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Abstract: Murine AIDS (MAIDS) is a pathology induced by the LP-BM5 murine leukaemia virus mixture in susceptible strains of mice such as C57BL/6J resulting in lymphoproliferation and progressive immunodeficiency. The etiologic agent of this pathology is BM5d, a replication defective virus. BM5e is a replication competent virus in the viral mixture that functions as a helper virus. This paper describes real time PCR and RT-PCR assays for quantitation of the proviral DNA and viral RNA of BM5d and BM5e. Data is presented describing the change in BM5d and BM5e proviral DNA levels and viral RNA levels in both blood and spleen in the first 8weeks of infection. Infected mice have increasing levels of BM5d and BM5e viral DNA and RNA detectable from as early as 2weeks post infection. Similar levels of proviral DNA was found for BM5d and BM5e in PBMC and spleen, however higher levels of BM5e viral RNA were observed in both tissues throughout infection. The assays described can be used as both a diagnostic tool and to investigate the direct effect of treatments on the BM5d and BM5e viruses and MAIDS development. [Copyright &y& Elsevier]

Surgical resection of cancer remains the frontline therapy for millions of patients annually, but post-operative recurrence is common, with a relapse rate of around 45% for non-small cell lung cancer. The tumour draining lymph nodes (dLN) are resected at the time of surgery for staging purposes, and this cannot be a null event for patient survival and future response to immune checkpoint blockade treatment. This project investigates cancer surgery, lymphadenectomy, onset of metastatic disease, and response to immunotherapy in a novel model that closely reflects the clinical setting. In a murine metastatic lung cancer model, primary subcutaneous tumours were resected with associated dLNs remaining intact, completely resected or partially resected. Median survival after surgery was significantly shorter with complete dLN resection at the time of surgery (49 days (95%CI)) compared to when lymph nodes remained intact (> 88 days; p < 0.05). Survival was partially restored with incomplete lymph node resection and CD8 T cell dependent. Treatment with aCTLA4 whilst effective against the primary tumour was ineffective for metastatic lung disease. Conversely, aPD-1/aCD40 treatment was effective in both the primary and metastatic disease settings and restored the detrimental effects of complete dLN resection on survival. In this pre-clinical lung metastatic disease model that closely reflects the clinical setting, we observe decreased frequency of survival after complete lymphadenectomy, which was ameliorated with partial lymph node removal or with early administration of aPD-1/aCD40 therapy. These findings have direct relevance to surgical lymph node resection and adjuvant immunotherapy in lung cancer, and perhaps other cancer, patients. [ABSTRACT FROM AUTHOR]

Aim: The scales of space and time over which biotic interactions influence distribution patterns remain an area of debate. Biotic interactions may be particularly influential in the ecology of mammalian carnivores, which engage in strong predator–prey and competitive interactions. Regional, multi‐scale data on distribution patterns of interacting carnivore species are key to informing our understanding of this issue. Location: Washington State, USA. Methods: Using a spatially extensive camera‐trapping array, we examined the factors influencing distribution patterns of seven carnivore species at multiple spatial and temporal scales. We used single‐species occupancy models to assess the relative influence of abiotic and biotic covariates on distribution of individual carnivore species, and two‐species occupancy models to assess how dominant carnivores influence subordinate carnivore occupancy and detection. Results: Carnivore occupancy patterns responded more strongly to abiotic than biotic covariates at both spatial grains and extents of analysis. The influence of biotic variables decreased as the grain of analysis increased, while constraining our study area extent substantially decreased the explanatory power of abiotic variables. Interspecific interactions among carnivores influenced occupancy and detection across spatial scales. However, there was little evidence that interactions were more pronounced at finer temporal scales. Main Conclusions: Results demonstrate that at broad spatial extents, species distributions are largely dictated by abiotic factors, particularly climate. Although biotic factors related to habitat and prey were important factors for some species, they generally declined in importance as grain size of analysis increased, suggesting these interactions play out at finer resolutions. However, competitive and mutualistic interactions exerted an independent influence on distribution over broad extents and coarse grains of analysis, suggesting that failure to account for interactions may limit our ability to accurately model distributions of species and their responses to future large‐scale disturbances. [ABSTRACT FROM AUTHOR]

Populations of species located at southern range edges may be particularly vulnerable to the effects of climate change as warming temperatures and subsequent changes to ecosystems exceed species‐specific tolerances. One such species is Canada lynx (Lynx canadensis), a cold‐adapted mesocarnivore that maintains a large core population in Alaska, USA, and Canada but exists within several peripheral populations in the contiguous United States. Increases in temperature, declines in snow pack, and climate‐influenced increases in fire frequency and intensity, could negatively affect lynx populations, threatening their long‐term persistence in the continental United States. Despite these threats, our understanding of broad‐scale effects on lynx occupancy and the extent of current lynx distribution in many of these peripheral populations is minimal. We conducted an occupancy survey of lynx in Washington, USA, using a spatially extensive camera‐trapping array covering 7,000 km2 of potential lynx habitat. We used the resulting database of detection data to develop single‐season occupancy models to examine the abiotic and biotic effects on current lynx occupancy and predict future lynx distribution based on climate change forecasts. Our results show lynx occupancy across the Washington landscape is restricted and dictated largely by abiotic factors, disturbance regimes, and distance from source populations in Canada. Predictions of future distribution suggest lynx will be increasingly challenged by climatic changes, particularly at the southern and lower elevation portions of their range in Washington. Our results paint an alarming picture for lynx persistence in Washington that is relevant to current deliberations regarding lynx delisting from the Endangered Species Act. Our simple camera design was a highly effective method for surveying lynx across broad spatial scales, and could be a key monitoring tool for lynx that is easy to implement by researchers and government agencies. © 2020 The Wildlife Society. Lynx occupancy across Washington state is highly restricted and dictated largely by abiotic factors, disturbance regimes, and distance from source populations in Canada with future predictions demonstrating a substantial potential northward retraction of lynx range in Washington by 2100. Understanding current occupancy patterns and factors influencing occupancy in Washington is particularly relevant in the broader context of ongoing discussions to remove lynx from listing in the Endangered Species Act. [ABSTRACT FROM AUTHOR]

