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Maegan M Whitworth,1 Krystal K Haase,1 David S Fike,1 Ravindra M Bharadwaj,2 Rodney B Young,3 Eric J MacLaughlin1,4 1Department of Pharmacy Practice, Texas Tech University Health Sciences Center (TTUHSC) School of Pharmacy, 2Department of Internal Medicine, 3Department of Family and Community Medicine, 4Departments of Family Medicine and Internal Medicine, TTUHSC School of Medicine, Amarillo, TX, USA Background: Scant literature exists evaluating utilization patterns for direct oral anticoagulants (DOACs). Objectives: The primary objective was to assess DOAC prescribing in patients with venous thromboembolism (VTE) and nonvalvular atrial fibrillation (NVAF) in outpatient clinics. Secondary objectives were to compare utilization between family medicine (FM) and internal medicine (IM) clinics, characterize potentially inappropriate use, and identify factors associated with adverse events (AEs). Methods: This was a retrospective cohort study of adults with NVAF or VTE who received a DOAC at FM or IM clinics between 10/19/2010 and 10/23/2014. Descriptive statistics were utilized for the primary aim. Fisher’s exact test was used to evaluate differences in prescribing using an adapted medication appropriateness index. Logistic regression evaluated factors associated with inappropriate use and AEs. Results: One-hundred twenty patients were evaluated. At least 1 inappropriate criterion was met in 72 patients (60.0%). The most frequent inappropriate criteria were dosage (33.0%), duration of therapy (18.4%), and correct administration (18.0%). Apixaban was dosed inappropriately most frequently. There was no difference in dosing appropriateness between FM and IM clinics. The odds of inappropriate choice were lower with apixaban compared to other DOACs (odds ratio [OR]=0.088; 95% confidence interval [CI] 0.008–0.964; p=0.047). Twenty-seven patients (22.5%) experienced an AE while on a DOAC, and the odds of bleeding doubled with each inappropriate criterion met (OR=1.949; 95% CI 1.190–3.190; p=0.008). Conclusion: Potentially inappropriate prescribing of DOACs is frequent with the most common errors being dosing, administration, and duration of therapy. These results underscore the importance of prescriber education regarding the appropriate use and management of DOACs. Keywords: rivaroxaban, dabigatran, apixaban, atrial fibrillation, venous thromboembolism, medication errors

Membrane distillation (MD) has received ample recognition for treating complex wastewater, including hypersaline oil and gas (O&G) produced water (PW). Rigorous water quality assessment is critical in evaluating PW treatment because PW consists of numerous contaminants beyond the targets listed in general discharge and reuse standards. This study evaluated a novel photocatalytic membrane distillation (PMD) process, with and without a UV light source, against a standard vacuum membrane distillation (VMD) process for treating PW, utilizing targeted analyses and a non-targeted chemical identification workflow coupled with toxicity predictions. PMD with UV light resulted in better removals of dissolved organic carbon, ammoniacal nitrogen, and conductivity. Targeted organic analyses identified only trace amounts of acetone and 2-butanone in distillates. According to non-targeted analysis, the number of suspects reduced from 65 in feed to 25-30 across all distillate samples. Certain physicochemical properties of compounds influenced contaminant rejection in different MD configurations. According to preliminary toxicity predictions, VMD, PMD with and without UV distillate samples, respectively contained 21, 22, and 23 suspects associated with critical toxicity concerns. Overall, non-targeted analysis together with toxicity prediction provides a competent supportive tool to assess treatment efficiency and potential impacts on public health and the environment during PW reuse., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Many gut microorganisms critical to human health rely on nutrients produced by each other for survival; however, these cross-feeding interactions are still challenging to quantify and remain poorly characterized. Here, we introduce a Metabolite Exchange Score (MES) to quantify those interactions. Using metabolic models of prokaryotic metagenome-assembled genomes from over 1600 individuals, MES allows us to identify and rank metabolic interactions that are significantly affected by a loss of cross-feeding partners in 10 out of 11 diseases. When applied to a Crohn's disease case-control study, our approach identifies a lack of species with the ability to consume hydrogen sulfide as the main distinguishing microbiome feature of disease. We propose that our conceptual framework will help prioritize in-depth analyses, experiments and clinical targets, and that targeting the restoration of microbial cross-feeding interactions is a promising mechanism-informed strategy to reconstruct a healthy gut ecosystem., (© 2023. Springer Nature Limited.)

Immune checkpoint inhibitors cause rare but potentially fatal neuromuscular complications, leading to a concern to use these agents in cancer patients with pre-existing autoimmune or inflammatory neuromuscular diseases. We report two such patients with paraneoplastic dermatomyositis and "seronegative" paraneoplastic demyelinating neuropathy, respectively, who have been successfully treated with immune checkpoint inhibitor monotherapy as well as maintenance intravenous immunoglobulin. While controlling the paraneoplastic or autoimmune neuromuscular diseases, the use of intravenous immunoglobulin did not compromise the anti-cancer effect of immune checkpoint inhibitor., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Xiong, Young, Chow, Maverakis, Maselli, Richman and Li.)

Globally, the anaerobic bacterium Clostridium perfringens causes severe disease in a wide array of hosts; however, C. perfringens strains are also carried asymptomatically. Accessory genes are responsible for much of the observed phenotypic variation and virulence within this species, with toxins frequently encoded on conjugative plasmids and many isolates carrying up to 10 plasmids. Despite this unusual biology, current genomic analyses have largely excluded isolates from healthy hosts or environmental sources. Accessory genomes, including plasmids, also have often been excluded from broader scale phylogenetic investigations. Here we interrogate a comprehensive collection of 464 C . perfringens genomes and identify the first putative non-conjugative enterotoxin (CPE)-encoding plasmids and a putative novel conjugative locus (Bcp) with sequence similarity to a locus reported from Clostridium botulinum . We sequenced and archived 102 new C. perfringens genomes, including those from rarely sequenced toxinotype B, C, D and E isolates. Long-read sequencing of 11 C . perfringens strains representing all toxinotypes (A-G) identified 55 plasmids from nine distinct plasmid groups. Interrogation of the 464 genomes in this collection identified 1045 plasmid-like contigs from the nine plasmid families, with a wide distribution across the C. perfringens isolates. Plasmids and plasmid diversity play an essential role in C. perfringens pathogenicity and broader biology. We have expanded the C. perfringens genome collection to include temporal, spatial and phenotypically diverse isolates including those carried asymptomatically in the gastrointestinal microbiome. This analysis has resulted in the identification of novel C. perfringens plasmids whilst providing a comprehensive understanding of species diversity.

