[68Ga]Ga-THP-tetrazine for bioorthogonal click radiolabelling: pretargeted PET imaging of liposomal nanomedicines.

Pretargeted PET imaging using bioorthogonal chemistry is a leading strategy for the tracking of long-circulating agents such as antibodies and nanoparticle-drug delivery systems with short-lived isotopes. Here, we report the synthesis, characterisation and in vitro/vivo evaluation of a new 68Ga-based radiotracer [68Ga]Ga-THP-Tetrazine ([68Ga]Ga-THP-Tz) for bioorthogonal click radiochemistry and in vivo labelling of agents with slow pharmacokinetics. THP-tetrazine (THP-Tz) can be radiolabelled to give [68/67Ga]Ga-THP-Tz at room temperature in less than 15 minutes with excellent radiochemical stability in vitro and in vivo. [68Ga]Ga-THP-Tz was tested in vitro and in vivo for pretargeted imaging of stealth PEGylated liposomes, chosen as a leading clinically-approved platform of nanoparticle-based drug delivery, and for their known long-circulating properties. To achieve this, PEGylated liposomes were functionalised with a synthesised transcyclooctene (TCO) modified phospholipid. Radiolabelling of TCO-PEG-liposomes with [68/67Ga]Ga-THP-Tz was demonstrated in vitro in human serum, and in vivo using both healthy mice and in a syngeneic cancer murine model (WEHI-164 fibrosarcoma). Interestingly in vivo data revealed that [68Ga]Ga-THP-Tz was able to in vivo radiolabel liposomes present in the liver and spleen, and not those in the blood pool or in the tumour. Overall, these results demonstrate the potential of [68Ga]Ga-THP-Tz for pretargeted imaging/therapy but also some unexpected limitations of this system.

Prediction of spontaneous combustion susceptibility of coal seams based on coal intrinsic properties using various machine learning tools.

Spontaneous combustion of coal leading to mine fire is a major problem in most of the coal mining countries in the world. It causes major loss to the Indian economy. The liability of coal to spontaneous combustion varies from place to place and mainly depends on the coal intrinsic properties and other geo-mining factors. Hence, the prediction of spontaneous combustion susceptibility of coal is of utmost importance for preventing the risk of fire in coal mines and utility sectors. Machine learning tools are pivotal in system improvements in relation to the statistical analysis of experimental results. Wet oxidation potential (WOP) of coal determined in the laboratory is one of the most relied indices used for assessing the spontaneous combustion susceptibility of coal. In this study, multiple linear regression (MLR) and five different machine learning (ML) techniques, such as Support Vector Regression (SVR), Artificial Neural Network (ANN), Random Forest (RF), Gradient Boosting (GB) and Extreme Gradient Boost (XGB) algorithms, were used to predict the spontaneous combustion susceptibility (WOP) of coal seams based on the coal intrinsic properties. The results derived from the models were compared with the experimental data. The results indicated excellent prediction accuracy and ease of interpretation of tree-based ensemble algorithms, like Random Forest, Gradient Boosting and Extreme Gradient Boosting. The MLR exhibited the least while XGB demonstrated the highest predictive performance. The developed XGB achieved R2 of 0.9879, RMSE of 4.364 and VAF of 84.28%. In addition, the results of sensitivity analysis showed that the volatile matter is most sensitive to the changes in WOP of coal samples considered in the study. Thus, during spontaneous combustion modelling and simulation, volatile matter can be used as the most influential parameter for assessing the fire risk of the coal samples considered in the study. Further, the partial dependence analysis was done to interpret the complex relationships between the WOP and intrinsic properties of coal.

Rapid short-pulses of focused ultrasound and microbubbles deliver a range of agent sizes to the brain.

Focused ultrasound and microbubbles can non-invasively and locally deliver therapeutics and imaging agents across the blood-brain barrier. Uniform treatment and minimal adverse bioeffects are critical to achieve reliable doses and enable safe routine use of this technique. Towards these aims, we have previously designed a rapid short-pulse ultrasound sequence and used it to deliver a 3 kDa model agent to mouse brains. We observed a homogeneous distribution in delivery and blood-brain barrier closing within 10 min. However, many therapeutics and imaging agents are larger than 3 kDa, such as antibody fragments and antisense oligonucleotides. Here, we evaluate the feasibility of using rapid short-pulses to deliver higher-molecular-weight model agents. 3, 10 and 70 kDa dextrans were successfully delivered to mouse brains, with decreasing doses and more heterogeneous distributions with increasing agent size. Minimal extravasation of endogenous albumin (66.5 kDa) was observed, while immunoglobulin (~ 150 kDa) and PEGylated liposomes (97.9 nm) were not detected. This study indicates that rapid short-pulses are versatile and, at an acoustic pressure of 0.35 MPa, can deliver therapeutics and imaging agents of sizes up to a hydrodynamic diameter between 8 nm (70 kDa dextran) and 11 nm (immunoglobulin). Increasing the acoustic pressure can extend the use of rapid short-pulses to deliver agents beyond this threshold, with little compromise on safety. This study demonstrates the potential for deliveries of higher-molecular-weight therapeutics and imaging agents using rapid short-pulses.

