Spectinamide MBX-4888A exhibits favorable lesion and tissue distribution and promotes treatment shortening in advanced murine models of tuberculosis.

The spectinamides are novel, narrow-spectrum semisynthetic analogs of spectinomycin, modified to avoid intrinsic efflux by Mycobacterium tuberculosis . Spectinamides, including lead MBX-4888A (Lee-1810), exhibit promising therapeutic profiles in mice, as single drugs and as partner agents with other anti-tuberculosis antibiotics including rifampin and/or pyrazinamide. To demonstrate that this translates to more effective cure, we first confirmed the role of rifampin, with or without pyrazinamide, as essential to achieve effective bactericidal responses and sterilizing cure in the current standard of care regimen in chronically infected C3HeB/FeJ mice compared to BALB/c mice. Thus, demonstrating added value in testing clinically relevant regimens in murine models of increasing pathologic complexity. Next we show that MBX-4888A, given by injection with the front-line standard of care regimen, is treatment shortening in multiple murine tuberculosis infection models. The positive treatment responses to MBX-4888A combination therapy in multiple mouse models including mice exhibiting advanced pulmonary disease can be attributed to favorable distribution in tissues and lesions, retention in caseum, along with favorable effects with rifampin and pyrazinamide under conditions achieved in necrotic lesions. This study also provides an additional data point regarding the safety and tolerability of spectinamide MBX-4888A in long-term murine efficacy studies.

The influence of diet on gut microbiome and body mass dynamics in a capital-breeding migratory bird.

Gut-associated microbial communities are known to play a vital role in the health and fitness of their hosts. Though studies investigating the factors associated with among-individual variation in microbiome structure in wild animal species are increasing, knowledge of this variation at the individual level is scarce, despite the clear link between microbiome and nutritional status uncovered in humans and model organisms. Here, we combine detailed observational data on life history and foraging preference with 16S rRNA profiling of the faecal microbiome to investigate the relationship between diet, microbiome stability and rates of body mass gain in a migratory capital-breeding bird, the light-bellied Brent goose (Branta bernicla hrota). Our findings suggest that generalist feeders have microbiomes that are intermediate in diversity and composition between two foraging specialisms, and also show higher within-individual plasticity. We also suggest a link between foraging phenotype and the rates of mass gain during the spring staging of a capital breeder. This study offers rare insight into individual-level temporal dynamics of the gut microbiome of a wild host. Further work is needed to uncover the functional link between individual dietary choices, gut microbiome structure and stability, and the implications this has for the reproductive success of this capital breeder.

High-pressure single-crystal diffraction at the Australian Synchrotron.

A new high-pressure single-crystal diffraction setup has been designed and implemented at the Australian Synchrotron for collecting molecular and protein crystal structures. The setup incorporates a modified micro-Merrill-Bassett cell and holder designed specifically to fit onto the horizontal air-bearing goniometer, allowing high-pressure diffraction measurements to be collected with little to no modification of the beamline setup compared with ambient data collections. Compression data for the amino acid, L-threonine, and the protein, hen egg-white lysozyme, were collected, showcasing the capabilities of the setup.

Pressure and guest-mediated pore shape modification in a small pore MOF to 1200 bar.

Guest-mediated pore-shape modification of the metal-organic framework, Sc2BDC3 upon adsorption of n-pentane and isopentane is examined from 50-1200 bar. Rotation of the BDC linker responsible for the change in pore shape occurs at much lower pressures than previously reported, with distinct adsorption behaviour observed between pentane isomers.

The development and clinical use of the Guy's and St Thomas' quality of life questionnaire (GAST) for adult cochlear implant patients.

OBJECTIVES: To Review the benefit of the GAST questionnaire (Tilston, S. 2003. Assessing the quality of life in adult cochlear implant users. MSc dissertation. London: City University.) to measure hearing related quality of life for adults pre and post-cochlear implantation. To develop a scoring methodology as to what score constitutes a significantly 'good' or 'poor' change to better target rehabilitation to those most in need. DESIGN: The GAST Questionnaire was developed using a robust cycle of validation and reliability analyses using the Statistical Package for the Social Sciences (Norusis, M. 1993. SPSS for windows: professional statistics. release 6.0. Chicago: SPSS Inc.). The scoring methodology was developed by dividing 83 patient full data sets into quintiles for the delta of quality of life improvement and the 9-12 month post-implantation GAST score. RESULTS: The GAST questionnaire design was deemed robust. The scoring methodology used led to the 20th percentile score highlighting individuals requiring further support and the 80th percentiles for those suitable for partial booking. CONCLUSION: The GAST questionnaire is a useful way of identifying the patients in need of support as well as to measure patient reported quality of life improvements following cochlear implantation.