We have developed a handheld $\gamma $ -ray spectrometer based on 1024 pixels, $2.8\,\,\text {mm} \times 2.8\,\,\text {mm} \times 6$ mm in size, of gadolinium–yttrium–gallium–aluminum garnet (GYGAG) ((Gd,Y,Ce)3(Ga,Al)5O12) ceramic scintillator (total of 48-cm3 detector volume) coupled to silicon photodiode (SiPD) arrays. The SiPD arrays and readout ASIC, originally developed for medical imaging applications, have been adapted for portability in a lightweight box with heatsink and thermoelectric cooling. Custom readout firmware for $\gamma $ -ray spectroscopy has been implemented, and a system user interface was developed that runs on an Android tablet. We have optimized the processing of the GYGAG(Ce), the pixel optical coupling, and electronics readout parameters to obtain single pixel energy resolution as good as $R (662\,\,\text {keV}) = 3.1$ % full-width at half-maximum and full device resolution with singles events from all pixels summed of 4.5%. When Compton-summed events are included, full-energy peak efficiency increases by $\sim 2\times $ , and $R (662\,\,\text {keV}) = 4.7$ % is obtained for the full device. The pixelated architecture is leveraged to locate point sources of radiological materials using Compton imaging and active masking techniques. Directional detection of a 1 mCi Cs-137 source at 10 m can be made to ±10° in ~2 s. [ABSTRACT FROM AUTHOR]

Interspecies foraging associations occur in a wide variety of vertebrate taxa and are maintained through gains in foraging efficiency and (or) predator avoidance. Despite their advantages, foraging associations often are variable in space and time and benefits may not accrue equally to all participants. In mammals, interspecies associations between solitary mammalian carnivores are rare. Coyotes ( Canis latrans Say, 1823) and American badgers ( Taxidea taxus (Schreber, 1777)) occasionally form hunting associations in pursuit of ground squirrels ( Spermophilus armatus Kennicott, 1863), yet spatiotemporal variation in this association may be substantial. Better documentation of coyote-badger interactions across space and time will improve our understanding of the environmental drivers of this relationship and its benefit to both species. We used a broad-scale camera trapping array to document coyote-badger hunting associations. Out of 46 detections of badgers, we found five instances of hunting associations with coyotes, all of which occurred in mid- to late summer when ground squirrels were most active. Given our high rate of detection, these interactions are likely common on our study area. Habitat characteristics of the regions where we document coyote-badger interactions may have increased the likelihood of hunting associations. Our study demonstrates the effectiveness of camera traps for documenting this association and suggests that the coyote-badger system may be ideal for studying drivers of spatiotemporal variation in foraging associations. [ABSTRACT FROM AUTHOR]

Abstract: Recent renewed emphasis placed on gamma-ray detectors for national security purposes has motivated researchers to identify and develop new scintillator materials capable of high energy resolution and growable to large sizes. We have discovered that SrI2(Eu) has many desirable properties for gamma-ray detection and spectroscopy, including high light yield of ∼90,000 photons/MeV and excellent light yield proportionality. We have measured <2.7% FWHM at 662keV with small detectors (<1cm3) in direct contact with a photomultiplier tube, and ∼3% resolution at 662keV is obtained for 1in.3 crystals. Due to the hygroscopic nature of SrI2(Eu), similar to NaI(Tl), proper packaging is required for field use. This work describes a systematic study performed to determine the key factors in the packaging process to optimize performance. These factors include proper polishing of the surface, the geometry of the crystal, reflector materials and windows. A technique based on use of a collimated 137Cs source was developed to examine light collection uniformity. Employing this technique, we found that when the crystal is packaged properly, the variation in the pulse height at 662keV from events near the bottom of the crystal compared to those near the top of the crystal could be reduced to <1%. This paper describes the design and engineering of our detector package in order to improve energy resolution of 1in.3-scale SrI2(Eu) crystals. [Copyright &y& Elsevier]

