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Abstract Background Late-onset Pompe disease (LOPD) is an autosomal recessive lysosomal storage disorder that results in severe progressive proximal muscle weakness. Over time, reductions in muscle strength result in respiratory failure and a loss of ambulation. Delayed diagnosis of LOPD deprives patients of treatments that can enhance quality of life and potentially slow disease progression. The objective of this study is to determine if patients with a nonspecific diagnosis, such as muscle weakness, may be at risk for LOPD using retrospective chart review of patients seen at two centers: an academic center and a community health system. Results Initial data pulls identified 80,070 patients with one of the ICD-10 codes of interest. Chart review found 551 of these patients also had at least one lab value commonly abnormal in individuals with LOPD and of these 110 scored as “at-risk”. After removing phenocopies/other confirmed unrelated diagnoses, 46 individuals were contacted either directly or through their healthcare provider for genetic counseling. Three patients had pretest genetic counseling and were tested for decreased levels of acid-α-glucosidase. One patient was found to have deficient acid-α-glucosidase. Additionally, a physician educated through the program ordered LOPD testing for their patient and diagnosed them with LOPD. Conclusion This study confirms that a symptom-based scoring tool and chart review combined with provider education can identify patients who are at increased likelihood to have a missed LOPD diagnosis.

Abstract Introduction Levels of plant-based aeroallergens are rising as growing seasons lengthen and intensify with anthropogenic climate change. Increased exposure to pollens could increase risk for mortality from respiratory causes, particularly among older adults. We determined short-term, lag associations of four species classes of pollen (ragweed, deciduous trees, grass pollen and evergreen trees) with respiratory mortality (all cause, chronic and infectious related) in Michigan, USA. Methods We obtained records for all Michigan deaths from 2006-2017 from the Michigan Department of Health and Human Services (MDHHS). Deaths from infectious and chronic respiratory-related causes were selected using International Classification of Diseases (ICD-10) codes. Pollen data were obtained from a prognostic model of daily pollen concentrations at 25 km resolution. Case-crossover models with distributed lag non-linear crossbases for pollen were used to estimate associations between lags of daily pollen concentrations with mortality and to explore effect modification by sex and racial groups. Results 127,163 deaths were included in the study. Cumulative daily high concentrations (90th percentile) of deciduous broadleaf, grass and ragweed were associated with all-cause respiratory mortality at early lags with e.g., a 1.81 times higher risk of all respiratory deaths at cumulative 7 day lag exposure to deciduous broadleaf pollen at the 90th percentile (95% confidence interval: 1.04, 3.15). Exposure to high concentrations of grass and ragweed pollens was associated with increased risk for death from chronic respiratory causes. No association was found for any pollen species with death from infectious respiratory causes though there was a positive but non-significant association of exposure to deciduous broadleaf and ragweed pollens. We found no evidence to suggest effect modification by race or sex. Conclusions Modelled exposures to high concentrations of pollen taxa were associated with increased all-cause and chronic respiratory mortality among older adults. Results suggest that pollen exposure may become more important to respiratory mortality as the temperatures increase and pollen seasons lengthen.

Abstract Background Anorexia nervosa (AN) is a polygenic, severe metabopsychiatric disorder with poorly understood aetiology. Eight significant loci have been identified by genome-wide association studies (GWAS) and single nucleotide polymorphism (SNP)-based heritability was estimated to be ~ 11–17, yet causal variants remain elusive. It is therefore important to define the full spectrum of genetic variants in the wider regions surrounding these significantly associated loci. The hypothesis we evaluate here is that unrecognised or relatively unexplored variants in these regions exist and are promising targets for future functional analyses. To test this hypothesis, we implemented a novel approach with targeted nanopore sequencing (Oxford Nanopore Technologies) for 200 kb regions centred on each of the eight AN-associated loci in 10 AN case samples. Our bioinformatics pipeline entailed base-calling and alignment with Dorado and minimap2 software, followed by variant calling with four separate tools, Sniffles2, Clair3, Straglr, and NanoVar. We then leveraged publicly available databases to characterise these loci in putative functional context and prioritise a subset of potentially relevant variants. Results Targeted nanopore sequencing effectively enriched the target regions (average coverage 14.64x). To test our hypothesis, we curated a list of 20 prioritised variants in non-coding regions, poorly represented in the current human reference genome but that may have functional consequences in AN pathology. Notably, we identified a polymorphic SINE-VNTR-Alu like sub-family D element (SVA-D), intergenic with IP6K2 and PRKAR2A, and a poly-T short tandem repeat (STR) in the 3ʹUTR of FOXP1. Conclusions Our results highlight the potential of targeted nanopore sequencing for characterising poorly resolved or complex variation, which may be initially obscured in risk-associated regions detected by GWAS. Some of the variants identified in this way, such as the polymorphic SVA-D and poly-T STR, could contribute to mechanisms of phenotypic risk, through regulation of several neighbouring genes implicated in AN biology, and affect post-transcriptional processing of FOXP1, respectively. This exploratory investigation was not powered to detect functional effects, however, the variants we observed using this method are poorly represented in the current human reference genome and accompanying databases, and further examination of these may provide new opportunities for improved understanding of genetic risk mechanisms of AN.

Abstract Recent trials have shown the efficacy of trastuzumab deruxtecan (T-DXd) in HER2-negative patients, but there is not yet a way to identify which patients will best respond, especially with the inability of current HER2 IHC and FISH assays to accurately determine HER2 expression in the unamplified setting. Here, we present a heavily pre-treated patient with triple-negative breast cancer (HER2 IHC 0 who had a complete response to T-DXd. In this case, we used a CLIA-certified reverse-phase protein array-based proteomic assay (RPPA) to determine that the patient had moderate HER2 protein expression (HER2Total 2+, 42%) and activation (HER2Y1248 1+, 23%). Using these results, we determined that the patient may benefit from T-Dxd despite being traditionally qualified as HER2 IHC 0. These findings highlight the potential for proteomics-based assays that may more accurately quantitate HER2 and (its activation) in the HER2 unamplified/IHC 0 setting to better select patients whose tumors are classically molecularly defined as HER2 IHC 0, but still could respond to HER2-directed therapy, and give patients access to therapies which for which they otherwise would not be eligible.

Abstract Inflammation and a metabolic shift from oxidative metabolism to glycolysis are common in the ischemic heart, the latter partly controlled by pyruvate kinase (muscle, PKM). We previously identified alternative splicing promoting the PKM2 isoform after myocardial infarction (MI). We examined the role of PKM2 physiological upregulation after MI, modeled by ligation of the left anterior descending coronary artery, using global PKM2 knockout (PKM2−/−) mice. Echocardiography showed similar cardiac function between PKM2−/− and control mice after MI. However, PKM2−/− infarcted hearts had increased abundances of transcripts associated with oxidative stress and immune responses. Immunohistochemistry revealed greater abundance of macrophages in PKM2−/− hearts prior to MI, with a small increase in CD86+ macrophages in PKM2−/− infarcted hearts. Elevated baseline plasma IL‐6, IL‐1β, and C‐reactive protein, and cardiac IL‐6, 3 days post‐MI, were observed in PKM2−/− mice. Oxidative lipid products were also elevated in baseline PKM2−/− hearts, while antioxidant glutathione peroxidase 4 was reduced. Greater fibrosis was seen in PKM2−/− hearts 28 days after MI. These findings suggest Pkm2 ablation primes the heart for increased oxidative stress, inflammation, and fibrosis post‐MI. The natural upregulation of PKM2 may mitigate fibrosis by reducing oxidative stress and inflammation, highlighting its protective role in the infarcted heart.

Triglyceride (TG)/HDL-C ratio (THR) is a surrogate predictor of hyperinsulinemia. To identify novel genetic loci for THR change over time (ΔTHR), we conducted genome-wide association study (GWAS) and genome-wide linkage scan (GWLS) among nondiabetic Europeans from the Long Life Family Study (n = 1,384). Subjects with diabetes or on dyslipidemia medications were excluded. ΔTHR was derived using growth curve modeling and adjusted for age, sex, field centers, and principal components. GWAS used a linear mixed model accounting for familial relatedness. GWLS employed haplotype-based identity-by-descent estimation with 0.5 cM average spacing. Heritability of ΔTHR was moderate (46%). Our GWAS identified a significant locus at the LPL (P = 1.58e-9) for ΔTHR; this locus has been reported before influencing baseline THR levels. Our GWLS found significant linkage with a logarithm of the odds exceeding 3 on 3q28 (logarithm of the odds = 4.1). Using a subset of 25 linkage-enriched families, we assessed sequence elements under 3q28 and identified two novel variants (EIF4A2 [eukaryotic translation initiation factor 4A2]/ADIPOQ-rs114108468, p = 5e-6, minor allele frequency = 1.8%; TPRG1-rs16864075, p = 3e-6, minor allele frequency = 8%; accounted for ∼28% and ∼29% of the linkage, respectively). While the former variant was associated with EIF4A2 (p = 7e-5)/ADIPOQ (P = 3.49e-2) transcriptional levels, the latter variant was not associated with TPRG1 (P = 0.23) transcriptional levels. Replication in the Framingham Heart Study Offspring Cohort observed modest effect of these loci on ΔTHR. Our approach discovered two novel gene variants EIF4A2/ADIPOQ-rs114108468 and TPRG1-rs16864075 on 3q28 for ΔTHR among subjects without diabetes. Our findings provided novel insights into the molecular regulation of insulin resistance.

