Citations

ABSTRACT Purpose We studied histologic Ewing sarcoma (ES) tumor response at local control surgery to explore associations between histologic response and chemotherapy practice changes over time and confirm its prognostic significance in the modern era. Methods We reviewed clinical data for 126 patients with ES treated at Mayo Clinic from 1977 to 2022, analyzing the percentage necrosis as a prognostic factor across chemotherapy regimens (including modern interval‐compressed VDC‐IE). Results In the historical cohort, 5‐year event‐free survival (EFS) and overall survival (OS) for patients with ≤5% versus greater than 5% viable tumor were 76% versus 59% ( p = 0.01) and 83% versus 62% ( p = 0.01). Review of 104 specimens by two pathologists identified the percentage remaining viable tumor as more reproducible than the percentage necrosis and/or fibrosis. On multivariable analysis of patients with localized disease ( n = 87), pelvic tumors and greater than 5% viable tumor remained independent risk factors for poor survival (EFS: hazard ratio [HR] 3.1, p < 0.01; OS: HR 2.1, p = 0.048). The modern cohort consisted of 22 patients. No patients with tumor size less than 8 cm at diagnosis and 100% necrosis after surgery had events; four of seven with tumors greater than 8 cm had events despite 100% necrosis. Conclusions Our results validate the historical threshold of greater than 5% viable tumor with older chemotherapy regimens but not with modern interval‐compressed VDC‐IE. In addition, inter‐rater reliability for the percentage viable is more reproducible than the percentage necrosis. Although numbers are small, tumors less than 8 cm with no viable tumor appear to be a favorable subgroup and suggest risk stratification using a combination of tumor size and any viable tumor should be evaluated in ES.

Background: Cocaine use and cardiovascular disease (CVD)-related mortality trends in the U.S. are underreported. Using the CDC’s Wide-Ranging Online Data for Epidemiologic Research database, we analyzed the relationship between cocaine use and CVD-related deaths. Methods: We analyzed death certificates from the Multiple Cause of Death dataset to identify CVD-related deaths involving cocaine use. Age-adjusted mortality rates (AAMRs) per 1 000 000 and annual percentage changes (APCs) were calculated using Joinpoint Regression. Results: From 1999 to 2022, 82 944 deaths were linked to cocaine use and CVDs. AAMRs rose from 9.6 in 1999 to 32 in 2022. Mortality trends varied by demographics and regions. Males and females showed early increases (1999–2006), declines (2006–2012), and sharp rises (2012–2022), with APCs of 11.8% and 12.8%, respectively. Non-Hispanic Black individuals exhibited consistently higher rates than non-Hispanic Whites, though both groups experienced sharp increases starting in 2014 (APCs: 12.5 and 12.7%). Geographic trends revealed early 2000s increases, mid-2000s declines, and rapid rises beginning in the 2010s. The Northeast and South had notable APCs of 13.5 and 12.6% (2013–2022). Rural areas initially showed steeper increases than urban areas but displayed steady upward trends since 2011 (APC: 9.5%). Age-wise, older adults experienced a particularly sharp rise from 2012 to 2022 (APC: 15.0%). Conclusion: Cocaine use and CVD-related mortality rose significantly from 1999 to 2022, with pronounced increases among men, non-Hispanic Black individuals, older adults, urban residents, and those in the South. These findings highlight a growing public health crisis.

ABSTRACT As a principal metabolite of ginsenosides, compound K (CK) exhibits remarkable antioxidant, anti‐inflammatory, antidiabetic, and anticancer bioactivities. However, CK is naturally scarce in ginseng and its industrial application is limited by the lack of efficient deglycosylating enzymes for large‐scale CK bioproduction. Here, we report a synthetic biology‐driven approach for high‐yield CK production via recombinant β‐glycosidase‐mediated bioconversion of the polar ginsenoside Rb1. The β‑glycosidase‐encoding gene was introduced into Escherichia coli BL21 (DE3), and the resulting engineered strain produced 2.1 g/L β‑glycosidase in shake flasks and 10.1 g/L in a 5‐L fermentor. The crude β‐glycosidase exhibited excellent enzymatic activity; at a concentration of 1.5 g/L, it converted 95% of the Rb1 substrate (initial concentration 10 g/L) to CK within 4 h. In vitro studies demonstrated CK's high cytocompatibility and potent anti‐aging effects, including suppression of reactive oxygen species ( via upregulation of superoxide dismutase, catalase, and glutathione peroxidase), enhanced collagen I, Ⅲ and elastin synthesis, and matrix metalloproteinase‐1 inhibition. These findings highlight CK as a promising candidate for anti‐wrinkle formulations, supported by the feasibility of scalable bioproduction.

