Koala populations are in catastrophic decline in certain eastern Australian regions. Spanning from 1997–2013, a database derived from wildlife hospitals in southeast Queensland with N = 20,250 entries was classified by causes of morbidity and mortality. A total of 11 aetiologies were identified, with chlamydiosis, trauma, and wasting being most common. The clinical diagnosis at submission varied significantly over the observation period. Combinations of aetiologies were observed in 39% of koalas submitted, with chlamydiosis frequently co-occurring. Urogenital (cystitis 26.8%, bursitis 13.5%) and ocular (conjunctivitis 17.2%) chlamydiosis were the most frequently diagnosed representations of the infection. Approximately 26% of submissions comprised koalas involved in vehicle accidents that were otherwise healthy. Age and sex of the koala as well as season and submission period were compared for the case outcomes of ‘dead on arrival’, ‘euthanized’, or ‘released’ for the four most common clinical diagnoses using multinomial logistic regression models. Exploratory space-time permutation scans were performed and overlapping space-time clusters for chlamydiosis, motor vehicle traumas and wasting unveiled high risk areas for koala disease and injury. Our results suggest that these aetiologies are acting jointly as multifactorial determinants for the continuing decline of koalas.

RNA interference (RNAi) may provide a therapeutic solution to many pulmonary epithelium diseases. However, the main barrier to the clinical use of RNAi remains the lack of efficient delivery vectors. Research has mainly concentrated on the intranasal route of delivery of short interfering RNA (siRNA) effector molecules for the treatment of respiratory diseases. However, this may be complicated in a diseased state due to the increased fluid production and tissue remodeling. Therefore, we investigated our hydration of a freeze-dried matrix (HFDM) formulated liposomes for systemic delivery to the lung epithelium. Here, we show that 45 ± 2% of epithelial murine lung cells receive siRNA delivery upon intravenous (IV) liposomal administration. Furthermore, we demonstrate that liposomal siRNA delivery resulted in targeted gene and protein knockdown throughout the lung, including lung epithelium. Taken together, this is the first description of lung epithelial delivery via cationic liposomes, and provides a proof of concept for the use of IV liposomal RNAi delivery to specifically knockdown targeted genes in the respiratory system. This approach may provide an attractive alternate therapeutic delivery strategy for the treatment of lung epithelium diseases.

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Abstract 19Failure of lamellar energy metabolism may contribute to the pathophysiology of equine 20 laminitis. Tissue microdialysis has the potential to dynamically monitor lamellar energy 21 balance over time. The objectives of this study were to develop a minimally invasive lamellar 22 microdialysis technique and use it to measure normal lamellar energy metabolite 23 concentrations over 24 h. Microdialysis probes were placed (through the white line) into 24 either the lamellar dermis (LAM) (n = 6) or the sublamellar dermis (SUBLAM) (n = 6) and 25 perfused continuously over a 24 h study period. Probes were placed in the skin dermis 26 (SKIN) for simultaneous comparison to LAM (n = 6). Samples were collected every 2 h and 27 analysed for glucose, lactate, pyruvate, urea and glycerol concentrations. LAM was further 28 compared with SUBLAM by simultaneous placement and sampling in four feet from two 29 horses over 4 h. Horses were monitored for lameness, and either clinically evaluated for 1 30 month after probe removal (n = 4) or subjected to histological evaluation of the probe site (n 31 = 10). 32 33 There were no deleterious clinical effects of probe placement and the histological 34 response was mild. Sample fluid recovery and metabolite concentrations were stable for 24 h. 35Glucose was lower (and lactate:glucose ratio higher) in LAM compared with SUBLAM and 36 SKIN (P < 0.05). Pyruvate was lower in SUBLAM than SKIN and urea was lower in LAM 37 than SKIN (P < 0.05). These differences suggest lower perfusion and increased glucose 38 consumption in LAM compared with SUBLAM and SKIN. In conclusion, lamellar tissue 39 microdialysis was well tolerated and may be useful for determining the contribution of 40 energy failure in laminitis pathogenesis. 41 42

This work investigates the influence of Ag (1 wt%) on the mechanical properties, in vitro and in vivo corrosion, and biocompatibility of Fe‐35Mn. The microstructure of Fe‐35Mn‐1Ag possesses a uniform dispersion of discrete silver particles. Slight improvements in compressive properties are attributed to enhanced density and low porosity volume. Fe‐35Mn‐1Ag exhibits good in vitro and in vivo corrosion rate of Fe‐35Mn due to an increase in microgalvanic corrosion. Gas pockets, which originate from an inflammatory response to the implants, are observed in the rats after 4 weeks implantation but are undetectable after 12 weeks. No chronic toxicity is observed with the Fe‐35Mn‐1Ag, suggesting acceptable in vivo biocompatibility. The high corrosion rate of the alloy triggers an increased level of nonadverse tissue inflammatory responses 4 weeks after implantation, which subsequently subsides at 12 weeks. The Fe‐35Mn‐1Ag displays properties that are suitable for orthopedic applications.