Background: Antibodies that target immune checkpoints such as cytotoxic T lymphocyte antigen 4 (CTLA-4), programmed cell death protein/ligand 1 (PD-1/PD-L1) are approved for treatment of multiple cancer types. Chemotherapy is often administered with immune checkpoint blockade (ICB) therapies that target CTLA-4 and/or PD-(L)1. ICB targeting other immune checkpoints such as lymphocyte activating gene-3 (LAG-3) has the potential to improve antitumor responses when combined with chemotherapy. Response to anti-PD-1 ICB is dependent on progenitor exhausted CD8 + T cells (T PEX ) in the tumor, but it is unclear how chemotherapy alters T PEX proportions and phenotype., Methods: Here we investigated whether sequential chemotherapy altered T PEX frequency and immune checkpoint expression in multiple murine tumor models., Results: Two doses of two different anti-metabolite chemotherapies increased tumor infiltrating CD4 + , and CD8 + T PEX expressing LAG-3 in multiple mouse models, which was not restricted to tumor antigen specific CD8 + T cells. To determine if LAG-3 + tumor infiltrating lymphocytes (TILs) could be targeted to improve tumor control, we administered anti-LAG-3 and anti-PD-1 ICB after two doses of chemotherapy and found combination therapy generated robust antitumor responses compared with each agent alone. Both anti-LAG-3 and anti-PD-1 ICB with chemotherapy were required for the complete tumor regression observed., Conclusions: Changes in immune checkpoint expression on TILs during chemotherapy administration informs selection of ICB therapies to combine with., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY. Published by BMJ.)

Mesothelioma is a lethal cancer. Despite promising outcomes associated with immunotherapy, durable responses remain restricted to a minority of patients, highlighting the need for improved strategies that better predict outcome. Here, we described the development of a mesothelioma-specific gene signature that accurately predicts survival. Comprehensive gene expression analysis of asbestos exposed MexTAg Collaborative Cross mouse tumors revealed distinct tumor clusters characterized by epithelial mesenchymal transition/extracellular matrix, or immune infiltrate related gene expression profiles. Weighted gene co-expression network analysis (WGCNA) identified 20 hub genes that drove differential gene expression. Human homologues of these 20 hub genes were refined through univariate Cox regression and least absolute shrinkage and selection operator (LASSO) regression analyses to identify a six-gene mesothelioma-specific prognostic signature that accurately predicted patient survival across four independent human mesothelioma datasets. Furthermore, this six-gene signature demonstrated the potential to predict treatment response, thus advancing the management of this challenging malignancy., Competing Interests: The authors declare no competing interests., (© 2024 The Author(s).)

Objectives: This study combines two innovative mouse models in a major gene discovery project to assess the influence of host genetics on asbestos related disease (ARD). Conventional genetics studies provided evidence that some susceptibility to mesothelioma is genetic. However, the identification of host modifier genes, the roles they may play, and whether they contribute to disease susceptibility remain unknown. Here we report a study designed to rapidly identify genes associated with mesothelioma susceptibility by combining the Collaborative Cross (CC) resource with the well-characterised MexTAg mesothelioma mouse model. Methods: The CC is a powerful mouse resource that harnesses over 90% of common genetic variation in the mouse species, allowing rapid identification of genes mediating complex traits. MexTAg mice rapidly, uniformly, and predictably develop mesothelioma, but only after asbestos exposure. To assess the influence of host genetics on ARD, we crossed 72 genetically distinct CC mouse strains with MexTAg mice and exposed the resulting CC-MexTAg (CCMT) progeny to asbestos and monitored them for traits including overall survival, the time to ARD onset (latency), the time between ARD onset and euthanasia (disease progression) and ascites volume. We identified phenotype-specific modifier genes associated with these traits and we validated the role of human orthologues in asbestos-induced carcinogenesis using human mesothelioma datasets. Results: We generated 72 genetically distinct CCMT strains and exposed their progeny (2,562 in total) to asbestos. Reflecting the genetic diversity of the CC, there was considerable variation in overall survival and disease latency. Surprisingly, however, there was no variation in disease progression, demonstrating that host genetic factors do have a significant influence during disease latency but have a limited role once disease is established. Quantitative trait loci (QTL) affecting ARD survival/latency were identified on chromosomes 6, 12 and X. Of the 97-protein coding candidate modifier genes that spanned these QTL, eight genes ( CPED1, ORS1, NDUFA1, HS1BP3, IL13RA1, LSM8, TES and TSPAN12 ) were found to significantly affect outcome in both CCMT and human mesothelioma datasets. Conclusion: Host genetic factors affect susceptibility to development of asbestos associated disease. However, following mesothelioma establishment, genetic variation in molecular or immunological mechanisms did not affect disease progression. Identification of multiple candidate modifier genes and their human homologues with known associations in other advanced stage or metastatic cancers highlights the complexity of ARD and may provide a pathway to identify novel therapeutic targets., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Fisher, Patrick, Hoang, Marcq, Behrouzfar, Young, Miller, Robinson, Bueno, Nowak, Lesterhuis, Morahan and Lake.)

Background: Chemotherapy is included in treatment regimens for many solid cancers, but when administered as a single agent it is rarely curative. The addition of immune checkpoint therapy to standard chemotherapy regimens has improved response rates and increased survival in some cancers. However, most patients do not respond to treatment and immune checkpoint therapy can cause severe side effects. Therefore, there is a need for alternative immunomodulatory drugs that enhance chemotherapy., Methods: We used gene expression data from cyclophosphamide (CY) responders and non-responders to identify existing clinically approved drugs that could phenocopy a chemosensitive tumor microenvironment (TME), and tested combination treatments in multiple murine cancer models., Results: The vitamin A derivative tretinoin was the top predicted upstream regulator of response to CY. Tretinoin pre-treatment induced an inflammatory, interferon-associated TME, with increased infiltration of CD8 + T cells, sensitizing the tumor to subsequent chemotherapy. However, while combination treatment significantly improved survival and cure rate in a CD4 + and CD8 + T cell dependent manner in AB1-HA murine mesothelioma, this effect was model-selective, and could not be replicated using other cell lines., Conclusions: Despite the promising data in one model, the inability to validate the efficacy of combination treatment in multiple cancer models deprioritizes tretinoin/cyclophosphamide combination therapy for clinical translation., (© 2024. The Author(s).)

