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Abstract A significant challenge in measurements of neutrino oscillations is reconstructing the incoming neutrino energies. While modern fully-active tracking calorimeters such as liquid argon time projection chambers in principle allow the measurement of all final state particles above some detection threshold, undetected neutrons remain a considerable source of missing energy with little to no data constraining their production rates and kinematics. We present the first demonstration of tagging neutrino-induced neutrons in liquid argon time projection chambers using secondary protons emitted from neutron-argon interactions in the MicroBooNE detector. We describe the method developed to identify neutrino-induced neutrons and demonstrate its performance using neutrons produced in muon-neutrino charged current interactions. The method is validated using a small subset of MicroBooNE’s total dataset. The selection yields a sample with $$60\%$$ 60 % of selected tracks corresponding to neutron-induced secondary protons. At this purity, the integrated efficiency is 8.4% for neutrons that produce a detectable proton.

IntroductionPrimary central nervous system lymphoma (PCNSL) is a rare type of non-Hodgkin’s lymphoma that affects brain parenchyma, eyes, cerebrospinal fluid, and spinal cord. Diagnosing PCNSL can be challenging because imaging studies often show similar patterns as other brain tumors, and stereotactic brain lesion biopsy conformation is invasive and not always possible. This study aimed to validate a previous proteomic profiling (PMID: 32610669) of cerebrospinal fluid (CSF) and develop a CSF-based proteomic panel for accurate PCNSL diagnosis and differentiation.MethodsCSF samples were collected from patients of 30 PCNSL, 30 other brain tumors, and 31 tumor-free/benign controls. Liquid chromatography tandem-mass spectrometry targeted proteomics analysis was used to establish CSF-based proteomic panels.ResultsFinal proteomic panels were selected and optimized to diagnose PCNSL from tumor-free controls or other brain tumor lesions with an area under the curve (AUC) of 0.873 (95%CI: 0.723-0.948) and 0.937 (95%CI: 0.807- 0.985), respectively. Pathways analysis showed diagnosis panel features were significantly enriched in pathways related to extracellular matrices-receptor interaction, focal adhesion, and PI3K-Akt signaling, while prion disease, mineral absorption and HIF-1 signaling were significantly enriched with differentiation panel features.DiscussionThis study suggests an accurate clinical test panel for PCNSL diagnosis and differentiation with CSF-based proteomic signatures, which may help overcome the challenges of current diagnostic methods and improve patient outcomes.

Effective therapy for liver fibrosis is lacking. Here, we examined whether LP340, the lead candidate of a new-generation of hydrazide-based HDAC1,2,3 inhibitors (HDACi), decreases liver fibrosis. Liver fibrosis was induced by CCl4 treatment and bile duct ligation (BDL) in mice. At 6 weeks after CCl4, serum alanine aminotransferase increased, and necrotic cell death and leukocyte infiltration occurred in the liver. Tumor necrosis factor-α and myeloperoxidase markedly increased, indicating inflammation. After 6 weeks, α-smooth muscle actin (αSMA) and collagen-1 expression increased by 80% and 575%, respectively, indicating hepatic stellate cell (HSC) activation and fibrogenesis. Fibrosis detected by trichrome and Sirius-red staining occurred primarily in pericentral regions with some bridging fibrosis in liver sections. 4-Hydroxynonenal adducts (indicator of oxidative stress), profibrotic cytokine transforming growth factor-β (TGFβ), and TGFβ downstream signaling molecules phospho-Smad2/3 also markedly increased. LP340 attenuated indices of liver injury, inflammation, and fibrosis markedly. Moreover, Ski-related novel protein-N (SnoN), an endogenous inhibitor of TGFβ signaling, decreased, whereas SnoN expression suppressor microRNA-23a (miR23a) increased markedly. LP340 (0.05 mg/kg, ig., daily during the last 2 weeks of CCl4 treatment) decreased 4-hydroxynonenal adducts and miR23a production, blunted SnoN decreases, and inhibited the TGFβ/Smad signaling. By contrast, LP340 had no effect on matrix metalloproteinase-9 expression. LP340 increased histone-3 acetylation but not tubulin acetylation, indicating that LP340 inhibited Class-I but not Class-II HDAC in vivo. After BDL, focal necrosis, inflammation, ductular reactions, and portal and bridging fibrosis occurred at 2 weeks, and αSMA and collagen-1 expression increased by 256% and 560%, respectively. LP340 attenuated liver injury, ductular reactions, inflammation, and liver fibrosis. LP340 also decreased 4-hydroxynonenal adducts and miR23a production, prevented SnoN decreases, and inhibited the TGFβ/Smad signaling after BDL. In vitro, LP340 inhibited immortal human hepatic stellate cells (hTERT-HSC) activation in culture (αSMA and collagen-1 expression) as well as miR23a production, demonstrating its direct inhibitory effects on HSC. In conclusions, LP340 is a promising therapy for both portal and pericentral liver fibrosis, and it works by inhibiting oxidative stress and decreasing miR23a.

Abstract Childhood acute lymphoblastic leukemia (ALL) genomes show that relapses often arise from subclonal outgrowths. However, the impact of clonal evolution on the actionable proteome and response to targeted therapy is not known. Here, we present a comprehensive retrospective analysis of paired ALL diagnosis and relapsed specimen. Targeted next generation sequencing and proteome analysis indicate persistence of actionable genome variants and stable proteomes through disease progression. Paired viably-frozen biopsies show high correlation of drug response to variant-targeted therapies but in vitro selectivity is low. Proteome analysis prioritizes PARP1 as a pan-ALL target candidate needed for survival following cellular stress; diagnostic and relapsed ALL samples demonstrate robust sensitivity to treatment with two PARP1/2 inhibitors. Together, these findings support initiating prospective precision oncology approaches at ALL diagnosis and emphasize the need to incorporate proteome analysis to prospectively determine tumor sensitivities, which are likely to be retained at disease relapse.

Abstract Congenital heart disease (CHD) represents a significant contributor to both morbidity and mortality in neonates and children. There’s currently no analogous dried blood spot (DBS) screening for CHD immediately after birth. This study was set to assess the feasibility of using DBS to identify reliable metabolite biomarkers with clinical relevance, with the aim to screen and classify CHD utilizing the DBS. We assembled a cohort of DBS datasets from the California Department of Public Health (CDPH) Biobank, encompassing both normal controls and three pre-defined CHD categories. A DBS-based quantitative metabolomics method was developed using liquid chromatography with tandem mass spectrometry (LC-MS/MS). We conducted a correlation analysis comparing the absolute quantitated metabolite concentration in DBS against the CDPH NBS records to verify the reliability of metabolic profiling. For hydrophilic and hydrophobic metabolites, we executed significant pathway and metabolite analyses respectively. Logistic and LightGBM models were established to aid in CHD discrimination and classification. Consistent and reliable quantification of metabolites were demonstrated in DBS samples stored for up to 15 years. We discerned dysregulated metabolic pathways in CHD patients, including deviations in lipid and energy metabolism, as well as oxidative stress pathways. Furthermore, we identified three metabolites and twelve metabolites as potential biomarkers for CHD assessment and subtypes classifying. This study is the first to confirm the feasibility of validating metabolite profiling results using long-term stored DBS samples. Our findings highlight the potential clinical applications of our DBS-based methods for CHD screening and subtype classification.