An investigation into the preparation of carbon black by partial oxidation of spent tyre pyrolysis oil.

To promote the use of recycled waste materials as an industrial feedstock, this study examined the preparation of carbon black (CB) by partial oxidation of a spent tyre pyrolysis oil using a drop tube furnace. The effect of reaction temperature, the residence time of gas in the reactor and inlet gas oxygen concentration on the yield and properties of the CB were evaluated. The surface chemistry, chemical composition, morphological and thermal properties of the CB samples were characterised using XPS, EA, TEM, BET, and TGA, respectively. The CB yield increased with increasing reaction temperature but decreased as the residence time or oxygen concentration increases. The CB primarily consisted of C (90.5-98.6%) and O (0.9-7.4%), with small traces of S (<1%), Si (<1%) and H (<2%). Hydroxyl, carbonyl, and carboxyl are the key functional groups found on the CB surface, with the hydroxyl groups being dominant. The CB were highly graphitic with a lattice spacing in the range of 0.338-0.350 nm and had BET surface areas of 4-22 m2g-1. The mean primary particle size ranged from 92 to 176 nm and decreased with increasing reaction temperature and oxygen concentration. The CB aggregate configuration became more complex with increasing reaction temperature, residence time and oxygen concentration. The results were not only comparable with commercial CB products from fossil fuel feedstocks but are expected to provide the needed motivation to move towards circular economy strategies, which have positive impacts from a sustainability perspective.

Performance of activated carbons prepared from spent tyres in the adsorption of rhodamine B in aqueous solutions.

Activated carbons were produced from spent tyre pyrolysis char by steam or CO2 activation and evaluated for their performance in rhodamine B (RhB) adsorption in aqueous solutions. The effect of RhB starting concentration (80-150 mg L-1), contact time (0-80 min), temperature (298-318 K) and initial pH on the adsorption process was examined. Pseudo-first-order and pseudo-second-order models were carried out to fit the experimental data to derive RhB adsorption kinetics. Langmuir, Freundlich and Temkin isotherm models were applied to depict RhB adsorption behaviour of the prepared activated carbons. Gibbs free energy (ΔG), enthalpy (ΔH) and entropy (ΔS) were calculated. It has been found that the activated carbons can effectively adsorb RhB due to high mesoporosity and RhB equilibrium adsorption capacity (qe) increased almost linearly with increasing total mesopore volumes, regardless of the activation agents. When BET surface areas are similar, CO2-activated carbon obtained higher qe than steam due to higher mesoporosity of CO2-activated carbon. The results show that pseudo-second-order well fitted the experimental data. RhB starting concentration increased from 80 to 150 mg L-1 causing qe increased from 158 to 251 mg g-1 but RhB removal decreased from 99.7 to 84.5%. The RhB adsorption process follows the Langmuir model and thermodynamic calculation, indicating RhB adsorption is an endothermic, spontaneous process, dominated by both chemisorption and physisorption.

Antibody-mediated depletion of CCR10+EphA3+ cells ameliorates fibrosis in IPF.

Idiopathic pulmonary fibrosis (IPF) is characterized by aberrant repair that diminishes lung function via mechanisms that remain poorly understood. CC chemokine receptor (CCR10) and its ligand CCL28 were both elevated in IPF compared with normal donors. CCR10 was highly expressed by various cells from IPF lungs, most notably stage-specific embryonic antigen-4-positive mesenchymal progenitor cells (MPCs). In vitro, CCL28 promoted the proliferation of CCR10+ MPCs while CRISPR/Cas9-mediated targeting of CCR10 resulted in the death of MPCs. Following the intravenous injection of various cells from IPF lungs into immunodeficient (NOD/SCID-γ, NSG) mice, human CCR10+ cells initiated and maintained fibrosis in NSG mice. Eph receptor A3 (EphA3) was among the highest expressed receptor tyrosine kinases detected on IPF CCR10+ cells. Ifabotuzumab-targeted killing of EphA3+ cells significantly reduced the numbers of CCR10+ cells and ameliorated pulmonary fibrosis in humanized NSG mice. Thus, human CCR10+ cells promote pulmonary fibrosis, and EphA3 mAb-directed elimination of these cells inhibits lung fibrosis.

Utilisation of spent tyre pyrolysis char as activated carbon feedstock: The role, transformation and fate of Zn.