Wildfires, which are increasing in frequency and severity in the western U.S., impact water quality through increases in erosion, and transport of nutrients and metals. Meanwhile, beaver populations have been increasing since the early 1900s, and the ponds they create slow or impound hydrologic and elemental fluxes, increase soil saturation, and have a high potential to transform redox active elements ( e.g. , oxygen, nitrogen, sulfur, and metals). However, it remains unknown how the presence of beaver ponds in burned watersheds may impact retention and transformation of chemical constituents originating in burned uplands ( e.g. , pyrogenic dissolved organic matter; pyDOM) and the consequences for downstream water quality. Here, we investigate the impact of beaver ponds on the chemical properties and molecular composition of dissolved forms of C and N, and the microbial functional potential encoded within these environments. The chemistry and microbiology of surface water and sediment changed along a stream sequence starting upstream of fire and flowing through multiple beaver ponds and interconnecting stream reaches within a burned high-elevation forest watershed. The relative abundance of N-containing compounds increased in surface water of the burned beaver ponds, which corresponded to lower C/N and O/C, and higher aromaticity as characterized by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). The resident microbial communities lack the capacity to process such aromatic pyDOM, though genomic analyses demonstrate their potential to metabolize various compounds in the anaerobic sediments of the beaver ponds. Collectively, this work highlights the role of beaver ponds as biological "hotspots" with unique biogeochemistry in fire-impacted systems.

Summary: Shotgun metagenomic sequencing provides the capacity to understand microbial community structure and function at unprecedented resolution; however, the current analytical methods are constrained by a focus on taxonomic classifications that may obfuscate functional relationships. Here, we present expam, a tree-based, taxonomy agnostic tool for the identification of biologically relevant clades from shotgun metagenomic sequencing., Availability and Implementation: expam is an open-source Python application released under the GNU General Public Licence v3.0. expam installation instructions, source code and tutorials can be found at https://github.com/seansolari/expam., Supplementary Information: Supplementary data are available at Bioinformatics online., (© The Author(s) 2022. Published by Oxford University Press.)

Background: Immune checkpoint inhibitor (ICI) therapy has improved survivals with a favorable toxicity profile in a variety of cancer patients. We hypothesized that hospitalized cancer patients who have acute or chronic comorbidities may have suppressed immune systems and poor clinical outcomes to ICIs. The objective of this study was to explore clinical outcomes and predictive factors of hospitalized cancer patients who received ICI therapy at an NCI-designated Comprehensive Cancer Center., Methods: A retrospective review of electronic medical records was conducted for adult cancer patients who received an FDA-approved ICI during admission from 08/2016 to 01/2022. For each patient we extracted demographics, cancer histology, comorbidities, reasons for hospitalization, ICI administered, time from treatment to discharge, time from treatment to progression or death, and complete blood counts. Progression-free survival (PFS) and overall survival (OS) were estimated using the Kaplan-Meier method and compared using the log-rank test. The 95% confidence interval for survival was calculated using the exact binomial distribution. Statistical significance was defined as 2-sided p <0.05., Results: Of 37 patients identified, 2 were excluded due to lack of complete blood counts on admission. Average hospital stay was 24.2 (95% CI 16.5, 31.9) days. Ten (27.0%) patients died during the same hospitalization as treatment. Of those who followed up, 22 (59.5%) died within 90 days of inpatient therapy. The median PFS was 0.86 (95% CI 0.43, 1.74) months and median OS was 1.55 (95% CI 0.76, 3.72) months. Patients with ≥3 comorbidities had poorer PFS (2.4 vs. 0.4 months; p=0.0029) and OS (5.5 vs. 0.6 months; p =0.0006). Pre-treatment absolute lymphocyte counts (ALC) <600 cells/µL were associated with poor PFS (0.33 vs. 1.35 months; p =0.0053) and poor OS (0.33 vs. 2.34 months; p =0.0236). Pre-treatment derived neutrophil to lymphocyte ratio (dNLR) <4 was associated with good median PFS (1.6 vs. 0.4 months; p=0.0157) and OS (2.8 vs. 0.9 months; p=0.0375)., Conclusions: Administration of ICI therapy was associated with poor clinical outcomes and high rates of both inpatient mortality and 90-day mortality after inpatient ICI therapy. The presence of ≥3 comorbidities, ALC <600/μL, or dNLR >4 in hospitalized patients was associated with poor survival outcomes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Young, Panchal, Ma, Chen, Steele, Iannucci and Li.)

Forest soil microbiomes have crucial roles in carbon storage, biogeochemical cycling and rhizosphere processes. Wildfire season length, and the frequency and size of severe fires have increased owing to climate change. Fires affect ecosystem recovery and modify soil microbiomes and microbially mediated biogeochemical processes. To study wildfire-dependent changes in soil microbiomes, we characterized functional shifts in the soil microbiota (bacteria, fungi and viruses) across burn severity gradients (low, moderate and high severity) 1 yr post fire in coniferous forests in Colorado and Wyoming, USA. We found severity-dependent increases of Actinobacteria encoding genes for heat resistance, fast growth, and pyrogenic carbon utilization that might enhance post-fire survival. We report that increased burn severity led to the loss of ectomycorrhizal fungi and less tolerant microbial taxa. Viruses remained active in post-fire soils and probably influenced carbon cycling and biogeochemistry via turnover of biomass and ecosystem-relevant auxiliary metabolic genes. Our genome-resolved analyses link post-fire soil microbial taxonomy to functions and reveal the complexity of post-fire soil microbiome activity., (© 2022. The Author(s).)

Fourier transform ion-cyclotron resonance mass spectrometry (FT-ICR MS) is the only mass analyzer that can resolve the molecular complexity of natural organic matter at the level of elemental composition assignment. Here, we leverage the high dynamic range, resolving power, resistance to peak coalescence, and maximum ion number and ion trapping duration in a custom built, 21 tesla hybrid linear ion trap /FT-ICR mass spectrometer for a dissolved organic matter standard (Suwanne River Fulvic Acid). We compare the effect of peak-picking threshold (3σ, 4σ, 5σ, and 6σ) on number of elemental composition assignments, mass measurement accuracy, and dynamic range for a 6.3 s transient across the mass range of m / z 200-1200 that comprises the highest achieved resolving power broadband FT-ICR mass spectrum collected to date. More than 36 000 species are assigned with signal magnitude greater than 3σ at root-mean-square mass error of 36 ppb, the most species identified reported to date for dissolved organic matter. We identify 18 O and 17 O isotopologues and resolve isobaric overlaps on the order of a few electrons across a wide mass range (up to m / z 1000) leveraging mass resolving powers (3 000 000 at m/z 200) only achievable by 21 T FT-ICR MS and increased by ∼30% through absorption mode data processing. Elemental compositions unique to the 3σ span a wide compositional range of aromaticity not detected at higher peak-picking thresholds. Furthermore, we leverage the high dynamic range at 21 T FT-ICR MS to provide a molecular catalogue of a widely utilized reference standard (SRFA) to the analytical community collected on the highest performing mass analyzer for complex mixture analysis to date. This instrument is available free of charge to scientists worldwide.