Abstract Motivation Ovarian cancer (OC) is a highly lethal gynecological malignancy. Extensive research has shown that OC cells undergo significant metabolic alterations during tumorigenesis. In this study, we aim to leverage these metabolic changes as potential biomarkers for assessing ovarian cancer. Methods A functional module-based approach was utilized to identify key gene expression pathways that distinguish different stages of ovarian cancer (OC) within a tissue biopsy cohort. This cohort consisted of control samples (n = 79), stage I/II samples (n = 280), and stage III/IV samples (n = 1016). To further explore these altered molecular pathways, minimal spanning tree (MST) analysis was applied, leading to the formulation of metabolic biomarker hypotheses for OC liquid biopsy. To validate, a multiple reaction monitoring (MRM) based quantitative LCMS/MS method was developed. This method allowed for the precise quantification of targeted metabolite biomarkers using an OC blood cohort comprising control samples (n = 464), benign samples (n = 3), and OC samples (n = 13). Results Eleven functional modules were identified as significant differentiators (false discovery rate, FDR

Abstract The ICARUS collaboration employed the 760-ton T600 detector in a successful 3-year physics run at the underground LNGS laboratory, performing a sensitive search for LSND-like anomalous $$\nu _e$$ ν e appearance in the CERN Neutrino to Gran Sasso beam, which contributed to the constraints on the allowed neutrino oscillation parameters to a narrow region around 1 eV $$^2$$ 2 . After a significant overhaul at CERN, the T600 detector has been installed at Fermilab. In 2020 the cryogenic commissioning began with detector cool down, liquid argon filling and recirculation. ICARUS then started its operations collecting the first neutrino events from the booster neutrino beam (BNB) and the Neutrinos at the Main Injector (NuMI) beam off-axis, which were used to test the ICARUS event selection, reconstruction and analysis algorithms. ICARUS successfully completed its commissioning phase in June 2022. The first goal of the ICARUS data taking will be a study to either confirm or refute the claim by Neutrino-4 short-baseline reactor experiment. ICARUS will also perform measurement of neutrino cross sections with the NuMI beam and several Beyond Standard Model searches. After the first year of operations, ICARUS will search for evidence of sterile neutrinos jointly with the Short-Baseline Near Detector, within the Short-Baseline Neutrino program. In this paper, the main activities carried out during the overhauling and installation phases are highlighted. Preliminary technical results from the ICARUS commissioning data with the BNB and NuMI beams are presented both in terms of performance of all ICARUS subsystems and of capability to select and reconstruct neutrino events.

A recently developed Integral Suspension Pressure (ISP) sedimentation-based particle size analysis technique was tested as an alternative to the hydrometer method. The ISP method determines the particle size distribution using an electronic device, namely the PARIO meter. The PARIO meter provides an alternative to collecting manual readings with the hydrometer, by automatically recording differences in suspension pressure during a settling period of up to 24 h at a depth of 18 cm below the surface of the test suspension. The particle size distribution of the samples is then determined by inverse modeling the recorded pressure data. The objective of this study was to validate the accuracy of the ISP method compared to the hydrometer method using a number of soil mixtures manufactured from materials with known particle size composition. Nine soil mixtures with known reference values were analysed by the hydrometer and ISP methods. Statistical comparison of the results revealed that both measurement methods overestimated clay content and underestimated silt content, the hydrometer method providing more accurate results compared to the ISP method. Thirty-eight mixtures of soil were tested using the ISP method to further validate the accuracy. The results indicated that the accuracy of the ISP method depends on the mass of clay-sized particles and the species composition of the clay minerals in suspension. The ISP method obtained more accurate results for soil mixtures containing expandable clay minerals than to those containing non-expandable clays.

Category: Midfoot/Forefoot; Basic Sciences/Biologics Introduction/Purpose: Plantar fibromatosis (PFi) is characterized by collagen-containing nodules on the foot along the plantar fascia that can cause pain and functional disability. Current treatments focus on symptom relief, which may become ineffective, or surgery, which can have lengthy recovery times. Collagenase clostridium histolyticum (CCH) is approved for Dupuytren’s and Peyronie’s disease treatment, which are both fibroproliferative disorders that may coexist in patients with PFi. This dose-ranging study assessed the efficacy and safety of CCH treatment of PFi. Methods: In this Phase 1, multicenter study, participants were randomly allocated (1:1:1) to receive 0.6, 1.2, or 2.25 mg/mL CCH. All participants underwent the initial treatment period (ITP) and received 1–2 intralesional injections per nodule on Day 1 with 5 follow-up assessments. Participants with palpable treated nodules could receive additional CCH doses (at 2.25 mg/mL; retreatment period, RP), with follow-up at specified intervals. Assessments included a 5-scale measure of nodule hardness (0 [nonpalpable] to 4 [hard]), and the Foot Function Index-Short Form23 (FFI-SF23) reported as a composite score (0 – 100) of 3 subscale scores (disability, difficulty, and pain subscales). Results: After ITP (n=24), 100% (8/8) of participants receiving 2.25 mg/mL CCH showed improvement in nodule hardness, compared with 87.5% (7/8) and 75% (6/8) of participants receiving 1.2 and 0.6 mg/mL, respectively. FFI-SF23 also showed a negative change from baseline across treatment groups, indicating improvement. Participants that received 2.25 mg/mL reported the largest percent improvement (-83.7%). After RP (n=11), 81.8% (9/11) of participants showed improvement in nodular hardness and an FFI-SF23 score change of -9.3%, indicating further improvement from baseline after ITP. CCH had a consistent safety profile in ITP/RP. Conclusion: CCH caused dose-dependent improvement of nodule hardness after treatment, with 2.25 mg/mL utilized for nodule retreatment; FFI-SF23 scores correlated with this finding, indicating CCH may be a potential nonsurgical intralesional option for PFi treatment.

Category: Midfoot/Forefoot; Basic Sciences/Biologics Introduction/Purpose: Currently, there is no safe and efficacious injectable therapy for symptomatic plantar fibromatosis (PFi) that addresses its pathophysiology. Patients with PFi nodules experience symptoms that reduce quality of life, causing functional disability leading to severe impairment in some patients. Collagenase clostridium histolyticum (CCH) is approved for Dupuytren’s and Peyronie’s disease treatment, which are both fibroproliferative disorders that may coexist in patients with PFi. This first study of CCH treatment for PFi evaluated participant satisfaction and investigator assessment of improvement. Methods: Participants were randomly allocated (1:1:1) in this Phase 1, multicenter study to receive 0.6, 1.2, or 2.25 mg/mL CCH. All participants underwent the initial treatment period (ITP) and received 1–2 intralesional injections per nodule on Day 1 with follow-up assessments at specified intervals. Participants with palpable treated nodules could enter the retreatment period (RP) and receive additional CCH doses (at 2.25 mg/mL), with follow-up at specified intervals. Assessments included the Subject Satisfaction with Treatment Rating Scale (+2: very satisfied; to -2: very dissatisfied), and the Investigator Assessment of Improvement Scale (+3: very much improvement; to -3: very much worse). Results: After ITP, ≥75% of participants reported “very” or “quite” satisfied across all treatment groups, with 2.25 mg/mL group reporting 100% satisfaction. For ≥72.7% of nodules, participants reported satisfaction with outcome. Investigators reported improvement (“minimal”, “much”, or “very much”) in ≥75% of participants and ≥80% of treated nodules. After RP, 63.6% of participants reported treatment satisfaction; participants were satisfied with 66.7% of treated nodules, indicating continued improvement from the ITP. Almost all participants (90.9%) were deemed by investigators to have shown improvement, with 86.7% of treated nodules being assessed as having improvement. Conclusion: CCH-treated PFi participants were satisfied with their improvement; physicians assessed improvement in >90% of participants. Results indicate CCH as a potential nonsurgical intralesional option for PFi treatment.