Utilisation and minimisation of spent tyre stockpiles has been made more viable by pyrolysis and activation to produce low-cost activated carbons. The unique chemical composition of spent tyre pyrolysis chars (STPC), particularly the high Zn content, has been shown to affect their activation and subsequent utilisation. Nonetheless, little research has examined exactly how these additives affect activation and, ultimately, what becomes of Zn during the activation process. This paper presents a systematic study of the effect of Zn, ZnO and ZnS on the physical properties of STPC and their transformation mechanisms during CO2 activation. Samples of acid-washed STPC with and without ZnO and ZnS addition were activated using a fixed-bed reactor in 66.7%v/v CO2 for 3 h at 850, 950, 1000 and 1050 °C. Under these conditions, both ZnO and ZnS were found to act as a catalyst during activation, increasing surface area, pore volume and burn-off. During the activation, ZnO was reduced by C to form elemental Zn and ZnS was thermally decomposed to release Zn and S. Thermogravimetric analysis of Zn and its compounds above 600 °C, separately and mixed with acid-washed char, under CO2 confirms that ZnO and ZnS dissociate to release Zn(v) that further reacts with CO2 or S to reform ZnO or ZnS. However, Zn is progressively removed from activated carbon at temperatures between 950 °C and 1050 °C. These results have direct implications for the utilisation of SPTC as a feedstock for activated carbon, and the production of Zn-loaded activated carbons.

Effect of biochar addition on microbial community and methane production during anaerobic digestion of food wastes: The role of minerals in biochar.

The effect of biochar addition on the microbial community and methane (CH4) production during anaerobic digestion was experimentally investigated, focusing on the role of minerals in biochar. The biochar was prepared from pine sawdust by pyrolysis at 650 °C and 900 °C, respectively, and a subsample was leached with citric acid. The cultures with the addition of biochar, leached biochar, Fe, and leached biochar combined with Fe, respectively, were placed in bench-scale bioreactors for anaerobic digestion. Daily biogas production was measured by volume displacement method and analysed for CH4 concentration, which allowed the cumulative CH4 yield (YM) and daily CH4 production rate (RM) to be determined. Culture samples were also taken daily for volatile fatty acids (VFAs) and microbial community analysis. Compared to the control without biochar addition, the addition of raw biochar significantly increased YM by 46.9% and RM by 43.0%, while leached biochar only increased the YM by 33.2% and RM by 18.2%, respectively. The Fe-containing minerals in biochar were found to enhance VFA degradation and increase population of Clostridia and Methanosaeta, improving the CH4 production.

Targeting MAP3K19 prevents human lung myofibroblast activation both in vitro and in a humanized SCID model of idiopathic pulmonary fibrosis.

Idiopathic Pulmonary Fibrosis (IPF) is a disease with a devastating prognosis characterized by unrelenting lung scarring. Aberrant activation of lung fibroblasts is a key feature of this disease, yet the key pathways responsible for this are poorly understood. Mitogen-activated protein kinase, kinase, kinase- 19 (MAP3K19) was recently shown to be upregulated in IPF and this MAPK has a key role in target gene transcription in the TGF-ß pathway. Herein, we further investigate the role of MAP3K19 in cultured normal and IPF fibroblasts and in a humanized SCID mouse model of IPF employing both short interfering (si) RNA and novel small-molecule inhibitors directed at this kinase. Targeting MAP3K19 had significant inhibitory effects on the expression of both alpha smooth muscle actin and extracellular matrix in cultured human IPF fibroblasts. Quantitative protein and biochemical assays, as well as histological analysis, showed that MAP3K19 was required for the development of lung fibrosis in SCID mice humanized with IPF lung fibroblasts. MAP3K19 was required for IPF myofibroblast differentiation, and targeting its activity attenuated the profibrotic activity of these cells both in vitro and in an adoptive transfer SCID model of pulmonary fibrosis.

DNA-PKcs modulates progenitor cell proliferation and fibroblast senescence in idiopathic pulmonary fibrosis.