While chemotherapy remains the first-line treatment for many cancers, it is still unclear what distinguishes responders from non-responders. Here, we characterize the chemotherapy-responsive tumor microenvironment in mice, using RNA sequencing on tumors before and after cyclophosphamide, and compare the gene expression profiles of responders with progressors. Responsive tumors have an inflammatory and highly immune infiltrated pre-treatment tumor microenvironment characterized by the enrichment of pathways associated with CD4 + T cells, interferons (IFNs), and tumor necrosis factor alpha (TNF-α). The same gene expression profile is associated with response to cyclophosphamide-based chemotherapy in patients with breast cancer. Finally, we demonstrate that tumors can be sensitized to cyclophosphamide and 5-FU chemotherapy by pre-treatment with recombinant TNF-α, IFNγ, and poly(I:C). Thus, a CD4 + T cell-inflamed pre-treatment tumor microenvironment is necessary for response to chemotherapy, and this state can be therapeutically attained by targeted immunotherapy., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Chemotherapy has historically been the mainstay of cancer treatment, but our understanding of what drives a successful therapeutic response remains limited. The diverse response of cancer patients to chemotherapy has been attributed principally to differences in the proliferation rate of the tumor cells, but there is actually very little experimental data supporting this hypothesis. Instead, other mechanisms at the cellular level and the composition of the tumor microenvironment appear to drive chemotherapy sensitivity. In particular, the immune system is a critical determinant of chemotherapy response with the depletion or knock-out of key immune cell populations or immunological mediators completely abrogating the benefits of chemotherapy in pre-clinical models. In this perspective, we review the literature regarding the known mechanisms of action of cytotoxic chemotherapy agents and the determinants of response to chemotherapy from the level of individual cells to the composition of the tumor microenvironment. We then summarize current work toward the development of dynamic biomarkers for response and propose a model for a chemotherapy sensitive tumor microenvironment., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Tilsed, Fisher, Nowak, Lake and Lesterhuis.)

Antibodies that target immune checkpoints such as cytotoxic T lymphocyte antigen 4 (CTLA-4) and the programmed cell death protein 1/ligand 1 (PD-1/PD-L1) are now a treatment option for multiple cancer types. However, as a monotherapy, objective responses only occur in a minority of patients. Chemotherapy is widely used in combination with immune checkpoint blockade (ICB). Although a variety of isolated immunostimulatory effects have been reported for several classes of chemotherapeutics, it is unclear which chemotherapeutics provide the most benefit when combined with ICB. We investigated 10 chemotherapies from the main canonical classes dosed at the clinically relevant maximum tolerated dose in combination with anti-CTLA-4/anti-PD-L1 ICB. We screened these chemo-immunotherapy combinations in two murine mesothelioma models from two different genetic backgrounds, and identified chemotherapies that produced additive, neutral or antagonistic effects when combined with ICB. Using flow cytometry and bulk RNAseq, we characterized the tumor immune milieu in additive chemo-immunotherapy combinations. 5-fluorouracil (5-FU) or cisplatin were additive when combined with ICB while vinorelbine and etoposide provided no additional benefit when combined with ICB. The combination of 5-FU with ICB augmented an inflammatory tumor microenvironment with markedly increased CD8 + T cell activation and upregulation of IFNγ, TNFα and IL-1β signaling. The effective anti-tumor immune response of 5-FU chemo-immunotherapy was dependent on CD8 + T cells but was unaffected when TNFα or IL-1β cytokine signaling pathways were blocked. Our study identified additive and non-additive chemotherapy/ICB combinations and suggests a possible role for increased inflammation in the tumor microenvironment as a basis for effective combination therapy., Competing Interests: LB was employed by JJP Biologics. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Principe, Aston, Hope, Tilsed, Fisher, Boon, Dick, Chin, McDonnell, Nowak, Lake, Chee and Lesterhuis.)

Background: It is well established that the experience of chronic pain significantly differs among ethnic-racial groups. There is mixed evidence to suggest that societal influences may contribute to pain prevalence among cultural groups and their treatment response. One possible explanation for differences in pain experience are the differences in socioeconomic status among patients with chronic pain., Objective: To determine whether there is any difference in pain scores or treatment responses among patients with different socioeconomic status., Study Design: Retrospective analysis., Setting: Outpatient pain clinic., Methods: After approval from the Advocate Healthcare Institutional Review Board, we included 1,149 patients treated for different chronic pain conditions who were followed for at least 12 months. Patients were stratified into quartiles determined by median income according to ZIP code., Results: Of the sampled patients, 207 patients lived in ZIP codes with median incomes > $51,294; 515 in ZIP codes with median incomes between $40,083 and $51,294; 332 in ZIP  codes with median incomes between $30,625 and $40,083; and 95 in ZIP codes with median incomes < $30,625. Groups differed in age (P = 0.047), race (P < 0.001), body mass index (BMI) (P = 0.019), utilization of opioid medications (P = 0.011), morphine milligram equivalents (MME) on first visit (P = 0.036), and utilization of membrane stabilizers such as gabapentin (P = 0.019). There were no significant differences among groups in terms of gender (P = 0.531), type of pain experienced (P = 0.679), or time since pain onset (P = 0.174). Groups were treated similarly, with no statistically significant differences in the proportions of patients who had taken various nonopioid medications throughout their treatment course other than membrane stabilizers, the number of patients who received interventional pain management procedures, or MME at last visit. Average pretreatment numeric rating scale pain scores were not significantly different among quartiles (P = 0.079), posttreatment pain scores (P = 0.767), and subjective percent improvement (P = 0.434)., Limitations: This is a single center study and may have limitations in extrapolating to the general population., Conclusion: The results of our study show that there are no differences in pain perception or treatment responses in patients from different socioeconomic statuses despite differences among groups in age, BMI, race, utilization of opioid medications, and MME at first visit. Patients at this pain practice appear to have been treated with similar modalities regardless of socioeconomic status.