Category: Bunion; Midfoot/Forefoot Introduction/Purpose: The minimally-invasive Chevron-Akin (MICA) procedure for hallux valgus correction is an increasingly popular technique that has yielded comparable radiographic and clinical outcomes to open procedures3–5. Intraoperative radiation dosages for certain foot and ankle surgeries have been reported relative to the international occupational radioprotection thresholds6. In particular, the mean intraoperative radiation exposure during the MIS Scarf osteotomy has been reported to be 14-times greater than open osteotomy7. However, no study to date has reported on the intraoperative radiation exposure during MICA procedures in relation to that of open procedures. Methods: This is a retrospective comparative study from a single institution. Patients underwent a MICA (n=25) or open (n=40) hallux valgus correction by a single foot and ankle fellowship trained surgeon. To account for the learning curve8, the number of intraoperative fluoroscopy shots for the first 50 consecutive MICA procedures were analyzed, and the first 25 procedures were excluded. Four data points were retrieved from the mini C-arm, including 1) number of fluoroscopy shots taken; 2) total fluoroscopy time (s); 3) total radiation dose (mGy); and 4) total dose area product (DAP; mGy*cm 2 ). Preoperative and postoperative radiographs were referenced to measure the hallux valgus (HVA) and intermetatarsal angles (IMA) as previously described.9 Categorical variables were compared using the chi-square test, and continuous variables were compared using Student’s t-test or the Mann-Whitney U test. Analysis of variance was used to determine the statistical significance of differences in the fluoroscopy data between groups. Results: The average total dose absorbed (4.00 vs. 1.34 mGy, p< 0.001), total fluoroscopy time (226 vs. 48.5 s, p< 0.001), and number of fluoroscopy shots (125 vs. 22.3, p< 0.001) were 2.9, 4.6, and 5.6 times greater in the MICA versus the open group, respectively. The total DAP (4.22 vs. 2.56 mGy*cm 2 , p=0.175) was 1.6 times greater in the MICA group but the difference was not statistically significant. The mean preoperative and postoperative HVA and IMA were greater in the MICA cohort compared to the open cohort. However, there was no statistical difference in change in HVA (MICA -17.43 vs. Open -20.05, p=0.125) or IMA (MICA -8.08 vs. open -9.31, p=0.11) between the two cohorts, with each resulting in normal postoperative angles. Conclusion: In this single surgeon series, the MICA procedure demonstrated significantly greater dose absorbed, total fluoroscopy time, and number of fluoroscopy shots compared to the open procedure. The extent of radiation exposure to the surgeon and the patient in relation to the radioprotection thresholds must be closely monitored to specify the health risks involved with this procedure.

Category: Midfoot/Forefoot; Basic Sciences/Biologics Introduction/Purpose: Plantar fibromatosis (PFi) is a functionally disabling disorder characterized by fibrous collagen-containing nodules along the plantar fascia. Collagenase clostridium histolyticum (CCH) is a nonsurgical treatment approved for Dupuytren’s and Peyronie’s disease; both can coexist with PFi. Composed of two collagenases, CCH specifically hydrolyzes certain collagen types and may disrupt the nodule when injected directly. This proof-of-concept study evaluated pharmacokinetics, immunogenicity, and assessed tolerability and safety of CCH treatment of PFi. Methods: In this Phase 1, multicenter study, participants were randomly allocated (1:1:1) to receive 0.6, 1.2, or 2.25 mg/mL CCH. All participants underwent the initial treatment period (ITP) and received 1–2 intralesional injections per nodule on Day 1 with pharmacokinetic samples collected over 24hrs, followed by 5 follow-up visits. Post-ITP, participants with a remaining, palpable nodule could enter the retreatment period (RP) and receive additional CCH doses (at 2.25 mg/mL) with follow-up at specified intervals. Immunogenicity test sampling occurred at screening and at the end of ITP/RP. Adverse events (AEs) were monitored throughout. Results: Twenty-four participants enrolled into ITP, with 11 continuing into RP. Pharmacokinetic and immunogenicity findings were consistent with previous CCH studies in other therapeutic areas. CCH treatment was generally well tolerated, with no serious treatment emergent adverse events (TEAEs), drug discontinuations, or study withdrawals by study end. Over 70% of participants during ITP (19/24) and RP (8/11) reported at least 1 TEAE. Eighty-six TEAEs occurred during ITP and 35 in RP; most were mild in severity (73%, ITP; 83%, RP), and 67% of ITP and 49% of RP TEAEs were treatment related. TEAEs were reportedly injection site- related, with injection site bruising, pain, and swelling frequently recorded. Conclusion: CCH had an expected pharmacokinetics/immunogenicity profile and was safe and well tolerated in participants with PFi. CCH may be a potential nonsurgical intralesional option for PFi treatment.

Category: Ankle; Other Introduction/Purpose: Operative management of acute Achilles tendon ruptures can be associated with a myriad of complications1,2. Demographics and comorbidities have been previously explored in relation to outcomes after operative management of Achilles tendon ruptures3,4. However, the relationship between socioeconomic factors and postoperative outcomes in these patients has not been explored. Distressed community index (DCI) is a score that represents a community’s level of well-being across the United States, with higher scores representing greater distress experienced in the community. Independent socioeconomic deprivation factors have been shown to be associated with suboptimal postoperative outcomes for patients undergoing certain orthopedic surgeries6–9. The purpose of this study is to determine if there is any association between DCI scores and short-term complication rates and functional outcomes following ruptured Achilles tendon repair. Methods: Patients who underwent surgical repair of ruptured Achilles tendon between 2015 and 2019 were identified. Patients’ residence zip codes were obtained and matched to corresponding DCI scores (0-100) based on data provided by the Economic Innovation Group for the same time period (2015-2019). Scores were divided into quintiles. Patient age, race, ethnicity, alcohol and smoking history, need for assistance with activities of daily living, ambulation status unrelated to injury, and access to transportation were also noted. Complications such as re-rupture, superficial or deep infection, wound complications, and deep vein thrombosis within 1-year since date of surgery were noted. Preoperative Foot and Ankle Ability Measure and Visual Analogy Pain scales were compared to postoperative scores at 6 months, 1 year, and 2 years. Analysis of variance (ANOVA) was used to determine the statistical significance of differences in the rate of complications and FAAM-VAS scores between quintiles. Results: Patients in the 2nd DCI quintile experienced the highest rate (9.25%) of total complications while patients in the 5th quintile experienced the lowest rate (3.33%). Difference in complication rates across DCI quintiles was not statistically significant (p=0.925). Out of the 17 patients who experienced complications, 12 (70.6%) patients belonged to the 1st and 2nd quintile while 2 patients (11.7%) belonged to the 4th and 5th quintiles. The differences in FAAM and VAS pain scores were not significantly different at preoperative and at 1-year and 2-year postoperative time points across quintiles. However, patients in the 5th and 4th quintiles experienced the greatest improvement in FAAM Overall scores at 1-year and 2-year postoperative time points, respectively. Conclusion: Patients living in more distressed communities did not experience greater postoperative complication rates or report lower patient-reported outcomes up to 2 years following operative repair of Achilles tendon rupture.