Category: Bunion; Midfoot/Forefoot Introduction/Purpose: Following Minimally Invasive Bunionectomy (MISB), removal of hardware rates are about 5% and commonly due to midfoot pain near the distal metatarsal screw site. This distal screw head site corresponds to the proximal midshaft width measurement location, which has shown to increase postoperatively. One explanation for this increased width is medial subluxation of the first metatarsal at the tarsal-metatarsal (TMT) joint. This “TMT slippage” occurs as the metatarsal base is shifted medially to accommodate the metatarsal head lateralization at the metatarsal phalangeal joint. The force across the TMT joint overcomes the ligamentous stabilizers, allowing the metatarsal to “slip.” The aim of this study was to investigate the relation between TMT slippage and removal of hardware (ROH) rates. Methods: A retrospective analysis of prospectively collected data identified 38 patients undergoing MISB for hallux valgus (HV) with subsequent distal chevron screw removal and a control group of 40 MISB cases without hardware removal. Demographic data and surgical details were obtained via chart review. Pre- and at least 6-month post-operative radiographs were measured to obtain transverse TMT slippage on anteroposterior (AP) radiographs (Figures 1). Additionally, distal foot width, midshaft foot width, hallux valgus angle (HVA) and intermetatarsal angle (IMA) were measured. Independent samples t-tests and chi-square analyses were used to compare continuous and categorical characteristics, respectively, between patients who underwent removal of hardware. Multivariable logistic regression modeling was used to identify risk factors for undergoing hardware removal. Results: Interobserver reliability, measured by intraclass correlation coefficients (ICCs), was high for all measurements (ICC > 0.70). The change in TMT slip was significantly higher in the hardware removal (ROH) group compared to the control group (+1.39 versus +0.80, p=0.004) (Table 1). A greater decrease in distal foot width (narrower distal width postoperatively) was associated with an increased likelihood of hardware removal (OR: 0.76), while smaller reductions in HVA and IMA were linked to a higher likelihood of requiring ROH (OR: 1.12 and 1.42, respectively). Notably, a larger increase in TMT slip correlated with the highest likelihood of being in the ROH cohort (OR: 2.87) (Table 2). TMT slippage did not correlate with any other measurements (all p > 0.05). Conclusion: Our study highlights the relationship between MISB outcomes and TMT slippage. Patients requiring distal chevron screw removal showed significantly higher transverse TMT slippage. Interestingly, a smaller distal foot width reduction was associated with decreased hardware removal likelihood, suggesting a potential protective mechanism. Conversely, smaller corrections of both HVA and IMA were associated with higher chances of hardware removal. Notably, for every 1-centimeter increase in TMT slippage postoperatively, there was a nearly 3-fold increase in the likelihood of distal chevron screw removal. Further research is needed to understand the mechanisms underlying TMT slippage and its impact on HV correction outcomes.

Category: Bunion; Midfoot/Forefoot Introduction/Purpose: Hallux Valgus (HV) is a common foot condition often requiring surgical intervention. Minimally invasive bunionectomy (MISB) has gained popularity for HV correction. While previous research has focused on distal foot width changes following surgery, this study introduces a novel midshaft measurement to assess proximal transverse midfoot width. The study aims to provide a comprehensive evaluation of foot width changes post-MISB. Methods: A retrospective review of prospectively collected data was conducted for 44 patients who underwent MISB for HV via an institutional outcome registry. Demographic data, surgical details, radiographs, and foot width measurements were collected. Changes in hallux valgus angle (HVA) and intermetatarsal angle (IMA) were also measured. The novel midshaft bony and soft tissue foot width measurements were included. Two independent observers conducted all preoperative and postoperative measurements. Results: Intraclass correlation coefficients (ICCs) demonstrated high interobserver reliability (all ICCs >0.70) for all measurements. MISB resulted in a consistent reduction in distal foot width, both bony (8.17%) and soft tissue (5.24%)(p 0.375, p

Category: Other; Basic Sciences/Biologics Introduction/ Purpose: Structural allografts have long been used in foot and ankle reconstructive surgery. Pre-contoured allograft wedges, manufactured in a variety of shapes and sizes for specific uses, have gained popularity in recent years due to their ease of use and ability to decrease operative time. A high rate of resorption of such pre-contoured allograft wedges was noted anecdotally by the senior authors, particularly in 2022. There is no known literature describing this phenomenon and thus, the purpose of this study was to identify the resorption and complication rates of pre-contoured allograft wedges over time. Methods: A retrospective review of patients who had a pre-contoured allograft wedge (Paragon28, Englewood, CO) implanted during foot and ankle surgery between 2017 and 2022 at a single institution by one of 10 surgeons was performed. Chart review was used to obtain demographic and implant information and confirm the surgical procedure(s) performed. Radiographic review was completed in order to identify patients with graft resorption and/or nonunion, confirm the site of allograft implantation, and determine the method of fixation used at the allograft site. A linear regression model was utilized to determine if there was a statistically significant association between the year and the number of cases of resorption. Results: In the 320 patients that met inclusion criteria, there were 334 wedges implanted. There were 24 grafts that resorbed and an additional 10 wedges that developed a non-union without resorption. Resorption was noted as early as 5.7 weeks post-operatively and as late as 36.6 weeks post-operatively (average 12.9 weeks). Of the allografts that resorbed, the majority were Lapidus wedges (38%) and Cotton wedges (56%) (Table 1). Resorption rates were highest in the last 2 years studied (Figure 1). Of the 78 wedges used in 2022, 11 wedges resorbed and there were 3 non-unions without resorption, for an overall complication rate of 18%. There was a statistically significant association between the year and the number of patients that had graft resorption (p = 0.03). Conclusion: While complications of allograft wedges in foot and ankle surgery have previously been described, including nonunion, resorption of pre-contoured wedges has not. Over a 5-year period at a single institution, there was a wedge resorption rate of 7% and an overall radiographic complication rate of 10%. There was an increase in allograft resorption cases as well as overall resorption rate and complication rate in 2022 compared to previous years. The majority of cases in which there was a complication occurred with the use of a Cotton or Lapidus wedge; however, no concrete identifiable patient or surgical risk factors were identified.

Category: Bunion Introduction/ Purpose: Hallux valgus deformity affects more than 35% of people over 65 years old. Surgical correction in this population can be more complicated due to the likelihood of poor bone quality, worse deformity, and post-operative recovery challenges associated with weight bearing restrictions. There is no consensus on which surgical option is the best in this age-group. Minimally invasive bunionectomy (MIS) has gained popularity, namely due to evidence showing reduced recovery time, earlier weight-bearing, and less post-operative pain which can eliminate the need for opioids, all of which are beneficial to older surgical patients. The purpose of this study was to compare the radiographic and clinical outcomes of patients age 65 and older who underwent either open Lapidus or minimally invasive chevron akin osteotomy for bunion correction. Methods: A retrospective review of prospectively collected data within an institutional registry was performed. We identified 62 patients age 65 or older who were treated surgically for hallux valgus (open Lapidus or minimally invasive chevron akin osteotomy) between January 2017 and August 2022 with at least 1-year post-operative PROMIS scores. Chart review was performed to obtain demographic information and to confirm the surgical procedures completed. Pre- and at least 6-month post-operative radiographs were measured to obtain the hallux valgus angle (HVA) and intermetatarsal angle (IMA). PROMIS scores were obtained pre-operatively and at one and/or two years post-operatively. Differences in demographic, clinical, and radiographic outcomes between cohorts were assessed using the Mann Whitney U test and p-values were adjusted for a false discovery rate of 5%. Results: Both the MIS and open cohorts showed significant improvement in radiographic measurements between pre- and post-operative radiographs (p < 0.001), though there was no significant difference between groups (p > 0.05). There was a clinically and statistically significant improvement within the PROMIS domain of Pain Interference for the open and MIS groups and Physical Function for the MIS group at 1 and 2-years post-operatively, however, there was only a significant difference at 2-years for the PROMIS Physical Function domain in the open group. No difference was noted in Physical Function at 1-year post-operatively for the open group or between groups (Table 1). Within the MIS and open groups, there was an overall complication rate of 26% in each group (p > 0.05). Conclusion: Patients in both the MIS and open surgical groups had improvement in radiographic measurements and PROMIS scores, though there was no clinical or statistical difference found between groups. Additionally, patients in the open group may take longer to improve as there was not a significant improvement in the PROMIS Physical Function domain until the 2-year post-operative time point. Further investigation of recovery metrics between techniques in the earlier post-operative time-point is warranted. Given the results, both MIS and open surgical techniques appear to be safe and effective in correcting hallux valgus in older patients.

Category: Other; Hindfoot Introduction/Purpose: The current classification system of progressive collapsing foot deformity (PCFD) is comprised of 5 classes that can occur simultaneously. Classes A-E indicate various deformities: hindfoot valgus, midfoot/forefoot abduction, forefoot varus/medial column instability, and ankle joint instability/valgus tilting in the ankle mortise, respectively. While the relationship between each of the PCFD deformities in the classification system to one another has been studied, to our knowledge, the correlation between the various classes and patient-reported outcomes has not been assessed. The purpose of this study was to evaluate whether the PCFD classification system is predictive of patient-reported outcomes and overall prognosis following surgery. The hypothesis was that patients with more severe deformity would have worse patient-reported outcomes. Methods: A retrospective review of prospectively collected data within an institutional registry was performed. We identified 74 patients aged 18 or older who were treated surgically for PCFD between October 2018 and October 2021 with preoperative, 1-year, and 2-year postoperative PROMIS scores. Chart review was performed to obtain demographic information and to confirm the surgical procedures completed. Using radiographs, weight-bearing computed tomography (WBCT), and office notes, patients were classified using the PCFD classification system. Patients were divided into two groups depending on whether subclasses D and/or E were present, indicating more severe deformity. Linear mixed models were used to assess longitudinal change in PROMIS scores between groups over time. Results: Patient demographic and classification characteristics are presented in Table 1. 47 patients were allocated to the Class D/E group and 27 patients were allocated to the No Class D/E group. Patients in the Class D/E group had worse pre-operative Global Physical Health (GPH) (p=0.018) and depression scores (p=0.012); however, post-operatively, there was only a difference in the PROMIS domain of Physical Function (PF) at both 1-Year (p=0.019) and 2-Year (p=0.038) time points (Table 2). There were no statistical differences between the two groups for any other domain at any timepoint (Table 2). Conclusion: Patients with more severe PCFD had clinically significant improvement in physical function at both 1- and 2-years post-operatively, but had worse outcomes compared to those with less severe deformity. The findings regarding preoperative GPH and depression suggest that while Class D/E patients did not perform as well functionally, they may be satisfied with their overall quality of life due to the similar scores postoperatively when compared to the patients with less severe deformity. Future studies are needed to better determine if the novel PCFD classification system is predictive of overall prognosis following surgery based on the various deformities present.

