Multiplexed Intact-Tissue Transcriptional Analysis at Cellular Resolution.

In recently developed approaches for high-resolution imaging within intact tissue, molecular characterization over large volumes has been largely restricted to labeling of proteins. But volumetric nucleic acid labeling may represent a far greater scientific and clinical opportunity, enabling detection of not only diverse coding RNA variants but also non-coding RNAs. Moreover, scaling immunohistochemical detection to large tissue volumes has limitations due to high cost, limited renewability/availability, and restricted multiplexing capability of antibody labels. With the goal of versatile, high-content, and scalable molecular phenotyping of intact tissues, we developed a method using carbodiimide-based chemistry to stably retain RNAs in clarified tissue, coupled with amplification tools for multiplexed detection. The resulting technology enables robust measurement of activity-dependent transcriptional signatures, cell-identity markers, and diverse non-coding RNAs in rodent and human tissue volumes. The growing set of validated probes is deposited in an online resource for nucleating related developments from across the scientific community.

Translation of small downstream ORFs enhances translation of canonical main open reading frames.

In addition to canonical open reading frames (ORFs), thousands of translated small ORFs (containing less than 100 codons) have been identified in untranslated mRNA regions (UTRs) across eukaryotes. Small ORFs in 5' UTRs (upstream (u)ORFs) often repress translation of the canonical ORF within the same mRNA. However, the function of translated small ORFs in the 3' UTRs (downstream (d)ORFs) is unknown. Contrary to uORFs, we find that translation of dORFs enhances translation of their corresponding canonical ORFs. This translation stimulatory effect of dORFs depends on the number of dORFs, but not the length or peptide they encode. We propose that dORFs represent a new, strong, and universal translation regulatory mechanism in vertebrates.

Smoking and vaping alter genes related to mechanisms of SARS-CoV-2 susceptibility and severity.

Evidence for the impact of smoking on COVID-19 is contradictory, and there is little research on vaping. Here we provide greater clarity on mechanisms perturbed by tobacco cigarette, electronic cigarette and nicotine exposures that may impact the risks of infection and/or disease severity.Following PRISMA guidelines, OVID and Web of Science databases were searched. Study design and exposure-induced gene expression changes were extracted. Each study was quality assessed and higher confidence scores were assigned to genes consistently changed across multiple studies following the same exposure. These genes were used to explore pathways significantly altered following exposure.125 studies provided data on 480 genes altered by exposure to tobacco cigarettes, e-cigarettes, nicotine or SARS-CoV-2. Genes involved in both SARS-CoV-2 viral-entry and inflammation were changed following exposure. Pathway analysis revealed that many of those genes with high confidence scores are involved in common cellular processes relating to hyperinflammatory immune responses.Exposure to tobacco cigarettes, e-cigarettes, or nicotine, may therefore impact initial host-pathogen interactions and disease severity. Smokers and vapers of e-cigarettes with nicotine, could potentially be at increased risk of SARS-CoV-2 infection, associated cytokine storm, and acute respiratory distress syndrome. However, further research is required, particularly on e-cigarettes, to determine the biological mechanisms involved in perturbation of viral-entry genes and host-pathogen interactions and subsequent responses within the respiratory tract. This will improve our physiological understanding of the impact of smoking and vaping on COVID-19, informing public health advice and providing improved guidance for management of SARS-CoV-2 and other respiratory viruses.

COVID-19 outcomes in haemopoietic stem cell transplant recipients in Western Australia: the value of vaccination and antiviral therapy.

Coronavirus-19 (COVID-19) mortality rates among haemopoietic stem cell transplant (HSCT) patients are high, ranging between 20% and 40%. We prospectively evaluated the mortality outcomes of COVID-19 in Western Australian HSCT patients. A total of 32/492 (6.5%) HSCT recipients contracted COVID-19 during the study, of whom 30/32 (94%) developed mild or asymptomatic disease. Two allogeneic HSCT patients were hospitalised for severe COVID-19; one patient died. Stringent healthcare, social isolation practices, aggressive vaccination programmes and rapid access to COVID-19 antivirals may have promoted mild COVID-19 illness in Western Australian HSCT patients, resulting in one of the lowest COVID-19 mortality rates in HSCT recipients worldwide.

Genome-wide chromosomal association of Upf1 is linked to Pol II transcription in Schizosaccharomyces pombe.

