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To facilitate hazard characterisation for inhaled engineered nanomaterials we have established an aerosol exposure air-liquid-interface (AE-ALI) system, incorporating a Cultex RFSTM cell exposure system. While offering potential benefits in terms of more realistic aerosol exposure mode over traditional submerged in vitro models and being in-line with the principles of the 3Rs, (replace, reduce and refine animal use), detailed system characterisation is required to produce reliable and reproducible results. As such, this work looked to fully characterise our AE-ALI system with a view to optimising cell viability of system controls and deposition quantity, uniformity, and reproducibility for engineered nanomaterials. Using a nano-CeO2 aerosol, the effects of exposure conditions and system parameters (e.g. temperature, electrostatic precipitator voltage and airflow) on deposition efficiency, pattern and reproducibility were investigated using ICP-MS. While results showed that appropriate choice of operating parameters can produce broadly uniform deposition, reproducibility was low, and dose monitoring during exposures is recommended. After initial optimisation of the system for deposition, the effect of exposure duration, well size and flowrate on cellular responses (e.g. cytotoxicity (LDH) and selected gene expression (IL-8, CXCL1, HMOX1, SPP1) of system controls (H2O aerosol) was investigated using human lung alveolar (A549) and primary small airway epithelial cells. Results indicate that even after optimisation for cell health, the system itself is “toxic” to cells in comparison to incubator controls, with increasing effects observed with exposure duration meaning exposure durations must be minimised. Results from this study highlight the need for detailed characterisation of AE-ALI systems prior to use.

Cerium oxide nanoparticles (CeO2NPs) from some diesel fuel additives and other applications have been detected in ambient air. Concerns have been raised over their potential human health impact in situations of inadvertent exposure. Oxidative mechanisms have been suggested as a common feature for pulmonary injury in response to airborne particulate matter, including engineered nanomaterials. To understand in depth how CeO2NPs may influence oxidative stress induced pulmonary inflammation and fibrotic events, we used both in vivo and in vitro bleomycin-induced lung injury models. Male Sprague-Dawley rats were intratracheally instilled with bleomycin or saline (control) followed by nose-only inhalation exposure to nano-sized CeO2NP aerosols (mass concentration 1.8 mg/m3) or water (controls) for 3 hours per day for 4 days per week for one or two weeks. At 3 days post exposure, animals were sacrificed and bronchoalveolar lavage (BAL) fluid, lung histopathology and global mRNA expression analysed. Bleomycin exposure resulted in an increase in total BAL cells, fibrotic staining and significant induction of inflammatory and oxidative stress on mRNA sequencing analysis. Modifications of these responses by one-week exposure to CeO2NPs included attenuation of fibrotic staining and gene expression markers of lung function, inflammation and epithelial-mesenchymal transition (EMT). CeO2NP alone resulted in increased inflammatory responses but did not appear to cause fibrotic changes. Interpretation of these responses at a cellular level was further explored using 3D human small airway epithelium cultures (SmallAirTM) in an aerosol exposure air-liquid-interface system. This also indicated that some bleomycin-induced cellular responses could be attenuated by exposure to CeO2NP aerosols.

Obtaining high resolution, quality formation evaluation data is still only possible with wireline logging. However, with the continued push into deeper and more complex drilling environments, many challenges have been placed in the way of wireline logging, including high tension, high deviation, and increased differential pressure. These factors contribute to an increased risk of tool sticking incidents and lost-in-hole scenarios. Several methods of mitigating these issues on surface (powered capstans, pipe conveyance, etc) have been implemented in the past, but none have been successful in reducing or eliminating the risk downhole without introducing further drawbacks. This paper describes how a new wireline conveyance system has eliminated these issues. The conveyance system consists of wheeled carriages that carry the toolstring off-centre. The mass of the toolstring acts as a counterweight to ensure correct tool orientation in the wellbore. This orientation feature also enables a "guide" device to help navigate ledges and washouts. Such a system eliminates toolstring hold ups, allows access to highly deviated wells without pipe conveyance or tractors, and significantly mitigates differential sticking hazards, while also offering additional benefits in operational efficiency and data quality. A case study from a particularly difficult well in New Zealand is presented. Data acquisition in this well was fraught with challenges: In addition to the 2000m tangent section at 67° deviation, well had severe borehole breakouts. Previous experience in similar scenarios with conventional data acquisition methods yeided poor results. The wheeled carriage system was deployed in multiple innovative configurations resulting in the acquisition of excellent quality data from five wireline descents in hole. This wireline conveyance system has been routinely deployed on multiple deepwater operations in the Gulf of Mexico. One such operation is presented where large gains in logging efficiency have been realised, particularly with the elimination of differential sticking risk and time-consuming pipe conveyed logging. The new technology takes a holistic approach to wireline tool conveyance: Prevent sticking issues using wheeled carriages and mitigate fishing risk using ultra-high strength wireline cables. Wheeled carriages greatly reduce the tool-borehole contact area, preventing the incidence of tool sticking. In addition, wheeled carriages reduce drag while ensuring optimum data quality by sensor position and orientation within the wellbore. Ultra-high strength cables provide ability to log at very high tensions and at the same time provide high overpull capability. The result is a safe, efficient, cost effective and complete Wireline data acquisition.