Objective: To evaluate the effects of a flotation vest (FV) and water flow rate (WFR) on limb kinematics of dogs swimming against a current., Animals: 7 (1 male and 6 female) healthy adult Siberian Huskies., Procedures: Dogs were habituated to swim with and without an FV beside an investigator in a continuous-flow pool against WFRs up to 2.9 km/h. During each of 4 experimental sessions in a repeated-measures study, markers were wrapped around the right carpus and tarsus, and a video was recorded while each dog swam with or without an FV for about 2 minutes at each of 7 WFRs between 0 and 2.9 km/h when the WFR was incrementally decreased or increased. Motion tracking software was used to measure stroke excursion and frequency., Results: Stroke excursion varied more than frequency among all dogs and in response to changes in experimental conditions. The male dog and 1 female dog were unable to complete the study. For the remaining 5 dogs across all experimental conditions, mean tarsus excursion was 30% that of the carpus. Mean total excursion (sum of the excursion-frequency products for the carpus and tarsus) decreased when an FV was worn and increased with WFR by 69% and 19% when WFR was incrementally increased and decreased, respectively., Conclusions and Clinical Relevance: In dogs, range of motion during swimming was greater for the carpus than tarsus, when an FV was not worn, and increased more with WFR when WFR was incrementally increased. Those factors should be considered during swimming-based rehabilitation.

Thoracic cancers pose a significant global health burden. Immune checkpoint blockade therapies have improved treatment outcomes, but durable responses remain limited. Understanding how the host immune system interacts with a developing tumor is essential for the rational development of improved treatments for thoracic malignancies. Recent technical advances have improved our understanding of the mutational burden of cancer cells and changes in cancer-specific gene expression, providing a detailed understanding of the complex biology underpinning tumor-host interactions. While there has been much focus on the genetic alterations associated with cancer cells and how they may impact treatment outcomes, how host genetics affects cancer development is also critical and will greatly determine treatment response. Genome-wide association studies (GWAS) have identified genetic variants associated with cancer predisposition. This approach has successfully identified host genetic risk factors associated with common thoracic cancers like lung cancer, but is less effective for rare cancers like malignant mesothelioma. To assess how host genetics impacts rare thoracic cancers, we used the Collaborative Cross (CC); a powerful murine genetic resource designed to maximize genetic diversity and rapidly identify genes associated with any biological trait. We are using the CC in conjunction with our asbestos-induced MexTAg mouse model, to identify host genes associated with mesothelioma development. Once genes that moderate tumor development and progression are known, human homologues can be identified and human datasets interrogated to validate their association with disease outcome. Furthermore, our CC-MexTAg animal model enables in-depth study of the tumor microenvironment, allowing the correlation of immune cell infiltration and gene expression signatures with disease development. This strategy provides a detailed picture of the underlying biological pathways associated with mesothelioma susceptibility and progression; knowledge that is crucial for the rational development of new diagnostic and therapeutic strategies. Here we discuss the influence of host genetics on developing an effective immune response to thoracic cancers. We highlight current knowledge gaps, and with a focus on mesothelioma, describe the development and application of the CC-MexTAg to overcome limitations and illustrate how the knowledge gained from this unique study will inform the rational design of future treatments of mesothelioma., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Behrouzfar, Burton, Mutsaers, Morahan, Lake and Fisher.)

Immune checkpoint therapy (ICT) results in durable responses in individuals with some cancers, but not all patients respond to treatment. ICT improves CD8 + cytotoxic T lymphocyte (CTL) function, but changes in tumor antigen-specific CTLs post-ICT that correlate with successful responses have not been well characterized. Here, we studied murine tumor models with dichotomous responses to ICT. We tracked tumor antigen-specific CTL frequencies and phenotype before and after ICT in responding and non-responding animals. Tumor antigen-specific CTLs increased within tumor and draining lymph nodes after ICT, and exhibited an effector memory-like phenotype, expressing IL-7R (CD127), KLRG1, T-bet, and granzyme B. Responding tumors exhibited higher infiltration of effector memory tumor antigen-specific CTLs, but lower frequencies of regulatory T cells compared to non-responders. Tumor antigen-specific CTLs persisted in responding animals and formed memory responses against tumor antigens. Our results suggest that increased effector memory tumor antigen-specific CTLs, in the presence of reduced immunosuppression within tumors is part of a successful ICT response. Temporal and nuanced analysis of T cell subsets provides a potential new source of immune based biomarkers for response to ICT., (Copyright © 2020 Principe, Kidman, Goh, Tilsed, Fisher, Fear, Forbes, Zemek, Chopra, Watson, Dick, Boon, Holt, Lake, Nowak, Lesterhuis, McDonnell and Chee.)