Although the RNA helicase Upf1 has hitherto been examined mostly in relation to its cytoplasmic role in nonsense mediated mRNA decay (NMD), here we report high-throughput ChIP data indicating genome-wide association of Upf1 with active genes in Schizosaccharomyces pombe. This association is RNase sensitive, correlates with Pol II transcription and mRNA expression levels. Changes in Pol II occupancy were detected in a Upf1 deficient (upf1Δ) strain, prevalently at genes showing a high Upf1 relative to Pol II association in wild-type. Additionally, an increased Ser2 Pol II signal was detected at all highly transcribed genes examined by ChIP-qPCR. Furthermore, upf1Δ cells are hypersensitive to the transcription elongation inhibitor 6-azauracil. A significant proportion of the genes associated with Upf1 in wild-type conditions are also mis-regulated in upf1Δ. These data envisage that by operating on the nascent transcript, Upf1 might influence Pol II phosphorylation and transcription.

Use of Absorbable Dermal Staples in Bilateral Breast Reduction Surgery: Effects on Operative Times, Surgical Outcomes, and Procedure Charges.

BACKGROUND: Use of the absorbable deep dermal stapler in wound closure has become more common in plastic surgery because of its possible reduction in operative times and subsequent decrease in operative room costs. In this study, we examine the effects of this stapler on operative times and postoperative complications in bilateral reduction mammaplasties. METHODS: A retrospective, observational cohort study was conducted via electronic chart review on patients who underwent bilateral reduction mammaplasties. Patients were stratified by wound closure method. One group was closed with sutures only, and in the other group, deep dermal staples were used during closure of the inframammary fold incision. Incidences of patient comorbidities and postoperative complications were compared. In addition, a financial cost analysis was performed. RESULTS: The final patient cohort included 62 patients. Operative time was reduced by an average of 21.8 minutes when using deep dermal staples during closure, compared with when closing solely with sutures (P = 0.032). When controlling for mass of breast tissue removed and type of pedicle, deep dermal staple closure still predicted a reduction of 26.5 (SE, 9.9) minutes in operative time (P = 0.010). Postoperative complications were not affected by wound closure method (odds ratio, 4.36; 95% confidence interval, 0.91-31.7, P = 0.087). Though not statistically significant, financial charge was decreased with usage of deep dermal staples (P = 0.34). CONCLUSIONS: Use of absorbable deep dermal staples produces a significant decrease in operative time for reduction mammaplasties with no increase in postoperative complication rates.

Accessing Mg-Ion Storage in V2PS10 via Combined Cationic-Anionic Redox with Selective Bond Cleavage.

Magnesium batteries attract interest as alternative energy-storage devices because of elemental abundance and potential for high energy density. Development is limited by the absence of suitable cathodes, associated with poor diffusion kinetics resulting from strong interactions between Mg2+ and the host structure. V2PS10 is reported as a positive electrode material for rechargeable magnesium batteries. Cyclable capacity of 100 mAh g-1 is achieved with fast Mg2+ diffusion of 7.2 × ${\times }$ 10-11-4 × ${\times }$ 10-14 cm2 s-1. The fast insertion mechanism results from combined cationic redox on the V site and anionic redox on the (S2)2- site; enabled by reversible cleavage of S-S bonds, identified by X-ray photoelectron and X-ray absorption spectroscopy. Detailed structural characterisation with maximum entropy method analysis, supported by density functional theory and projected density of states analysis, reveals that the sulphur species involved in anion redox are not connected to the transition metal centres, spatially separating the two redox processes. This facilitates fast and reversible Mg insertion in which the nature of the redox process depends on the cation insertion site, creating a synergy between the occupancy of specific Mg sites and the location of the electrons transferred.

New Methodology for Determining Plasma Protein Binding Kinetics Using an Enzyme Reporter Assay Coupling with High-Resolution Mass Spectrometry.

Determination of drug binding kinetics in plasma is important yet extremely challenging. Accordingly, we introduce "dynamic free fraction" as a new binding parameter describing drug-protein binding kinetics. We demonstrate theoretically and experimentally that the dynamic free fraction can be determined by coupling the drug binding assay with a reporter enzyme in combination with high-resolution mass spectrometry measuring the relative initial steady-state rates of enzymatic reactions in the absence and presence of matrix proteins. This novel and simple methodology circumvents a long-standing challenge inherent in existing methods for determining binding kinetics constants, such as kon and koff, and enables assessment of the impact of protein binding kinetics on pharmaceutical properties of drugs. As demonstrated with nine model drugs, the predicted liver extraction ratio, a measure of efficiency of drug removal by the liver, correlates significantly better to the observed extraction ratio when using the dynamic free fraction (fD) in place of the unbound fraction (fu) of the drug in plasma. Similarly, the in vivo hepatic clearance of these drugs, a measure of liver drug elimination, is highly comparable to the clearance values calculated with the dynamic free fraction (fD), which is markedly better than those calculated with the unbound fraction (fu). In contrast to the prevailing view, these results indicate that protein binding kinetics is an important pharmacokinetic property of a drug. As plasma protein binding is one of the most important drug properties, this new methodology may represent a breakthrough and could have a real impact on the field.

