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.

Koalas are an iconic Australian marsupial undergoing precipitous population reduction in South-East Queensland from complex interacting threats. To investigate the causes of death and the interaction of comorbidities with demography in South-East Queensland koalas, a large scale, high-throughput prospective necropsy survey was conducted spanning 2013–2016. During this period, 519 necropsies were conducted in 155 young/subadult koalas, 235 mature, 119 old koalas and 10 of unknown age. Similar numbers of males and females were assessed. Trauma and infectious disease at were the most common single diagnoses. However, comorbidity was frequent, including multicentric infection or infectious disease in combination with trauma or senescence. Female koalas had proportionally more reproductive chlamydiosis compared to males in which the ocular and urinary systems were more commonly affected. Comorbidity and disease were strongly associated with poor body condition, and trauma was associated with good body condition. Animals affected by motor vehicle trauma were often in better body condition than those affected by animal attack, tree fall or other causes of trauma. This study identified a higher frequency of infections and comorbidity then previously reported, confirming the complex nature of interacting threats to the koala population.

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 toMycobacterium tuberculosis, microarray analysis was used to examine gene regulation in murine bone marrow-derived macrophages in response to 90-μm-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 anin vivoMatrigel granuloma model, the presence and activity of MMP-9 were examined by immunohistochemistry andin situzymography (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.