This article is meant to serve as a general guide to perform quantitative fecal egg counts on grazing small ruminants, specifically sheep and goats. This tool is utilized to estimate the extent of pasture parasite burden as well as individual animal parasite burden, and to determine the efficacy of dewormer or anthelmintic treatment. Written by Bridget Stice, Laura H. Bennett, Christa L. Kirby, Allison L. Williams, and Brittany N. Diehl, and published by the UF College of Veterinary Medicine, UF/IFAS Extension, December 2024.

Category: Ankle Arthritis; Hindfoot Introduction/Purpose: Hindfoot (subtalar and/or talonavicular) fusion is a commonly used procedure to treat arthritis and/or deformity. As a result, some patients with ankle arthritis who undergo total ankle replacement (TAR) have a history of prior hindfoot fusion or undergo concomitant hindfoot fusion at the time of TAR. Previous studies have demonstrated hindfoot fusion as a risk factor for failure of TAR and associated with worse clinical outcomes. This study seeks to determine if these findings are reproducible by evaluating clinical and patient-reported outcomes (PROs) of primary TAR with ipsilateral hindfoot fusion at a minimum 2-years follow-up from primary TAR. We hypothesized that TAR patients with ipsilateral hindfoot fusion would have higher rates of revision and worse PROs compared to controls. Methods: A retrospective review of prospectively collected data within an institutional registry was performed. Inclusion criteria for this study were patients undergoing primary TAR (Cadence, Inbone, Invision, Infinity, Salto, Vantage, or Zimmer) between October 2015 and January 2022, with or without prior/concomitant ipsilateral hindfoot fusion, with preoperative and 2-year postoperative PROMIS scores. 400 TARs met inclusion criteria for analysis. Demographic, clinical, patient-reported outcomes, and radiographs were collected. Radiographic complications (including cysts, lucency, subsidence) were recorded. Multivariate linear regression models were used to evaluate the associations of hindfoot fusion, implant type, reoperations (defined as any surgical procedure following primary TAA, including retention of metal implants with polyethylene explant), revisions (defined as exchange of tibial and/or talar components), and demographics with 2-year postoperative PROMIS scores. Finally, proportions of patients meeting PROMIS thresholds for the patient-acceptable symptom state (PASS) at 2-years postoperative was compared between groups via Pearson Chi square tests. Results: Of 400 ankles included, 49 (12.3%) had ipsilateral hindfoot fusion. In multivariate analysis, hindfoot fusion patients had significantly worse 2-year postoperative PROMIS scores in all domains (except depression) versus patients without hindfoot fusion. Preoperative PROMIS domains were predictive of 2-year postoperative PROMIS scores for each respective domain (except pain intensity) (Figure). Hindfoot fusion patients had a significantly higher rate of revision (10.2%) than those without hindfoot fusion (2.6%) (P=0.038). However, reoperations were similar between groups (9.7% versus 12.2%, P=0.460). Additionally, the incidence of radiographic complications was not significantly different between groups (P=0.075). Hindfoot fusion patients were equally as likely to meet PASS thresholds at 2-years postoperatively in all domains except global mental health (P=0.015) and depression (P=0.024). Conclusion: This study demonstrates that TAR patients with ipsilateral hindfoot fusion experience worse pain, function, and physical and mental health postoperatively compared to their counterparts without ipsilateral hindfoot fusion. Although hindfoot fusion may be critical to achieving a stable, neutrally aligned ankle and hindfoot, hindfoot fusion may increase the risk for worse patient-reported TAR outcomes. As indications for TAR continue to expand, it is imperative to understand the role of hindfoot fusion in the survivorship and function of TAR to guide clinical decision making and patient education for establishment of postoperative expectations.

ABSTRACTWith recent advances in U.S. clinical trials for norovirus vaccines, it is an opportune time to examine what is known about the public receptivity to this novel vaccine. From October 2016–September 2017, we surveyed Kaiser Permanente Northwest members in Portland, Oregon, to ask their level of agreement on a 5-point scale with statements about the need for and willingness to get a potential norovirus vaccine for themselves or their child and analyzed their responses according to age, occupational status, prior vaccine uptake, and history of prior norovirus diagnoses. The survey response rate was 13.5% (n = 3,894); 807 (21%) responded as legal guardians, on behalf of a child

ABSTRACT Human noroviruses (HuNoVs) are a significant cause of epidemic and sporadic acute gastroenteritis worldwide. The lack of a reproducible culture system hindered the study of HuNoV replication and pathogenesis for almost a half-century. This barrier was overcome with our successful cultivation of multiple HuNoV strains in human intestinal enteroids (HIEs), which has significantly advanced HuNoV research. We optimized culture media conditions and generated genetically modified HIE cultures to enhance HuNoV replication in HIEs. Building upon these achievements, we now present new insights into this culture system, which involve testing different media, unique HIE lines, and additional virus strains. HuNoV infectivity was evaluated and compared in new HIE models, including HIEs generated from different intestinal segments of individual adult organ donors, HIEs from human intestinal organoids produced from directed differentiation of human embryonic stem cells that were then transplanted and matured in mice before making enteroids (H9tHIEs), genetically engineered (J4FUT2 knock-in [KI], J2STAT1 knockout [KO]) HIEs, as well as HIEs derived from a patient with common variable immunodeficiency (CVID) and from infants. Our findings reveal that small intestinal HIEs, but not colonoids, from adults, H9tHIEs, HIEs from a CVID patient, and HIEs from infants support HuNoV replication with segment and strain-specific differences in viral infection. J4FUT2-KI HIEs exhibit the highest susceptibility to HuNoV infection, allowing the cultivation of a broader range of genogroup I and II HuNoV strains than previously reported. Overall, these results contribute to a deeper understanding of HuNoVs and highlight the transformative potential of HIE cultures in HuNoV research.IMPORTANCEHuman noroviruses (HuNoVs) cause global diarrheal illness and chronic infections in immunocompromised patients. This paper reports approaches for cultivating HuNoVs in secretor positive human intestinal enteroids (HIEs). HuNoV infectivity was compared in new HIE models, including ones from (i) different intestinal segments of single donors, (ii) human embryonic stem cell-derived organoids transplanted into mice, (iii) genetically modified lines, and (iv) a patient with common variable immunodeficiency disease. HIEs from small intestine, but not colon, support HuNoV replication with donor, segment, and strain-specific variations. Unexpectedly, HIEs from one donor are resistant to GII.3 infection. The genetically modified J4FUT2 knock-in (KI) HIEs enable cultivation of a broad range of GI and GII genotypes. New insights into strain-specific differences in HuNoV replication in HIEs support this platform for advancing understanding of HuNoV biology and developing potential therapeutics.

Melanoma arising in association with a blue nevus (BN) is rare but has molecular similarities to uveal melanoma (UM), including GNAQ/11 mutations. Tebentafusp was recently approved for UM based on improved overall survival in a phase 3 study. We hypothesized that tebentafusp may be active in BN-associated melanoma. Here, we present a case of metastatic BN-associated melanoma with rapid response and ~1 year of disease control on tebentafusp. We also explore molecular and histological features of secondary resistance. Our case highlights that PD-1-resistant melanomas should be screened for GNAQ/11 mutations, as tebentafusp may be a treatment option in this extremely rare disease.

Abstract Determining movement parameters for pest insects such as tephritid fruit flies is critical to developing models which can be used to increase the effectiveness of surveillance and control strategies. In this study, harmonic radar was used to track wild-caught male Queensland fruit flies (Qflies), Bactrocera tryoni, in papaya fields. Experiment 1 continuously tracked single flies which were prodded to induce movement. Qfly movements from this experiment showed greater mean squared displacement than predicted by both a simple random walk (RW) or a correlated random walk (CRW) model, suggesting that movement parameters derived from the entire data set do not adequately describe the movement of individual Qfly at all spatial scales or for all behavioral states. This conclusion is supported by both fractal and hidden Markov model (HMM) analysis. Lower fractal dimensions (straighter movement paths) were observed at larger spatial scales (> 2.5 m) suggesting that Qflies have qualitatively distinct movement at different scales. Further, a two-state HMM fit the observed movement data better than the CRW or RW models. Experiment 2 identified individual landing locations, twice a day, for groups of released Qflies, demonstrating that flies could be tracked over longer periods of time.

When women receive a diagnosis of a gynecologic malignancy, they can have questions about their diagnosis or treatment that can result in voice queries to virtual assistants for more information. Recent advancement in artificial intelligence (AI) has transformed the landscape of medical information accessibility. The Google virtual assistant (VA) outperformed Siri, Alexa and Cortana in voice queries presented prior to the explosive implementation of AI in early 2023. The efforts presented here focus on determining if advances in AI in the last 12 months have improved the accuracy of Google VA responses related to gynecologic oncology. Previous questions were utilized to form a common basis for queries prior to 2023 and responses in 2024. Correct answers were obtained from the UpToDate medical resource. Responses related to gynecologic oncology were obtained using Google VA, as well as the generative AI chatbots Google Bard/Gemini and Microsoft Bing-Copilot. The AI narrative responses varied in length and positioning of answers within the response. Google Bard/Gemini achieved an 87.5% accuracy rate, while Microsoft Bing-Copilot reached 83.3%. In contrast, the Google VA’s accuracy in audible responses improved from 18% prior to 2023 to 63% in 2024. While the accuracy of the Google VA has improved in the last year, it underperformed Google Bard/Gemini and Microsoft Bing-Copilot so there is considerable room for further improved accuracy.