Reducing Peri-implant Capsule Thickness in Submuscular Rodent Model of Breast Reconstruction With Delayed Radiotherapy.

INTRODUCTION: Capsular contracture remains the most common complication following device-based breast reconstruction, occurring in up to 50% of women who also undergo adjuvant radiotherapy either before or after device-based reconstruction. While certain risk factors for capsular contracture have been identified, there remains no clinically effective method of prevention. The purpose of the present study is to determine the effect of coating the implant with the novel small molecule Met-Z2-Y12, with and without delayed, targeted radiotherapy, on capsule thickness and morphologic change around smooth silicone implants placed under the latissimus dorsi in a rodent model. METHODS: Twenty-four female Sprague Dawley rats each had 2 mL smooth round silicone breast implants implanted bilaterally under the latissimus dorsi muscle. Twelve received uncoated implants and twelve received implants coated with Met-Z2-Y12. Half of the animals from each group received targeted radiotherapy (20 Gray) on postoperative day ten. At three and 6 months after implantation, the tissue surrounding the implants was harvested for analysis of capsular histology including capsule thickness. Additionally, microCT scans were qualitatively analyzed for morphologic change. RESULTS: Capsules surrounding Met-Z2-Y12-coated implants were significantly thinner (P = 0.006). The greatest difference in capsule thickness was seen in the irradiated 6-month groups, where mean capsule thickness was 79.1 ± 27.3 µm for uncoated versus 50.9 ± 9.6 µm for Met-Z2-Y12-coated implants (P = 0.038). At the time of explant, there were no capsular morphologic differences between the groups either grossly or per microCT. CONCLUSIONS: Met-Z2-Y12 coating of smooth silicone breast implants significantly reduces capsule thickness in a rodent model of submuscular breast reconstruction with delayed radiotherapy.

Filling a nick in NIK: Extending the half-life of a NIK inhibitor through structure-based drug design.

Inhibition of NF-κB inducing kinase (NIK) has been pursued as a promising therapeutic target for autoimmune disorders due to its highly regulated role in key steps of the NF-κB signaling pathway. Previously reported NIK inhibitors from our group were shown to be potent, selective, and efficacious, but had higher human dose projections than desirable for immunology indications. Herein we report the clearance-driven optimization of a NIK inhibitor guided by metabolite identification studies and structure-based drug design. This led to the identification of an azabicyclo[3.1.0]hexanone motif that attenuated in vitro and in vivo clearance while maintaining NIK potency and increasing selectivity over other kinases, resulting in a greater than ten-fold reduction in predicted human dose.

Letermovir for pre-emptive cytomegalovirus therapy after allogeneic hematopoietic cell transplantation.

BACKGROUND: Cytomegalovirus (CMV) is a common cause of morbidity after allogeneic haematopoietic cell transplantation (alloHCT). Pre-emptive therapy (PET) with valganciclovir (VGC) is associated with haematological toxicity. METHODS: We included alloHCT patients from 2018 to 2021 where letermovir (LTV) was used for CMV PET because of cytopenias. RESULTS: Ten patients were included. Six received VGC prior to LTV. VGC was commenced at median d42, given for median 40 days. LTV was commenced at median d90, given for median 54 days. At commencement of antiviral, CMV viral load was higher for VGC at 3.7 log10 IU/mL, compared to LTV at 2.9 log10 IU/mL. Viral load reduction occurred at 0.18 log10 IU/mL per week for VGC, compared to 0.17 log10 IU/mL per week for LTV. There was no clinically significant CMV viremia after stopping LTV. Cytopenias improved on LTV. CONCLUSION: LTV was effective in controlling CMV viremia when it was given at a lower starting CMV viral load and later post alloHCT than VGC. Further study is required of LTV as upfront PET following alloHCT.

A quasi-one-dimensional model for ion-aerosol interactions and aerosol charge state downwind of corona-producing alternating current (AC) HVPL under stable atmospheric conditions.