Objective: There is substantial evidence that exercise can safely reduce the risk of cancer and improve survival in different human cancer populations. Long latency periods associated with carcinogen-induced cancers like asbestos induced mesothelioma provide an opportunity to implement exercise as an intervention to delay or prevent disease development. However, there are limited studies investigating the ability of exercise to prevent or delay cancer, and exercise as a preventive strategy has never been assessed in models with a known carcinogen. We investigated the potential of voluntary exercise (VE) to delay development of asbestos related disease (ARD) in our well-characterised, asbestos induced MexTAg model of mesothelioma., Results: Asbestos exposed MexTAg mice were given continuous or delayed access to VE and ARD assessed over time. We found that the addition of VE did not affect ARD development in asbestos exposed MexTAg mice. However, non-asbestos exposed, aged matched control mice participated in significantly more VE behaviours, suggesting subclinical development of ARD after asbestos exposure had a greater impact on VE participation than age alone. These data highlight the importance of model choice and the potential limitation that some pre-clinical studies may not accurately represent the clinical paradigm, particularly in the context of prevention studies.

Objectives: To demonstrate coronary sinus (CS) retrograde catheterization as a practicable technique for delivering biologics into the heart., Background: There are many options to deliver biologics into the heart. However, there is no single optimal technique when considering safety, biologic retention, and reproducibility. Retrograde delivery has the potential to address many of these concerns. This study evaluated retrograde CS infusion of luciferase-expressing plasmid in a porcine model using the Advance® CS Coronary Sinus Infusion Catheter and bioluminescence imaging to track the expression of the infused biological markers., Methods: Plasmid was delivered retrograde into the CS in one of three infusion volumes. Twenty-four hours post-infusion, hearts were excised and underwent bioluminescence imaging to characterize the expression of the infusates. Heart and lung biopsies were also assessed for luciferase expression using RT-qPCR., Results: Retrograde infusion was safe and successful in all nine test subjects. Luciferase detection was inconsistent in the low volume group. Bioluminescence was confined predominantly along the posterolateral left ventricle for medium volume infusions and was more broadly dispersed along the anterior side of the heart for high volume infusions. Tissue mRNA analysis corroborated the bioluminescence results, with the highest concentration of luciferase expression localized in the left ventricle., Conclusions: Retrograde CS infusion is a promising technique for delivering biological molecules to the heart. Specifically, this study demonstrated that the low pressure coronary venous system accommodates a wide range of infusion volumes and that biological infusates can be maintained in situ following the resumption of coronary venous flow., (© 2019 Wiley Periodicals, Inc.)

Objectives: The global burden of asbestos-related diseases (ARDs) is significant, and most of the world's population live in countries where asbestos use continues. We examined the gaps between ARD research and suggestions of WHO and the International Labour Organization on prevention., Methods: From the Web of Science, we collected data on all articles published during 1991-2016 and identified a subset of ARD-related articles. We classified articles into three research areas-laboratory, clinical and public health-and examined their time trends. For all and the top 11 countries publishing ARD-related articles, we calculated the proportions of all ARD-related articles that were in each of the three areas, the average rates of ARD-related articles over all articles, and the average annual per cent changes of rates., Results: ARD-related articles (n=14 284) accounted for 1.3‰ of all articles in 1991, but this had declined to 0.8‰ by 2016. Among the three research areas, the clinical area accounted for the largest proportion (65.0%), followed by laboratory (26.5%) and public health (24.9%). The public health area declined faster than the other areas, at -5.7% per year. Discrepancies were also observed among the top 11 countries regarding emphasis on public health research, with Finland and Italy having higher, and China and the Netherlands lower, emphases., Conclusions: There is declining emphasis on the public health area in the ARD-related literature. Under the ongoing global situation of ARD, primary prevention will remain key for some time, warranting efforts to rectify the current trend in ARD-related research., Competing Interests: Competing interests: RL testifies in asbestos litigation., (© Author(s) (or their employer(s)) 2018. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Introduction: Regulatory T cells (Treg) play an important role in suppressing anti- immunity and their depletion has been linked to improved outcomes. To better understand the role of Treg in limiting the efficacy of anti-cancer immunity, we used a Diphtheria toxin (DTX) transgenic mouse model to specifically target and deplete Treg., Methods: Tumor bearing BALB/c FoxP3.dtr transgenic mice were subjected to different treatment protocols, with or without Treg depletion and tumor growth and survival monitored., Results: DTX specifically depleted Treg in a transient, dose-dependent manner. Treg depletion correlated with delayed tumor growth, increased effector T cell (Teff) activation, and enhanced survival in a range of solid tumors. Tumor regression was dependent on Teffs as depletion of both CD4 and CD8 T cells completely abrogated any survival benefit. Severe morbidity following Treg depletion was only observed, when consecutive doses of DTX were given during peak CD8 T cell activation, demonstrating that Treg can be depleted on multiple occasions, but only when CD8 T cell activation has returned to base line levels. Finally, we show that even minimal Treg depletion is sufficient to significantly improve the efficacy of tumor-peptide vaccination., Conclusions: BALB/c.FoxP3.dtr mice are an ideal model to investigate the full therapeutic potential of Treg depletion to boost anti-tumor immunity. DTX-mediated Treg depletion is transient, dose-dependent, and leads to strong anti-tumor immunity and complete tumor regression at high doses, while enhancing the efficacy of tumor-specific vaccination at low doses. Together this data highlight the importance of Treg manipulation as a useful strategy for enhancing current and future cancer immunotherapies.