Objective: To evaluate the heterogeneity in treatment effect in posttraumatic stress disorder (PTSD) trials. Patients and Methods: We downloaded data from a publicly available repository that captured PTSD trials published from January 1988 through February 2023. We applied restricted maximum-likelihood random-effect meta-analyses and meta-regression to explore potential moderators of treatment effect including methodologic study features (risk of bias domains and control group response rate), characteristics of the population, and intervention features following the theme, intensity, and platform framework. Results: We included 199 PTSD trials that reported the outcomes of diagnosis resolution (122 trials, 8437 patients) and clinically meaningful improvement (133 trials, 9895 patients). Multiple treatments demonstrated effectiveness but with significant heterogeneity. Statistically significant moderators included risk of bias domains of randomization sequence and outcome measurement, control group response rate reflecting severity of PTSD in the enrolled population, and whether the psychotherapeutic approach was trauma focused (P values

Abstract Background Hypoxia is a detrimental factor in solid tumors, leading to aggressiveness and therapy resistance. OMX, a tunable oxygen carrier from the heme nitric oxide/oxygen-binding (H-NOX) protein family, has the potential to reduce tumor hypoxia. [18F]Fluoromisonidazole ([18F]FMISO) positron emission tomography (PET) is the most widely used and investigated method for non-invasive imaging of tumor hypoxia. In this study, we used [18F]FMISO PET/CT (computed tomography) to assess the effect of OMX on tumor hypoxia in spontaneous canine tumors. Results Thirteen canine patients with various tumors (n = 14) were randomly divided into blocks of two, with the treatment groups alternating between receiving intratumoral (IT) OMX injection (OMX IT group) and intravenous (IV) OMX injection (OMX IV group). Tumors were regarded as hypoxic if maximum tumor-to-muscle ratio (TMRmax) was greater than 1.4. In addition, hypoxic volume (HV) was defined as the region with tumor-to-muscle ratio greater than 1.4 on [18F]FMISO PET images. Hypoxia was detected in 6/7 tumors in the OMX IT group and 5/7 tumors in the OMX IV injection group. Although there was no significant difference in baseline hypoxia between the OMX IT and IV groups, the two groups showed different responses to OMX. In the OMX IV group, hypoxic tumors (n = 5) exhibited significant reductions in tumor hypoxia, as indicated by decreased TMRmax and HV in [18F]FMISO PET imaging after treatment. In contrast, hypoxic tumors in the OMX IT group (n = 6) displayed a significant increase in [18F]FMISO uptake and variable changes in TMRmax and HV. Conclusions [18F]FMISO PET/CT imaging presents a promising non-invasive procedure for monitoring tumor hypoxia and assessing the efficacy of hypoxia-modulating therapies in canine patients. OMX has shown promising outcomes in reducing tumor hypoxia, especially when administered intravenously, as evident from reductions in both TMRmax and HV in [18F]FMISO PET imaging.

Adult-onset diabetes increases one’s risk of neurodegenerative disease including Alzheimer’s disease (AD); however, the risk associated with youth-onset diabetes (Y-DM) remains underexplored. We quantified plasma biomarkers of neurodegeneration and AD in participants with Y-DM from the SEARCH cohort at adolescence and young adulthood (Type 1, n = 25; Type 2, n = 25; 59% female; adolescence, age = 15 y/o [2.6]; adulthood, age = 27.4 y/o [2.2]), comparing them with controls (adolescence, n = 25, age = 14.8 y/o [2.7]; adulthood, n = 21, age = 24.9 y/o [2.8]). Plasma biomarkers, including glial fibrillary acidic protein (GFAP), neurofilament light chain protein (NfL), phosphorylated tau-181 (pTau181), and amyloid beta (Aβ40, Aβ42), were measured via Simoa. A subset of participants (n = 7; age = 27.5 y/o [5.7]) and six controls (age = 25.1 y/o [4.5]) underwent PET scans to quantify brain amyloid and tau densities in AD sensitive brain regions. Y-DM adolescents exhibited lower plasma levels of Aβ40, Aβ42, and GFAP, and higher pTau181 compared to controls (p < 0.05), a pattern persisting into adulthood (p < 0.001). All biomarkers showed significant increases from adolescence to adulthood in Y-DM (p < 0.01), though no significant differences in brain amyloid or tau were noted between Y-DM and controls in adulthood. Preliminary evidence suggests that preclinical AD neuropathology is present in young people with Y-DM, indicating a potential increased risk of neurodegenerative diseases.

Background and purpose: Cardiac implanted electronic devices (CIED) require dose monitoring during each fraction of radiotherapy, which can be time consuming and may have delayed read-out times. This study explores the potential of Cherenkov imaging combined with scintillation dosimetry as an alternative verification system. Methods and materials: Time-gated, complementary metal–oxide–semiconductor (iCMOS) cameras were used to collect video images of anthropomorphic phantoms and patients undergoing radiation treatment near chest wall cardiac devices. Scintillator discs and optically stimulated luminescence dosimeters (OSLDs) were used for dose measurement. Accuracy of spatial delivery was assessed by overlaying predicted surface dose outlines derived from the treatment planning system (TPS) with the Cherenkov images. Dose measurements from OSLDs and scintillators were compared. Results: In phantom studies, Cherenkov images visibly indicated when dose was delivered to the CIED as compared to non-overlapping dose deliveries. Comparison with dose overlays revealed congruence at the planned position and non-congruence when the phantom was shifted from the initial position. Absolute doses derived from scintillator discs aligned well with the OSLD measurements and TPS predictions for three different positions, measuring within 10 % for in-field positions and within 5 % for out-of-field positions. For two patients with CIEDs imaged over 18 fractions, Cherenkov imaging confirmed positional accuracy for all fractions, and dose measured by scintillator discs deviated by

Climate change is a complex, global issue that is impacting human health in various ways, with healthcare being a significant contributor to carbon emissions in the United States. This review discusses the environmental impact of important aspects of gynecologic oncology care, including surgery, anesthesia care, radiology, chemotherapy, and radiation oncology. Operating room energy and material use is highlighted, with a focus on the environmental impact of robotic surgery. The contribution of certain anesthetic gases in increasing greenhouse gas emissions is addressed. Additionally, the environmental impacts of radiologic imaging, chemotherapy, and radiation oncology are also discussed. Despite the complexity of climate change, there are multiple strategies on the individual and institutional level that can help mitigate the environmental impact of gynecologic oncology care. Individual efforts include practicing red bag stewardship, limiting single use-supplies, decreasing the use of potentially deleterious anesthetics, and supporting research into alternative dosing for chemotherapy and radiation which requires less patient travel. Institutional strategies include investing in efficient HVAC systems, utilizing reusable and reprocessed materials and devices, and purchasing renewable energy sources. Both individuals and institutions can advocate with industry and government at all levels for practices and policies that support lower carbon emissions. By recognizing our role in reducing carbon emissions, we can work towards improving the well-being of our patients and the larger community.

Dark chocolate produced on equipment used to manufacture milk chocolate can contain milk due to cross-contact. This study evaluated the use of dry cleaning methods for removing milk chocolate residue from a butterfly or ball valve attached to a stainless steel pipe and from pilot-scale equipment used in chocolate manufacture. Milk-free dark chocolate (40 °C) was pumped through a milk chocolate-contaminated valve/pipe assembly after no cleaning, use of a pig purging treatment, or a 40 °C cocoa butter flush. Dark chocolate samples were collected at 7-sec intervals. Treatments investigated for the removal of residual milk chocolate from a conche and a ball mill included no cleaning, a 40 °C cocoa butter rinse, and wet cleaning. After cleaning, three batches of dark chocolate (40 °C) were processed in the ball mill and conche, and each batch was collected. Milk chocolate was processed on a 3-roll refiner, followed by push-through with dark chocolate (∼9 kg) with 0.3 kg samples collected at 5-min intervals. Dark chocolate samples were analyzed for milk concentrations by ELISA. Trials and analyses were completed in triplicate. Dark chocolate push-through alone resulted in milk concentrations ≥4,500 µg/g in samples obtained from the contaminated valve/pipe combinations within the first few seconds of collection, and ≥16.2 kg of dark chocolate was needed to obtain milk concentrations below the ELISA LOQ (2.5 µg/g). A pig purging treatment of the ball valve/pipe assembly resulted in milk concentrations below the ELISA LOQ. A cocoa butter flush of the butterfly valve/pipe decreased initial milk concentrations, but milk was detected until ≥18.7 kg dark chocolate purge. Milk concentrations in first batches of dark chocolate processed in a ball mill and conche without cleaning were ≥17,000 µg/g while the use of a cocoa butter rinse reduced milk levels in dark chocolate by ≥89%. Some dry cleaning treatments were effective at reducing levels of milk in dark chocolate due to cross-contact.

BACKGROUND While issues related to lesbian, gay, bisexual, and transgender (LGBT) health are increasingly incorporated into medical training, there remains limited guidance and opportunities for continuing medical education in LGBT health. It is unclear how participation in LGBT-specific training is distributed across physician specialties and practice regions. Additionally, national data assessing cultural competency training for physicians are scarce and do not delineate LGBT-specific training or training completed prior to, during, or after graduate medical education. METHODS Using data from the 2016 National Culturally and Linguistically Appropriate Services Physician Survey, this study evaluated patterns of post-residency cultural competency training, as well as associations between LGBT-specific training and provider perceptions of patient care outcomes. RESULTS Provider specialty, practice region, and receiving cultural competency training as a trainee were associated with post-residency LGBT-specific training. Surgical providers (odds ratio [OR]: 0.42; confidence interval [CI] 0.25–0.73; p = .002) and those practicing in the South (OR: 0.49; CI: 0.26–0.92; p = .025) had lower odds of completing LGBT-specific cultural competency training while in independent practice. Post-residency LGBT-specific training was associated with provider agreement that cultural competency training improves the quality of care (OR: 2.76; CI: 1.44–5.28; p = .002), patient satisfaction (OR: 2.55; CI: 1.32–4.93; p = .005), and patient comprehension (OR: 2.03; CI: 1.05–3.90; p = .034). CONCLUSIONS Our findings provide disaggregated analyses that nuance the assessment of cultural competency interventions and support a broader effort to increase attention to LGBT health in continuing medical education.

