Inactivation of Salmonella Enteritidis PT30 on black peppercorns in thermal treatments with controlled relative humidities.

Traditional method utilizes steam to pasteurize low-moisture ingredients like black peppercorns and almonds. Exposure to steam results in direct condensation on the product, unfavorable for a broader range of food ingredients such as dried herbs, fruits, and ground materials. Recent studies on the thermal inactivation of Salmonella in low-moisture foods suggest that the relative humidity in treatment chambers is an important factor, besides temperature, that determines the death rate of bacteria. Thus, thermal treatments with controlled high relative humidity can be an effective method to replace steam pasteurization. No condensation will occur when the products are preheated to above the dew-point temperature of the hot air in the treatment chamber, thus eliminating the need for post-treatment drying. To prove this concept, a special device was developed that preheated samples in a dry environment before exposing them to a controlled relative humidity (RH) at a high temperature. Using this device, the death rate of Salmonella Enteritidis PT30 (S. Enteritidis) in black peppercorns was determined at 80 °C and three different RH levels (60, 70, or 80 %) after the innoculated samples were heated to 78oC. The results indicate that the treatments at 80 °C and 80 % RH for 3 min, 70 % RH for 9 min, and 60 % RH for 25 min caused 5.4 ± 0.2, 6.2 ± 0.6, and 6.1 ± 1.0 log reductions, respectively. No condensation was observed on all of the treated samples. The moisture content (wet basis) of fully pasteurized (5-log reduction) black peppercorns at 60, 70, and 80 %RH reduced from 9.7 ± 0.4 % (untreated) to 8.7 ± 0.5 %, 9.2 ± 0.4 %, and 9.2 ± 0.2 %, respectively, indicating that post-drying is not required after the treatments. This study demonstrated the potential of using short-time high-RH treatments to control pathogens in low-moisture foods without the need for post-treatment drying.

Three-dimensional structure of kinetochore-fibers in human mitotic spindles.

During cell division, kinetochore microtubules (KMTs) provide a physical linkage between the chromosomes and the rest of the spindle. KMTs in mammalian cells are organized into bundles, so-called kinetochore-fibers (k-fibers), but the ultrastructure of these fibers is currently not well characterized. Here, we show by large-scale electron tomography that each k-fiber in HeLa cells in metaphase is composed of approximately nine KMTs, only half of which reach the spindle pole. Our comprehensive reconstructions allowed us to analyze the three-dimensional (3D) morphology of k-fibers and their surrounding MTs in detail. We found that k-fibers exhibit remarkable variation in circumference and KMT density along their length, with the pole-proximal side showing a broadening. Extending our structural analysis then to other MTs in the spindle, we further observed that the association of KMTs with non-KMTs predominantly occurs in the spindle pole regions. Our 3D reconstructions have implications for KMT growth and k-fiber self-organization models as covered in a parallel publication applying complementary live-cell imaging in combination with biophysical modeling (Conway et al., 2022). Finally, we also introduce a new visualization tool allowing an interactive display of our 3D spindle data that will serve as a resource for further structural studies on mitosis in human cells.

Self-organization of kinetochore-fibers in human mitotic spindles.

During eukaryotic cell division, chromosomes are linked to microtubules (MTs) in the spindle by a macromolecular complex called the kinetochore. The bound kinetochore microtubules (KMTs) are crucial to ensuring accurate chromosome segregation. Recent reconstructions by electron tomography (Kiewisz et al., 2022) captured the positions and configurations of every MT in human mitotic spindles, revealing that roughly half the KMTs in these spindles do not reach the pole. Here, we investigate the processes that give rise to this distribution of KMTs using a combination of analysis of large-scale electron tomography, photoconversion experiments, quantitative polarized light microscopy, and biophysical modeling. Our results indicate that in metaphase, KMTs grow away from the kinetochores along well-defined trajectories, with the speed of the KMT minus ends continually decreasing as the minus ends approach the pole, implying that longer KMTs grow more slowly than shorter KMTs. The locations of KMT minus ends, and the turnover and movements of tubulin in KMTs, are consistent with models in which KMTs predominately nucleate de novo at kinetochores in metaphase and are inconsistent with substantial numbers of non-KMTs being recruited to the kinetochore in metaphase. Taken together, this work leads to a mathematical model of the self-organization of kinetochore-fibers in human mitotic spindles.