Case Presentation: A 70-year-old man was admitted to the hospital for planned chemotherapy for recently diagnosed CNS lymphoma. His medical history included follicular lymphoma (achieved remission 1 year prior with chemotherapy) and tonic-clonic seizure 1 month prior to admission, which led to his eventual biopsy-confirmed diagnosis of CNS lymphoma. Physical examination revealed temperature 36.4 °C, heart rate of 60 beats/min, BP of 160/81 mm Hg, and 98% oxygen saturation on room air. Neurologic condition, including mental status examination, was normal. His cardiac examination revealed regular rate and rhythm with normal first and second heart sounds without murmurs, rubs, or gallops. The remainder of the examination was unremarkable. Review of systems noted progressive and intermittent confusion prior to his seizure. He denied any shortness of breath, dyspnea on exertion, orthopnea, lower extremity edema, palpitations, or syncope. Laboratory data were unremarkable., (Copyright © 2022 American College of Chest Physicians. Published by Elsevier Inc. All rights reserved.)

Background: From consumption of fermented foods and probiotics to emerging applications of faecal microbiota transplantation, the health benefit of manipulating the human microbiota has been exploited for millennia. Despite this history, recent technological advances are unlocking the capacity for targeted microbial manipulation as a novel therapeutic., Aim: This review summarises the current developments in microbiome-based medicines and provides insight into the next steps required for therapeutic development., Methods: Here we review current and emerging approaches and assess the capabilities and weaknesses of these technologies to provide safe and effective clinical interventions. Key literature was identified through Pubmed searches with the following key words, 'microbiome', 'microbiome biomarkers', 'probiotics', 'prebiotics', 'synbiotics', 'faecal microbiota transplant', 'live biotherapeutics', 'microbiome mimetics' and 'postbiotics'., Results: Improved understanding of the human microbiome and recent technological advances provide an opportunity to develop a new generation of therapies. These therapies will range from dietary interventions, prebiotic supplementations, single probiotic bacterial strains, human donor-derived faecal microbiota transplants, rationally selected combinations of bacterial strains as live biotherapeutics, and the beneficial products or effects produced by bacterial strains, termed microbiome mimetics., Conclusions: Although methods to identify and refine these therapeutics are continually advancing, the rapid emergence of these new approaches necessitates accepted technological and ethical frameworks for measurement, testing, laboratory practices and clinical translation., (© 2022 The Authors. Alimentary Pharmacology & Therapeutics published by John Wiley & Sons Ltd.)

Background: The Patient Assessment of Chronic Illness Care (PACIC) tool measures patient satisfaction with chronic disease care. Objective: A modified PACIC tool (PACIC-RxFM) was used to assess patient satisfaction in a pharmacist-led chronic disease state management clinic. The secondary outcome compared satisfaction with pharmacist-led and physician-led visits. Methods: This cross-sectional study surveyed individuals with ≥ 1 chronic disease who saw a pharmacist (pharmacotherapy) or primary care provider (usual care) in the Texas Tech Physicians Family Medicine Clinic. The PACIC-RxFM survey included 15 items rated on a five-point Likert scale (5 = "always satisfied"). Results: A total of 107 patients with no significant differences in demographics or complexity between groups were surveyed. All mean domain scores indicated a high level of satisfaction in the pharmacotherapy group with statements regarding perceptions of care organization and encouragement to go to group classes yielding statistically significantly higher scores than the usual care group. Conclusion: Patients are satisfied with both providers' and pharmacists' involvement in chronic illness care. This involvement leads to significant improvement in patient perception of care organization., Competing Interests: Declaration of Conflicting Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2022.)

Wildfires in forested watersheds dramatically alter stored and labile soil organic matter (SOM) pools and the export of dissolved organic matter (DOM). Ecosystem recovery after wildfires depends on soil microbial communities and revegetation and therefore is limited by the availability of nutrients, such as nitrogen-containing and labile, water-soluble compounds. However, SOM byproducts produced at different wildfire intensities are poorly understood, leading to difficulties in assessing wildfire severity and predicting ecosystem recovery. In this work, water-extractable organic matter (WEOM) from laboratory microcosms of soil burned at discrete temperatures was characterized by ultrahigh-resolution Fourier transform ion cyclotron resonance mass spectrometry to study the impacts of fire temperature on SOM and DOM composition. The molecular composition derived from different burn temperatures indicated that nitrogen-containing byproducts were enriched with heating and composed of a wide range of aromatic features and oxidation states. Mass difference-based analysis also suggested that products formed during heating could be modeled using transformations along the Maillard reaction pathway. The enrichment of N-containing SOM and DOM at different soil burning intensities has important implications for ecosystem recovery and downstream water quality.

Per- and polyfluoroalkyl substances (PFASs) are a large family of thousands of chemicals, many of which have been identified using nontargeted time-of-flight and Orbitrap mass spectrometry methods. Comprehensive characterization of complex PFAS mixtures is critical to assess their environmental transport, transformation, exposure, and uptake. Because 21 tesla (T) Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS) offers the highest available mass resolving power and sub-ppm mass errors across a wide molecular weight range, we developed a nontargeted 21 T FT-ICR MS method to screen for PFASs in an aqueous film-forming foam (AFFF) using suspect screening, a targeted formula database (C, H, Cl, F, N, O, P, S; ≤865 Da), isotopologues, and Kendrick-analogous mass difference networks (KAMDNs). False-positive PFAS identifications in a natural organic matter (NOM) sample, which served as the negative control, suggested that a minimum length of 3 should be imposed when annotating CF 2 -homologous series with positive mass defects. We putatively identified 163 known PFASs during suspect screening, as well as 134 novel PFASs during nontargeted screening, including a suspected polyethoxylated perfluoroalkane sulfonamide series. This study shows that 21 T FT-ICR MS analysis can provide unique insights into complex PFAS composition and expand our understanding of PFAS chemistries in impacted matrices.

Wildfires affect soils through the formation of pyrogenic organic matter (pyOM) (e.g., char and soot). While many studies examine the connection between pyOM persistence and carbon (C) composition, nitrogen (N) transformation in wildfire-impacted systems remains poorly understood. Thermal reactions in wildfires transform biomass into a highly complex, polyfunctional, and polydisperse organic mixture that challenges most mass analyzers. High-field Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) is the only mass analyzer that achieves resolving powers sufficient to separate species that differ in mass by the mass of an electron across a wide molecular weight range ( m / z 150-1500). We report enhanced speciation of organic N by positive-ion electrospray ionization (ESI) that leverages ultrahigh resolving power ( m /Δ m 50% = 1 800 000 at m / z 400) and mass accuracy (<10-100 ppb) achieved by FT-ICR MS at 21 T. Isobaric overlaps, roughly the mass of an electron ( M e - = 548 μDa), are resolved across a wide molecular weight range and are more prevalent in positive ESI than negative ESI. The custom-built 21 T FT-ICR MS instrument identifies previously unresolved mass differences in C c H h N n O o S s formulas and assigns more than 30 000 peaks in a pyOM sample. This is the first molecular catalogue of pyOM by positive-ion ESI 21 T FT-ICR MS and presents a method to provide new insight into terrestrial cycling of organic carbon and nitrogen in wildfire impacted ecosystems.