BACKGROUND: Recent studies have highlighted the contribution of senescent mesenchymal and epithelial cells in Idiopathic Pulmonary Fibrosis (IPF), but little is known regarding the molecular mechanisms that regulate the accumulation of senescent cells in this disease. Therefore, we addressed the hypothesis that the loss of DNA repair mechanisms mediated by DNA protein kinase catalytic subunit (DNA-PKcs) in IPF, promoted the accumulation of mesenchymal progenitors and progeny, and the expression of senescent markers by these cell types. METHODS: Surgical lung biopsy samples and lung fibroblasts were obtained from patients exhibiting slowly, rapidly or unknown progressing IPF and lung samples lacking any evidence of fibrotic disease (i.e. normal; NL). The expression of DNA-Pkcs in lung tissue was assessed by quantitative immunohistochemical analysis. Chronic inhibition of DNA-PKcs kinase activity was mimicked using a highly specific small molecule inhibitor, Nu7441. Proteins involved in DNA repair (stage-specific embryonic antigen (SSEA)-4+ cells) were determined by quantitative Ingenuity Pathway Analysis of transcriptomic datasets (GSE103488). Lastly, the loss of DNA-PKc was modeled in a humanized model of pulmonary fibrosis in NSG SCID mice genetically deficient in PRKDC (the transcript for DNA-PKcs) and treated with Nu7441. RESULTS: DNA-PKcs expression was significantly reduced in IPF lung tissues. Chronic inhibition of DNA-PKcs by Nu7441 promoted the proliferation of SSEA4+ mesenchymal progenitor cells and a significant increase in the expression of senescence-associated markers in cultured lung fibroblasts. Importantly, mesenchymal progenitor cells and their fibroblast progeny derived from IPF patients showed a loss of transcripts encoding for DNA damage response and DNA repair components. Further, there was a significant reduction in transcripts encoding for PRKDC (the transcript for DNA-PKcs) in SSEA4+ mesenchymal progenitor cells from IPF patients compared with normal lung donors. In SCID mice lacking DNA-PKcs activity receiving IPF lung explant cells, treatment with Nu7441 promoted the expansion of progenitor cells, which was observed as a mass of SSEA4+ CgA+ expressing cells. CONCLUSIONS: Together, our results show that the loss of DNA-PKcs promotes the expansion of SSEA4+ mesenchymal progenitors, and the senescence of their mesenchymal progeny.

CCR10+ epithelial cells from idiopathic pulmonary fibrosis lungs drive remodeling.

Idiopathic pulmonary fibrosis (IPF) is a devastating fibrotic lung disease of unknown etiology and limited therapeutic options. In this report, we characterize what we believe is a novel CCR10+ epithelial cell population in IPF lungs. There was a significant increase in the percentage of CCR10+ epithelial cells in IPF relative to normal lung explants and their numbers significantly correlated to lung remodeling in humanized NSG mice. Cultured CCR10-enriched IPF epithelial cells promoted IPF lung fibroblast invasion and collagen 1 secretion. Single-cell RNA sequencing analysis showed distinct CCR10+ epithelial cell populations enriched for inflammatory and profibrotic transcripts. Consistently, cultured IPF but not normal epithelial cells induced lung remodeling in humanized NSG mice, where the number of CCR10+ IPF, but not normal, epithelial cells correlated with hydroxyproline concentration in the remodeled NSG lungs. A subset of IPF CCR10hi epithelial cells coexpress EphA3 and ephrin A signaling induces the expression of CCR10 by these cells. Finally, EphA3+CCR10hi epithelial cells induce more consistent lung remodeling in NSG mice relative to EphA3-CCR10lo epithelial cells. Our results suggest that targeting epithelial cells, highly expressing CCR10, may be beneficial in IPF.

Targeting of TAM Receptors Ameliorates Fibrotic Mechanisms in Idiopathic Pulmonary Fibrosis.

RATIONALE: Idiopathic pulmonary fibrosis (IPF) is characterized by aberrant lung remodeling, which progressively abolishes lung function in an RTK (receptor tyrosine kinase)-dependent manner. Gas6 (growth arrest-specific 6) ligand, Tyro3 (TYRO3 protein tyrosine kinase 3), and Axl (anexelekto) RTK expression and activity are increased in IPF. OBJECTIVES: To determine if targeting these RTK pathways would inhibit fibroblast activation and the development of pulmonary fibrosis. METHODS: Quantitative genomic, proteomic, and functional analyses were used to determine Gas6/TAM (Tyro3, Axl, and Mertk [MER proto-oncogene, tyrosine kinase]) RTK expression and activation in tissues and fibroblasts from normal and IPF lungs. The profibrotic impact of these RTK pathways were also examined in bleomycin-induced pulmonary fibrosis and in SCID/Bg mice that developed pulmonary fibrosis after the intravenous administration of primary IPF fibroblasts. MEASUREMENTS AND MAIN RESULTS: Gas6, Axl, and Tyro3 were increased in both rapidly and slowly progressive IPF compared with normal lung samples and fibroblasts. Targeting these pathways with either specific antibodies directed at Gas6 or Axl, or with small-molecule TAM inhibitors indicated that the small molecule-mediated targeting approach was more efficacious in both in vitro and in vivo studies. Specifically, the TAM receptor inhibitor R428 (also known as BGB324) significantly inhibited the synthetic, migratory, and proliferative properties of IPF fibroblasts compared with the other Gas6/TAM receptor targeting agents. Finally, loss of Gas6 expression decreased lung fibrotic responses to bleomycin and treatment with R428 inhibited pulmonary fibrosis in humanized SCID/Bg mice. CONCLUSIONS: Gas6/TAM receptor activity contributes to the activation of pulmonary fibroblasts in IPF, suggesting that targeting this RTK pathway might be an effective antifibrotic strategy in this disease.