ABSTRACT Background and Objectives Surgical management of ipsilateral breast tumor recurrence (IBTR) is often mastectomy. No guidelines specifically address contralateral prophylactic mastectomy (CPM) for this population. We assessed factors associated with CPM at the time of IBTR mastectomy. Methods Patients undergoing mastectomy for IBTR (10/2008‐5/2024) were identified. Demographics, tumor characteristics, and treatment details were assessed. Results Of the 381 eligible IBTR patients, 221 (58.0%) had a unilateral therapeutic mastectomy and 160 (42.0%) underwent mastectomy with CPM. CPM patients were younger ( p < 0.0001), with lower clinical T category tumors ( p < 0.001) and less HR + HER2‐ invasive disease ( p = 0.004). CPM patients were more likely to undergo germline genetic testing (63.1% vs 41.6%, p < 0.0001) and were twice as likely to have immediate breast reconstruction ( p < 0.0001). On multivariable analysis, younger age, HR‐HER2 + IBTR subtype, pathogenic variants known at the time of surgery (relative to negative testing and/or no genetic testing), immediate breast reconstruction, and prior radiation for index cancer were each associated with CPM (all p < 0.05). Conclusion In our series of 381 patients treated with mastectomy for IBTR, 42% also had a CPM. Multiple factors, including age, tumor stage and biology, immediate breast reconstruction, and genetic testing may impact CPM choice.

G protein-coupled receptors (GPCRs) are among the most common drug targets, with numerous agonists and antagonists approved for clinical use. More recently, it has been appreciated that drugs can also modulate the action of natural agonists of these receptors, thus providing unique clinical advantages. Here, we describe the identification and characterization of a small molecule positive allosteric modulator (PAM) of secretin action at the class B G protein-coupled secretin receptor. This phenyl(trifluoromethyl)-pyrimidine can occupy the secretin receptor without stimulating its internalization, yet priming it to enhance both the potency and efficacy of the action of natural secretin. This is also shown to exhibit its effects on cells expressing low numbers of these receptors, without enhancing the effects of other structurally related hormones acting at other class B GPCRs. The mechanism responsible for this PAM effect is the slowing of the off-rate of receptor-bound secretin. This compound can serve as a lead to the development of other drugs that enhance the action of natural endogenous secretin and can be utilized to explore the potential therapeutic utility of such compounds.

Background: Atezolizumab plus bevacizumab (A + B) and durvalumab plus tremelimumab (D + T) are approved first-line systemic therapy options for advanced hepatocellular carcinoma (HCC), with no head-to-head comparison. In this propensity-matched analysis, we compared the survival of patients with advanced HCC who received either A + B or D + T therapy. Methods: Patients with advanced HCC who received A + B or D + T treatment in the first-line setting were identified using the TriNetX platform, a health research network that provides access to electronic health record data from 112 healthcare organizations. Propensity-score-matched (PSM) analysis was conducted, and the median overall survival (OS) was estimated using the Kaplan–Meier method. Results: We identified 2819 patients with HCC who were treated with A + B or D + T: 2031 patients received A + B and 788 received D + T. Compared to patients in the A + B cohort, those who received D + T were more likely to be older (median age: 68.4 vs. 66.9 years), have lower platelet counts (186.8 vs. 201.3), higher prevalence of hypoalbuminemia with levels < 2.7 g/dL (35.2% vs. 28.4%), higher prevalence of bilirubin levels between 2.0 and 2.9 mg/dL (28.8% vs. 23.8%), and decreased INR with levels between 0.0 and 1.6 (92.1% vs. 86.3%). After PSM, 1536 patients were included in the survival analyses, with all variables adequately matched. There was no significant difference in the median OS between those receiving A + B or D + T [16.3 vs. 22.5 months, hazard ratio (HR) 0.89, 95% confidence interval (CI) 0.76–1.0]. Higher rates of immune-mediated colitis were noted in the D + T group. Conclusions: Similar survival rates were observed among patients with advanced HCC who received A + B and D + T in the first-line setting. Our study suggests that both A + B and D + T are valid treatment options, and that therapy can be tailored based on comorbidities, adverse effects, and patient preferences.