Purpose: To determine whether a pharmacist-driven protocol improves guideline-concordant prescribing of diabetes medications in patients with atherosclerotic cardiovascular disease (ASCVD)., Methods: A retrospective pre- and post-intervention study was conducted at a university-based family medicine clinic. A pharmacist-driven protocol was implemented which involved the creation of an algorithm recommending specific diabetes medications in patients with ASCVD. An in-service presentation reviewing the algorithm and process for referral of eligible patients to an appointment with a clinical pharmacist was delivered to providers. Clinical pharmacist appointments focus was on improving diabetes management and initiating cardiovascular risk-reducing medications if appropriate., Results: A total of 234 patients were screened, and 108 met inclusion criteria. Upon completion of patient outreach, 34% were scheduled with a pharmacist. Forty-three percent of patients (16 of 37) attended the appointment. Of those, 31% were initiated on an evidence-based regimen indicated for diabetes and ASCVD. In comparing pre- to post-implementation of the pharmacist-driven protocol, the rate of guideline-concordant prescribing increased by 48% (3.8% to 5.6%)., Conclusion: Implementation of a pharmacist-driven protocol can increase guideline-concordant prescribing. However, further exploration of patient- and system-level barriers is necessary to implement such a program more broadly.

Biochar, a low-density yet carbon-rich material derived from different organic materials pyrolyzed under low or no oxygen conditions, has been widely studied as a soil amendment, for greenhouse gas mitigation and in remediation of trace element-contaminated soils. Molecular speciation of biochar compounds has been challenging due to low solubility, aggregation, and immense compositional polydispersity that challenges nearly all mass spectrometry methods routinely applied to carbon-based organic materials. Through a combined technique approach that applies advanced analytical strategies, we provide bulk and molecular characterization of Kentucky bluegrass biochar that can be applied to any biomass or biochar sample. First, we characterize Kentucky bluegrass biochar chemical functional groups by solid-state magic-angle spinning dynamic nuclear polarization NMR (MAS-DNP NMR) and resolve aromatic and aliphatic signals from the pyrogenic material and intact plant material. Next, we isolate water-soluble biochar species by solid-phase extraction followed by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and identify highly polar, oxygen species across a wide carbon number range. Solvent fractionation of biochar further expands the compositional range and identifies condensed polycyclic aromatic species across nonpolar and polar classes detected by two ionization modes (-ESI and +APPI) by FT-ICR MS. Plotting biochar species with DBE versus carbon number highlights the pericondensed molecular structural motif that persists across numerous heteroatom classes and ionization modes. To the best of our knowledge, this is the first molecular level identification of nonfunctionalized PAHs in biochar extracts by APPI FT-ICR MS. Thus, we identify biochar species that span the same compositional space as coal, heavy oil asphaltenes, and coal tar and correspond to condensed ring PAHs.

(1) Background: Endometriosis is characterized by the presence of endometrial glands and stroma outside of the uterus and is often associated with severe pelvic pain and infertility. Our study explored the utilization of B-Cell Lymphoma 6 (BCL6) and Sirtuin 1 (SIRT1) as potential biomarkers in serum, plasma, urine, and cervical mucus for a non-invasive diagnostic test for endometriosis. BCL6 was chosen based on its previously reported elevated expression in endometrial biopsies, and SIRT1 is co-expressed and upregulated in the endometrium of women with endometriosis. (2) Methods: BCL6 and SIRT1 levels were measured using enzyme-linked immunoassay (ELISA) in samples from 20 women with endometriosis (ten with stages I/II and ten with stages III/IV) and ten women without endometriosis. (3) Results: Levels of SIRT1 in sera showed a statistically significant elevation in advanced stages III/IV compared to controls and stages I/II. No significant differences were found in other bodily fluids for SIRT1 or any bodily fluids tested for BCL6. (4) Conclusions: These results suggest some potential of SIRT1 expression within serum as a predictor of advanced asymptomatic stages of endometriosis. Using immunohistochemistry (IHC) staining and H-SCORE values for the elevated BCL6 (and potentially SIRT1) levels in endometrial biopsy samples seems to have higher diagnostic potential based on the previously published studies.

The biogeochemical cycling of soil organic matter (SOM) plays a central role in regulating soil health, water quality, carbon storage, and greenhouse gas emissions. Thus, many studies have been conducted to reveal how anthropogenic and climate variables affect carbon sequestration and nutrient cycling. Among the analytical techniques used to better understand the speciation and transformation of SOM, Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) is the only technique that has sufficient mass resolving power to separate and accurately assign elemental compositions to individual SOM molecules. The global increase in the application of FTICR MS to address SOM complexity has highlighted the many challenges and opportunities associated with SOM sample preparation, FTICR MS analysis, and mass spectral interpretation. Here, we provide a critical review of recent strategies for SOM characterization by FTICR MS with emphasis on SOM sample collection, preparation, analysis, and data interpretation. Data processing and visualization methods are presented with suggested workflows that detail the considerations needed for the application of molecular information derived from FTICR MS. Finally, we highlight current research gaps, biases, and future directions needed to improve our understanding of organic matter chemistry and cycling within terrestrial ecosystems.

A growing number of experimental and computational approaches are illuminating the "microbial dark matter" and uncovering the integral role of commensal microbes in human health. Through this work, it is now clear that the human microbiome presents great potential as a therapeutic target for a plethora of diseases, including inflammatory bowel disease, diabetes and obesity. The development of more efficacious and targeted treatments relies on identification of causal links between the microbiome and disease; with future progress dependent on effective links between state-of-the-art sequencing approaches, computational analyses and experimental assays. We argue determining causation is essential, which can be attained by generating hypotheses using multi-omic functional analyses and validating these hypotheses in complex, biologically relevant experimental models. In this review we discuss existing analysis and validation methods, and propose best-practice approaches required to enable the next phase of microbiome research., Competing Interests: SCF is a consultant to BiomeBank, Australia. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Young, Marcelino, Chonwerawong, Gulliver and Forster.)

Oncogenic mutations and amplifications in the erythroblastic oncogene B ( ERBB2 ), or human epidermal growth factor receptor 2 ( HER2 ), have emerged as distinct oncogenic drivers and drug targets in non-small cell lung cancer (NSCLC). Each genomic alteration occurs in 2-4% of NSCLC by next generation sequencing and is associated with constitutive HER2 activation. The most common HER2 mutations in NSCLC are exon 20 mutation A775&#95;G776insYVMA mutation in the kinase domain and S310F mutation in the extracellular domain. Unlike in breast and gastric cancer, HER2 protein overexpression in NSCLC is not validated to be a biomarker predictive of clinical response to HER2-targeted agents. High HER2 protein overexpression by immunohistochemistry (3 + ) only occurs in 2-4% of NSCLC. Until now HER2-targeted agents (such as afatinib and ado-trastuzumab emtansine) only demonstrate modest clinical activity in patients with HER2 -mutant NSCLC. Retrospective studies show concern for inferior clinical benefit of immune checkpoint inhibitors in HER2 -mutated NSCLC. Therefore, platinum-based chemotherapy with or without an anti-angiogenesis inhibitor remains the first line standard treatment for this patient population. In May 2020 trastuzumab deruxtecan (T-DXd) received the U.S. Food and Drug Administration breakthrough therapy designation for HER2 -mutant metastatic NSCLC, and was added as an option for HER2 -mutant NSCLC to the NCCN guidelines V1.2021. A global phase III study of pyrotinib compared to docetaxel as a second line therapy for advanced NSCLC harboring HER2 exon 20 mutations was just opened for enrollment in September 2020. In this review, we highlight the current knowledge and perspectives on targeting- HER2 genomic alterations in NSCLC., Competing Interests: Dr. Li reports personal fees from Eisai, grants from Pfizer, grants from Merck, grants from Eureka, grants from OncoImmune, grants from Hengrui, grants from Tempus, outside the submitted work.

