Surgical excision and grafting of deep partial-thickness (DPT) and full-thickness (FT) burns is a cornerstone of wound care. The use of commercially available topical enzymatic agents has been limited due to slower and less complete eschar removal than surgical excision. Using a porcine model of DPT and FT burns, we compared the eschar removal efficacy of a bromelain-enriched enzymatic agent derived from the stems of pineapple plants and a commercially available collagenase. We created 40 DPT and 40 FT burns on four anesthetized Yorkshire pigs. Eschar removal was initiated 24 hours later. Two pigs each were randomly assigned to collagenase or the bromelain-enriched agent. The bromelain-enriched agent was applied topically once for 4 hours followed by a 2-hour soaking. The collagenase was applied topically daily until complete removal of eschar or for up to 14 days. All bromelain-enriched treated FT burns underwent complete removal of the eschar after a single application while none of the collagenase-treated FT burns underwent complete removal of the eschar even after 14 days of treatment. All bromelain-enriched treated DPT burns had complete eschar removal after the single application. None of the collagenase-treated DPT burns experienced complete removal of eschar after 10 days; by day 14, 35% had complete eschar removal, 30% had >50% eschar removed, and 35% had <50% eschar removed. We conclude that eschar removal is quicker and more complete with the bromelain-enriched compared with collagenase debriding agent.

CRISPR/Cas9-mediated genome editing provides potential for therapeutic development. Efficacy and long-term safety represent major concerns that remain to be adequately addressed in preclinical studies. Here we show that CRISPR/Cas9-mediated genome editing in two distinct SOD1-amyotrophic lateral sclerosis (ALS) transgenic mouse models prevented the development of ALS-like disease and pathology. The disease-linked transgene was effectively edited, with rare off-target editing events. We observed frequent large DNA deletions, ranging from a few hundred to several thousand base pairs. We determined that these large deletions were mediated by proximate identical sequences in Alu elements. No evidence of other diseases was observed beyond 2 years of age in these genome edited mice. Our data provide preclinical evidence of the efficacy and long-term safety of the CRISPR/Cas9 therapeutic approach. Moreover, the molecular mechanism of proximate identical sequences-mediated recombination provides mechanistic information to optimize therapeutic targeting design, and to avoid or minimize unintended and potentially deleterious recombination events.

Background: Burn injury progression in the secondary zone of ischemia is common leading to delayed healing and increased scarring. We hypothesized that a topical surfactant, would reduce burn injury progression in a validated rat comb burn model compared with topical antibiotic ointment. Methods: We created 40 comb burns on 20 rats which were randomized to daily topical application of the surfactant or a triple antibiotic ointment. The comb burns consisted of 4 full thickness burns with 3 unburned interspaces between the 4 burns. These unburned interspaces represented the zone of ischemia, and when left untreated, generally progress to full thickness necrosis within several days. Comb burns were assessed daily for the presence of gross necrosis of the interspaces. At 7 days the comb burns were excised and blindly evaluated for the presence of histological evidence of necrosis. The study had 80% power to detect a 25% difference in the percentages of necrotic interspaces on day 7. Results: There were no differences in the percentages of histologically necrotic interspaces at 7 days in burns treated with the surfactant or antibiotic ointment (85% [95%CI, 74 to 92] vs. 75% [95%CI, 63 to 84]; mean difference 10% [95%CI -4 to 24]). There were also no between group differences in the percentages of grossly necrotic interspaces on any of the seven days of the experiment. The surfactant remained intact and adherent while the antibiotic had been absorbed at each daily dressing change. Conclusions: A topical surfactant did not reduce injury progression in the rat comb burn model when compared with antibiotic ointment. The surfactant was more durable than the antibiotic ointment. (C) 2020 Elsevier Ltd and ISBI. All rights reserved.

Whether the depth and healing of scalds and contact burns are similar is controversial. Due to water's greater heat capacity, we hypothesized that when exposed to similar temperatures and durations of exposure, burns caused by hot water would be deeper than those caused by contact with hot metal. Forty standardized burns were created in two anesthetized female domestic pigs using a brass bar or circulating heated water. In one pig, the temperature was kept constant (95°C) while the duration of exposure varied (5, 10, 15 seconds) In the second pig, the exposure time was kept constant (10 seconds) while the temperature of exposure varied (70°C, 80°C, 98°C). Periodic punch biopsies were taken to determine burn depth immediately after injury, percentage burns reepithelialized within 21 days, and depth of scar at 28 days. The analysis was performed using analysis of variance. When the temperature was held constant, duration of exposure (5, 10, and 15 seconds) was associated with scar depth (2.1 vs 3.8 vs 5.0 mm, respectively, P = 0.001) but not with burn depth (2.0 vs 2.2 vs 2.3 mm, respectively, P = 0.10). When exposure duration was held constant, temperature (70°C, 80°C, 98°C) was associated with scar depth (0.6 vs 1.7 vs 3.6, P < 0.001) but not with burn depth (1.2 vs 1.5 vs 1.7 mm, respectively, P = 0.21). Burn depths were greater for scald than contact burns although not significantly greater. After controlling for temperature, the difference in scar depth between scalds and contact burns was statistically significant (marginal means 3.0 for contact burns, 4.3 for scalds, P = 0.008). We conclude that burns created in swine with circulating hot water result in deeper scars than those created by contact with a brass bar when controlling for temperature and duration of exposure.