Corona ions produced by high voltage power lines (HVPL) can alter the local atmospheric electrical environment downwind, potentially increasing electrostatic charge on airborne particulates via ion-aerosol attachment. However, previous epidemiological assessments attempting to assess this 'corona ion hypothesis' have used proxies e.g. ion concentration or distance from HVPL, rather than aerosol charge state directly, due to difficulties in modeling this quantity. We present a quasi-1D model incorporating both Gaussian plume dynamics and ion-aerosol and ion-ion interaction microphysics which could be applied to future studies of charged aerosol near HVPL. The response of the model to changes in a range of input parameters is characterized, and validation is attempted by means of comparison with previous work where ion- and aerosol concentrations and properties (including electrical mobility and electric charge states) upwind and downwind of HVPL are measured.

Overhead AC powerlines and rain can alter the electric charge distribution on airborne particles - Implications for aerosol dispersion and lung deposition.

Corona ions from high voltage power lines (HVPL) can increase electrostatic charge on airborne pollutant particulates, possibly increasing received dose upon inhalation. To investigate the potential increased risk of childhood leukemia associated with residence near alternating current (AC) HVPL, we measured the particle charge state and atmospheric electricity parameters upwind, downwind and away from HVPL. Although we observed noticeable charge state alteration from background levels, most HVPL do not significantly increase charge magnitude. Particular HVPL types are shown to have most effect, increasing net charge to 15 times that at background. However, the magnitude of charge alteration during rainfall is comparable with the most extreme HVPL measurement. On current evidence, based on the current adult lung model, we suggest that although charge is sometimes enhanced to levels which may alter atmospheric particle dynamics, increased lung deposition is unlikely.

Potential for cardiac toxicity with methylimidazolium ionic liquids.

Methylimidazolium ionic liquids (MILs) are solvent chemicals used in industry. Recent work suggests that MILs are beginning to contaminate the environment and lead to exposure in the general population. In this study, the potential for MILs to cause cardiac toxicity has been examined. The effects of 5 chloride MIL salts possessing increasing alkyl chain lengths (2 C, EMI; 4 C, BMI; 6 C; HMI, 8 C, M8OI; 10 C, DMI) on rat neonatal cardiomyocyte beat rate, beat amplitude and cell survival were initially examined. Increasing alkyl chain length resulted in increasing adverse effects, with effects seen at 10-5 M at all endpoints with M8OI and DMI, the lowest concentration tested. A limited sub-acute toxicity study in rats identified potential cardiotoxic effects with longer chain MILs (HMI, M8OI and DMI) based on clinical chemistry. A 5 month oral/drinking water study with these MILs confirmed cardiotoxicity based on histopathology and clinical chemistry endpoints. Since previous studies in mice did not identify the heart as a target organ, the likely cause of the species difference was investigated. qRT-PCR and Western blotting identified a marked higher expression of p-glycoprotein-3 (also known as ABCB4 or MDR2) and the breast cancer related protein transporter BCRP (also known as ABCG2) in mouse, compared to rat heart. Addition of the BCRP inhibitor Ko143 - but not the p-glycoproteins inhibitor cyclosporin A - increased mouse cardiomyocyte HL-1 cell sensitivity to longer chain MILs to a limited extent. MILs therefore have a potential for cardiotoxicity in rats. Mice may be less sensitive to cardiotoxicity from MILs due in part, to increased excretion via higher levels of cardiac BCRP expression and/or function. MILs alone, therefore may represent a hazard in man in the future, particularly if use levels increase. The impact that MILs exposure has on sensitivity to cardiotoxic drugs, heart disease and other chronic diseases is unknown.

Elite North American soccer performance in thermally challenging environments: An explorative approach to tracking outcomes.

AIMS: The physiologic challenges related to performances in hot conditions calls for dedicated consideration when planning athlete training, although complete amelioration of the effects of heat may not be possible. We aimed to quantify within-subject correlations between different measures of environmental temperature and performance changes over multiple elite soccer competitions. METHODS: Thirty-seven elite male soccer players (age: 26 ± 3.4 years, height: 171 ± 2 cm, body mass: 78 ± 7.1 kg) competed in North America over four seasons (range: 3 to 98 matches). Players wore global positioning system devices during games and reported differential-RPE immediately post game. Temperatures at kick-off, week average temperature, the difference between game-day and week average (DiffTemp), and heat index at kick-off were obtained. Within-player correlations were calculated using general linear models to quantify associations between fluctuations in temperature measures and physical and perceived outputs. RESULTS: Correlations between total distance and the various temperature measures were trivial to small (range: -0.08 to 0.13, p=<0.001-0.02). Small negative correlations were found between all temperature measures except DiffTemp and high-speed running (HSR) (range: -0.17 to -0.14, p=<0.001). Most correlations between differential-RPE and temperature measures were trivial to small and not significant (r = 0.06 to 0.18 p = 0.03-0.92) although breathlessness-RPE and heat index showed a small significant association (P = 0.018). CONCLUSION: Decrements in HSR appear to be associated with increased environmental temperature, however, these associations are small in magnitude.