Galectins are a family of β-galactoside-binding proteins that contribute to multiple cellular functions, including immune surveillance and apoptosis. Human galectins are also important regulators of inflammation, making them a research target for various inflammatory diseases and tumorigenesis associated with pro-inflammatory conditions. This review focuses on the involvement of human galectins in modulation of inflammation and in the pathophysiology of endometriosis and endometriosis-associated neoplasms. Endometriosis is a chronic inflammatory disease with unknown etiology. Galectins -1, -3 and -9 were found to be overexpressed in ectopic and eutopic endometrium of females with endometriosis compared to those without endometriosis. These findings suggest galectins' role in the progression on endometriotic lesions and their potential use as diagnostic biomarkers and/or targets for therapeutic approaches. Galectins -1, -3, and -9 have also been implicated in the development of endometriosis-associated neoplasms. Furthermore, galectin-3 has been shown to interact with KRAS protein and contribute to cellular growth, proliferation, inflammation, and the uptake of nutrients in endometriotic lesions and may be involved in the maintenance and propagation of endometriosis. These galectins have been shown to be upregulated in certain forms of cervical, ovarian, endometrial, and colon cancer associated with endometriosis and have become a potential target for anti-cancer therapies., Competing Interests: The authors declare no conflict of interest.

The ACC/AHA's lower BP goals are supported by previously unavailable evidence, the strongest of which is for patients with CVD. But others can benefit, too.

Amending soil with biochar (pyrolized biomass) is suggested as a globally applicable approach to address climate change and soil degradation by carbon sequestration, reducing soil-borne greenhouse-gas emissions and increasing soil nutrient retention. Biochar was shown to promote plant growth, especially when combined with nutrient-rich organic matter, e.g., co-composted biochar. Plant growth promotion was explained by slow release of nutrients, although a mechanistic understanding of nutrient storage in biochar is missing. Here we identify a complex, nutrient-rich organic coating on co-composted biochar that covers the outer and inner (pore) surfaces of biochar particles using high-resolution spectro(micro)scopy and mass spectrometry. Fast field cycling nuclear magnetic resonance, electrochemical analysis and gas adsorption demonstrated that this coating adds hydrophilicity, redox-active moieties, and additional mesoporosity, which strengthens biochar-water interactions and thus enhances nutrient retention. This implies that the functioning of biochar in soil is determined by the formation of an organic coating, rather than biochar surface oxidation, as previously suggested.

Background: Assuring quality in cancer care through peer review has become increasingly important in radiation oncology. In 2012, our department implemented an automated electronic system for managing radiation treatment plan peer review. The purpose of this study was to compare the overall impact of this electronic system to our previous manual, paper-based system., Methods and Materials: In an effort to improve management, an automated electronic system for case finding and documentation of review was developed and implemented. The rates of missed initial reviews, late reviews, and missed re-reviews were compared for the pre- versus postelectronic system cohorts using Pearson χ 2 test and relative risk. Major and minor changes or recommendations were documented and shared with the assigned clinical provider., Results: The overall rate of missed reviews was 7.6% (38/500) before system implementation versus 0.4% (28/6985) under the electronic system (P < .001). In terms of relative risk, courses were 19.0 times (95% confidence interval, 11.8-30.7) more likely to be missed for initial review before the automated system. Missed re-reviews occurred in 23.1% (3/13) of courses in the preelectronic system cohort and 6.6% (10/152) of courses in the postelectronic system cohort (P = .034). Late reviews were more frequent during high travel or major holiday periods. Major changes were recommended in 2.2% and 2.8% in the pre- versus postelectronic systems, respectively. Minor changes were recommended in 5.3% of all postelectronic cases., Conclusions: The implementation of an automated electronic system for managing peer review in a large, complex department was effective in significantly reducing the number of missed reviews and missed re-reviews when compared to our previous manual system., (Copyright © 2016 American Society for Radiation Oncology. Published by Elsevier Inc. All rights reserved.)

Background and Objectives: Recent legislative and regulatory initiatives have emphasized preventive medicine and team-based health care delivery and education. Influenced by these initiatives, the investigators created an interprofessional teaching clinic to provide preventive care services (PCS) structured around Medicare's Annual Wellness Visit (AWV). The primary objective of this pilot study was to determine if PCS status improved for participating patients., Methods: AWV-naïve Medicare beneficiaries aged 66--74 years were recruited for the interprofessional teaching clinic, which involved physicians, pharmacists, and nurses. Patients were screened for 11 PCS variables, underwent medication review, and received recommendations to address identified PCS deficiencies prior to completing a satisfaction survey. Follow-up telephone visits were completed to determine recommendation outcomes and final PCS status for each variable. Descriptive statistics were used to characterize patients, the medication review, PCS status, and satisfaction scores. McNemar tests were used to assess the PCS status of patients before and after participation, and Fisher's Exact tests were used to compare baseline PCS status between the pilot cohort and a comparator group., Results: Thirty-four patients were enrolled in the pilot intervention, and one patient was lost to follow-up. Ninety-one percent (10/11) of PCS variables improved following participation. Significant improvements were observed for pneumococcal vaccination, mammography screening, fecal occult blood testing, and bone mineral density scanning. Patient satisfaction was high (mean scores for all items ?4.7)., Conclusions: This interprofessional teaching clinic provides a promising mechanism to improve patients' PCS status, medication utilization, and satisfaction while training students to function effectively as a team.

Soils sampled from different locations of China were used to manipulate soil microbial diversity and to assess the effect of the diversity of the soil nitrifying community on the recovery of the soil nitrification to metal stress (zinc). Ten treatments were either or not amended with ZnCl2. Subsequently, a spike-on-spike assay was set up to test for the tolerance of soil nitrification to zinc (Zn) and copper (Cu). Initially, Zn amendment completely inhibited nitrification. After a year of Zn exposure, recovery of the potential nitrification rate in Zn-amended soils ranged from 28 to 126% of the potential nitrification rate in the corresponding Zn-nonamended soils. This recovery was strongly related to the potential nitrification rate before Zn amendment and soil pH. Increased Zn tolerance of the soil nitrification was consistently observed in response to corresponding soil contamination. Co-tolerance to Cu was obtained in all 1,000-mg kg(-1) Zn-amended soils. This tolerance was also strongly related to the potential nitrification rate before Zn amendment and soil pH. Our data indicate that inherently microbial activity can be a significant factor for the recovery of soil functioning derived from metal contamination.