Treatment of lung cancer remains a challenge, and lung cancer is still the leading cause of cancer-related mortality. Immunotherapy has previously failed in lung cancer but has recently emerged as a very effective new therapy, and there is now growing worldwide enthusiasm in cancer immunotherapy. We summarize why immune checkpoint blockade therapies have generated efficacious and durable responses in clinical trials and why this has reignited interest in this field. Cancer vaccines have also been explored in the past with marginal success. Identification of optimal candidate neoantigens may improve cancer vaccine efficacy and may pave the way to personalized immunotherapy, alone or in combination with other immunotherapy such as immune checkpoint blockade. Understanding the steps in immune recognition and eradication of cancer cells is vital to understanding why previous immunotherapies failed and how current therapies can be used optimally. We hold an optimistic view for the future prospect in lung cancer immunotherapy., (© 2016 Asian Pacific Society of Respirology.)

Background: Stromal cell-derived factor-1 (SDF-1) promotes tissue repair through mechanisms of cell survival, endogenous stem cell recruitment, and vasculogenesis. Stromal Cell-Derived Factor-1 Plasmid Treatment for Patients with Heart Failure (STOP-HF) is a Phase II, double-blind, randomized, placebo-controlled trial to evaluate safety and efficacy of a single treatment of plasmid stromal cell-derived factor-1 (pSDF-1) delivered via endomyocardial injection to patients with ischaemic heart failure (IHF)., Methods: Ninety-three subjects with IHF on stable guideline-based medical therapy and left ventricular ejection fraction (LVEF) ≤40%, completed Minnesota Living with Heart Failure Questionnaire (MLWHFQ) and 6-min walk distance (6 MWD), were randomized 1 : 1 : 1 to receive a single treatment of either a 15 or 30 mg dose of pSDF-1 or placebo via endomyocardial injections. Safety and efficacy parameters were assessed at 4 and 12 months after injection. Left ventricular functional and structural measures were assessed by contrast echocardiography and quantified by a blinded independent core laboratory. Stromal Cell-Derived Factor-1 Plasmid Treatment for Patients with Heart Failure was powered based on change in 6 MWD and MLWHFQ at 4 months., Results: Subject profiles at baseline were (mean ± SD): age 65 ± 9 years, LVEF 28 ± 7%, left ventricular end-systolic volume (LVESV) 167 ± 66 mL, N-terminal pro brain natriuretic peptide (BNP) (NTproBNP) 1120 ± 1084 pg/mL, MLWHFQ 50 ± 20 points, and 6 MWD 289 ± 99 m. Patients were 11 ± 9 years post most recent myocardial infarction. Study injections were delivered without serious adverse events in all subjects. Sixty-two patients received drug with no unanticipated serious product-related adverse events. The primary endpoint was a composite of change in 6 MWD and MLWHFQ from baseline to 4 months follow-up. The primary endpoint was not met (P = 0.89). For the patients treated with pSDF-1, there was a trend toward an improvement in LVEF at 12 months (placebo vs. 15 mg vs. 30 mg ΔLVEF: -2 vs. -0.5 vs. 1.5%, P = 0.20). A pre-specified analysis of the effects of pSDF-1 based on tertiles of LVEF at entry revealed improvements in EF and LVESV from lowest-to-highest LVEF. Patients in the first tertile of EF (<26%) that received 30 mg of pSDF-1 demonstrated a 7% increase in EF compared with a 4% decrease in placebo (ΔLVEF = 11%, P = 0.01) at 12 months. There was also a trend towards improvement in LVESV, with treated patients demonstrating an 18.5 mL decrease compared with a 15 mL increase for placebo at 12 months (ΔLVESV = 33.5 mL, P = 0.12). The change in end-diastolic and end-systolic volume equated to a 14 mL increase in stroke volume in the patients treated with 30 mg of pSDF-1 compared with a decrease of -11 mL in the placebo group (ΔSV = 25 mL, P = 0.09). In addition, the 30 mg-treated cohort exhibited a trend towards improvement in NTproBNP compared with placebo at 12 months (-784 pg/mL, P = 0.23)., Conclusions: The blinded placebo-controlled STOP-HF trial demonstrated the safety of a single endocardial administration of pSDF-1 but failed to demonstrate its primary endpoint of improved composite score at 4 months after treatment. Through a pre-specified analysis the STOP-HF trial demonstrates the potential for attenuating LV remodelling and improving EF in high-risk ischaemic cardiomyopathy. The safety profile supports repeat dosing with pSDF-1 and the degree of left ventricular remodelling suggests the potential for improved outcomes in larger future trials., (© The Author 2015. Published by Oxford University Press on behalf of the European Society of Cardiology.)

