittaci infection have been reported in Phascolarctos cinereus (koala), it is still unclear whether one or more chlamydial strains are responsible. In this study, we provide evidence, obtained by restriction enzyme and gene probe analysis, that two quite distinct strains of C. psittaci infect koalas; one strain was isolated from the conjunctivae, and the other was isolated from the urogenital tract and the rectum. A gene probe, pFEN207, containing the coding sequence for an enzyme involved in the biosynthesis of the chlamydial genus-specific lipopolysaccharide antigen, and a separate probe, pCPML-4N, prepared from a DNA fragment of a koala-infecting strain of C. psittaci, were used to determine the patterns of hybridization in the koala-infecting strains; these patterns were found to be quite distinct from those observed with C. psittaci isolates from other animals. We also demonstrated by hybridization analysis with an avian strain plasmid that all three koala urogenital isolates contain a plasmid and that there is no evidence for the presence of a homologous plasmid in any of the ocular isolates

Chlamydia trachomatis responds to heat shock, penicillin induced persistence, and IFN-gamma persistence by altering levels of the extracytoplasmic stress response protease HtrA

Background Chlamydia trachomatis, an obligate intracellular human pathogen, is the most prevalent bacterial sexually transmitted infection worldwide and a leading cause of preventable blindness. HtrA is a virulence and stress response periplasmic serine protease and molecular chaperone found in many bacteria. Recombinant purified C. trachomatis HtrA has been previously shown to have both activities. This investigation examined the physiological role of Chlamydia trachomatis HtrA. Results The Chlamydia trachomatis htrA gene complemented the lethal high temperature phenotype of Escherichia coli htrA- (>42°C). HtrA levels were detected to increase by western blot and immunofluorescence during Chlamydia heat shock experiments. Confocal laser scanning microscopy revealed a likely periplasmic localisation of HtrA. During penicillin induced persistence of Chlamydia trachomatis, HtrA levels (as a ratio of LPS) were initially less than control acute cultures (20 h post infection) but increased to more than acute cultures at 44 h post infection. This was unlike IFN-γ persistence where lower levels of HtrA were observed, suggesting Chlamydia trachomatis IFN-γ persistence does not involve a broad stress response. Conclusion The heterologous heat shock protection for Escherichia coli, and increased HtrA during cell wall disruption via penicillin and heat shock, indicates an important role for HtrA during high protein stress conditions for Chlamydia trachomatis

Twenty years of research into Chlamydia-like organisms: a revolution in our understanding of the biology and pathogenicity of members of the phylum Chlamydiae

Chlamydiae are obligate intracellular bacteria that share a unique but remarkably conserved biphasic developmental cycle that relies on a eukaryotic host cell for survival. Although the phylum was originally thought to only contain one family, the Chlamydiaceae, a total of nine families are now recognized. These so-called Chlamydia-like organisms (CLOs) are also referred to as ‘environmental chlamydiae', as many were initially isolated from environmental sources. However, these organisms are also emerging pathogens, as many, such as Parachlamydia sp., Simkania sp. and Waddlia sp., have been associated with human disease, and others, such as Piscichlamydia sp. and Parilichlamydia sp., have been documented in association with diseases in animals. Their strict intracellular nature and the requirement for cell culture have been a confounding factor in characterizing the biology and pathogenicity of CLOs. Nevertheless, the genomes of seven CLO species have now been sequenced, providing new information on their potential ability to adapt to a wide range of hosts. As new isolation and diagnostic methods advance, we are able to further explore the richness of this phylum with further research likely to help define the true pathogenic potential of the CLOs while also providing insight into the origins of the ‘traditional' chlamydia