Simple Summary This review presents and summarizes the key components and outcomes of a novel, unconventional radiation approach aimed to exploit immune-stimulatory radiation effects which, being added to direct radiation tumor cell killing, may improve the therapeutic ratio of radiotherapy. This technique, as a product of translational oncology research, was intentionally developed for the induction of immune-mediated bystander and abscopal effects in the treatment of unresectable bulky tumors which have much fewer therapeutic options and show poor prognoses after conventional treatments. This review offers insights into a unique unconventional radiotherapy technique which, due to its higher immunogenic potential, may improve the prognosis of patients affected by highly complex malignancies, providing additional opportunities for future research in terms of combining novel immuno-modulating agents with more modern radiotherapy approaches. Abstract Radiation-induced immune-mediated abscopal effects (AE) of conventional radiotherapy are very rare. Whole-tumor irradiation leads to lymphopenia due to killing of immune cells in the tumor microenvironment, resulting in immunosuppression and weak abscopal potential. This limitation may be overcome by partial tumor irradiation sparing the peritumoral immune-environment, and consequent shifting of immune-suppressive to immune-stimulatory effect. This would improve the radiation-directed tumor cell killing, adding to it a component of immune-mediated killing. Our preclinical findings showed that the high-single-dose irradiation of hypoxic tumor cells generates a stronger bystander effect (BE) and AE than the normoxic cells, suggesting their higher “immunogenic potential”. This led to the development of a novel Stereotactic Body RadioTherapy (SBRT)-based PArtial Tumor irradiation targeting HYpoxic segment (SBRT-PATHY) for induction of the immune-mediated BE and AE. Encouraging SBRT-PATHY-clinical outcomes, together with immunohistochemical and gene-expression analyses of surgically removed abscopal-tumor sites, suggested that delivery of the high-dose radiation to the partial (hypoxic) tumor volume, with optimal timing based on the homeostatic fluctuation of the immune response and sparing the peritumoral immune-environment, would significantly enhance the immune-mediated anti-tumor effects. This review discusses the current evidence on the safety and efficacy of SBRT-PATHY in the treatment of unresectable hypoxic bulky tumors and its bystander and abscopal immunomodulatory potential.

Background A novel unconventional SBRT-based PArtial Tumor irradiation targeting HYpoxic clonogenic cells (SBRT-PATHY) for induction of the tumoricidal bystander (BE) and abscopal effects (AE) was developed by translating our preclinical findings to a clinic in 2016. In order to further improve BE/AE response rate, SBRT-PATHY was upgraded in 2018 by the sparing of peritumoral immune microenvironment as a new OAR, defined by its own dose-constraints. Considering the anti-tumor immune response homeostatic fluctuation, which is cyclically suppressed and incited (“switched off and on”), we synchronized SBRT-PATHY with its most excitable phase, in order to overcome tumor tolerance locally and systemically. The aim of this study, therefore, was to report on the initial results of our latest innovation aimed to further improve BE/AE response rate by testing the effectiveness of the time-synchronized immune-guided SBRT-PATHY. Materials and methods In order to serially map the homeostatic anti-tumor immune response-fluctuations, High Sensitive C-Reactive Protein (HS-CRP), Lactate Dehydrogenase (LDH) and Lymphocyte/Monocyte Ratio (LMR) were analyzed using high-order polynomial trend analysis as surrogate of immune system response. After the biomarker data analysis detected the immune fluctuations and related idiosyncratic immune cycle periodicity, we determined the “most favourable” and “least favourable” treatment time-positions in the immune cycle. In order to evaluate the impact of an idiosyncratic immune cycle on treatment outcomes, our first consecutive four patients were treated on the “most favourable” while the remaining four on the “least favourable” day. Results The median follow-up was 11.8 months. The biomarker data analysis showed periodic immune response fluctuations of regular frequency. The “right” synchronization of SBRT-PATHY with the “most favorable day” of anti-tumor immune response was accompanied with improved clinical outcomes in terms of BE/AE-response rate. Conclusion We believe the right synchronization of radiotherapy with the homeostatically oscillating immune response may improve the probability of inducing BE/AE. Present study has been retrospectively registered on 18th of October 2019 by the ethic committee for Austrian region „Kärnten “in Klagenfurt (AUT), under study number A 37/19.