One of the world’s largest “blue carbon” ecosystems, Louisiana’s tidal wetlands on the US Gulf of Mexico coast, is rapidly being lost. Louisiana’s strong legal, regulatory, and monitoring framework, developed for one of the world’s largest tidal wetland systems, provides an opportunity for a programmatic approach to blue carbon accreditation to support restoration of these ecologically and economically important tidal wetlands. Louisiana’s coastal wetlands span ∼1.4 million ha and accumulate 5.5–7.3 Tg yr−1 of blue carbon (organic carbon), ∼6%–8% of tidal marsh blue carbon accumulation globally. Louisiana has a favorable governance framework to advance blue carbon accreditation, due to centralized restoration planning, long term coastal monitoring, and strong legal and regulatory frameworks around carbon. Additional restoration efforts, planned through Louisiana’s Coastal Master Plan, over 50 years are projected to create, or avoid loss of, up to 81,000 ha of wetland. Current restoration funding, primarily from Deepwater Horizon oil spill settlements, will be fully committed by the early 2030s and additional funding sources are required. Existing accreditation methodologies have not been successfully applied to coastal Louisiana’s ecosystem restoration approaches or herbaceous tidal wetland types. Achieving financial viability for accreditation of these restoration and wetland types will require expanded application of existing blue carbon crediting methodologies. It will also require expanded approaches for predicting the future landscape without restoration, such as numerical modeling, to be validated. Additional methodologies (and/or standards) would have many common elements with those currently available but may be beneficial, depending on the goals and needs of both the state of Louisiana and potential purchasers of Louisiana tidal wetland carbon credits. This study identified twenty targeted needs that will address data and knowledge gaps to maximize financial viability of blue carbon accreditation for Louisiana’s tidal wetlands. Knowledge needs were identified in five categories: legislative and policy, accreditation methodologies and standards, soil carbon flux, methane flux, and lateral carbon flux. Due to the large spatial scale and diversity of tidal wetlands, it is expected that progress in coastal Louisiana has high potential to be generalized to similar wetland ecosystems across the northern Gulf of Mexico and globally.

Abstract Cardiac metabolism ensures a continuous ATP supply, primarily using fatty acids in a healthy state and favoring glucose in pathological conditions. Pyruvate kinase muscle (PKM) controls the final step of glycolysis, with PKM1 being the main isoform in the heart. PKM2, elevated in various heart diseases, has been suggested to play a protective role in cardiac stress, but its function in basal cardiac metabolism remains unclear. We examined hearts from global PKM2 knockout (PKM2−/−) mice and found reduced intracellular glucose. Isotopic tracing of U‐13C glucose revealed a shift to biosynthetic pathways in PKM2−/− cardiomyocytes. Total ATP content was two‐thirds lower in PKM2−/− hearts, and functional analysis indicated reduced mitochondrial oxygen consumption. Total reactive oxygen species (ROS) and mitochondrial superoxide were also increased in PKM2−/− cardiomyocytes. Intriguingly, PKM2−/− hearts had preserved ejection fraction compared to controls. Mechanistically, increased calcium/calmodulin‐dependent kinase II activity and phospholamban phosphorylation may contribute to higher sarcoendoplasmic reticulum calcium ATPase 2 pump activity in PKM2−/− hearts. Loss of PKM2 led to altered glucose metabolism, diminished mitochondrial function, and increased ROS in cardiomyocytes. These data suggest that cardiac PKM2 acts as an important rheostat to maintain ATP levels while limiting oxidative stress. Although loss of PKM2 did not impair baseline contractility, its absence may make hearts more sensitive to environmental stress or injury.

Background In this open-label, randomized, non-comparative, multicenter phase II study (Alliance A091401) we report on three expansion cohorts treated with nivolumab (N) with and without ipilimumab (N+I) and provide a multi-omic correlative analysis of actionable biomarkers.Methods Patients were randomized (non-comparative) to receive either N or N+I. The primary endpoint was a 6-month confirmed response rate (CRR) defined by Response Evaluation Criteria in Solid Tumors version 1.1. Secondary endpoints included treatment-related adverse events (TRAEs), progression-free survival, and overall survival. Multi-omic correlative analyses were conducted using samples from both the primary and expansion cohorts.Results A total of 66 patients were evaluated for the primary endpoint with disease including gastrointestinal stromal tumor (GIST, n=18), undifferentiated pleomorphic sarcoma (UPS, n=24), and dedifferentiated liposarcoma (DDLPS, n=24). Neither N nor N+I achieved a complete or partial response in the GIST expansion cohort. In DDLPS and UPS, the primary response endpoint of CRR was met with N+I (both 16.6%, 2/12) but not with N alone (both 8.3%, 1/12). In the GIST cohort, TRAE was higher with N+I treatment, halting enrollment as required per protocol. In a correlative analysis of patients for the expansion cohort and the original cohort (n=86), traditional biomarkers of immunotherapy response were not correlated with response in any histological subtype. Markers of genomic instability including the presence of gene fusions and increased subclonal mutations correlated with improved clinical outcomes.Conclusions This expansion cohort reaffirms the outcomes of A091401. There remains a pressing need to determine the role of and predictive biomarkers for immunotherapy in sarcoma.Trial registration number NCT02500797.

Not available.

ABSTRACT Human intestinal enteroids (HIEs) are gaining recognition as physiologically relevant models of the intestinal epithelium. While HIEs from adults are used extensively in biomedical research, few studies have used HIEs from infants. Considering the dramatic developmental changes that occur during infancy, it is important to establish models that represent infant intestinal characteristics and physiological responses. We established jejunal HIEs from infant surgical samples and performed comparisons to jejunal HIEs from adults using RNA sequencing (RNA-Seq) and morphologic analyses. We then validated differences in key pathways through functional studies and determined whether these cultures recapitulate known features of the infant intestinal epithelium. RNA-Seq analysis showed significant differences in the transcriptome of infant and adult HIEs, including differences in genes and pathways associated with cell differentiation and proliferation, tissue development, lipid metabolism, innate immunity, and biological adhesion. Validating these results, we observed a higher abundance of cells expressing specific enterocyte, goblet cell, and enteroendocrine cell markers in differentiated infant HIE monolayers, and greater numbers of proliferative cells in undifferentiated 3D cultures. Compared to adult HIEs, infant HIEs portray characteristics of an immature gastrointestinal epithelium including significantly shorter cell height, lower epithelial barrier integrity, and lower innate immune responses to infection with an oral poliovirus vaccine. HIEs established from infant intestinal tissues reflect characteristics of the infant gut and are distinct from adult cultures. Our data support the use of infant HIEs as an ex vivo model to advance studies of infant-specific diseases and drug discovery for this population.IMPORTANCETissue or biopsy stem cell-derived human intestinal enteroids are increasingly recognized as physiologically relevant models of the human gastrointestinal epithelium. While enteroids from adults and fetal tissues have been extensively used for studying many infectious and non-infectious diseases, there are few reports on enteroids from infants. We show that infant enteroids exhibit both transcriptomic and morphological differences compared to adult cultures. They also differ in functional responses to barrier disruption and innate immune responses to infection, suggesting that infant and adult enteroids are distinct model systems. Considering the dramatic changes in body composition and physiology that begin during infancy, tools that appropriately reflect intestinal development and diseases are critical. Infant enteroids exhibit key features of the infant gastrointestinal epithelium. This study is significant in establishing infant enteroids as age-appropriate models for infant intestinal physiology, infant-specific diseases, and responses to pathogens.

Abstract FAM111A, a serine protease, plays roles in DNA replication and antiviral defense. Missense mutations in the catalytic domain cause hyper-autocleavage and are associated with genetic disorders with developmental defects. Despite the enzyme’s biological significance, the molecular architecture of the FAM111A serine protease domain (SPD) is unknown. Here, we show that FAM111A is a dimerization-dependent protease containing a narrow, recessed active site that cleaves substrates with a chymotrypsin-like specificity. X-ray crystal structures and mutagenesis studies reveal that FAM111A dimerizes via the N-terminal helix within the SPD. This dimerization induces an activation cascade from the dimerization sensor loop to the oxyanion hole through disorder-to-order transitions. Dimerization is essential for proteolytic activity in vitro and for facilitating DNA replication at DNA-protein crosslink obstacles in cells, while it is dispensable for autocleavage. These findings underscore the role of dimerization in FAM111A’s function and highlight the distinction in its dimerization dependency between substrate cleavage and autocleavage.

Abstract Reversion mutations that restore wild-type function of the BRCA gene have been described as a key mechanism of resistance to Poly(ADP-ribose) polymerase (PARP) inhibitor therapy in BRCA-associated cancers. Here, we report a case of a patient with metastatic castration-resistant prostate cancer (mCRPC) with a germline BRCA2 mutation who developed acquired resistance to PARP inhibition. Extensive genomic interrogation of cell-free DNA (cfDNA) and tissue at baseline, post-progression, and postmortem revealed ten unique BRCA2 reversion mutations across ten sites. While several of the reversion mutations were private to a specific site, nine out of ten tumors contained at least one mutation, suggesting a powerful clonal selection for reversion mutations in the presence of therapeutic pressure by PARP inhibition. Variable cfDNA shed was seen across tumor sites, emphasizing a potential shortcoming of cfDNA monitoring for PARPi resistance. This report provides a genomic portrait of the temporal and spatial heterogeneity of prostate cancer under the selective pressure of a PARP inhibition and exposes limitations in the current strategies for detection of reversion mutations.