Metabolic state of human blastocysts measured by fluorescence lifetime imaging microscopy.

STUDY QUESTION: Can non-invasive metabolic imaging via fluorescence lifetime imaging microscopy (FLIM) detect variations in metabolic profiles between discarded human blastocysts? SUMMARY ANSWER: FLIM revealed extensive variations in the metabolic state of discarded human blastocysts associated with blastocyst development over 36 h, the day after fertilization and blastocyst developmental stage, as well as metabolic heterogeneity within individual blastocysts. WHAT IS KNOWN ALREADY: Mammalian embryos undergo large changes in metabolism over the course of preimplantation development. Embryo metabolism has long been linked to embryo viability, suggesting its potential utility in ART to aid in selecting high quality embryos. However, the metabolism of human embryos remains poorly characterized due to a lack of non-invasive methods to measure their metabolic state. STUDY DESIGN, SIZE, DURATION: We conducted a prospective observational study. We used 215 morphologically normal human embryos from 137 patients that were discarded and donated for research under an approved institutional review board protocol. These embryos were imaged using metabolic imaging via FLIM to measure the autofluorescence of two central coenzymes, nicotinamide adenine (phosphate) dinucleotide (NAD(P)H) and flavine adenine dinucleotide (FAD+), which are essential for cellular respiration and glycolysis. PARTICIPANTS/MATERIALS, SETTING, METHODS: Here, we used non-invasive FLIM to measure the metabolic state of human blastocysts. We first studied spatial patterns in the metabolic state within human blastocysts and the association of the metabolic state of the whole blastocysts with stage of expansion, day of development since fertilization and morphology. We explored the sensitivity of this technique in detecting metabolic variations between blastocysts from the same patient and between patients. Next, we explored whether FLIM can quantitatively measure metabolic changes through human blastocyst expansion and hatching via time-lapse imaging. For all test conditions, the level of significance was set at P < 0.05 after correction for multiple comparisons using Benjamini-Hochberg's false discovery rate. MAIN RESULTS AND THE ROLE OF CHANCE: We found that FLIM is sensitive enough to detect significant metabolic differences between blastocysts. We found that metabolic variations between blastocyst are partially explained by both the time since fertilization and their developmental expansion stage (P < 0.05), but not their morphological grade. Substantial metabolic variations between blastocysts from the same patients remain, even after controlling for these factors. We also observe significant metabolic heterogeneity within individual blastocysts, including between the inner cell mass and the trophectoderm, and between the portions of hatching blastocysts within and without the zona pellucida (P < 0.05). And finally, we observed that the metabolic state of human blastocysts continuously varies over time. LIMITATIONS, REASONS FOR CAUTION: Although we observed significant variations in metabolic parameters, our data are taken from human blastocysts that were discarded and donated for research and we do not know their clinical outcome. Moreover, the embryos used in this study are a mixture of aneuploid, euploid and embryos of unknown ploidy. WIDER IMPLICATIONS OF THE FINDINGS: This work reveals novel aspects of the metabolism of human blastocysts and suggests that FLIM is a promising approach to assess embryo viability through non-invasive, quantitative measurements of their metabolism. These results further demonstrate that FLIM can provide biologically relevant information that may be valuable for the assessment of embryo quality. STUDY FUNDING/COMPETING INTEREST(S): Supported by the Blavatnik Biomedical Accelerator Grant at Harvard University. Becker and Hickl GmbH and Boston Electronics sponsored research with the loaning of equipment for FLIM. D.J.N. is an inventor on patent US20170039415A1. TRIAL REGISTRATION NUMBER: N/A.