Objective: To determine if asthma control improves in patients who receive physician-pharmacist collaborative management (PPCM) during visits to primary care medical offices., Design: Prospective pre-post study of patients who received the intervention in primary care offices for 9 months. The primary outcome was the sum of asthma-related emergency department (ED) visits and hospitalizations at 9 months before, 9 months during, and 9 months after the intervention. Events were analyzed using linear mixed-effects regression. Secondary analysis was conducted for patients with uncontrolled asthma (Asthma Control Test [ACT] less than 20). Additional secondary outcomes included the ACT, the Asthma Quality of Life Questionnaire by Marks (AQLQ-M) scores, and medication changes., Intervention: Pharmacists provided patients with an asthma self-management plan and education and made pharmacotherapy recommendations to physicians when appropriate., Results: Of 126 patients, the number of emergency department (ED) visits and/or hospitalizations decreased 30% during the intervention (p=0.052) and then returned to preenrollment levels after the intervention was discontinued (p=0.83). Secondary analysis of patients with uncontrolled asthma at baseline (ACT less than 20), showed 37 ED visits and hospitalizations before the intervention, 21 during the intervention, and 33 after the intervention was discontinued (p=0.019). ACT and AQLQ-M scores improved during the intervention (ACT mean absolute increase of 2.11, AQLQ-M mean absolute decrease of 4.86, p<0.0001) and sustained a stable effect after discontinuation of the intervention. Inhaled corticosteroid use increased during the intervention (p=0.024)., Conclusions: The PPCM care model reduced asthma-related ED visits and hospitalizations and improved asthma control and quality of life. However, the primary outcome was not statistically significant for all patients. There was a significant reduction in ED visits and hospitalizations during the intervention for patients with uncontrolled asthma at baseline. Our findings support the need for further studies to investigate asthma outcomes achievable with the PPCM model., (© 2014 Pharmacotherapy Publications, Inc.)

Background: The Student Perceptions of Physician-Pharmacist Interprofessional Clinical Education (SPICE) instrument contains 10 items, 3 factors (interprofessional teamwork and team-based practice, roles/responsibilities for collaborative practice, and patient outcomes from collaborative practice), and utilizes a five-point response scale (1 = strongly disagree, 5 = strongly agree). Given the SPICE instrument's demonstrated validity and reliability, the objective of this study was to evaluate whether it was capable of measuring changes in medical (MS) and pharmacy students' (PS) perceptions following an interprofessional education (IPE) experience., Methods: In this prospective cohort study, MS and PS completed the SPICE instrument before and after participation in a predefined IPE experience. Descriptive statistics were used to characterize students and pre-post responses. Independent samples t tests and Fisher's Exact tests were used to assess group difference in demographic variables. Mann Whitney U tests were used to assess between-group differences in item scores. Wilcoxon Signed-Rank tests were used to evaluate post-participation changes in item scores. Spearman correlations were calculated to assess associations between ordinal demographic variables and item scores, and whether the number of clinic visits completed was associated with post-test responses. Paired samples t tests were used to calculate mean score changes for each of the factors., Results: Thirty-four MS and 15 PS were enroled. Baseline differences included age (25.3. ± 1.3 MS vs. 28.7 ± 4.4 PS; p = 0.013), years full-time employment (0.71 ± 0.97 MS vs. 4.60 ± 4.55 PS; p < 0.001), and number of prior IPE rotations (1.41 ± 1.74 MS vs. 3.13 ± 2.1 PS; p < 0.001). Two items generated baseline differences; 1 persisted post-participation: whether MS/PS should be involved in teamwork (3.91 MS vs. 4.60 PS; p < 0.001). For all students, significant mean score increases were observed for role clarity ("my role" [3.72 vs. 4.11; p = 0.001] and "others' roles" [3.87 vs. 4.17; p = 0.001]), impact of teamwork on patient satisfaction (3.72 vs. 4.34; p < 0.001), and ideal curricular location for IPE (4.06 vs. 4.34; p = 0.002). Significant increases were observed for all three factors (teamwork, p = 0.003; roles/responsibilities and patient outcomes, p < 0.001)., Conclusions: This study demonstrated the SPICE instrument's ability to measure changes in perception for medical and pharmacy students exposed to an IPE experience, both at the individual item level and at the factor level.

Lamotrigine is an antiepileptic and mood stabilizing drug that has been detected in wastewater, groundwater, surface water and drinking water, at frequencies in surface water ranging from 47 to 97%. Because lamotrigine is a weak base (pKa = 5.7) that appears in two protonation states in natural waters, this study examined the direct photodegradation of lamotrigine (11.4 to 12.0 mg L(-1)) in simulated sunlight using liquid chromatography-UV diode array detection and buffered aqueous solutions at pH 3.3, 5.3, and 7.7. Lamotrigine's half-life varied little (100 ± 3 to 112 ± 2 h) with solution pH, but its specific light absorption rate was 12 times higher, and its reaction quantum yield was 13 times lower, at pH 7.7 versus pH 3.3. In the estimated midday, midsummer sunlight in Denver, CO, USA (latitude 39.8617 °N), lamotrigine's estimated photodegradation rate was more than twice as fast at pH 7.7 versus pH 3.3. Lamotrigine's photoproducts were detected by liquid chromatography-UV diode array detection and time-of-flight mass spectrometry. Solution pH was shown to affect the identities and relative abundances of lamotrigine's photoproducts. Some photoproducts appeared only in solutions containing protonated lamotrigine, and others appeared only in solutions containing neutral lamotrigine. As a result, different reaction mechanisms were proposed. Finally, lamotrigine's reaction quantum yield (2.51 ± 0.07 × 10(-5) mol einstein(-1) at pH 7.7) and other results suggested that lamotrigine and three photoproducts are approximately as resistant to direct photodegradation as carbamazepine, a frequently detected pharmaceutical in surface waters.

Objectives: To describe the development and validation of an instrument designed to assess student perceptions of physician-pharmacist interprofessional clinical education (SPICE)., Methods: Faculty members from pharmacy and medical schools developed items for the instrument, and 179 medical and pharmacy students completed the scale. Psychometric properties, including reliability and construct validity, were assessed using confirmatory factor analysis., Results: The final instrument consisted of 10 items with 3 subscales measuring student perceptions of interprofessional teamwork and team-based practice, roles/responsibilities for collaborative practice, and patient outcomes from collaborative practice. Validity and reliability of the instrument were demonstrated., Conclusion: The SPICE instrument demonstrated promise as a valid and reliable measure of pharmacy and medical student perceptions of interprofessional clinical education. SPICE may serve as a useful instrument for educational researchers in assessing the impact of interprofessional educational experiences.