Category: Ankle Arthritis; Ankle Introduction/Purpose: Socioeconomic deprivation factors such as income levels, race and ethnicity, low education level, and insurance status have been shown to be associated with suboptimal postoperative outcomes for patients undergoing orthopedic surgeries, including certain foot and ankle procedures1–3. Total ankle arthroplasty (TAA) is a surgical treatment for end-stage ankle joint disease that has seen a significantly increased utilization over the past two decades4. Distressed community index (DCI) is a score that represents a community’s level of well-being across the United States5, with higher scores representing greater distress experienced in the community. The purpose of this study was to determine if there is any association between DCI scores and complications rates along with functional outcomes following primary TAA. Methods: 566 patients who underwent primary TAA between 2015 and 2019 at our institution were identified. Patients’ residence zip codes at the time of surgery were obtained and the corresponding DCI scores (0-100) were determined using DCI data provided by the Economic Innovation Group for the same time period (2015-2019). Scores were divided into quintiles. Patient age, sex, body mass index, race, ethnicity, rheumatic disease status, Osteoporosis status, Charlson comorbidity index, alcohol use, smoking status, length of hospital stay, discharge to rehabilitation facility, depression history, anxiety history, and marital status were obtained. Postoperative FAAM-VAS scores at 6 months, 1-year, and 2-years were compared to their preoperative scores. Revision, infection, steroid injection, loose body or hardware removal, irrigation and debridement, periprosthetic fracture, and tibial/talar exostectomy were noted. Analysis of variance (ANOVA) was used to determine the statistical significance of differences in the rate of subsequent complications and interventions related to index TAA. Results: 88/566 (15.5%) of all patients required at least one operative intervention related to the index TAA. Patients in the 4th DCI quintile experienced the highest rate of subsequent procedures at 18.3% while those in the 5th DCI quintile experienced the lowest rate at 10.6%. The differences in rate of subsequent operative intervention across DCI quintiles was not statistically significant (p=0.775). However, the rates of revision and infection were the highest for patients in the 4th and 5th DCI quintiles, respectively. The differences in improvement of FAAM Overall and VAS Pain scores at 6 months, 1-year, and 2-years from index surgery were not statistically significant. Those in the 5th quintile experienced the least improvement in FAAM Overall at the 1- year and 2-year postoperative time points. Conclusion: DCI score was not associated with increased rates of postoperative complication requiring operative intervention and FAAM-VAS scores in patients undergoing primary TAA. However, patients in the 5th quintile of DCI experienced non-significantly lesser improvement in FAAM-VAS scores up to 2-years from surgery.

Category: Ankle; Other Introduction/Purpose: Obesity has shown to be associated with postoperative wound complications, infection, VTE, and wound dehiscence in patients undergoing surgical repair of the Achilles tendon1–3. Obesity is often defined as body mass index (BMI) greater than 30kg/m 2 . The Center for Disease Control defines adult BMI ranges as follows: underweight ( < 18.5kg/m 2 ), healthy (18.5 to 25 kg/m 2 ), overweight (25 to 30kg/m 2 ), Class I obesity (30 to 35 kg/m 2 ), Class II obesity (35 to 40 kg/m 2 ), Class III obesity (40 kg/m 2 and above)4. The purpose of this study is to compare the postoperative complications and FAAM-VAS scores between patients with surgically-repaired Achilles tendon according to their BMI ranges. Methods: Patients with mid-substance Achilles tendon rupture surgically repaired between 2016 and 2021 were identified. Chart notes were reviewed up to the 2-year postoperative period for wound complications and infection. Age, sex, Type II diabetes, rheumatoid arthritis, smoking status, and body mass index at the time of surgery were collected. Foot and Ankle Ability Measure and Visual Analog Pain Scales (FAAM-VAS) were collected preoperatively and at 6 months, 1 year, and 2 years postoperative time points. A total of 116 patients were identified and stratified into four groups based on their body mass index4: 18.5kg/m 2 - 25kg/m 2 (Group 1; n=24), 25kg/m 2 - 30kg/m 2 (Group 2; n=50), 30kg/m 2 - 35kg/m 2 (Group 3; n=20), and >35kg/m 2 (Group 4;n=22). Analysis of Variance Testing or Kruskal-Wallis Testing was used to calculate p-values for continuous data and Chi-Square testing was used to calculate p-values for categorical data. Pairwise testing was also conducted when appropriate. Results: The difference in rates of infection resulting in wound dehiscence or abscess drainage was not statistically significant across groups (p=0.362). No patient experienced re-rupture of the ipsilateral Achilles tendon. Group 4 patients reported the highest preoperative FAAM Overall, FAAM-ADL, and VAS scores. Their postoperative FAAM-Overall and FAAM-ADL scores were the lowest at all postoperative time points. Compared to patients in Group 2, patients in Group 4 demonstrated significantly less improvement in FAAM Overall score at 6 months (p= 0.029), 1-year (p=0.006), and 2-years (p=0.039) from surgery. The improvement in FAAM-ADL and VAS pain scores were not significantly different across BMI groups. However, the magnitude of improvement in FAAM-ADL was the lowest for patients in Group 4 at all postoperative time points. Conclusion: BMI was not significantly correlated with postoperative wound complication, infection, or re-rupture rates, but patients with BMI >35kg/m 2 reported significantly less improvement in FAAM Overall scores at the 6-month, 1-year, and 2-year postoperative periods compared to those with BMI between 25 and 30kg/m 2 .