Training Volume and High-Speed Loads Vary Within Microcycle in Elite North American Soccer Players.

ABSTRACT: Ishida, A, Draper, G, Wright, M, Emerson, J, and Stone, MH. Training volume and high-speed loads vary within microcycle in elite North American soccer players. J Strength Cond Res 37(11): 2229-2234, 2023-The purposes of this study were to reduce dimensionality of external training load variables and examine how the selected variables varied within microcycle in elite North American soccer players. Data were collected from 18 players during 2018-2020 in-seasons. Microcycle was categorized as 1, 2, 3, 4, 5 days before match day (MD-1, MD-2, MD-3, MD-4, and MD-5, respectively). Training load variables included total distance, average speed, maximum velocity, high-speed running distance (HSR), average HSR, HSR efforts, average HSR efforts, sprint distance, average sprint distance, sprint efforts, average sprint efforts, total PlayerLoad, and average PlayerLoad. The first principal component (PC) can explain 66.0% of the variances and be represented by "high-speed load" (e.g., HSR and sprint-related variables) with the second PC relating to "volume" (e.g., total distance and PlayerLoad) accounting for 17.9% of the variance. Average sprint distance and total distance were selected for further analysis. Average sprint distance was significantly higher at MD-3 than at MD-2 (p = 0.01, mean difference = 0.36 m•minute-1, 95% confidence intervals [CIs] = 0.07-0.65 m•minute-1) and MD-4 (p = 0.012, mean difference = 0.26 m•minute-1, 95% CIs = 0.10-0.41 m•minute-1). Total distance was significantly higher at MD-3 than at MD-1 (p < 0.001, mean difference = 1,465 m, 95% CIs = 1,003-1926 m), and MD-2 (p < 0.001, mean difference = 941 m, 95% CIs = 523-1,360 m). Principal component analysis may simplify reporting process of external training loads. Practitioners may need to choose "volume" and "high-speed load" variables. Elite North American Soccer players may accumulate higher average sprint distance at MD-3 than at other training days.

A Cohort Comparison of Midlife Marital Quality: A Quarter Century of Change.

Marital quality has been declining among recent cohorts, but whether this pattern characterizes middle-aged and older married adults is largely unknown. The doubling of the divorce rate among persons over the age of 50 years foretells poorer quality marriages for today's midlife adults than a generation ago. Combining data on married individuals aged 50-65 years from the 1987-88 National Survey of Families and Households (NSFH) and the 2013 Families and Relationships Study, we conduct a cohort comparison of five dimensions of midlife marital quality. Today's older adults report more marital disagreement and instability as well as less fairness and interaction with their spouses than their counterparts did a generation ago. The two cohorts report comparable levels of marital happiness. Consistent with the upward trend in divorce during the second half of life, the quality of midlife marriages appears to have declined over the past quarter century.

Physiologically Based Pharmacokinetic Models Can Be Used to Predict the Unique Nonlinear Absorption Profiles of Vismodegib.