Study Objective: To categorize institutional review board (IRB) challenges and solutions encountered in a multicenter practice-based research network (PBRN) study and to assess the impact of IRB requirements on the willingness of individual principal investigators (PIs) to participate in future PBRN studies., Design: Descriptive analysis of IRB challenges and solutions encountered in the Collaboration Among Pharmacists and Physicians to Improve Outcomes Now (CAPTION) trial, a multicenter prospective cluster-randomized study conducted by the National Interdisciplinary Primary Care PBRN, and a correlational analysis from a survey of individual site PIs., Measurements and Main Results: IRB barriers encountered and solutions were categorized for study sites. A survey of study-site PIs was conducted with a correlational analysis assessing the impact of various IRB requirements and the willingness of individual PIs to participate in future PBRN studies; of 31 study sites participating in the CAPTION study, 28 study-site PIs were surveyed. IRBs posed a number of challenges including bias regarding the source of the application, issues regarding study design, study instruments, access to patient records, study procedures, Spanish-only speaking subjects, role of clinic physicians, interdepartmental concerns, and updates at continuing review. Responses from the PI survey (21 of 28 PIs surveyed [75% response rate]) indicated that the willingness of an individual to serve as a PI in the future was inversely related to the perceived difficulty of obtaining initial (rS  = -0.599, p=0.004) and continuing (rS  = -0.464, p=0.034) IRB approval., Conclusion: Significant time and resources were required to address various challenges associated with IRB approval, which had a negative impact on an individual PI's willingness to participate in future PBRN projects. A revision of current rules and regulations regarding the protection of human subjects for practice-based studies, improvement in IRB processes, and support from coordinating centers may decrease the burden associated with IRB approval and increase participation in practice-based research., (© 2013 Pharmacotherapy Publications, Inc.)

In surface waters, two of the most commonly observed androgenic steroid hormones are androstenedione (AD) and testosterone (T). This study compares the photodegradation of dilute (<10 μg L(-1)) aqueous solutions of AD and T in natural sunlight, and evaluates the endocrine-disrupting potential of the resulting solutions. This study also examines the effect of dissolved organic matter (DOM) on AD photodegradation. During spring and summer at Henderson, NV, USA (latitude 36.04°N), AD and T underwent direct photodegradation, with half-lives ranging from 3.7 to 10.8 h. In three model DOM solutions, AD's half-life increased by 11% to 35%. Using screening factors to eliminate DOM's inner filter effect, quantum yield calculations suggested that light screening was primarily responsible for AD's increased half-life, and that physical quenching further inhibited AD's photodegradation in two out of three DOM solutions. In vitro androgenic activity of the AD and T solutions decreased approximately as fast as AD and T were removed, suggesting that solar photodegradation reduces the risk of endocrine disruption in surface waters impacted by AD or T, subject to continuing inputs. Reduced in vitro androgenic activity appears to be related to steroid ring cleavage and the formation of highly oxidized photoproducts.

The artificial sweetener sucralose has recently been shown to be a widespread of contaminant of wastewater, surface water, and groundwater. In order to understand its occurrence in drinking water systems, water samples from 19 United States (U.S.) drinking water treatment plants (DWTPs) serving more than 28 million people were analyzed for sucralose using liquid chromatography tandem mass spectrometry (LC-MS/MS). Sucralose was found to be present in source water of 15 out of 19 DWTPs (47-2900 ng/L), finished water of 13 out of 17 DWTPs (49-2400 ng/L) and distribution system water of 8 out of the 12 DWTPs (48-2400 ng/L) tested. Sucralose was only found to be present in source waters with known wastewater influence and/or recreational usage, and displayed low removal (12% average) in the DWTPs where finished water was sampled. Further, in the subset of DWTPs with distribution system water sampled, the compound was found to persist regardless of the presence of residual chlorine or chloramines. In order to understand intra-DWTP consistency, sucralose was monitored at one drinking water treatment plant over an 11 month period from March 2010 through January 2011, and averaged 440 ng/L in the source water and 350 ng/L in the finished water. The results of this study confirm that sucralose will function well as an indicator compound for anthropogenic influence on source, finished drinking and distribution system (i.e., tap) water, as well as an indicator compound for the presence of other recalcitrant compounds in finished drinking water in the U.S.

Environmental releases and fate of steroid sex hormones from livestock and wastewater treatment plants are of increasing regulatory concern. Despite the detection of these hormones in manures, biosolids, and the environment, little attention has been paid to characterization of fecal bacteria capable of hormone degradation. The enrichments of (swine) manure-borne bacteria capable of aerobic testosterone degradation were prepared and the testosterone mineralization pathway was elucidated. Six DNA sequences of bacteria from the Proteobacteria phylum distributed among the genera Acinetobacter, Brevundimonas, Comamonas, Sphingomonas, Stenotrophomonas, and Rhodobacter were identified in a testosterone-degrading enriched culture with testosterone as the sole carbon source. Three degradation products of testosterone were identified as androstenedione, androstadienedione, and dehydrotestosterone using commercially available reference standards, liquid chromatography-UV diode array detection, and liquid chromatography-time-of-flight mass spectrometry (LC-TOF/MS). Three additional degradation products of testosterone were tentatively identified as 9α-hydroxytestosterone, 9α-hydroxyandrostadienedione or 3-hydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione, and 9α-hydroxydehydrotestosterone or 9α-hydroxyandrostenedione using LC-TOF/MS. When (14)C-testosterone was introduced to the enriched culture, 49-68% of the added (14)C-testosterone was mineralized to (14)CO(2) within 8 days of incubation. The mineralization of (14)C-testosterone followed pseudo-first-order reaction kinetics in the enriched culture with half-lives (t(1/2)) of 10-143 h. This work suggests that Proteobacteria play an important environmental role in degradation of steroid sex hormones and that androgens have the potential to be mineralized during aerobic manure treatment or after land application to agricultural fields by manure-borne bacteria.

Land application of manure may contribute endocrine disrupting compounds (EDCs) such as steroid hormones to the environment. Little attention has been paid to the potential for degradation of steroid hormones by manure-borne bacteria and their degradation kinetics and pathways. In a laboratory study, the potential for biodegradation of testosterone, 17beta-estradiol (E2) and progesterone by swine (Sus scrofa) manure-borne bacteria was examined. In addition, the impact of temperature, pH (6, 7, and 7.5), glucose amendments (0, 3, and 22 mmol L(-1)), and presence of oxygen on testosterone degradation kinetics was determined. Testosterone, 17beta-estradiol and progesterone were biodegraded within 25 h of reaction initiation under aerobic conditions. The degradation of testosterone followed pseudo first-order and zero-order reaction kinetics under aerobic and anaerobic conditions, respectively, in tryptic soy broth (TSB) pre-enriched systems. The half-life (t1/2) for the degradation of testosterone under anaerobic conditions was six times longer than aerobic conditions. Testosterone degradation was found to significantly increase (- 17%) when incubated at 37 degrees C vs. 22 degrees C. The impact of pH (t1/2 ranged from 4.4-4.9 h) and glucose amendments (t1/2 ranged from 4.6-5.1 h) on the testosterone degradation rate were found to be small. Testosterone was transformed to dehydrotestosterone (DHT) (major degradation product), androstenedione (AD), and androstadienedione (ADD) under aerobic conditions as revealed by liquid chromatography-time-of-flight mass spectrometry (LC/TOF-MS). These results indicate that testosterone is rapidly degraded by manure-borne bacteria under a wide range of environmentally relevant conditions. However, the formed degradation products are still of potential concern due to their endocrine disrupting potential.