Category: Midfoot/Forefoot; Other Introduction/Purpose: Acute fractures of the proximal fifth metatarsal are one of the most common injuries to the foot, occurring in up to 70% of all metatarsal fractures1. Displaced zone 1 fractures of the fifth metatarsal with greater than 2 mm fracture gap or those comprising greater than 30% of the cubometatarsal joint warrant operative treatment with open reduction internal fixation (ORIF)2,7, as satisfactory bone healing and patient-reported outcomes have been reported previously8–13. More recently, a few studies have demonstrated good functional outcomes after conservative treatment of zone 1 fractures with initial displacement greater than 2 mm7,13. Nevertheless, the lack of comparative studies and conflicting available evidence of outcomes warrant further investigation on this topic. Methods: A chart review was conducted to identify patients with displaced zone 1 fractures of the fifth metatarsal. Age at time of injury, gender, smoking and diabetes history, were collected. Degree of fracture displacement were measured (mm) on AP, lateral, and oblique view radiographs obtained at the initial encounter. Non-operatively treated patients were instructed to weightbear as tolerated in a Controlled Ankle Motion (CAM) boot for 6 weeks. Surgically-treated patients were placed in a short-leg splint for 2 weeks, and remained non-weightbearing in a CAM boot until 6 week period. They transitioned to normal shoe wear at the 8 week postoperative period. Chart notes were reviewed to identify the time to complete healing and incidences of delayed or nonunion. Means were compared using a one-way ANOVA or Kruskal Wallis test depending on normality, and categorical variables were compared using Chi-square or Fisher’s exact test. Results: The average time to complete healing was greater in non-operatively treated patients by 3 weeks (17.1 vs. 14.1, p=0.282). 4/37 (10.8%) non-operatively treated patients experienced delayed or non-union whereas 4/38 (13.2%) operatively-treated patients experienced delayed or non-union. This difference was statistically non-significant. The average fracture displacement (mm) on the oblique view was significantly greater in operatively-treated patients (2.68 vs. 5.40, p< 0.001). For patients with fracture gaps greater than 2 mm on the oblique view, non-operatively treated patients took 20.9 weeks whereas operatively- treated patients took 11.8 weeks to reach complete healing (p < 0.01). Conclusion: Patients who were treated non-operatively for displaced zone 1 fractures of the fifth metatarsal took on average 3 weeks longer to reach complete healing than operatively-treated patients. For patients with fracture gap greater than 2 mm on the oblique view, operative treatment was associated with significantly reduced time to complete healing.

Category: Ankle Arthritis; Ankle Introduction/Purpose: Total ankle arthroplasty (TAA) has become an accepted alternative to ankle arthrodesis for end-stage ankle arthritis and has seen significantly increased utilization in the past decade1. The Salto Talaris total ankle prosthesis has shown excellent short-term and mid-term survivorship results since its FDA approval in the U.S. in 20062,3. At a minimum of five-year follow-up, the currently reported midterm survivorship has ranged from 93.3% to 100% with equally satisfactory clinical outcomes as evidenced in short-term outcome studies4–8. The purpose of this study is to report the outcomes of one of the largest cohort of patients who underwent a Salto Talaris total ankle arthroplasty with a minimum of five-year follow-up. Methods: 103 Salto Talaris TAA patients with 5-view weight-bearing series of radiographs and minimum follow-up time of 5 years since index surgery were included. Age at time of surgery, gender, body mass index, diagnosis, diabetes status, and tobacco use were collected through chart review. Radiographic imaging was assessed at the most recent follow-up for evidence of radiolucency and osteolysis as previously described9. Range of motion was assessed on plantarflexion and dorsiflexion views on sagittal radiographs as previously described10. The Foot and Ankle Ability Measure (FAAM) and Visual Analog Scale for pain (VAS) scores were obtained at the most recent follow up visit and compared with preoperative scores. Baseline characteristics were compared between groups using Wilcoxon rank-sum test for continuous data or Fisher exact test for discrete data. Survivorship probability was calculated using a Kaplan-Meier analysis for revisions and reoperations. Revisions were defined as exchange or removal of metal component. Results: The survivorship for revision surgery was 93.2% (95% CI [89.8,98.8]). 5 out of the 7 patients requiring revision surgery experienced talar subsidence, 1 patient experienced an infection, and 1 patient experienced lateral impingement. The survivorship for reoperation was 90.2% (95% CI [83.4-95.4]). 5 out of the 10 patients requiring reoperation experienced pain from impingement, 2 experienced periprosthetic fractures, 1 experienced infection, and 2 experienced osteolytic cysts. At last follow- up, the FAAM-ADL, FAAM-Sport, and VAS pain scores were improved from preoperatively. The average range of motion at last follow-up was 31.6 degrees. On the AP view, radiolucency and osteolysis were most frequently detected in zones 3 and 4, while on the lateral view they were most frequently seen in zones 2 and 6. Conclusion: Patients undergoing Salto Talaris prosthesis showed improvements in patient-reported outcomes with satisfactory survival rates.

Category: Ankle Arthritis; Ankle Introduction/Purpose: The Cadence total ankle arthroplasty (TAA) system first entered clinical use in 2016. Short-term outcomes with a minimum of two-year follow-up time have demonstrated survivorship between 93.7% and 100% with improvements in patient-reported outcomes measures, coronal alignment, and range of motion1–4. The purpose of this study is to report the midterm survivorship and complications of Cadence TAA patients with a minimum of 4.5-year radiographic follow-up. Methods: A retrospective chart review was conducted to identify a total of 22 TAA patients with the Cadence implant and a minimum follow-up time of 4.5-years. Patient age, sex, body mass index, diabetes mellitus status, rheumatoid arthritis status, smoking status, laterality, and diagnosis for TAA were collected from patient chart reviews. Preoperative FAAM-ADL subscore, FAAM-Sport subscore, and VAS pain scores and the postoperative scores at the last follow-up visit was also collected. 13 out of 22 patients had preoperative FAAM-VAS scores available while 10 out of 22 patients had postoperative FAAM-VAS scores available. Postoperative complications in the form of wound-related issues and those requiring reoperation or revision were noted, with revision defined as removal or exchange of metal component. 5-view radiographs obtained preoperatively and at the last follow-up visit were used to measure coronal alignment, range of motion, evidence of subsidence or loosening, and osteolysis as previously described1. Results: 3 out of 22 patients required a revision surgery, 1 for loosening of tibial component and 2 for infection. 4 out of 22 patients underwent a reoperation, 2 for gutter debridement due to impingement or heterotopic ossification, 1 for periprosthetic medial malleolar fracture, and 1 for removal of painful medial malleolar screws. The average FAAM-ADL, FAAM-Sport, and VAS pain scores improved from 50.6, 26.5, and 59.9 to 69.5, 35.5, and 23.6, respectively. The coronal alignment angle decreased from 5.96 degrees from neutral to 2.31 degrees. The average dorsiflexion and plantarflexion angles were 14.74 and 17.18 degrees, respectively, for an average range of motion of 31.9 degrees. 3 out of 22 patients had evidence of osteolysis while no patients had evidence of loosening or subsidence. Conclusion: The midterm survivorship of Cadence TAA system in 22 patients was 86.4%. Excluding the two cases of infection which are typically not attributed to implant performance, the survivorship was 95.4%. The reasons for reoperation included impingement, heterotopic ossification, periprosthetic fracture, and removal of painful hardware. The average FAAM-ADL, FAAM- Sport, VAS pain scores and coronal alignment improved. Analysis of longer follow-up time in greater patient cohorts is necessary for clearer understanding of this fixed-bearing implant’s performance.