The mucosal addressin cell adhesion molecule (MAdCAM) and vascular cell adhesion molecule (VCAM) appear to play roles in the recruitment of leukocytes to specialized endothelium lining the gastrointestinal tract. The purpose of this study was to clarify the role of MAdCAM and VCAM in the central nervous system by comparing protein expression in patients with multiple sclerosis (MS) and control subjects by immunohistochemistry. Specific antibodies to human VCAM and MAdCAM were used to confirm expression in control and MS nervous system specimens by immunohistochemistry. VCAM immunoreactivity was detected in endothelial cells, perivascular tissue, and in some cases, leukocytes within the meninges, gray, and white matter, of both controls and MS patients. VCAM immunoreactivity was maximal in a patient with acute active plaques, but of lower intensity and reduced distribution in controls and those with chronic active or inactive MS plaques. In contrast, MAdCAM immunoreactivity could not be detected in brain tissue from unaffected or MS patients. Taken together, these data support a role of VCAM, but not MAdCAM in the development of MS. (Am J Pathol 2010,

g Trehalose 6,6=-dimycolate (TDM) is a cell wall glycolipid and an important virulence factor of mycobacteria. In order to study the role of TDM in the innate immune response to Mycobacterium tuberculosis, microarray analysis was used to examine gene regulation in murine bone marrow-derived macrophages in response to 90-m-diameter polystyrene microspheres coated with TDM. A large number of genes, particularly those involved in the immune response and macrophage function, were up-or downregulated in response to these TDM-coated beads compared to control beads. Genes involved in the immune response were specifically upregulated in a myeloid differentiation primary response gene 88 (MyD88)-dependent manner. The complexity of the transcriptional response also increased greatly between 2 and 24 h. Matrix metalloproteinases (MMPs) were significantly upregulated at both time points, and this was confirmed by quantitative real-time reverse transcription-PCR (RT-PCR). Using an in vivo Matrigel granuloma model, the presence and activity of MMP-9 were examined by immunohistochemistry and in situ zymography (ISZ), respectively. We found that TDM-coated beads induced MMP-9 expression and activity in Matrigel granulomas. Macrophages were primarily responsible for MMP-9 expression, as granulomas from neutrophil-depleted mice showed staining patterns similar to that for wild-type mice. The relevance of these observations to human disease is supported by the similar induction of MMP-9 in human caseous tuberculosis (TB) granulomas. Given that MMPs likely play an important role in both the construction and breakdown of tuberculous granulomas, our results suggest that TDM may drive MMP expression during TB pathogenesis. Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), infects approximately one-third of the human population and is the leading bacterial cause of human mortality worldwide, killing 1.45 million individuals per year (1). In the majority of cases, however, the bacteria are sequestered within a well-organized granuloma, where they can remain in a poorly characterized, "latent" state for decades. The granuloma structure is typically composed of centrally located, infected macrophages in various stages of degeneration and necrosis, surrounded by epithelioid macrophages, foamy macrophages, and occasional multinucleated giant cells, all bordered by a mixed population of lymphocytes and a fibrous capsule. This capsule consists of a wall of collagen that is laid down by fibroblasts and must be broken down in order for transmission to occur. Infected individuals who are immunocompetent and/or treated can resolve granulomas, with disaggregation of the accumulated leukocytes, dissolution of the extracellular matrix, and either scar formation or a return to the normal pulmonary architecture. In progressively infected individuals, on the other hand, the granuloma liquefies, the capsule wall cavitates and ruptures into an adjacent airway, and the bacilli multiply and are released (2). While some causes of tubercu...

Vincristine is an antineoplastic substance that is part of many chemotherapy regimens, used especially for the treatment of a variety of pediatric cancers including leukemias and brain tumors. Unfortunately, many vincristine-treated patients develop peripheral neuropathy, a side effect characterized by sensory, motoric, and autonomic symptoms. The sensory symptoms include pain, in particular hypersensitivity to light touch, as well as loss of sensory discrimination to detect vibration and touch. The symptoms of vincristine-induced neuropathy are only poorly controlled by currently available analgesics and therefore often necessitate dose reductions or even cessation of treatment. The aim of this study was to identify new therapeutic targets for the treatment of vincristine-induced peripheral neuropathy (VIPN) by combining behavioral experiments, histology, and pharmacology after vincristine treatment. Local intraplantar injection of vincristine into the hind paw caused dose- and time-dependent mechanical hypersensitivity that developed into mechanical hyposensitivity at high doses, and lead to a pronounced, dose-dependent infiltration of immune cells at the site of injection. Importantly, administration of minocycline effectively prevented the development of mechanical hypersensitivity and infiltration of immune cells in mouse models of vincristine induce peripheral neuropathy (VIPN) based on intraperitoneal or intraplantar administration of vincristine. Similarly, Toll-like receptor 4 knockout mice showed diminished vincristine-induced mechanical hypersensitivity and immune cell infiltration, while treatment with the anti-inflammatory meloxicam had no effect. These results provide evidence for the involvement of Toll-like receptor 4 in the development of VIPN and suggest that minocycline and/or direct Toll-like receptor 4 antagonists may be an effective preventative treatment for patients receiving vincristine.

The active collection of wildlife sighting data by trained observers is expensive, restricted to small geographical areas and conducted infrequently. Reporting of wildlife sightings by members of the public provides an opportunity to collect wildlife data continuously over wider geographical areas, at lower cost. We used individual koala sightings reported by members of the public between 1997 and 2013 in South-East Queensland, Australia ( n = 14,076 koala sightings) to describe spatial and temporal trends in koala presence, to estimate koala sighting density and to identify biases associated with sightings. Temporal trends in sightings mirrored the breeding season of koalas. Sightings were high in residential areas (63%), followed by agricultural (15%), and parkland (12%). The study area was divided into 57,780 one-square kilometer grid cells and grid cells with no sightings of koalas decreased over time (from 35% to 21%) indicative of a greater level of spatial overlap of koala home ranges and human activity areas over time. The density of reported koala sightings decreased as distance from primary and secondary roads increased, indicative of a higher search effort near roads. Our results show that koala sighting data can be used to refine koala distribution and population estimates derived from active surveying, on the condition that appropriate bias correction techniques are applied. Collecting koala absence and search effort information and conducting repeated searches for koalas in the same areas are useful approaches to improve the quality of sighting data in citizen science programs.