Effects of Compliance With the Early Management Bundle (SEP-1) on Mortality Changes Among Medicare Beneficiaries With Sepsis: A Propensity Score Matched Cohort Study.

BACKGROUND: US hospitals have reported compliance with the SEP-1 quality measure to Medicare since 2015. Finding an association between compliance and outcomes is essential to gauge measure effectiveness. RESEARCH QUESTION: What is the association between compliance with SEP-1 and 30-day mortality among Medicare beneficiaries? STUDY DESIGN AND METHODS: Studying patient-level data reported to Medicare by 3,241 hospitals from October 1, 2015, to March 31, 2017, we used propensity score matching and a hierarchical general linear model (HGLM) to estimate the treatment effects associated with compliance with SEP-1. Compliance was defined as completion of all qualifying SEP-1 elements including lactate measurements, blood culture collection, broad-spectrum antibiotic administration, 30 mL/kg crystalloid fluid administration, application of vasopressors, and patient reassessment. The primary outcome was a change in 30-day mortality. Secondary outcomes included changes in length of stay. RESULTS: We completed two matches to evaluate population-level treatment effects. In standard match, 122,870 patients whose care was compliant were matched with the same number whose care was noncompliant. Compliance was associated with a reduction in 30-day mortality (21.81% vs 27.48%, respectively), yielding an absolute risk reduction (ARR) of 5.67% (95% CI, 5.33-6.00; P < .001). In stringent match, 107,016 patients whose care was compliant were matched with the same number whose care was noncompliant. Compliance was associated with a reduction in 30-day mortality (22.22% vs 26.28%, respectively), yielding an ARR of 4.06% (95% CI, 3.70-4.41; P < .001). At the subject level, our HGLM found compliance associated with lower 30-day risk-adjusted mortality (adjusted conditional OR, 0.829; 95% CI, 0.812-0.846; P < .001). Multiple elements correlated with lower mortality. Median length of stay was shorter among cases whose care was compliant (5 vs 6 days; interquartile range, 3-9 vs 4-10, respectively; P < .001). INTERPRETATION: Compliance with SEP-1 was associated with lower 30-day mortality. Rendering SEP-1 compliant care may reduce the incidence of avoidable deaths.

Central-spindle microtubules are strongly coupled to chromosomes during both anaphase A and anaphase B.

Spindle microtubules, whose dynamics vary over time and at different locations, cooperatively drive chromosome segregation. Measurements of microtubule dynamics and spindle ultrastructure can provide insight into the behaviors of microtubules, helping elucidate the mechanism of chromosome segregation. Much work has focused on the dynamics and organization of kinetochore microtubules, that is, on the region between chromosomes and poles. In comparison, microtubules in the central-spindle region, between segregating chromosomes, have been less thoroughly characterized. Here, we report measurements of the movement of central-spindle microtubules during chromosome segregation in human mitotic spindles and Caenorhabditis elegans mitotic and female meiotic spindles. We found that these central-spindle microtubules slide apart at the same speed as chromosomes, even as chromosomes move toward spindle poles. In these systems, damaging central-spindle microtubules by laser ablation caused an immediate and complete cessation of chromosome motion, suggesting a strong coupling between central-spindle microtubules and chromosomes. Electron tomographic reconstruction revealed that the analyzed anaphase spindles all contain microtubules with both ends between segregating chromosomes. Our results provide new dynamical, functional, and ultrastructural characterizations of central-spindle microtubules during chromosome segregation in diverse spindles and suggest that central-spindle microtubules and chromosomes are strongly coupled in anaphase.

Do adjunctive art therapies reduce symptomatology in schizophrenia? A meta-analysis.

