This study reports a novel nanoparticle system with simple and modular one-step assembly, which can respond intelligently to biologically relevant variations in pH. Importantly, these particles also show the ability to induce escape from the endosomal/lysosomal compartments of the cell, which is integral to the design of efficient polymeric delivery systems. The nanoparticles were formed by the nanoprecipitation of pH-responsive poly(2-(diethylamino)ethyl methacrylate) (PDEAEMA) and poly(2-(diethylamino)ethyl methacrylate)-b-poly(ethylene glycol) (PDEAEMA-b-PEG). Rhodamine B octadecyl ester perchlorate was successfully encapsulated within the hydrophobic core of the nanoparticle upon nanoprecipitation into PBS at pH 8. These particles disassembled when the pH was reduced below 6.8 at 37 °C. Cellular experiments showed the successful uptake of the nanoparticles into the endosomal/lysosomal compartments of 3T3 fibroblast cells. The ability to induce escape from the endosomes was demonstrated by the use of calcein, a membrane-impermeable fluorophore. The modular nature of these particles combined with promising endosomal escape capabilities make these dual component PDEAEMA nanoparticles useful for drug and gene delivery applications.

The effective escape of nanocarriers from endosomal compartments of the cell remains a major hurdle in nanomedicine. The endosomal escape of pH-responsive, self-assembled, dual component particles based on poly[2-(diethylamino)ethyl methacrylate)(PDEAEMA) and poly(ethylene glycol)-b-poly[2-(diethylamino)ethyl methacrylate) (PEG-b-PDEAEMA) has been recently reported. Herein, we report that polymer molecular weight (M ) can be used to tune endosomal escape of nanoparticle delivery systems. PDEAEMA of M 7 kDa, 27 kDa, 56 kDa and 106 kDa was synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization and co-assembled with PEG-b-PDEAEMA (16 kDa) via nanoprecipitation. All particles had similar size, displayed pH-responsive behaviour, and low toxicity regardless of molecular weight. Ovalbumin was loaded in the particles to demonstrate loading and release capabilities and as a marker to study internalization and endosomal escape. Association and endosomal escape was found to depend on molecular weight, with enhanced escape observed for high M PDEAEMA: 42% of cells with particle induced endosomal escape for 106 kDa nanoparticles, compared to minimal escape for 7 kDa particles. The results show that a simple variation in molecular weight can enhance the endosomal escape of polymeric carriers, and thus improve their effectiveness for intracellular delivery of therapeutics.

Targeting nanoparticles to specific cellular receptors has the potential to deliver therapeutic compounds to target sites while minimizing side effects. To this end, we have conjugated a targeting protein, holo-transferrin (holo-Tf), to pH-responsive polymers, poly­(2-(diethylamino)­ethyl methacrylate) (PDEAEMA) and poly­(2-(diethylamino)­ethyl methacrylate)-ran-poly­(2-(diisopropylamino)­ethyl methacrylate (PDEAEMA-r-PDPAEMA). These protein–polymer hybrid materials were observed to self-assemble when the pH is increased above the pK a of the polymer. We demonstrate that their response to pH could be tuned depending on the polymer constituent attached to holo-Tf. Importantly, the targeting behavior of these nanoparticles could be maximized by tuning the density of holo-Tf on the nanoparticle surface by the introduction of a (PDEAEMA-r-PDPAEMA)-b-poly­(ethylene glycol) (PEG) copolymer.

Background: Proximal sesamoid bone fractures are common catastrophic injuries in racehorses. Understanding the response of proximal sesamoid bones to race training can inform fracture prevention strategies.Objectives: To describe proximal sesamoid bone microstructure of racehorses and to investigate the associations between microstructure and racing histories. Study design: Cross-sectional.Methods: Proximal sesamoid bones from 63 Thoroughbred racehorses were imaged using micro-computed tomography. Bone volume fraction (BVTV) and bone material density (BMD) of the whole bone and four regions (apical, midbody dorsal, midbody palmar and basilar) were determined. Generalised linear regression models were used to identify the associations between bone parameters and race histories of the horses. Results:The mean sesamoid BVTV was 0.79 ± 0.08 and BMD was 806.02 ± 24.66 mg HA/ccm. BVTV was greater in medial sesamoids compared with lateral sesamoids (0.80 ± 0.07 vs 0.79 ± 0.08; P < .001) predominantly due to differences in the apical region (medial-0.76 ± 0.08 vs lateral-0.72 ± 0.07; P < .001). BVTV in the midbody dorsal region (0.86 ± 0.06) was greater than other regions (midbody palmar-0.79 ± 0.

