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Adenovirus-mediated combination gene therapies have shown promising results in vaccination or treating malignant and genetic diseases. Nevertheless, an efficient system for the rapid assembly and incorporation of therapeutic genes into high-capacity adenoviral vectors (HCAdVs) is still missing. In this study, we developed the iMATCH (integrated modular assembly for therapeutic combination HCAdVs) platform, which enables the generation and production of HCAdVs encoding therapeutic combinations in high quantity and purity within 3 weeks. Our modular cloning system facilitates the efficient combination of up to four expression cassettes and the rapid integration into HCAdV genomes with defined sizes. Helper viruses (HVs) and purification protocols were optimized to produce HCAdVs with distinct capsid modifications and unprecedented purity (0.1 ppm HVs). The constitution of HCAdVs, with adapters for targeting and a shield of trimerized single-chain variable fragment (scFv) for reduced liver clearance, mediated cell- and organ-specific targeting of HCAdVs. As proof of concept, we show that a single HCAdV encoding an anti PD-1 antibody, interleukin (IL)-12, and IL-2 produced all proteins, and it led to tumor regression and prolonged survival in tumor models, comparable to a mixture of single payload HCAdVs in vitro and in vivo. Therefore, the iMATCH system provides a versatile platform for the generation of high-capacity gene therapy vectors with a high potential for clinical development., Graphical Abstract, The iMATCH platform enables an efficient assembly of up to four orthogonal expression cassettes on a single plasmid from which high-capacity adenoviral vectors can be generated within 3 weeks in unprecedented purity. iMATCH vectors encoding immunotherapeutic combinations led to tumor regression and prolonged survival, comparable to mixtures of single payload HCAdVs.

Mastic asphalt (MA) is an important construction material mainly used for infrastructure pavements as well as industrial and garage floors. As mastic asphalt construction sites are often limited in space, the prevalent form of construction is by manual paving. Therefore, the emissions caused by mastic asphalt are not only relevant for the environment, but also highly relevant for workers' health. In this study, we examine four mastic asphalt mixtures in terms of their emissions and worker exposure at various construction temperatures, ranging from 195 °C to 245 °C. We conducted three laboratory studies and a field study to evaluate the emissions away from the construction site, emissions in close proximity to the construction site, direct worker exposure and, for the very first time, also the absolute emissions in mass PM10 a.d. per mass of MA. The experiments show that a big part (80 wt%) of the PM10 a.d. emissions consists of particles smaller than 0.8 μm a.d. and are therefore very health relevant. Furthermore, we could show consistently in all our experiments that decreasing the paving temperature by 50 °C (by using a MA mixture workable below 200 °C) leads to an emission reduction of up to 70%.

Summary Purpose Several operative approaches and various implants for osseous fixation have been described to achieve four-corner fusion of the wrist. Given the discordance and to aid in further standardizing the technique, this study directly compares the outcomes of K-wire, fusion plate, and headless retrograde compressive screw fixations to achieve four-corner arthrodesis. Methods Sixty-four patients underwent four-corner fusion over a period of 5 years and were reviewed retrospectively. Twenty-one patients underwent bone fixation with conventional K-wires, 26 with locking plates, and 17 patients were treated by headless retrograde compressive screw fixations. Patients of the different groups were comparable regarding age, sex, hand dominance, and stage of disease. Results All study groups showed significant improvements in grip strength, decrease in pain (NRS) at rest and with activity, range-of-motion of the wrist, and wrist function (measured by the DASH-score). When evaluating the three groups amongst each other, overall complication and nonunion rates were low and revealed no significant differences between the groups of patients. However, regarding postoperative NRS at activity, dorsal flexion, and DASH-scores, the "screw" group showed significantly better results than the "wire" group. Conclusion The results show that all examined techniques of four-corner fusion can improve wrist function when compared to preoperative baseline (NRS at rest and activity, postoperative DASH-scores). However, headless retrograde compressive screw fixation had significant better results regarding pain relief (NRS) at activity and postoperative DASH-scores.

It has been shown that pre- and postconditioning of ischemically challenged tissue with erythropoietin (EPO) is able to reduce necrosis in a dose-dependent manner. The aim of this study was to determine the tissue-protective effects of different EPO dosages and administration regimes. Three groups of six C57Bl/6-mice each were analyzed: (1) pre- and postconditioning with initial high doses of EPO (starting at 2500 I.U./kg bw i.p.) followed by low doses of EPO (125 I.U./kg bw i.p.) (EPO-high-dose); (2) pre- and postconditioning with low doses of EPO (125 I.U./kg bw i.p.) (EPO-low-dose); and (3) untreated control group. Randomly perfused musculocutaneous flaps were mounted on dorsal skinfold chambers undergoing acute persistent ischemia and developing ∼50% necrosis without treatment. Intravital epifluorescence microscopy was performed at days 1, 3, 5, 7, and 10 after surgery, assessing flap necrosis, microcirculation, and angiogenesis. The hematocrit was measured at days 0, 3, 7, and 10. Only the EPO-low-dose regimen was associated with a significant reduction of necrosis when compared to untreated controls. EPO-low-dose showed a higher increase in both arteriolar diameter and velocity, thereby resulting in a significantly increased arteriolar blood flow and a hence higher functional capillary density (FCD) of the critically perfused zone. EPO-induced angiogenesis was significantly increased in EPO-low-dose at days 7 and 10. Only EPO-high-dose reached a significant hematocrit increase by day 10. Tissue pre- and postconditioning with low doses of EPO protects the critically perfused musculocutaneous tissue by maintaining capillary perfusion because of increased arteriolar blood flow mediated by nitric oxide (NO) expression.