Background: Despite the recent advances in cardiac mapping, ablation of scar-related ventricular tachycardia (VT) still remains a clinical challenge. A detailed electroanatomical map is a prerequisite for accurate localization and ablation of the VT substrate., Objective: The purpose of this study was to evaluate the feasibility and accuracy of integrating the positron emission tomography (PET)/computed tomography (CT) with the electroanatomical map and compare the accuracy of the voltage-based scar with the biological scar., Methods: Patients undergoing radiofrequency ablation (n = 19) for scar-related VT were enrolled. CT angiography and PET scans were performed for all patients. Tomographic and volumetric data from both images were processed and coregistered using internally designed software. That image was segmented in an electrophysiology mapping system and registered to the electroanatomical map. Eight different thresholds were applied on the voltage map to define the scar. The surface areas of the biological and electrical dense scars at different thresholds were measured and compared., Results: The PET/CT image was well integrated with the electroanatomical map with a mean surface registration error of 5.1 +/- 2.1 mm. Of the eight different thresholds defining the scar, the surface area of the scar at a threshold of 0.9 mV (68.6 +/- 49.2 cm(2)) correlated best with the surface area of the PET-based scar (70.4 +/- 49.3 cm(2)) and had the least total area error (4.8 +/- 1.8 cm(2)) compared with the 0.5 threshold (29.7 +/- 23.9 cm(2))., Conclusion: Integrating PET/CT with the electroanatomical map is feasible and accurate. Based on the biological scar, readjustment of the voltage scar threshold to 0.9 mV is suggested. In view of the better accuracy of PET/CT in defining scar, the need for acquiring detailed voltage maps may be obviated.

Broiler chickens at 35 d of age were fed 1 ppm clenbuterol for 14 d. This level of dietary clenbuterol led to 5-7% increases in the weights of leg and breast muscle tissue. At the end of the 14-d period, serum was prepared from both control and clenbuterol-treated chickens, and was then employed as a component of cell culture media at a final concentration of 20% (v/v). Muscle cell cultures were prepared from both the leg and the breast muscle groups of 12-d chick embryos. Treatment groups included control chicken serum to which 10 nM, 50 nM, and 1 microM clenbuterol had been added, as well as cells grown in media containing 10% horse serum. Cultures were subjected to each treatment for 3 d, beginning on the seventh d in culture. Neither the percent fusion nor the number of nuclei in myotubes was significantly affected by any of the treatments. The quantity of myosin heavy chains (MHCs) was not increased by serum from clenbuterol-treated chickens in either breast or leg muscle cultures; however, the MHC quantity was 50-150% higher in cultures grown in control chicken serum to which 10 and 50 nM clenbuterol had also been added. The beta-adrenergic receptor (betaAR) population was 4000-7000 betaARs per cell in cultures grown in chicken serum, with leg muscle cultures having approximately 25-30% more receptors than breast muscle cultures. Receptor population was not significantly affected by the presence of clenbuterol or by the presence of serum from clenbuterol-treated chickens. In contrast, the betaAR population in leg and breast muscle cultures grown in the presence of 10% horse serum was 16,000-18,000 betaARs per cell. Basal concentration of cyclic adenosine 3':5'monophosphate (cAMP) was not significantly affected by the treatments. When cultures grown in chicken serum were stimulated for 10 min with 1 microM isoproterenol, limited increases of 12-20% in cAMP concentration above the basal levels were observed. However, when cultures grown in the presence of horse serum were stimulated with 1 microM isoproterenol, cAMP concentration was stimulated 5- to 9-fold above the basal levels. Thus, not only did cells grown in horse serum have a higher betaAR population, but also each receptor had a higher capacity for cAMP synthesis following isoproterenol stimulation. Finally, the hypothesis that clenbuterol exerts its action on muscle protein content by changes in cAMP concentration was tested. No correlation was apparent between basal cAMP concentration and MHC content.

Skeletal muscle hypertrophy is promoted in vivo by administration of beta-adrenergic receptor (betaAR) agonists. Chicken skeletal muscle cells were treated with 1 microM isoproterenol, a strong betaAR agonist, between days 7 and 10 in culture. betaAR population increased by approximately 40% during this treatment; however, the ability of the cells to synthesize cyclic adenosine monophosphate (cAMP) was diminished by twofold. Neither the basal concentration of cAMP nor the quantity of myosin heavy chain (MHC) was affected by the 3-d exposure to isoproterenol. To understand further the relationship between intracellular cAMP levels, betaAR population, and muscle protein accumulation, intracellular cAMP levels were artificially elevated by treatment with 0-10 betaM forskolin for 3 d. The basal concentration of cAMP in forskolin-treated cells increased up to sevenfold in a dose-dependent manner. Increasing concentrations of forskolin also led to an increase in betaAR population, with a maximum increase of approximately 40-60% at 10 microM forskolin. A maximum increase of 40-50% in the quantity of MHC was observed at 0.2 microM forskolin, but higher concentrations of forskolin reduced the quantity of MHC back to control levels. At 0.2 microM forskolin, intracellular levels of cAMP were higher by approximately 35%, and the betaAR population was higher by approximately 30%. Neither the number of muscle nuclei fused into myotubes nor the percentage of nuclei in myotubes was affected by forskolin at any of the concentrations studied.

Expression of the beta-adrenergic receptor (betaAR) and its coupling to cyclic AMP (cAMP) synthesis are important components of the signaling system that controls muscle atrophy and hypertrophy, and the goal of this study was to determine if electrical stimulation in a pattern simulating slow muscle contraction would alter the betaAR response in primary cultures of avian and mammalian skeletal muscle cells. Specifically, chicken skeletal muscle cells and rat skeletal muscle cells that had been grown for 7 d in culture were subjected to electrical stimulation for an additional 2 d at a pulse frequency of 0.5 pulses/sec and a pulse duration of 200 msec. In chicken skeletal muscle cells, the betaAR population was not significantly affected by electrical stimulation; however, the ability of these cells to synthesize cyclic AMP was reduced by approximately one-half. In contrast, the betaAR population in rat muscle cells was increased slightly but not significantly by electrical stimulation, and the ability of these cells to synthesize cyclic AMP was increased by almost twofold. The basal levels of intracellular cyclic AMP in neither rat muscle cells nor chicken muscle cells were affected by electrical stimulation.