Abstract Deep mutational scanning (DMS) has proven effective for mapping protein–protein interactions (PPIs), but it cannot provide complete coverage of the mutation landscape, particularly for multi‐mutant variants. To address this limitation, we trained machine‐learning (ML) models on previously generated DMS data for a stabilized amyloid precursor protein inhibitor (APPI) binding to either of two serine proteases, mesotrypsin and kallikrein‐6 (KLK6), which are implicated in various human disorders. We combined the models to accurately predict the binding selectivity of APPI variants, including double‐mutant variants, for the two serine proteases. We achieved a Pearson correlation of 0.937 between predicted log 2 selectivity enrichment ratios and DMS‐derived values. We further validated the predictions of our combined model by yeast‐surface‐display measurements and inhibition assays of purified APPI variants and revealed epistatic interactions that shape protease selectivity. Guided by binding selectivity predictions, we identified highly selective APPI variants, including the most selective mesotrypsin inhibitor reported to date. Together, these findings support the use of DMS and ML as a framework for predicting PPI selectivity and prioritizing selective therapeutic protein variants.

Abstract The mutation status of isocitrate dehydrogenase (IDH) in glioma patients has diagnostic and prognostic significance but is available only after surgery. Rapid and sensitive intraoperative molecular diagnosis is essential for distinguishing tumor from infiltrative and from normal brain tissue to maximize safe resection. IDH-mutant (mut)tumors produce high concentrations of 2-hydroxyglutarate (2HG), which serves as a surrogate marker of IDH mutation. Here, we use desorption electrospray ionization-multiple reaction monitoring (DESI-MRM), a form of mass spectrometry (MS), for rapid prediction of IDH mutation status. A set of 30 human brain smears was analyzed using DESI-MRM experiments to predict the mutation status of IDH and quantify 2HG in glioma samples within 2 min (endogenous reference molecule) or 7 min (external standard). Nanoelectrospray ionization (nESI) was employed as a validation method. An isotopically labeled surrogate internal standard allowed quantitative measurement of 2HG and clear separation between IDH-mut and IDH-wildtype (wt) samples, showing 100% agreement with clinical diagnosis. 2HG concentrations in IDH-mut samples ranged from 88 to 1445 ng mg–1, while IDH-wt samples exhibited 2HG concentrations of 9–15 ng mg–1. Calibration curves showed method accuracy within ≤15%. The quantitative results obtained using isotopically labeled standards showed a strong correlation between IDH mutation score and 2HG concentrations, as did an earlier semiquantitative (ratiometric, endogenous standard) method. These findings highlight the standalone capability of DESI-MRM for making measurements to justify assessments for intraoperative diagnosis and to support surgical decision-making to improve patient outcomes.

Abstract INTRODUCTION Although cerebrospinal fluid (CSF) biomarkers reflect neurodegeneration in Alzheimer's disease (AD), it remains unclear whether these biomarkers track neurodegeneration in early‐onset Alzheimer's disease (EOAD). METHODS In 80 EOAD patients, we examined correlations between eight CSF biomarker levels and cortical thickness decreases within the EOAD cortical atrophy signature. Significant correlations were then entered into a multiple regression analysis. We also examined contributions of EOAD atrophy and CSF biomarkers to cognitive impairment. RESULTS The levels of four CSF biomarkers correlated to EOAD signature atrophy. Stepwise regression analyses revealed that CSF neurofilament light chain (NfL) levels best predicted EOAD signature atrophy. Multiple regression showed EOAD signature atrophy combined with CSF NfL levels explained more variance in cognitive impairment than either factor alone. DISCUSSION Within EOAD patients, CSF NfL levels relate to the magnitude of cortical atrophy, and a combination of EOAD signature atrophy and CSF NfL levels most robustly predict cognitive impairment.