Reducing dairy fat intake is a common dietary guideline to limit energy and saturated fatty acid intake for the promotion of cardiometabolic health. However, research utilizing a holistic, food-based approach to assess the consumption of the fat found in dairy, a broad and diverse food group, may provide new insight into these guidelines. Dairy fat is comprised of a diverse assembly of fatty acids, triacylglycerols, sterols, and phospholipids, all uniquely packaged in a milk fat globule. The physical structure of this milk fat globule and its membrane is modified through different processing methods, resulting in distinctive dairy-fat matrices across each dairy product. The objectives of this narrative review were to first define and compare the dairy-fat matrix in terms of its unique composition, physical structure, and fat content across common dairy products (cow’s milk, yogurt, cheese, and butter). With this information, we examined observational studies and randomized controlled trials published within the last 10 years (2013–2023) to assess the individual effects of the dairy-fat matrix in milk, yogurt, cheese, and butter on cardiometabolic health and evaluate the implications for nutrition guidance. Searches conducted on Ovid MEDLINE and PubMed® utilizing search terms for cardiometabolic health, both broadly and regarding specific disease outcomes and risk factors, yielded 59 studies that were analyzed and included in this review. Importantly, this review stratifies by both dairy product and fat content. Though the results were heterogeneous, most studies reported no association between intake of these individual regular-fat dairy products and cardiometabolic outcome measures, thus, the current body of evidence suggests that regular-fat dairy product consumption may be incorporated within overall healthy eating patterns. Research suggests that there may be a beneficial effect of regular-fat milk and yogurt intake on outcome measures related to body weight and composition, and an effect of regular-fat cheese intake on outcome measures related to blood lipids, but more research is necessary to define the directionality of this relationship. Lastly, we identify methodological research gaps and propose future research directions to bolster the current evidence base available for ascertaining the role of dairy fat in a healthy diet.

Background: Hallux valgus deformity affects more than 35% of people aged ≥65 years. Surgical correction in this population can be more complicated because of poor bone quality, worse deformity, and postoperative recovery challenges. The purpose of this study was to compare the radiographic and clinical outcomes of patients aged ≥65 years who underwent either open Lapidus or minimally invasive chevron Akin osteotomy for bunion correction. Methods: A retrospective review identified 62 patients aged ≥65 years who were treated surgically for hallux valgus with at least 1-year postoperative Patient-Reported Outcomes Measurement Information System (PROMIS) scores (physical function and pain interference). Preoperative and at least 6-month postoperative radiographs were measured for the hallux valgus angle and intermetatarsal angle. PROMIS scores were obtained preoperatively and at 1 and/or 2 years postoperatively. Differences in demographic, clinical, and radiographic outcomes were assessed using the Mann Whitney U test and P values were adjusted for a false discovery rate of 5%. Results: There was no difference between the MIS and open cohorts in pre- or postoperative radiographic measurements or clinical outcomes at any time point. At 1 year postoperatively, both groups had statistically significant improvements in the PROMIS pain interference domain but only the MIS group had a statistically significant improvement in the PROMIS physical function domain. Clinical significance was equivocal. At 2 years postoperatively, there were clinically and statistically significant improvements in the PROMIS pain interference and physical function domains for the open and MIS groups. Conclusion: Patients in both surgical groups had improvement in radiographic measurements and 2-year PROMIS scores, although there was no clinical or statistical difference found between groups. MIS and open surgical techniques appear to be safe and effective in correcting hallux valgus in older patients; however, patients may need to be counseled that maximum improvement after surgery may take more than 1 year. Level of Evidence: Level III, retrospective cohort study.

The prevalence of obesity has increased dramatically worldwide and has become a critical public health priority. Obesity is associated with many co-morbid conditions, including hypertension, diabetes, and cardiovascular disease. Although the physiology of obesity is complex, a healthy diet and sufficient exercise are two elements known to be critical to combating this condition. Years of research on the Mediterranean diet, which is high in fresh fruits and vegetables, nuts, fish, and olive oil, have demonstrated a reduction in numerous non-communicable chronic diseases associated with this diet. There is strong evidence to support an anti-inflammatory effect of the diet, and inflammation is a key driver of obesity. Changes in diet alter the gut microbiota which are intricately intertwined with human physiology, as gut microbiota-derived metabolites play a key role in biological pathways throughout the body. This review will summarize recent published studies that examine the potential role of gut metabolites, including short-chain fatty acids, bile acids, trimethylamine-N-oxide, and lipopolysaccharide, in modulating inflammation after consumption of a Mediterranean-like diet. These metabolites modulate pathways of inflammation through the NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome, toll-like receptor 4 signaling, and macrophage driven effects in adipocytes, among other mechanisms.

Antimicrobial peptides (AMPs) show promise in enhancing resistance against pathogens. Previously, we integrated two AMP genes, cathelicidin (Cath) from alligator (Alligator mississippiensis or Alligator sinensis) and cecropin (Cec) from moth (Hyalophora cecropia), into the channel catfish (Ictalurus punctatus) genome. This study examines the efficacy of exogenous AMP gene integration in improving bacterial resistance in transgenic channel catfish and assesses the direct and pleiotropic effects of gene replacement/knockout on survival and growth based on insertion site. Transgenic Cath- and Cec-expressing fish exhibited similar or higher survival rates (P ​> ​0.05) compared to controls during the initial culture. Integration of the Cec transgene doubled the survival rate when challenged with Edwardsiella ictaluri, with knock-in (KI) of Cath further increasing bacterial resistance. Coupling Cec KI with mstn knockout (KO) increased survival 3-fold after E. ictaluri infection and growth by 50% at 4 months post-fertilization (mpf). However, random integration of Cec had a minimal effect on disease resistance and did not enhance growth. Random integration of Cath increased survival 2.5-fold and 4-fold against E. ictaluri and Flavobacterium covae, respectively, without affecting growth. Cath KI at the lh locus increased survival 4-fold when challenged with F. covae and reduced growth by 10% (P ​> ​0.05) at 24 mpf, whereas Cath KI coupled with mc4r KO resulted in a 2.5-fold increase in survival following F. covae infection compared with controls, and increased growth by 80% at 3 mpf. Simultaneous KI of Cath and Cec, along with KO of mc4r and mstn, increased survival 4-fold against E. ictaluri, while increasing growth by 50% at 3 mpf. Dual insertion of AMP genes yielded the greatest resistance to disease. These direct and pleiotropic effects may increase comprehension and societal acceptance of genetic engineering in aquaculture.

IntroductionLanguage skills, such as the ability to understand words (receptive language), develop during infancy and are built through interactions with the environment, including eating. Exposure to complementary foods also begins in infancy and may play a significant role in language development, especially in understanding of food-related words. However, the relationship between the complementary foods to which a child is exposed and early language acquisition has not been previously studied. We hypothesized that young children’s food-related receptive language (FRL) would reflect the complementary foods to which they were frequently offered by caregivers.MethodsCaregivers of young children (4-26 months; n = 408) in the Approaching Eating through Language (APPEAL) Study in the US were surveyed via Qualtrics. FRL was assessed by caregiver-report via a modified MacArthur-Bates Communicative Development Inventory. Complementary foods offered (CFO) by caregivers were assessed using a modified Food Frequency Questionnaire. Latent Class Analysis (LCA) was implemented to identify, 1) groupings of foods frequently offered (>1x/week) and 2) groupings of food-related words understood by the young children.ResultsA 5-class best fit LCA model was identified for CFO (-log likelihood [-llik]=-8727) and for FRL (-llik=-5476). Cross-classification of the CFO and FRL derived classes revealed that children with higher exposure to complementary foods were perceived by caregivers to be most likely to also understand a greater number of food-related words (Probability=0.48). As expected, children having been offered a greater number of complementary foods and who understood a greater number of food-related words were older, compared to those with less complementary food exposure and food-related language acquisition (p

Abstract Repeat vaccination with egg-based influenza vaccines could preferentially boost antibodies targeting the egg-adapted epitopes and reduce immunogenicity to circulating viruses. In this randomized trial (Clinicaltrials.gov: NCT03722589), sera pre- and post-vaccination with quadrivalent inactivated egg-based (IIV4), cell culture-based (ccIIV4), and recombinant (RIV4) influenza vaccines were collected from healthcare personnel (18-64 years) in 2018−19 (N = 723) and 2019−20 (N = 684) influenza seasons. We performed an exploratory analysis. Vaccine egg-adapted changes had the most impact on A(H3N2) immunogenicity. In year 1, RIV4 induced higher neutralizing and total HA head binding antibodies to cell- A(H3N2) virus than ccIIV4 and IIV4. In year 2, among the 7 repeat vaccination arms (IIV4-IIV4, IIV4-ccIIV4, IIV4-RIV4, RIV4-ccIIV4, RIV4-RIV4, ccIIV4-ccIIV4 and ccIIV4-RIV4), repeat vaccination with either RIV4 or ccIIV4 further improved antibody responses to circulating viruses with decreased neutralizing antibody egg/cell ratio. RIV4 also had higher post-vaccination A(H1N1)pdm09 and A(H3N2) HA stalk antibodies in year 1, but there was no significant difference in HA stalk antibody fold rise among vaccine groups in either year 1 or year 2. Multiple seasons of non-egg-based vaccination may be needed to redirect antibody responses from immune memory to egg-adapted epitopes and re-focus the immune responses towards epitopes on the circulating viruses to improve vaccine effectiveness.