Category: Midfoot/Forefoot; Other Introduction/Purpose: First metatarsolphalangeal joint (MTPJ) arthrodesis is a commonly performed surgical procedure for end- stage arthritis. Various fixation techniques have been described. The most commonly used are screw-only fixation (either crossed or parallel) or plate fixation (with or without independent lag-screws). A recent systematic review reported that union rates for screw fixation were lower compared to plate constructs. The aim of this study was to compare union rates, complication rates and functional outcomes between screw-only fixation and plate fixation in a recent cohort of patients undergoing first MTPJ arthrodesis at a tertiary foot ankle orthopaedic institution. Methods: This was a retrospective cohort study. After institutional review board approval, a database search was conducted to identify patients who underwent primary first MTPJ arthrodesis between 2015 and 2022. All patients had a dorsal approach, preparation of joint surfaces with cup and cone reamers and fixation with either crossed cannulated screws or a dorsal pre- contoured locking plate (with or without an independent lag-screw). Demographic, operative (fixation type) and outcome data was collected. The primary outcome was the incidence of arthrodesis at 6 months postoperatively. Secondary outcomes included complications at 6 months postoperatively and change in functional outcome scores from preoperatively to 6 months postoperatively as assessed by the Functional Ankle Ability Measure (FAAM) and Visual Analog Pain scores (VAS). Statistical analyses included descriptive statistics as well as univariable and multivariable analyses to determine if one fixation method was associated with a higher incidence of arthrodesis. Results: There were 108 patients identified, 54 in the screw-only group (mean age 62.9 years, 79.6% female, mean body-mass index (BMI) 28.6 kg/m 2 ) and 54 in the plate-fixation group (mean age 63.2 years, 81.5% female, mean BMI 30.0kg/m 2 ). Arthrodesis at 6 months in the screw-only group occurred in 52 of 54 patients (96.3%) and in 50 of 54 patients (92.6%) in the plate-fixation group, p=0.68. Fixation type was not a predictor of non-union on multivariable analysis. The complication rate was 11.1% (n=6) in screw-only patients (four hardware removals, two non-unions) and 14.8% (n=8) in plate-fixation patients (four non-unions, two hardware removals, two wound infections), p=0.57. Both groups reported increases in VAS and FAAM scores at 6 months from baseline, with no significant between-group differences. Conclusion: The arthrodesis rate for screw-only fixation was higher than patients receiving plate-fixation, however, this difference did not reach statistical significance. Similar complication rates were observed in each group. These results provide evidence for the safety and efficacy of screw-only fixation constructs, however, further adequately powered prospective studies are required. Future studies should also include a cost effectiveness analysis and a comparison of radiation exposure and operative time between groups to determine the optimal fixation method for patients undergoing this procedure.

Background: Due to the poor prognosis and rising occurrence, there is a crucial need to improve the diagnosis of Primary Central Nervous System Lymphoma (PCNSL), which is a rare type of non-Hodgkin’s lymphoma. This study utilized targeted metabolomics of cerebrospinal fluid (CSF) to identify biomarker panels for the improved diagnosis or differential diagnosis of primary central nervous system lymphoma (PCNSL).Methods: In this study, a cohort of 68 individuals, including patients with primary central nervous system lymphoma (PCNSL), non-malignant disease controls, and patients with other brain tumors, was recruited. Their cerebrospinal fluid samples were analyzed using the Ultra-high performance liquid chromatography - tandem mass spectrometer (UHPLC-MS/MS) technique for targeted metabolomics analysis. Multivariate statistical analysis and logistic regression modeling were employed to identify biomarkers for both diagnosis (Dx) and differential diagnosis (Diff) purposes. The Dx and Diff models were further validated using a separate cohort of 34 subjects through logistic regression modeling.Results: A targeted analysis of 45 metabolites was conducted using UHPLC-MS/MS on cerebrospinal fluid (CSF) samples from a cohort of 68 individuals, including PCNSL patients, non-malignant disease controls, and patients with other brain tumors. Five metabolic features were identified as biomarkers for PCNSL diagnosis, while nine metabolic features were found to be biomarkers for differential diagnosis. Logistic regression modeling was employed to validate the Dx and Diff models using an independent cohort of 34 subjects. The logistic model demonstrated excellent performance, with an AUC of 0.83 for PCNSL vs. non-malignant disease controls and 0.86 for PCNSL vs. other brain tumor patients.Conclusion: Our study has successfully developed two logistic regression models utilizing metabolic markers in cerebrospinal fluid (CSF) for the diagnosis and differential diagnosis of PCNSL. These models provide valuable insights and hold promise for the future development of a non-invasive and reliable diagnostic tool for PCNSL.

Abstract Background To inform the development of updated World Health Organization (WHO) guidelines on simplified service delivery for HCV infection, a global survey was undertaken among people affected or infected by HCV. The objective of this analysis is to identify specific needs and preferences among people who inject drugs. Methods A multi-country, anonymous, self-administered online survey conducted in 2021 was developed by Coalition PLUS and the World Hepatitis Alliance in partnership with the WHO. Preferences for test and treat locations and simplifying HCV care were collected among people affected or infected by HCV. Chi-square tests were used to compare respondents who identified with current or former injection drug users through identification with key population to other respondents who did not identify with this key population. Results Among 202 respondents, 62 (30.7%) identified with current/former injection drug users. Compared to other respondents, they were: older [median (IQR): 48 (36–57) vs. 39 (31–51) years, p = 0.003]; more likely to have been tested for HCV (90.2% vs. 64.3%, p = 0.001); more likely to prefer testing in a community-based centre (CBC) (55.4% vs. 33.3%, p = 0.005); or in a support centres for people who use drugs (SCPUD)(50.0% vs. 9.8%, p

Forkhead box transcription factor, FOXQ1 is reported to promote epithelial-mesenchymal transition (EMT) and cancer metastasis. Here the authors show that FOXQ1 recruits the KMT2/MLL histone methyltransferase complex as a transcriptional coactivator to activate EMT programme in breast cancer.

Summary: Low-grade and high-grade serous carcinomas have unique clinical, morphological, underlying molecular alterations, and vastly different biologic behavior (Prat et al., 2018, Vang et al., 2009). The differentiation into high and low-grade serous carcinoma is important for clinical management and prognosis and is easily recognized by practicing pathologists. High-grade serous carcinoma is characterized by marked nuclear atypia and pleomorphism, frequent, often atypical mitosis with papillary or three-dimensional clusters, p53 mutation, and block-like p16 staining. In contrast, low-grade serous carcinomas have a different morphologic appearance with micropapillary formation, small nests of tumor cells having low to intermediate grade nuclei, and absence of significant mitosis. Low-grade serous carcinoma is often associated with micropapillary variant of ovarian serous borderline tumor. The low-grade serous carcinoma shows wild type p53 expression, patchy p16 staining, and often K-RAS, N-RAS, and/or B-RAF mutation. Here we report a case of mullerian high grade serous with a deceptive morphology resembling low-grade serous carcinoma with micropapillary features and moderate nuclear atypia. However, the tumor is simultaneously p53 and K-RAS mutated. This case illustrates three critical issues; a) potential to be mistaken as a low-grade serous carcinoma because of morphologic appearance and relative uniform cytologic feature. b). raise the question of true progression of low-grade to high-grade serous carcinoma, a rare phenomenon as described in the literature, and c). whether the biologic behavior and/or response to therapy would differ from the classic forms.

Abstract Primary challenges for current and future precision neutrino experiments using liquid argon time projection chambers (LArTPCs) include understanding detector effects and quantifying the associated systematic uncertainties. This paper presents a novel technique for assessing and propagating LArTPC detector-related systematic uncertainties. The technique makes modifications to simulation waveforms based on a parameterization of observed differences in ionization signals from the TPC between data and simulation, while remaining insensitive to the details of the detector model. The modifications are then used to quantify the systematic differences in low- and high-level reconstructed quantities. This approach could be applied to future LArTPC detectors, such as those used in SBN and DUNE.