BACKGROUND: Art therapies are advocated by national bodies, such as the United Kingdom's National Institute for Health and Care Excellence, to alleviate the negative symptoms associated with schizophrenia. The last decade has however, seen several new larger well-controlled trials published suggesting an update is timely. AIM: To asses randomised controlled trials (RCT) of art therapies for reducing the symptoms of schizophrenia - particularly negative symptoms. METHODS: Searches of PubMed and Scopus were conducted until May 2019 for RCTs examining the impact of art therapies on psychosis (positive, negative and total) symptoms in people diagnosed with schizophrenia. Study quality was assessed using the Cochrane risk of bias tool. Random effects meta-analyses were used to derive overall effect sizes. Moderator analyses were conducted using both meta-regression and categorical comparisons. RESULTS: We identified 133 articles, of which 9 RCTs involving 948 participants (475 assigned to art therapies and 473 controls) met our inclusion criteria. Using random effects models, we calculated pooled effect sizes (Hedges g) for end-of-trial symptomatic outcomes. Effect sizes both for total symptoms [g = -0.27, 95% confidence interval (CI) -0.60 to 0.05, k = 6] and for positive symptoms (g = -0.10, 95%CI -0.35 to 0.15, k = 6) were non-significant; however, we did find significant reduction of negative symptoms (g = -0.42, 95%CI -0.70 to -0.14, k = 9). Meta-regression revealed that negative symptom reduction was larger in trials with a greater proportion of women and in trials with younger patients. Crucially, the negative symptom reduction following art therapies was limited to lower quality trials and did not emerge in trials that used blind assessment of outcomes. CONCLUSION: This review presents a comprehensive meta-analysis of art therapies in schizophrenia in terms of both studies included and participant numbers. We found that art therapies did not significantly reduce total or positive symptoms. A "small" therapeutic effect was found for negative symptoms, but we show that the effect is not present in blind trials and may be subject to publication bias.

Response to survey directed to patient portal members differs by age, race, and healthcare utilization.

Health care systems are increasingly utilizing electronic medical record-associated patient portals to facilitate communication with patients and between providers and their patients. These patient portals are growing in recognition as potentially valuable research tools. While there is much information about the response rates and demographics of internet-based surveys as well as the demographics of patients who are portal members, not much is known about the response rate of internet-based surveys directed to a group of patient portal members or the demographics of which portal members respond to internet-based surveys issued within that specific population. The objective of these analyses was to determine the demographics of patient portal users who respond to an internet-based survey request. We hypothesized that respondents would more likely be: (1) older (65+), (2) European American, (3) married, (4) female, (5) college educated, (6) have higher medical care utilization, (7) have more comorbidities, and (8) have a private practice primary care physician (as opposed to a salaried group practice primary care physician). We found that our respondents tended to be older, of European geographic ancestry, and more frequent users of healthcare. While patient portal members are an easily identifiable and contactable group that are potentially valuable participants for research, it is important to understand that respondents to surveys solicited from this sampling frame may not be entirely representative. It will be important to develop strategies to more fully engage populations that represent the target population in order to increase overall and subgroup response rates.

Multiagent neoadjuvant chemotherapy and tumor response are associated with improved survival in pancreatic cancer.