Methane migration and explosive fringe localisation in retreating longwall panel under varied ventilation scenarios: a numerical simulation approach.

Methane-based inflammable underground coal mine environment has led to catastrophic losses in the past. Migration of methane from the working seam and desorption region above and below the seam causes explosion hazard. In this study, the computational fluid dynamics (CFD)-based simulations of a longwall panel in a methane-rich inclined coal seam of the Moonidih mine in India established that the ventilation parameters greatly influence the methane flow in the longwall tailgate and porous medium of the goaf. The field survey and CFD analysis revealed that methane accumulation on the "rise side" wall of the tailgate is attributable to the geo-mining parameters. Further, the turbulent energy cascade was observed to impact the distinct dispersion pattern along the tailgate. The numerical code was used to investigate the changes in ventilation parameters made to dilute the methane concentration in the longwall tailgate. Methane concentration in the tailgate outlet decreased from 2.4 to 1.5% as the inlet air velocity increased from 2 to 4 m/s. The oxygen ingress into the goaf increased from 0.5 to 4.5 lps as the velocity was increased, causing the explosive zone in the goaf to expand from 5 to 100 m. Amongst all velocity variations, the lowest level of gas hazard was observed at an inlet air velocity of 2.5 m/s. This study, thus, demonstrated the ventilation-based numerical method to assess the coexistence of gas hazard in the goaf and longwall workings. Moreover, it provided impetus to the necessity of novel strategies to monitor and mitigate the methane hazard in U-type longwall mine ventilation.

Non-invasive radionuclide imaging of trace metal trafficking in health and disease: "PET metallomics".

Several specific metallic elements must be present in the human body to maintain health and function. Maintaining the correct quantity (from trace to bulk) and location at the cell and tissue level is essential. The study of the biological role of metals has become known as metallomics. While quantities of metals in cells and tissues can be readily measured in biopsy and autopsy samples by destructive analytical techniques, their trafficking and its role in health and disease are poorly understood. Molecular imaging with radionuclides - positron emission tomography (PET) and single photon emission computed tomography (SPECT) - is emerging as a means to non-invasively study the acute trafficking of essential metals between organs, non-invasively and in real time, in health and disease. PET scanners are increasingly widely available in hospitals, and methods for producing radionuclides of some of the key essential metals are developing fast. This review summarises recent developments in radionuclide imaging technology that permit such investigations, describes the radiological and physicochemical properties of key radioisotopes of essential trace metals and useful analogues, and introduces current and potential future applications in preclinical and clinical investigations to study the biology of essential trace metals in health and disease.

Liposome delivery to the brain with rapid short-pulses of focused ultrasound and microbubbles.

Liposomes are clinically used drug carriers designed to improve the delivery of drugs to specific tissues while minimising systemic distribution. However, liposomes are unable to cross the blood-brain barrier (BBB) and enter the brain, mostly due to their large size (ca. 100 nm). A noninvasive and localised method of delivering liposomes across the BBB is to intravenously inject microbubbles and apply long pulses of ultrasound (pulse length: >1 ms) to a targeted brain region. Recently, we have shown that applying rapid short pulses (RaSP) (pulse length: 5 µs) can deliver drugs with an improved efficacy and safety profile. However, this was tested with a relatively smaller 3-kDa molecule (dextran). In this study, we examine whether RaSP can deliver liposomes to the murine brain in vivo. Fluorescent DiD-PEGylated liposomes were synthesized and injected intravenously alongside microbubbles. The left hippocampus of mice was then sonicated with either a RaSP sequence (5 µs at 1.25 kHz in groups of 10 ms at 0.5 Hz) or a long pulse sequence (10 ms at 0.5 Hz), with each pulse having a 1-MHz centre frequency (0.35 and 0.53 MPa). The delivery and distribution of the fluorescently-labelled liposomes were assessed by fluorescence imaging of the brain sections. The safety profile of the sonicated brains was assessed by histological staining. RaSP was shown to locally deliver liposomes across the BBB at 0.53 MPa with a more diffused and safer profile compared to the long pulse ultrasound sequence. Cellular uptake of liposomes was observed in neurons and microglia, while no uptake within astrocytes was observed in both RaSP and long pulse-treated brains. This study shows that RaSP allows a targeted and safe delivery of liposomal drugs into the murine brain with potential to deliver drugs into neuronal and glial targets.