Preferences are fundamental in decision-making, so understanding preference management is key in developing systems that guide the choices of the users. These choices can be decided through argument(s) which are known to have various strengths, as one argument can rely on more certain or vital information than the other. We explored argumentation technique from a previous study, and validated its potentials by applying to it several real-life scenarios. The exploration demonstrates the usefulness of argumentation in handling conflicting preferences and inconsistencies, and provides effective ways to manage, reason and represents users’ preferences. Using argumentation, we provide a practical implementation of a system to manage conflicting situations, and a simple interface that aids the flow of preferences from users to the system. We illustrated using the interface, how the changes in users’ preferences can effect system output in a smart home. This article describes the functionalities of the implemented system, and illustrates the functions by solving some of the complexities in users’ preferences in a real smart home. The system detects potential conflicts, and tries solve them using a redefined precedence order among some preference criteria. We also show how our system is capable of interacting with external sources data. The system was used to access and use live data of a UK supermarket chain store, through their application programming interface (API) and provide users suggestions on their eating habits, based on their set preference(s). The system was used to filter-specific products from the live data, and check the product description, before advising the user accordingly.

Abstract The MicroBooNE liquid argon time projection chamber located at Fermilab is a neutrino experiment dedicated to the study of short-baseline oscillations, the measurements of neutrino cross sections in liquid argon, and to the research and development of this novel detector technology. Accurate and precise measurements of calorimetry are essential to the event reconstruction and are achieved by leveraging the TPC to measure deposited energy per unit length along the particle trajectory, with mm resolution. We describe the non-uniform calorimetric reconstruction performance in the detector, showing dependence on the angle of the particle trajectory. Such non-uniform reconstruction directly affects the performance of the particle identification algorithms which infer particle type from calorimetric measurements. This work presents a new particle identification method which accounts for and effectively addresses such non-uniformity. The newly developed method shows improved performance compared to previous algorithms, illustrated by a 93.7% proton selection efficiency and a 10% muon mis-identification rate, with a fairly loose selection of tracks performed on beam data. The performance is further demonstrated by identifying exclusive final states in ν μ CC interactions. While developed using MicroBooNE data and simulation, this method is easily applicable to future LArTPC experiments, such as SBND, ICARUS, and DUNE.

The impact of gamma radiation on a commercial off the shelf microcontroller board has been investigated. Three different tests have been performed to ascertain the radiation tolerance of the device from a nuclear decommissioning deployment perspective. The first test analyses the effect of radiation on the output voltage of the on-board voltage regulator during irradiation. The second test evaluated the effect of gamma radiation on the voltage characteristics of analogue and digital inputs and outputs. The final test analyses the functionality of the microcontroller when using an external, shielded voltage regulator instead of the on-board voltage regulator. The results suggest that a series of latch-ups occurs in the microcontroller during irradiation, causing increased current drain which can damage the voltage regulator if it does not have short-circuit protection. The analogue to digital conversion functionality appears to be more sensitive to gamma radiation than digital and analogue output functionality. Using an external, shielded voltage regulator can prove beneficial when used for certain applications. The collected data suggests that detaching the voltage regulator can extend the lifespan of the platform up to approximately 350 Gy.

BackgroundKawasaki disease (KD) is the leading cause of acquired heart disease in children. The major challenge in KD diagnosis is that it shares clinical signs with other childhood febrile control (FC) subjects. We sought to determine if our algorithmic approach applied to a Taiwan cohort.MethodsA single center (Chang Gung Memorial Hospital in Taiwan) cohort of patients suspected with acute KD were prospectively enrolled by local KD specialists for KD analysis. Our previously single-center developed computer-based two-step algorithm was further tested by a five-center validation in US. This first blinded multi-center trial validated our approach, with sufficient sensitivity and positive predictive value, to identify most patients with KD diagnosed at centers across the US. This study involved 418 KDs and 259 FCs from the Chang Gung Memorial Hospital in Taiwan.FindingsOur diagnostic algorithm retained sensitivity (379 of 418; 90.7%), specificity (223 of 259; 86.1%), PPV (379 of 409; 92.7%), and NPV (223 of 247; 90.3%) comparable to previous US 2016 single center and US 2020 fiver center results. Only 4.7% (15 of 418) of KD and 2.3% (6 of 259) of FC patients were identified as indeterminate. The algorithm identified 18 of 50 (36%) KD patients who presented 2 or 3 principal criteria. Of 418 KD patients, 157 were infants younger than one year and 89.2% (140 of 157) were classified correctly. Of the 44 patients with KD who had coronary artery abnormalities, our diagnostic algorithm correctly identified 43 (97.7%) including all patients with dilated coronary artery but one who found to resolve in 8 weeks.InterpretationThis work demonstrates the applicability of our algorithmic approach and diagnostic portability in Taiwan.

In situ (AIS) and invasive endocervical adenocarcinoma have two broad categories, HPV-associated (HPV) and HPV-independent groups. (1) These entities show various types of cell morphology. Tubal and tubo-endometrioid type metaplasia of the cervix is a benign finding (Suh and Silverberg, 1990). Tubal metaplasia is also encountered in benign and malignant endometrial lesions. During cervical biopsy interpretations, differentiating the site of origin of the tissue is often tricky. We intend to document three cases of the sparsely reported hrHPV-associated ciliated/tubal-type endocervical AIS and invasive adenocarcinoma and bring it to the attention of readers how to avoid any misinterpretation during routine sign-out. Only three of fifty-three cases of hrHPV-associated AIS and invasive adenocarcinoma were of ciliated/tubal type in our department over a 5-year time. The presence of tubal-type epithelium should not automatically trigger the assumption of endometrial origin of the lesion. These cases are red herrings as tubal/ciliated type dysplasia, and carcinoma is rare and have potential to escape accurate diagnosis.

Aims: Mesonephric-like adenocarcinoma (MLA) is a recently described histologic tumor subtype of the Müllerian tract. MLA can arise in association with Müllerian lesions that share common mutations. We report three MLAs and hypothesize that concurrent endometriosis and cystadenofibroma with focal borderline changes might also carry common mutations. Methods and results: We searched “mesonephric” in our database from 2015 to mid-2021 to retrieve MLA cases. Somatic mutation analysis was performed on tumors and on associated benign proliferative lesions. All MLAs (2 ovarian and 1 uterine) harbored KRAS G12D or G12 V mutations. A PIK3CA alteration (H1047Q) was detected in one MLA and in the associated cystadenofibroma with focal borderline changes. The molecular profile of MLA-associated Müllerian lesions (endometriosis and seromucinous cystadenofibroma with focal borderline changes) was similar to concurrent adenocarcinoma. However, tumor contamination could not be excluded in the endometriotic lesion. Patients presented at various stages, with no evidence of post-operative recurrence after 15 months (FIGO IC) and 33 months (FIGO IIA2). One patient (FIGO IIIA1) died of disease 32 months after surgery. Conclusions: KRAS mutations commonly characterize MLA. At least some MLA-associated Müllerian lesions show MLA-like genetic profiles, suggesting a precursor role. As far as we are aware, we describe for the first time in MLA the potentially actionable H1047Q variant of PIK3CA.