BACKGROUND: Neoadjuvant therapy (NT) for borderline resectable pancreatic cancer (BRPC) has evolved to include multi-agent regimens and chemoradiation. We report our experience and compare outcomes of initially resectable pancreatic cancer (IRPC) vs BRPC receiving NT across two eras of chemotherapy regimens. METHODS: Data were collected retrospectively on pancreaticoduodenectomy patients between January 2008 and October 2015. Outcomes and survival were compared based on patient, laboratory and treatment factors. RESULTS: 195 patients were included and 133 had IRPC and 62 BRPC. IRPC operations were shorter (449 min vs 520 min, p = 0.003), had less blood loss (663 ml vs 954 ml, p = 0.002) and involved fewer vascular resections (29% vs 76%, p = 0.002). The rate of R0 resection was identical (82%, p = 1) and the IRPC group had higher node-positive ratio (19.3% vs 7.2% p < 0.0001). 15 patients received a single agent regimen while 47 received multi-agent regimens with 90% receiving radiation.Survival was similar between BRPC and IRPC (log-rank p = 0.7). Histopathologic response (CAP grade 0 or 1) was not associated with survival (p = 0.13), but completion of ≥4 cycles of multi-agent pre-operative chemotherapy (p = 0.001) and complete response to NT (p = 0.04) were significant predictors of survival. CONCLUSIONS: BRPC patients treated with NT have similar morbidity and survival to their IRPC counterparts. Pathologic response and modern NT are associated with improved survival.

Mechanism Investigation of Advanced Metal-Ion-Mediated Amine Regeneration: A Novel Pathway to Reducing CO2 Reaction Enthalpy in Amine-Based CO2 Capture.

The high energy consumption of CO2 and absorbent regeneration is one of the most critical challenges facing commercial application of amine-based postcombustion CO2 capture. Here, we report a novel approach of metal-ion-mediated amine regeneration (MMAR) to advance the process of amine regeneration. MMAR uses the dual ability of amine to reversibly react with CO2 and reversibly complex with metal ions to reduce the enthalpy of the CO2 reaction, thus decrease the overall heat requirement for amine regeneration. To elucidate the mechanistic effects behind MMAR's ability to reduce CO2 reaction enthalpy, we developed a comprehensive chemical model describing the chemistry of Me(II)-monoethanolamine(MEA)-CO2-H2O system. The model was then validated using experimentally determined CO2 partial pressures via vapor-liquid equilibrium (VLE) measurements. We used the validated chemical model to gain insight into VLE behavior and solution chemistry, and to identify the specific changes in CO2 reaction enthalpy with and without metal ions. Two metals and five amines were evaluated in detail, which revealed that metal-ions with high complexation enthalpy and amines with large carbamate stability constant are preferred in MMAR, owing to their large reduction in reaction enthalpy and regeneration duty. We anticipate that MMAR could provide an alternative pathway to reducing the energy consumption of absorbent regeneration, ultimately making amine-based processes more technically and economically viable.

Measuring NDC80 binding reveals the molecular basis of tension-dependent kinetochore-microtubule attachments.

Proper kinetochore-microtubule attachments, mediated by the NDC80 complex, are required for error-free chromosome segregation. Erroneous attachments are corrected by the tension dependence of kinetochore-microtubule interactions. Here, we present a method, based on fluorescence lifetime imaging microscopy and Förster resonance energy transfer, to quantitatively measure the fraction of NDC80 complexes bound to microtubules at individual kinetochores in living human cells. We found that NDC80 binding is modulated in a chromosome autonomous fashion over prometaphase and metaphase, and is predominantly regulated by centromere tension. We show that this tension dependency requires phosphorylation of the N-terminal tail of Hec1, a component of the NDC80 complex, and the proper localization of Aurora B kinase, which modulates NDC80 binding. Our results lead to a mathematical model of the molecular basis of tension-dependent NDC80 binding to kinetochore microtubules in vivo.

Insights into the Chemical Mechanism for CO2(aq) and H+ in Aqueous Diamine Solutions - An Experimental Stopped-Flow Kinetic and 1H/13C NMR Study of Aqueous Solutions of N,N-Dimethylethylenediamine for Postcombustion CO2 Capture.