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Background: Certain stride characteristics have been shown to affect changes in biomechanical factors that are associated with injuries in human athletes. Determining the relationship between stride characteristics and musculoskeletal injury (MSI) may be key in limiting injury occurrence in the racehorse.Objectives: This study aimed to determine whether changes in race day speed and stride characteristics over career race starts are associated with an increased risk of MSI in racehorses. Study design: Case-control study. Methods: Speed, stride length, and stride frequency data were obtained from the final 200 m sectional of n = 5660 race starts by n = 584 horses (case n = 146, control n = 438). Multivariable joint models, combining longitudinal and survival (time to injury) analysis, were generated. Hazard ratios and their 95% confidence intervals (CI) are presented.Results: The risk of MSI increased by 1.18 (95% CI 1.09, 1.28; P < 0.001) for each 0.1 m/s decrease in speed and by 1.11 (95% CI 1.02, 1.21; P = 0.01) for each 10 cm decrease in stride length over time (career race starts). A more marked rate of decline in speed and stride length was observed approximately 6 races prior to injury. Risk of MSI was highest early in the horse's racing career. Main limitations: Only final sectional stride characteristics were assessed in the model. The model did not account for time between race starts. Conclusions: Decreasing speed and stride length over multiple races is associated with MSI in racehorses. Monitoring stride characteristics over time may be beneficial for the early detection of MSI.

Currently, there is a paucity of data on the barriers for Australian Thoroughbred horses transitioning from stud farm to racetrack. This paper reports the reasons why horses failed to enter race training and documents their exit destinations. Biographical records of Australian Thoroughbred horses born in 2014 were investigated to determine the number of horses that had not officially entered race training by the start of the 4-year old racing season (1 August 2018). Of the 13,677 foals born in 2014, 66% had commenced training and 51% had raced before the beginning of their 4-year-old season in Australia. A sampling frame based on the post code of the premises where foals were born and records from Racing Australia were used to select a geographically representative sample of the 2014 Australian Thoroughbred foal crop (n = 4,124). From the population eligible for sampling 1,275 horses that had not entered training were enrolled in the survey and their breeders were sent an online questionnaire with follow-up phone calls for those who had not responded. Of the 633 responses (50% of 1275) the most frequent outcomes for horses were: death (38%, n = 239), participation in the racing industry in their 4-year old racing season (24%, n = 154) and retirement (16%, n = 100) either as Australian Stud Book (ASB) bloodstock (n = 17), or as horses rehomed outside the Thoroughbred industry (n = 83). Illness or injury was the most frequent reason for horses not entering race training that were ASB bloodstock, rehomed or deceased. There was a loss of traceability at the point of sale with most horses sold at 1 year of age. This study provides important information on the reasons, alternative outcomes and gaps in traceability for horses not entering training prior to the 4-year-old racing season.

Background With each stride, galloping horses generate large skeletal loads which influence bone physiology, and may contribute to musculoskeletal injury. Horse speed and stride characteristics are related, but the usefulness of using horse speed and distance travelled as a proxy for stride characteristics is unknown. Objectives We aimed to determine stride characteristics, their variance and their relationship with speed in horses performing maximally. Study design Retrospective cross‐sectional analysis of archived data. Methods Stride characteristics obtained using GPS and inertial sensors in Thoroughbred horses were retrieved. Data per 200 m race segment (‘sectionals’) for horses competing in races (N = 25,259 race starts) were analysed to determine if speed predicted stride parameters. Multivariable mixed‐effects linear regression models were fitted. Results Mean (±SD) stride length, stride count (number of strides per 200 m), duration and speed were 7.08 ± 0.39 m, 28.32 ± 1.56 strides/200 m, 0.43 ± 0.02 s/stride and 16.63 ± 1.04 m/s across all sectionals and starts. Speed and stride length decreased, and stride count increased with race progression (P < 0.001). Male sex, greater race distance, better finishing position and firmer track surfaces were associated with less strides per 200 m and longer stride durations. Main limitations Lack of an independent party validation of the measurement system used in this study. Conclusions There was a substantial inter‐horse variation in stride parameters, with speed predicting half or less of this variation. Speed alone does not fully explain stride characteristics in horses. Future studies aimed at investigating the impact of gait on bone biology and pathology would benefit from accounting for stride characteristics (eg length and duration).