Predicting human pharmacokinetics (PK) during the drug discovery phase is valuable to assess doses required to reach therapeutic exposures. For orally administered compounds, however, this can be especially difficult, since the absorption process is complex. Vismodegib is a compound with unique nonlinear oral PK characteristics in humans. Oral physiologically based pharmacokinetic (PBPK) models were built using preclinical in vitro and in vivo data and successfully predicted the oral PK profiles in rats, dogs, and monkeys. Simulated drug exposures (area under the concentration-time curve from time 0 to infinity and Cmax) following oral administration were within twofold of observed values for dogs and monkeys, and close to twofold for rats, providing validation to the model structure. Adaptation of this oral PBPK model to humans, using human physiologic parameters coupled with predicted human PK, resulted in underpredictions of vismodegib exposure following both single and multiple doses. When observed human PK was used to drive the oral PBPK model, oral PK profiles in humans were well predicted, with fold errors in predicted versus observed drug exposures being close to 1. Importantly, the oral PBPK model captured the unique nonlinear, nondose-dependent PK of vismodegib at a steady state. The mechanism responsible for nonlinearity was consistent with oral absorption being influenced by nonsink permeation conditions. We introduce a new parameter, the permeation gradient factor, to characterize the effect of nonsink conditions on permeation. Using vismodegib as an example, we demonstrate the value of using oral PBPK models in drug discovery to predict the oral PK of compounds with nonlinear absorption characteristics in human. SIGNIFICANCE STATEMENT: A physiologically based pharmacokinetic (PBPK) model was built to demonstrate the value of these models early in the drug discovery stage for the prediction of human pharmacokinetics for compounds with unusual oral pharmacokinetics. In this study, our PBPK model could successfully capture the unique steady-state oral pharmacokinetics of our model compound, vismodegib. The mechanism for nonlinearity can be attributed to nonsink permeation conditions in vivo. We introduce the permeation gradient factor as a parameter to assess this effect.

QuantiFERON-cytomegalovirus to predict clinically significant cytomegalovirus infection after allogeneic hematopoietic stem cell transplantation.

BACKGROUND: Controlling cytomegalovirus (CMV) infection through prophylaxis or pre-emptive therapy remains an important contributor to outcomes after allogeneic hematopoetic stem cell transplant (alloHCT). Predicting clinically significant CMV infection (csCMVi) after day 100 remains a challenge. METHODS: We examined the abilty of the QuantiFERON-CMV assay (QFN-CMV) at day 100 (d100) and day 150 (d150) after alloHCT to predict csCMVi after these time points, with median follow-up of 3.1 years (range 1.3-4.3 years). RESULTS: In 46 transplants (donor seropositive (D+) recipient seronegative (R-) = 12, D+R+ = 25, D-R+ = 9; matched related = 13, unrelated donor = 32, haploidentical = 1), for the prediction of freedom from csCMVi >d100, QFN-CMVd100 (positive compared to negative/indeterminate) had sensitivity 62% (23/37), specificity 100% (9/9), positive predictive value 100% (23/23), and negative predictive value 39% (9/23). For the prediction of freedom from csCMVi >d150, QFN-CMVd150 (positive compared to negative/indeterminate) had sensitivity 62% (18/29), specificity 83% (5/6), positive predictive value 95% (18/19), and negative predictive value 31% (5/16). CONCLUSION: Positive QFN-CMV at d100 and d150 strongly predicted freedom from csCMVi after these time points. QFN-CMV could be utilized to predict the need for pre-emptive therapy and CMV viral load monitoring after day 100 post-alloHCT.

Topical α-Gal Nanoparticles Enhance Wound Healing in Radiated Skin.

PURPOSE: Surgery within radiated tissue is associated with increased complication rates. It is hypothesized that impaired wound healing may result from aberrant inflammatory responses that occur in previously radiated tissues. Previous work has demonstrated that the topical application of naturally occurring antigen α-gal (Galα1-3Galß1-(3)4GlcNAc-R) nanoparticles (AGNs) within wounds accelerates macrophage recruitment and subsequent healing in both normal and diabetic wounds. Herein, we hypothesize that application of this antigen would similarly enhance wound healing in irradiated tissues. METHODS: To simulate human physiology, α-1,3-galactosyltransferase knockout (KO) mice were exposed to the antigen to produce anti-α-gal antibodies (anti-Gal). Ten days prior to wounding, the dorsal skin was irradiated with 1 session of 40 Gy. Bilateral dorsal 6-mm splinted full-thickness wounds were created within the radiated skin and treated with 50 µL of AGNs (50 mg/mL) immediately after wounding and again on postoperative day 1. A control KO group underwent similar irradiation and wounding protocols but was treated with phosphate-buffered saline (PBS) vehicle. Wild-type (WT) mice, which do not produce anti-Gal, went through the same irradiation and wounding. RESULTS: Histologic analysis demonstrated enhanced epithelial migration in the radiated/AGN-treated KO wounds, which was significantly elevated in comparison to radiated/PBS-treated KO wounds beginning by day 15 and continuing until the end of the study (p < 0.01). In WT mice, treatment with AGNs showed no effect on epithelial migration. CONCLUSIONS: Topical application of AGNs onto irradiated wounds significantly ameliorates the delayed wound healing classically seen in radiated skin and results in faster wound closure with only transient application.