In an effort to advance the understanding of multiamine based CO2 capture process absorbents, we report here the determination of the kinetic and equilibrium constants for a simple linear diamine N,N-dimethylethylenediamine (DMEDA) via stopped-flow spectrophotometric kinetic measurements and 1H/13C NMR titrations at 25.0 °C. From the kinetic data, the formation of monocarbamic acid (DMEDACOOH) from the reaction of DMEDA with CO2(aq) is the dominant reaction at high pH > 9.0 (k7 = 6.99 × 103 M-1·s-1). Below this pH, the formation of protonated monocarbamic acid (DMEDACOOH2) via the pathway involving DMEDAH+ and CO2(aq) becomes active and contributes to the kinetics despite the 107-fold decrease in the rate constant between the two pathways. 1H and 13C NMR spectra as a function of decreasing pH (increasing HCl concentration) at 25.0 °C have been evaluated here to confirm the protonation events in DMEDA. Calculations of the respective DMEDA nitrogen partial charges have also been undertaken to support the NMR protonation study. A comparison of the DMEDA kinetic constants with the corresponding data for piperazine (PZ) reveals that despite the larger basicity of DMEDA, the enhanced and superior kinetic performance of PZ with CO2(aq) above its predicted Bronsted reactivity is not observed in DMEDA.

Evaluation and Modeling of Vapor-Liquid Equilibrium and CO2 Absorption Enthalpies of Aqueous Designer Diamines for Post Combustion Capture Processes.

Novel absorbents with improved characteristics are required to reduce the existing cost and environmental barriers to deployment of large scale CO2 capture. Recently, bespoke absorbent molecules have been specifically designed for CO2 capture applications, and their fundamental properties and suitability for CO2 capture processes evaluated. From the study, two unique diamine molecules, 4-(2-hydroxyethylamino)piperidine (A4) and 1-(2-hydroxyethyl)-4-aminopiperidine (C4), were selected for further evaluation including thermodynamic characterization. The solubilities of CO2 in two diamine solutions with a mass fraction of 15% and 30% were measured at different temperatures (313.15-393.15 K) and CO2 partial pressures (up to 400 kPa) by thermostatic vapor-liquid equilibrium (VLE) stirred cell. The absorption enthalpies of reactions between diamines and CO2 were evaluated at different temperatures (313.15 and 333.15 K) using a CPA201 reaction calorimeter. The amine protonation constants and associated protonation enthalpies were determined by potentiometric titration. The interaction of CO2 with the diamine solutions was summarized and a simple mathematical model established that could make a preliminary but good prediction of the VLE and thermodynamic properties. Based on the analyses in this work, the two designer diamines A4 and C4 showed superior performance compared to amines typically used for CO2 capture and further research will be completed at larger scale.

Computational Modeling and Simulation of CO2 Capture by Aqueous Amines.

We review the literature on the use of computational methods to study the reactions between carbon dioxide and aqueous organic amines used to capture CO2 prior to storage, reuse, or sequestration. The focus is largely on the use of high level quantum chemical methods to study these reactions, although the review also summarizes research employing hybrid quantum mechanics/molecular mechanics methods and molecular dynamics. We critically review the effects of basis set size, quantum chemical method, solvent models, and other factors on the accuracy of calculations to provide guidance on the most appropriate methods, the expected performance, method limitations, and future needs and trends. The review also discusses experimental studies of amine-CO2 equilibria, kinetics, measurement and prediction of amine pKa values, and degradation reactions of aqueous organic amines. Computational simulations of carbon capture reaction mechanisms are also comprehensively described, and the relative merits of the zwitterion, termolecular, carbamic acid, and bicarbonate mechanisms are discussed in the context of computational and experimental studies. Computational methods will become an increasingly valuable and complementary adjunct to experiments for understanding mechanisms of amine-CO2 reactions and in the design of more efficient carbon capture agents with acceptable cost and toxicities.

Pilot Study to Show the Feasibility of High-Resolution Sagittal Ultrasound Imaging of the Temporomandibular Joint.