Age-related macular degeneration (AMD) causes blindness due to loss of retinal pigment epithelium (RPE) and photoreceptors (PRs), which comprise the two outermost layers of the retina. Given the small size of the macula and the importance of direct contact between RPE and PRs, the use of scaffolds for targeted reconstruction of the outer retina in later stage AMD and other macular dystrophies is particularly attractive. We developed microfabricated, honeycomb-patterned, biodegradable poly(glycerol sebacate) (PGS) scaffolds to deliver organized, adjacent layers of RPE and PRs to the subretinal space. Furthermore, an optimized process was developed to photocure PGS, shortening scaffold production time from days to minutes. The resulting scaffolds robustly supported the seeding of human pluripotent stem cell-derived RPE and PRs, either separately or as a dual cell-layered construct. These advanced, economical, and versatile scaffolds can accelerate retinal cell transplantation efforts and benefit patients with AMD and other retinal degenerative diseases.

Trapped-ion systems are a leading platform for quantum information processing, but they are currently limited to 1D and 2D arrays, which imposes restrictions on both their scalability and their range of applications. Here, we propose a path to overcome this limitation by demonstrating that Penning traps can be used to realize remarkably clean bilayer crystals, wherein hundreds of ions self-organize into two well-defined layers. These bilayer crystals are made possible by the inclusion of an anharmonic trapping potential, which is readily implementable with current technology. We study the normal modes of this system and discover salient differences compared to the modes of single-plane crystals. The bilayer geometry and the unique properties of the normal modes open new opportunities—in particular, in quantum sensing and quantum simulation—that are not straightforward in single-plane crystals. Furthermore, we illustrate that it may be possible to extend the ideas presented here to realize multilayer crystals with more than two layers. Our work increases the dimensionality of trapped-ion systems by efficiently utilizing all three spatial dimensions, and it lays the foundation for a new generation of quantum information processing experiments with multilayer 3D crystals of trapped ions.

Intravaginal rings (IVRs) represent a well-established, woman-controlled and sustained vaginal drug delivery system suitable for a wide range of applications. Here, we sought to investigate the differences in etonogestrel (ENG) and ethinyl estradiol (EE) release from a 3D-printed IVR utilizing continuous liquid interface production (CLIP™) (referred to as CLIPLOW for low drug loading and CLIPHIGH IVRs for high drug loading) and NuvaRing, a commercially available injection molded IVR. We conducted in vitro release studies in simulated vaginal fluid to compare the release of ENG and EE from CLIPLOW IVRs and NuvaRing. CLIPLOW IVRs had a similar hormone dose to NuvaRing and exhibited slightly slower ENG release and greater EE release in vitro compared to NuvaRing. When administered to female sheep, NuvaRing demonstrated greater ENG/EE levels in plasma, vaginal tissue and vaginal fluids compared to CLIPLOW IVR despite similar drug loadings. Leveraging observed hormones levels in sheep from NuvaRing as an effective contraceptive benchmark, we developed a long-acting CLIPHIGH IVR with increased ENG and EE doses that demonstrated systemic and local hormone levels greater than the NuvaRing for 90 days in sheep. No signs of toxicity were noted regarding general health, colposcopy, or histological analysis in sheep after CLIPHIGH IVR administration. Our results provided (1) a comparison of ENG and EE release between a 3D-printed IVR and NuvaRing in vitro and in vivo, (2) a preclinical pharmacokinetic benchmark for vaginally delivered ENG and EE and (3) the generation of a 90-day CLIP IVR that will be utilized in future work to support the development of a long-acting ENG/EE IVR combined with an antiretroviral for the prevention of HIV and unplanned pregnancy.

The use of mathematical models to make predictions about tumor growth and response to treatment has become increasingly prevalent in the clinical setting. The level of complexity within these models ranges broadly, and the calibration of more complex models requires detailed clinical data. This raises questions about the type and quantity of data that should be collected and when, in order to maximize the information gain about the model behavior while still minimizing the total amount of data used and the time until a model can be calibrated accurately. To address these questions, we propose a Bayesian information-theoretic procedure, using an adaptive score function to determine the optimal data collection times and measurement types. The novel score function introduced in this work eliminates the need for a penalization parameter used in a previous study, while yielding model predictions that are superior to those obtained using two potential pre-determined data collection protocols for two different prostate cancer model scenarios: one in which we fit a simple ODE system to synthetic data generated from a cellular automaton model using radiotherapy as the imposed treatment, and a second scenario in which a more complex ODE system is fit to clinical patient data for patients undergoing intermittent androgen suppression therapy. We also conduct a robust analysis of the calibration results, using both error and uncertainty metrics in combination to determine when additional data acquisition may be terminated.

Diagnosis of portosystemic shunts (PSS) in dogs often requires multiple diagnostic tests, and available clinicopathologic tests have limitations in sensitivity and specificity. The objective of this study was to train and validate a machine learning model (MLM) that can accurately predict the presence of a PSS utilizing routinely collected demographic data and clinicopathologic features. Dogs diagnosed with PSS or control dogs tested for PSS but had the condition ruled out (non-PSS) were identified. Dogs were included if a complete blood count and serum chemistry panel were available from PSS diagnostic testing. Dogs with a PSS were subcategorized as having a single intrahepatic PSS, a single extrahepatic PSS, or multiple extrahepatic PSS. An extreme gradient boosting (XGboost) MLM was trained with data from 70% of the cases, and MLM performance was determined on the test set, comprising the remaining 30% of the case data. Two MLMs were created. The first was designed to predict the presence of any PSS (PSS MLM), and the second to predict the PSS subcategory (PSS SubCat MLM). The trained PSS MLM had a sensitivity of 94.3% (95% CI 90.1–96.8%) and specificity of 90.5% (95% CI 85.32–94.0%) for dogs in the test set. The area under the receiver operator characteristic curve (AUC) was 0.976 (95% CI; 0.964–0.989). The mean corpuscular hemoglobin, lymphocyte count, and serum globulin concentration were most important in prediction classification. The PSS SubCat MLM had an accuracy of 85.7% in determining the subtype of PSS of dogs in the test set, with variable sensitivity and specificity depending on PSS subtype. These MLMs have a high accuracy for diagnosing PSS; however, the prediction of PSS subclassification is less accurate. The MLMs can be used as a screening tool to increase or decrease the index of suspicion for PSS before confirmatory diagnostics such as advanced imaging are pursued.

Introduction/Purpose: In an ever-changing healthcare landscape, patient-reported outcomes (PROs) are becoming more important for reimbursement and evaluating the success of surgical procedures. Unfortunately, it is unclear which PROs correlate best with clinical improvement. The patient acceptable symptom state (PASS) asks the patient whether or not their outcome is acceptable at a particular post-operative time point. Recently, the use of this metric has gained traction within the foot and ankle literature. To our knowledge, this is the first study that aims to establish PASS thresholds for Patient-Reported Outcome Measurement Information System (PROMIS) scores in patients who underwent operative intervention for hallux rigidus. Methods: A retrospective review of prospectively collected data within an institutional registry was performed. We identified 174 patients treated for hallux rigidus between February 2019 and March 2021 with at least 2-year post-operative PROMIS scores. Chart review was performed to obtain demographic information and to confirm the surgical procedures that were completed. Two-years post-operatively, patients answered two PASS anchor questions (Satisfaction, Delighted-Terrible scale) with Likert- scale responses, which was collected along with pre-operative and 2-year post-operative PROMIS scores via the registry. After patient’s answers to the Satisfaction and Delighted-Terrible scales were recategorized into binary responses, PASS thresholds were determined using the maximum Youden Index and a 95% confidence interval was quantified using 2000 bootstrapped iterations. Differences in patient and surgical characteristics between patients who met or did not meet the PASS threshold were compared using independent samples t-test and Pearson chi square. Statistical significance was established at an alpha of 0.05. Results: There was excellent association between PASS thresholds and the PROMIS domains of Physical Function (48.6, AUC=0.82) and Pain Interference (52.1, AUC =0.86). Overall, 125/174 (71.8%) and 105/174 (60.3%) patients met the threshold for Physical Function (PF) and Pain Interference (PI), respectively. For the PF and PI domains, age, BMI, method of cheilectomy and use of Akin/Moberg osteotomy did not impact a patient’s likelihood of meeting the PASS threshold. For the PF domain and the PI domain stratified by the Delighted-Terrible scale, women were more likely to meet the PASS threshold than men (63.2% vs. 36.8%, p = 0.057 and 73% vs. 45%, p =0.023, respectively). Lastly, patients with a higher pre-operative PF score had a higher chance of meeting the PASS threshold (p < 0.001). Conclusion: Following operative intervention to treat hallux rigidus, pre-operative PROMIS scores were strongly associated with a patient’s likelihood of meeting the PASS threshold. After surgical intervention for hallux rigidus, less than 75% of the patients felt their outcome was acceptable; however, the PASS threshold for PF was lower than population average, so it may not be necessary for patients to reach normal physical function levels post-operatively in order to have an acceptable outcome.