Gut microbial communities are vital for maintaining host health, and are sensitive to diet, environment, and chemical exposures. Wastewater treatment plants (WWTPs) release effluents containing antimicrobials, pharmaceuticals, and other contaminants that may negatively affect the gut microbiome of downstream organisms. This study investigated changes in the diversity and composition of the digestive gland microbiome of flutedshell mussels (Lasmigona costata) from upstream and downstream of two large (service >100,000) WWTPs. Mussel digestive gland microbiome was analyzed following the extraction, PCR amplification, and sequencing of bacterial DNA using the V3-V4 hypervariable regions of the 16 S rRNA gene. Bacterial alpha diversity decreased at sites downstream of the second WWTP and these sites were dissimilar in beta diversity from sites upstream and downstream of the first upstream WWTP. The microbiomes of mussels collected downstream of the first WWTP had increased relative abundances of Actinobacteria, Bacteroidetes, and Firmicutes, with a decrease in Cyanobacteria, compared to upstream mussels. Meanwhile, those collected downstream of the second WWTP increased in Proteobacteria and decreased in Actinobacteria, Bacteroidetes, and Tenericutes. Increased Proteobacteria has been linked to adverse effects in mammals, but their functions in mussels is currently unknown. Finally, effluent-derived bacteria were found in the microbiome of mussels downstream of both WWTPs but not in those from upstream. Overall, results show that the digestive gland microbiome of mussels collected upstream and downstream of WWTPs differed, which has implications for altered host health and the transport of WWTP-derived bacteria through aquatic ecosystems.

Fibroblasts, or their homolog stromal cells, are present in most tissues and play an essential role in tissue homeostasis and regeneration. As a result, fibroblast-based strategies have been widely employed in tissue engineering. However, while considered to have immunosuppressive properties, the survival and functionality of allogeneic fibroblasts after transplantation remain controversial. Here, we evaluated innate and adaptive immune responses against allogeneic fibroblasts following intradermal injection into different immune-deficient mouse strains. While allogeneic fibroblasts were rejected 1 week after transplantation in immunocompetent mice, rejection did not occur in immunodeficient γ chain–deficient NOD-SCID (NSG) mice. T-cell- and B-cell-deficient RAG1 knockout mice showed greater loss of fibroblasts by day 5 after transplantation compared with NSG mice ( P ≤ 0.05) but prolonged persistence compared with wild-type recipient ( P ≤ 0.005). Loss of fibroblasts correlated with the expression of proinflammatory chemokine genes and infiltration of myeloid cells in the transplantation site. Depletion of macrophages and neutrophils delayed rejection, revealing the role of innate immune cells in an early elimination of fibroblasts that is followed by T-cell-mediated rejection in the second week. These findings indicate that the application of allogeneic fibroblasts in tissue engineering products requires further improvements to overcome cell rejection by innate and adaptive immune cells.

The DNA-sensing cGAS-STING pathway promotes the senescence-associated secretory phenotype (SASP) and mediates type-I interferon inflammatory responses to foreign viral and bacterial DNA as well as self-DNA. Studies of the intervertebral disc in humans and mice demonstrate associations between aging, increased cell senescence, and disc degeneration. Herein we assessed the role of STING in SASP promotion in STING gain- (N153S) and loss-of-function mouse models. N153S mice evidenced elevated circulating levels of proinflammatory markers including IL-1β, IL-6, and TNF-α, showed elevated monocyte and macrophage abundance in the vertebral marrow, and exhibited a mild trabecular and cortical bone phenotype in caudal vertebrae. Interestingly, despite systemic inflammation, the structural integrity of the disc and knee articular joint remained intact, and cells did not show a loss of their phenotype or elevated SASP. Transcriptomic analysis of N153S tissues demonstrated an upregulated immune response by disc cells, which did not closely resemble inflammatory changes in human tissues. Interestingly, STING-/- mice also showed a mild vertebral bone phenotype, but the absence of STING did not reduce the abundance of SASP markers or improve the age-associated disc phenotype. Overall, the analyses of N153S and STING-/- mice suggest that the cGAS-STING pathway is not a major contributor to SASP induction and consequent disc aging and degeneration but may play a minor role in the maintenance of trabecular bone in the vertebrae. This work contributes to a growing body of work demonstrating that systemic inflammation is not a key driver of disc degeneration.

Abstract Janus Kinases (JAKs) have emerged as an important drug target for the treatment of a number of immune disorders due to the central role that they play in cytokine signalling. 4 isoforms of JAKs exist in mammalian cells and the ideal isoform profile of a JAK inhibitor has been the subject of much debate. JAK3 has been proposed as an ideal target due to its expression being largely restricted to the immune system and its requirement for signalling by cytokine receptors using the common γ-chain. Unlike other JAKs, JAK3 possesses a cysteine in its ATP binding pocket and this has allowed the design of isoform selective covalent JAK3 inhibitors targeting this residue. We report here that mutating this cysteine to serine does not prevent JAK3 catalytic activity but does greatly increase the IC50 for covalent JAK3 inhibitors. Mice with a Cys905Ser knockin mutation in the endogenous JAK3 gene are viable and show no apparent welfare issues. Cells from these mice show normal STAT phosphorylation in response to JAK3 dependent cytokines but are resistant to the effects of covalent JAK3 inhibitors. These mice therefore provide a chemical-genetic model to study JAK3 function.

Abstract Background Murine xenografts of pediatric leukemia accurately recapitulate genomic aberrations. How this translates to the functional capacity of cells remains unclear. Here, we studied global protein abundance, phosphorylation, and protein maturation by proteolytic processing in 11 pediatric B- and T- cell ALL patients and 19 corresponding xenografts. Methods Xenograft models were generated for each pediatric patient leukemia. Mass spectrometry-based methods were used to investigate global protein abundance, protein phosphorylation, and limited proteolysis in paired patient and xenografted pediatric acute B- and T- cell lymphocytic leukemia, as well as in pediatric leukemia cell lines. Targeted next-generation sequencing was utilized to examine genetic abnormalities in patients and in corresponding xenografts. Bioinformatic and statistical analysis were performed to identify functional mechanisms associated with proteins and protein post-translational modifications. Results Overall, we found xenograft proteomes to be most equivalent with their patient of origin. Protein level differences that stratified disease subtypes at diagnostic and relapse stages were largely recapitulated in xenografts. As expected, PDXs lacked multiple human leukocyte antigens and complement proteins. We found increased expression of cell cycle proteins indicating a high proliferative capacity of xenografted cells. Structural genomic changes and mutations were reflected at the protein level in patients. In contrast, the post-translational modification landscape was shaped by leukemia type and host and only to a limited degree by the patient of origin. Of 201 known pediatric oncogenic drivers and drug-targetable proteins, the KMT2 protein family showed consistently high variability between patient and corresponding xenografts. Comprehensive N terminomics revealed deregulated proteolytic processing in leukemic cells, in particular from caspase-driven cleavages found in patient cells. Conclusion Genomic and host factors shape protein and post-translational modification landscapes differently. This study highlights select areas of diverging biology while confirming murine patient-derived xenografts as a generally accurate model system.