Abstract Introduction Whether the depth and healing of scalds and contact burns are similar is controversial. Prior studies have not compared scalds and contact burns using identical temperatures and durations of exposure limiting direct comparison. We hypothesized that when exposed to similar temepratures and durations of exposure, burns caused by hot water and contact with hot metal would be similar. Methods Multiple 2.5 or 5 cm diameter standardized cutaneous burns were created in two anesthetized female domestic pigs (~45kg) using a brass bar or circulating heated water in replicates. The temperatures were controlled and kept constant using thermosensors. In one pig, the temperature was kept constant (95 degrees C) while the duration of exposure varied (5, 10, 15 sec.) In the second pig, the exposure time was kept constant (10 sec.) while the temperature of exposure varied (70, 80, 98 degrees C). Burns were covered with a topical antibiotic ointment and frequent dressing changes were performed for 4 weeks. Periodic punch biospies were taken to determine burn depth immediately after injury, percentage burns reepithelialized within 21 days, and depth of scar at 28 days. Analysis was performed using analysis of variance. Results When temperature was held constant, duration of exposure (5, 10, and 15 sec.) was associated with scar depth (2.1 vs 3.8 vs 5.0 mm respectively, P=0.001) and there was a trend for an association with burn depth (2.0 vs 2.2 vs 2.3 mm respectively, P=0.10). When exposure duration was held constant, temperature (70, 80, 98 degrees C) was associated with scar depth (0.6 vs 1.7 vs 3.6, P< 0.001) but not significantly associated with burn depth (1.2 vs 1.5 vs 1.7 mm respectively, P=0.21). For each pig a comparison of burn and scar depth by device generally showed that depths were greater for scald than contact burns although not significantly greater. Only scar depth in the first pig showed a difference with a significance of less than 0.10 (2.9 for contact vs 4.1 mm for scalds, P=0.08). After controlling for temperature the difference between scalds and contact burns was statistically significant (marginal means 3.0 for contact burns, 4.3 for scalds, P=0.008). When combining data from both pigs and controlling for temperatures and duration, scar depths for scalds (2.7 mm) vs contacts (1.9 mm) showed a significant difference (P=0.02) while burn depth was not quite significant (1.7 vs 1.5 mm, P=0.07). Almost all burns were reepithelialized by day 21. Conclusions Burns created in swine with circulating hot water tend to be deeper and result in deeper scars than those created by contact with a brass bar when controlling for temperature and duration of exposure. Applicability of Research to Practice Scalds of similar temperature and exposure times result in deeper scars than contact burns.

Purpose:The objective of this study was to understand the molecular and physiologic mechanisms behind the lens cataract differences in Aquaporin 0-knockout-Heterozygous (AQP0-Htz) mice developed in C57 and FVB (lacks beaded filaments [BFs]) strains.Methods:Lens transparency was studied using dark field light microscopy. Water permeability (Pf) was measured in fiber cell membrane vesicles. Western blotting/immunostaining was performed to verify expression of BF proteins and connexins. Microelectrode-based intact lens intracellular impedance was measured to determine gap junction (GJ) coupling resistance. Lens intracellular hydrostatic pressure (HP) was determined using a microelectrode/manometer system.Results:Lens opacity and spherical aberration were more distinct in AQP0-Htz lenses from FVB than C57 strains. In either background, compared to wild type (WT), AQP0-Htz lenses showed decreased Pf (approximately 50%), which was restored by transgenic expression of AQP1 (TgAQP1/AQP0-Htz), but the opacities and differences between FVB and C57 persisted. Western blotting revealed no change in connexin expression levels. However, in C57 AQP0-Htz and TgAQP1/AQP0-Htz lenses, GJ coupling resistance decreased approximately 2.8-fold and the HP gradient decreased approximately 1.9-fold. Increased Pf in TgAQP1/AQP0-Htz did not alter GJ coupling resistance or HP.Conclusions:In C57 AQP0-Htz lenses, GJ coupling resistance decreased. HP reduction was smaller than the coupling resistance reduction, a reflection of an increase in fluid circulation, which is one reason for the less severe cataract in C57 than FVB. Overall, our results suggest that AQP0 modulates GJs in the presence of BF proteins to maintain lens transparency and homeostasis.