In this study intestinal and blood-brain barrier (BBB) permeability of ginkgolides A, B, C, J and bilobalide, isolated from Ginkgo biloba (Ginkgoaceae), was evaluated in Caco-2 and MDR1-MDCK cell monolayer models. The transport was examined for 2 hours in both absorptive and secretory directions. Quantitation was performed by UPLC-MS. In the Caco-2 model, each compound (100 microM) displayed a pH-dependent transport in the absorptive direction. A low permeability of ginkgolides was observed across the MDR1-MDCK model in the absorptive direction. An efflux was observed for all compounds in both the models. The efflux ratio was much higher in the MDR1-MDCK cell model (> 10) compared to the Caco-2 cell model (1.5-3.6). In comparison to ginkgolides, the permeability of bilobalide was much higher across the Caco-2 monolayer in both directions. However, a poor transport of bilobalide was observed in the MDR1-MDCK model in the absorptive direction. A high efflux was observed for all compounds in the mixture form as compared to their isolated forms. In rats, a single dose of bilobalide (8 mg/kg) administered intravenously resulted in a significant level of bilobalide in both plasma and brain. A brain-to-plasma partition coefficient of 0.56 at 120 min indicated its possibility of brain uptake., (Georg Thieme Verlag KG Stuttgart.New York.)

Purpose: Although androgens are known to protect bone, side effects and poor oral bioavailability have limited their use. We previously reported that S-3-(4-acetylamino-phenoxy)-2-hydroxy-2-methyl-N-(4-nitro-3-trifluoromethyl-phenyl)-propionamide (S-4) is a potent and tissue-selective androgen receptor modulator (SARM). This study was designed to evaluate the skeletal effects of S-4 in an osteopenic model., Methods: Aged female rats were gonadectomized or sham operated on day 1 and assigned to treatment groups. Dosing was initiated on day 90 and continued daily until day 210. Whole animal bone mineral density (BMD), body weight, and fat mass were determined by dual energy x-ray absorptiometry (DEXA). Regional analysis of excised bones was performed using DEXA or computed tomography. Femur strength was evaluated by 3-point bending., Results: S-4 restored whole body and lumbar vertebrae (L5-L6) BMD to the level of intact controls. Significant increases in cortical bone quality were observed at the femoral midshaft, resulting in increased load bearing capacity., Conclusions: S-4 demonstrated partial/complete recovery of bone parameters to age-matched intact levels. Increased efficacy observed in cortical bone sites is consistent with reported androgen action in bone. The ability of S-4 to promote bone anabolism, prevent bone resorption, and increase skeletal muscle mass/strength positions these drugs as promising new alternatives for the treatment of osteoporosis.

Purpose: This study was conducted to examine the bone and body composition effects of S-4, an aryl-propionamide derived Selective Androgen Receptor Modulator (SARM) in an ovariectomy induced model of accelerated bone loss., Methods: One hundred twenty female Sprague-Dawley rats aged to twenty-three weeks were randomly assigned to twelve treatment groups. Drug treatment was initiated immediately following ovariectomy and continued for one hundred twenty days. Whole body bone mineral density (BMD), body composition, and lumbar vertebrae BMD were measured by dual energy x-ray absorptiometry. More stringent regional pQCT and biomechanical strength testing was performed on excised femurs., Results: We found that S-4 treatment maintained whole body and trabecular BMD, cortical content, and increased bone strength while decreasing body fat in these animals., Conclusions: The data presented herein show the protective skeletal effects of S-4. Our previous reports have shown the tissue selectivity and muscle anabolic activity of S-4. Together these data suggest that S-4 could reduce the incidence of fracture via two different mechanisms (i.e., via direct effects in bone and reducing the incidence of falls through increased muscle strength). This approach to fracture reduction would be advantageous over current therapies in these patients which are primarily antiresorptive in nature.

We recently reported two nonsteroidal androgen receptor (AR) ligands that demonstrate tissue-selective pharmacological activity, identifying these S-3-(phenoxy)-2-hydroxy-2-methyl-N-(4-nitro-3-trifluoromethyl-phenyl)-propionamide analogs as the first members of a new class of drugs known as selective androgen receptor modulators. The purpose of these studies was to explore additional structure-activity relationships of selective androgen receptor modulators to enhance their AR binding affinity, AR-mediated transcriptional activation, and in vivo pharmacological activity. The AR binding affinity (K(i)) of 29 novel synthetic AR ligands was determined by a radioligand competitive binding assay and ranged from 1.0-51 nM. Compounds with electron-withdrawing substituents at the para- and meta-positions of the B-ring demonstrated the highest AR binding affinity. The AR-mediated transcriptional activation was determined using a cotransfection assay in CV-1 cells. Most compounds with two substituents in the B-ring maintained or improved their functional activity in vitro. However, compounds with three halogen substituents exhibited significant regioselectivity. Fifteen compounds were selected to examine their pharmacological activity in castrated rats. In vivo pharmacological activity and selectivity were significantly changed by structural modification in the B-ring. Compounds with halogen groups at the para- and meta-positions of the B-ring displayed the highest pharmacological activity. Incorporating substituents at the ortho-position of the B-ring resulted in poor pharmacological activity. In vitro and in vivo agonist activities were partially correlated. In conclusion, novel selective androgen receptor modulators with improved in vivo pharmacological activity can be designed and synthesized based on the structure-activity relationship identified in these studies.

Liver fibrosis was correlated with immunological parameters. Peripheral blood mononuclear cells (PBMCs) from patients with low fibrosis scores had more [corrected] interferon (IFN)-gamma-producing cells than did patients with higher fibrosis scores, when stimulated with hepatitis C virus (HCV) core antigen. Irrespective of liver fibrosis score, cells from all cytomegalovirus (CMV)-seropositive patients had similar IFN-gamma responses, when stimulated by CMV antigen, so patients with fibrosis did not have a broad-spectrum immunodeficiency. IFN-gamma response by PBMCs to HCV core antigen may provide a useful marker of the severity of liver disease in patients with hepatitis C.