Introduction/Purpose: Surgical interventions to correct hallux valgus have been shown to improve patient reported outcomes (PROs); however, many of these instruments do not measure a patient’s subjective outcome experience. The patient acceptable symptom state (PASS) is defined as the symptom threshold that a patient must reach to be satisfied with the outcome of their surgery. PASS thresholds have been defined for hallux valgus correction using American Orthopaedic Foot & Ankle Society (AOFAS) scores; however, no studies have used a validated PRO metric. This is the first study that aims to establish PASS thresholds for Patient-Reported Outcome Measurement Information System (PROMIS) scores in patients following operative intervention for hallux valgus. Methods: A retrospective review of prospectively collected data within an institutional registry was performed. We identified 291 patients treated for hallux valgus with or without second hammertoe correction between February 2019 and March 2021 with at least 2-year post-operative PROMIS scores. Chart review was performed to obtain demographic information and to confirm the surgical procedures completed. Two-years post-operatively, patients answered two PASS anchor questions (Satisfaction, Delighted-Terrible scale) with Likert-scale responses, which was collected along with pre-operative and 2-year post-operative PROMIS scores via the registry. After patient’s answers to the Satisfaction and Delighted-Terrible scales were recategorized into binary responses; PASS thresholds were determined using the maximum Youden Index and a 95% confidence interval was quantified using 2000 bootstrapped iterations. Differences in patient and surgical characteristics between patients who met or did not meet the PASS threshold were compared using independent samples t-test and Pearson chi square. Results: There was excellent association between PASS thresholds and the PROMIS domains of Physical Function (50.3, AUC=0.86) and Pain Interference (51.5, AUC=0.86). Overall, 204/291 and 205/291 patients met the threshold for Physical Function (PF) and Pain Interference (PI), respectively. For both PROMIS domains, a lower BMI was associated with a higher likelihood of meeting the PASS threshold (p=0.002 for PF, p=0.032 for PI). For the PF domain, Lapidus patients were more likely to meet the PASS threshold (p=0.05), and patients with first MTP fusion were less likely to meet the PASS threshold (p=0.004). Meeting the PASS threshold wasn’t impacted by the concomitant correction of a second hammertoe. Lastly, patients with a higher pre-operative PF score had a greater chance of meeting the PASS threshold (p < 0.001). Conclusion: This is the first study to define a PASS threshold for hallux valgus correction using PROMIS scores, a validated outcomes measure. Pre-operative PROMIS scores, patient BMI, and the type of procedure performed impacted a patient’s likelihood of meeting the PASS threshold. These results may be helpful in counseling and educating patients on their chances of obtaining a satisfactory result following hallux valgus correction.

Introduction/Purpose: Current literature shows similar clinical outcomes between first metatarsophalangeal joint arthrodesis and polyvinyl alcohol hydrogel (synthetic cartilage implant or SCI) hemiarthroplasty in the treatment of hallux rigidus; however, prior studies have not reported validated patient-reported outcome measures. To our knowledge, this is the first study to compare patient-reported outcome measures using the validated Patient Reported Outcomes Measurement Information System (PROMIS) in patients treated for hallux rigidus with metatarsophalangeal joint arthrodesis and SCI hemiarthroplasty. In addition, this novel study provides comparative data of the complication and revision rates for each procedure. Methods: A retrospective review of prospectively collected data within an institutional registry identified 101 patients who underwent metatarsophalangeal joint arthrodesis and 82 patients who underwent SCI hemiarthroplasty for treatment of hallux rigidus between January 2016 and June 2022. Chart review was performed to obtain demographic information, complications, and revision rates. Preoperative, 1-year, and 2-year PROMIS scores were obtained from the registry and confirmed via chart review. Equivalence testing was performed using two one sided t-tests (TOST) and setting a +/-5 unit margin of difference as the clinically meaningful difference to determine if there was a difference in PROMIS scores between groups. Linear regression models were also utilized to compare adjusted postoperative PROMIS scores between the two cohorts. For the equivalence testing, a p-value > 0.05 indicated a statistically significant result. Results: Demographic information and preoperative hallux rigidus severity between the cohorts showed no statistically significant difference (Table 1). The SCI cohort had no intra-operative or post-operative wound complications, but one patient within the MTP fusion cohort was treated for a superficial wound infection. The SCI group had six patients revised to MTP fusions within 3- years due to continued pain and/or implant subsidence. The MTP fusion group had 2 patients revised due to non-union and 13 patients that had symptomatic hardware requiring removal. Equivalence comparison of PROMIS scores showed the SCI cohort had significantly worse pain intensity scores at 2 years and significantly less improvement in pain intensity scores from pre- operative to 1-year post-op (Table 2). There were no differences found between cohorts for other PROMIS domains. Conclusion: For the treatment of hallux rigidus, the SCI hemiarthroplasty and metatarsophalangeal joint arthrodesis have equivalent outcomes for all PROMIS domains except the pain intensity domain. While patients in both cohorts had improvement in pain from pre-operative to post-operative time points, SCI was not as effective as first MTP fusion at relieving pain intensity at a follow-up of 2 years. While SCI is a motion-sparing procedure, patients with a primary goal of improving pain may be better suited for first MTP joint arthrodesis. Tables demonstrating the comparisons of both the preoperative demographic data as well as demonstrating the PROMIS scores across different time periods (preoperative, 1 year and 2 years).

Introduction/Purpose: Physical activity and returning to sports are vital for athletes after foot and ankle surgery. While joint preservation is preferred, joint arthrodesis becomes necessary in certain injuries or deformities. Common procedures include Lisfranc arthrodesis, modified Lapidus for hallux valgus (HV), and first metatarsophalangeal (MTP) arthrodesis for hallux rigidus (HR), addressing pain and deformities. Prior studies show promising outcomes individually, yet lack comparisons, impeding understanding of postoperative differences in function. Comparative insights can guide patient expectations and shared decisions pre-surgery. Thus, this retrospective study aims to compare clinical outcomes and return to sports in the three procedure groups. Methods: Between 2006 and 2014, a study assessed 48 modified Lapidus procedures for HV (average age 37.3 years, average follow-up 2.8 years, 95.8% female), 38 Lisfranc injuries (average age 31.8 years, average follow-up 5.2 years, 47.4% female), and 50 MTP fusion for HR cases (average age 54.9 years, average follow-up 5.1 years, 66% female). The assessment focused on physical activity and return to sport, utilizing sports-specific, patient-administered questionnaires. Clinical outcomes were evaluated through the Foot and Ankle Outcome Score (FAOS), encompassing pain, symptoms, stiffness, daily function, sports function, and quality of life. Comparison among the surgical groups included return-to-activity difficulty, participation levels, postoperative FAOS scores, and patient satisfaction. ANOVA analysis gauged significance in FAOS subscale differences among procedures, followed by the Tukey HSD test to pinpoint specific procedure variations if significant differences arose. Results: Regarding FAOS scores, the procedures yielded similar outcomes in the pain subscale. No differences between Lapidus- Lisfranc in the symptom subscale (P=0.6). However, MTP fusion displayed poorer scores than Lapidus (P=0.04) and Lisfranc (P=0.005). No differences between Lapidus-Lisfranc (P=0.64) and Lapidus-MTP Fusion in Activities of Daily Living (P=0.18). Yet, MTP fusion exhibited notably worse outcomes than Lisfranc (P=0.03). For Sports, no differences between Lapidus-Lisfranc (P=0.94), while MTP fusion showed inferior outcomes to Lapidus (P=0.002) and Lisfranc (P=0.001). Although means varied, no differences were observed for Lapidus vs. Lisfranc (P=0.95), Lapidus vs. MTP Fusion (P=0.09), or Lisfranc vs. MTP fusion (P=0.06) in Quality of Life [Table 1]. Collectively, patients engaged in 40 activities. Return to sport post-surgery percentage, compared among procedure groups, yielded similar outcomes [Table 2]. Conclusion: Although all procedure groups had similar overall return to sports and physical activities patient satisfaction rates, there were significant differences in regard to FAOS scores. Lapidus and Lisfranc procedures displayed similar postoperative outcomes in all FAOS domains. MTP Fusion showed similar postoperative outcomes to both Lapidus and Lisfranc procedures in the FAOS pain and quality of life domains, however displayed significantly worse outcomes in symptoms, activities of daily living, and sports domains. Therefore, it appears that an MTP fusion results in more functional limitations when compared to Lapidus and Lisfranc procedures. Table 1: Comparison of FAOS Scores in Patients who Underwent Lapidus, Lisfranc, and MTP Fusion Procedures; Table 2: Postoperative Changes in Activity Difficulty and Participation Levels

Summary: Numerous multi-omic investigations of cancer tissue have documented varying and poor pairwise transcript:protein quantitative correlations, and most deconvolution tools aiming to predict cell type proportions (cell admixture) have been developed and credentialed using transcript-level data alone. To estimate cell admixture using protein abundance data, we analyzed proteome and transcriptome data generated from contrived admixtures of tumor, stroma, and immune cell models or those selectively harvested from the tissue microenvironment by laser microdissection from high grade serous ovarian cancer (HGSOC) tumors. Co-quantified transcripts and proteins performed similarly to estimate stroma and immune cell admixture (r ≥ 0.63) in two commonly used deconvolution algorithms, ESTIMATE or ConsensusTME. We further developed and optimized protein-based signatures estimating cell admixture proportions and benchmarked these using bulk tumor proteomic data from over 150 patients with HGSOC. The optimized protein signatures supporting cell type proportion estimates from bulk tissue proteomic data are available at https://lmdomics.org/ProteoMixture/.

Abstract Invariant Natural Killer T (iNKT) cells undergo immune exhaustion during chronic activation caused by cancer and viral infections, such as HIV. Exhaustion is marked by cell dysfunction and increased expression of immune checkpoint proteins programmed cell-death-1 (PD-1) and lymphocyte-activation-gene-3 (LAG-3). We hypothesize that blockade of PD-1 and/or LAG-3 will enhance iNKT cell function. Utilizing peripheral blood mononuclear cells from healthy donors, LAG-3 and PD-1 expression on iNKT cells was assessed using flow cytometry following in vitro stimulation with iNKT-specific stimulant α-galactosylceramide (n = 4). Efficacy of anti-LAG-3 and/or anti-PD-1 antibody blockades in enhancing iNKT cell function was assessed by determining proliferative capacity and IFN-γ production (n = 9). LAG-3 and PD-1 expression on iNKT cells peaked at Day 4 (98.8%; p ≤ 0.0001 and 98.8%; p = 0.005, respectively), followed by steep decrease by Day 10, coinciding with peak iNKT cell proliferation. In a 10-day blocking assay, both the anti-PD-1 alone and dual anti-PD-1 and anti-LAG-3 significantly increased iNKT proliferation (6 and 6.29 log2 fold-change respectively) compared to the no blockade control (ANOVA-p = 0.0005) with the dual blockade system being more effective (t-test-p = 0.013). This provides proof-of-concept for LAG-3 and PD-1 as immunotherapeutic targets to enhance human iNKT cell function, with the long-term goal of addressing immune exhaustion.