PURPOSE: To show the feasibility of acquiring high-resolution sagittal ultrasound (US) images of the temporomandibular joint (TMJ). PATIENTS AND METHODS: We used commercially available US probes to assess the TMJ via a transoral soft tissue window to acquire sagittal images. Magnetic resonance imaging and clinical correlation were compared with the US findings by the consensus assessment of 2 of the senior investigators. RESULTS: The sample was composed of 10 TMJs (6 participants) with an age range of 34 to 71 years and a male-female ratio of 3:1. The condyle and subcondylar surface were visible in 10 of 10 joints (100%), the disc in 7 of 10 joints (70%), and the pterygoid muscles in 6 of 10 joints (60%). In the 5 joints with magnetic resonance correlation, disc position and configuration were confirmed in all cases. CONCLUSIONS: We show the first sagittal transoral sonograms of the TMJ disc and associated joint components.

Evaluation of Rural vs Urban Trauma Patients Served by 9-1-1 Emergency Medical Services.

Importance: Despite a large rural US population, there are potential differences between rural and urban regions in the processes and outcomes following trauma. Objectives: To describe and evaluate rural vs urban processes of care, injury severity, and mortality among injured patients served by 9-1-1 emergency medical services (EMS). Design, Setting, and Participants: This was a preplanned secondary analysis of a prospective cohort enrolled from January 1 through December 31, 2011, and followed up through hospitalization. The study included 44 EMS agencies transporting to 28 hospitals in 2 rural and 5 urban counties in Oregon and Washington. A population-based, consecutive sample of 67 047 injured children and adults served by EMS (1971 rural and 65 076 urban) was enrolled. Among the 53 487 patients transported by EMS, a stratified probability sample of 17 633 patients (1438 rural and 16 195 urban) was created to track hospital outcomes (78.9% with in-hospital follow-up). Data analysis was performed from June 12, 2015, to May 20, 2016. Exposures: Rural was defined at the county level by 60 minutes or more driving proximity to the nearest level I or II trauma center and/or rural designation in the Centers for Medicare & Medicaid Services ambulance fee schedule by zip code. Main Outcomes and Measures: Mortality (out-of-hospital and in-hospital), need for early critical resources, and transfer rates. Results: Of the 53 487 injured patients transported by EMS (17 633 patients in the probability sample), 27 535 were women (51.5%); mean (SD) age was 51.6 (26.1) years. Rural vs urban sensitivity of field triage for identifying patients requiring early critical resources was 65.2% vs 80.5%, and only 29.4% of rural patients needing critical resources were initially transported to major trauma centers vs 88.7% of urban patients. After accounting for transfers, 39.8% of rural patients requiring critical resources were cared for in major trauma centers vs 88.7% of urban patients. Overall mortality did not differ between rural and urban regions (1.44% vs 0.89%; P = .09); however, 89.6% of rural deaths occurred within 24 hours compared with 64% of urban deaths. Rural regions had higher transfer rates (3.2% vs 2.7%) and longer transfer distances (median, 97.4 km; interquartile range [IQR], 51.7-394.5 km; range, 47.8-398.6 km vs 22.5 km; IQR, 11.6-24.6 km; range, 3.5-97.4 km). Conclusions and Relevance: Most high-risk trauma patients injured in rural areas were cared for outside of major trauma centers and most rural trauma deaths occurred early, although overall mortality did not differ between regions. There are opportunities for improved timeliness and access to major trauma care among patients injured in rural regions.

Integrated Patient-Derived Models Delineate Individualized Therapeutic Vulnerabilities of Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDAC) harbors the worst prognosis of any common solid tumor, and multiple failed clinical trials indicate therapeutic recalcitrance. Here, we use exome sequencing of patient tumors and find multiple conserved genetic alterations. However, the majority of tumors exhibit no clearly defined therapeutic target. High-throughput drug screens using patient-derived cell lines found rare examples of sensitivity to monotherapy, with most models requiring combination therapy. Using PDX models, we confirmed the effectiveness and selectivity of the identified treatment responses. Out of more than 500 single and combination drug regimens tested, no single treatment was effective for the majority of PDAC tumors, and each case had unique sensitivity profiles that could not be predicted using genetic analyses. These data indicate a shortcoming of reliance on genetic analysis to predict efficacy of currently available agents against PDAC and suggest that sensitivity profiling of patient-derived models could inform personalized therapy design for PDAC.

Preoperative Modified FOLFIRINOX Treatment Followed by Capecitabine-Based Chemoradiation for Borderline Resectable Pancreatic Cancer: Alliance for Clinical Trials in Oncology Trial A021101.

IMPORTANCE: Although consensus statements support the preoperative treatment of borderline resectable pancreatic cancer, no prospective, quality-controlled, multicenter studies of this strategy have been conducted. Existing studies are retrospective and confounded by heterogeneity in patients studied, therapeutic algorithms used, and outcomes reported. OBJECTIVE: To determine the feasibility of conducting studies of multimodality therapy for borderline resectable pancreatic cancer in the cooperative group setting. DESIGN, SETTING, AND PARTICIPANTS: A prospective, multicenter, single-arm trial of a multimodality treatment regimen administered within a study framework using centralized quality control with the cooperation of 14 member institutions of the National Clinical Trials Network. Twenty-nine patients with biopsy-confirmed pancreatic cancer preregistered, and 23 patients with tumors who met centrally reviewed radiographic criteria registered. Twenty-two patients initiated therapy (median age, 64 years [range, 50-76 years]; 55% female). Patients registered between May 29, 2013, and February 7, 2014. INTERVENTIONS: Patients received modified FOLFIRINOX treatment (85 mg/m2 of oxaliplatin, 180 mg/m2 of irinotecan hydrochloride, 400 mg/m2 of leucovorin calcium, and then 2400 mg/m2 of 5-fluorouracil for 4 cycles) followed by 5.5 weeks of external-beam radiation (50.4 Gy delivered in 28 daily fractions) with capecitabine (825 mg/m2 orally twice daily) prior to pancreatectomy. MAIN OUTCOMES AND MEASURES: Feasibility, defined by the accrual rate, the safety of the preoperative regimen, and the pancreatectomy rate. RESULTS: The accrual rate of 2.6 patients per month was superior to the anticipated rate. Although 14 of the 22 patients (64% [95% CI, 41%-83%]) had grade 3 or higher adverse events, 15 of the 22 patients (68% [95% CI, 49%-88%]) underwent pancreatectomy. Of these 15 patients, 12 (80%) required vascular resection, 14 (93%) had microscopically negative margins, 5 (33%) had specimens that had less than 5% residual cancer cells, and 2 (13%) had specimens that had pathologic complete responses. The median overall survival of all patients was 21.7 months (95% CI, 15.7 to not reached) from registration. CONCLUSIONS AND RELEVANCE: The successful completion of this collaborative study demonstrates the feasibility of conducting quality-controlled trials for this disease stage in the multi-institutional setting. The data generated by this study and the logistical elements that facilitated the trial's completion are currently being used to develop cooperative group trials with the goal of improving outcomes for this subset of patients. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT01821612.

RNA Polymerase II cluster dynamics predict mRNA output in living cells.

Protein clustering is a hallmark of genome regulation in mammalian cells. However, the dynamic molecular processes involved make it difficult to correlate clustering with functional consequences in vivo. We developed a live-cell super-resolution approach to uncover the correlation between mRNA synthesis and the dynamics of RNA Polymerase II (Pol II) clusters at a gene locus. For endogenous ß-actin genes in mouse embryonic fibroblasts, we observe that short-lived (~8 s) Pol II clusters correlate with basal mRNA output. During serum stimulation, a stereotyped increase in Pol II cluster lifetime correlates with a proportionate increase in the number of mRNAs synthesized. Our findings suggest that transient clustering of Pol II may constitute a pre-transcriptional regulatory event that predictably modulates nascent mRNA output.