Medical Research Foundation Research Articles

Clinical questionnaire filling based on question answering framework

BackgroundElectronic Health Records (EHR) are the foundation of much medical research. However, analyzing the text data of EHRs directly is an challenging task. Therefore, physicians often use questionnaires to first convert text data to structured data. Filling in these questionnaires requires a considerable amount of time and medical knowledge. It is a significant task to develop an algorithm to make computers fill out these questionnaires automatically. ObjectiveThis research aims to build a deep learning model that can automatically complete questionnaires with given medical text. MethodsThis task is a part of Information Extraction (IE), but it differs from the existing tasks of medical IE. Because of the questions in questionnaires are closed-end type, which refers to making a selection among given options, we could treat this task as a classification problem. However, conventional classification algorithms are resource-consuming when filling out one entire questionnaire with one model. They also could not use the question information to guide the questionnaire filling task. To handle these issues, we propose a neural network model based on question answering (QA) framework in this paper. With this framework, our neural network can fill out one complete questionnaire using only one model. ResultsWe perform experiments on three real-world Chinese medical datasets and related clinical questionnaires. Our model respectively achieves F1 scores of 0.9273, 0.8834, and 0.9846. The results outperform several baseline models. ConclusionThe strong performance of our QA model will allow us to build a system which can help physicians to fill out questionnaires and convert text data to structured data. This system can reduce the workload of physicians.

A Foundation for the future: How the Medical Research Foundation is looking long-term in its medical research funding

A Foundation for the future: How the Medical Research Foundation is looking long-term in its medical research funding

2310-PUB: Chronic Western Diets Initiate Fibrosis Progression in Nonalcoholic Fatty Liver of Male Mice Modifying the PPARγ Pathway via Mir-27b-5p Regulation

Increased fructose intake in Western diets contributes to metabolic diseases. However, the epigenetic mechanisms are mostly unknown. Here, six week-old C57BL/6J male and female mice were fed with a low-fat diet (LFD), high-fat diet (HFD), and high-fat high-fructose diet (HFD-HF) for 20 weeks. We determined that HFD-HF or HFD mice experienced significant metabolic dysregulation compared to the LFD. Metabolic changes in male mice were significantly dysregulated compared to female mice. HFD-HF and HDF male-specific responses included physiological/metabolic changes such as significant increased body weight, increased liver size, increased fasting glucose levels and downregulated PPARγ (P < 0.001), SCD1 (P < 0.05 to P < 0.01), and FAS (P < 0.05) protein expression. In contrast, female mice were less affected by HFD and HFD-HF. Interestingly, none of the groups exhibited less insulin sensitivity after 20 weeks of feeding when compared to LFD. As miR-27b contains a seed sequence in PPARγ, it was discovered that these changes are accompanied by male-specific upregulation of miR-27b-5p, which is even more pronounced in the HFD-HF group (P < 0.01 vs. LFD) compared to the HFD group (P < 0.05 vs. LFD). Other miR-27 subtypes were increased but not significantly. Fat ballooning in hepatocytes were increased in HFD fed mice compared to HFD-HF fed mice, however, the HFD-HF liver showed an increase in the number of small cells. Here, we concluded that our chronic western diet-composition administered for 20 weeks surpassed the nonalcoholic fatty liver (NAFL) stage and initiated liver fibrosis via miR-27b-5p-induced PPARγ downregulation. Disclosure J. Zhang: None. C. Powell: None. M.K. Kay: None. R. Sonkar: None. S. Meruvu: None. M. Choudhury: None. Funding Morris L. Lichtenstein, Jr. Medical Research Foundation

4013 Does dietary fat composition predict short-term elevations in lipid levels in adults on a modified Atkins diet?

OBJECTIVES/GOALS: The modified Atkins diet (MAD) is used in the management of drug-resistant epilepsy in adults. Some patients on MAD show an increase in serum levels of total cholesterol and low-density lipoprotein (LDL) cholesterol. We explored whether dietary fat composition predicts short-term elevations in serum lipid levels in diet-naïve adults who begin MAD. METHODS/STUDY POPULATION: Participants self-reported their diet intake with 3-day food records at baseline, 1 month and 2 months. Food records were analyzed using Nutrition Data System for Research software. Fasting serum levels of total cholesterol (TC), high-density lipoprotein (HDL) cholesterol, and triglycerides were also collected and LDL level calculated at baseline, 1 month, and 2 months. RESULTS/ANTICIPATED RESULTS: 38 patients submitted complete food records at each study visit (baseline, 1 month, and 2 month). Compared to baseline diet intake, there was a significant reduction in daily carbohydrate intake at 1 and 2 months months (p<0.001). There was also a significant increase in daily saturated fatty acid (SFA) intake at 1 and 2 months (p<0.001), daily mono-unsaturated fatty acid (MUFA) intake at 1 and 2 months (p<0.001), and daily cholesterol intake at 1 month (p<0.05) and 2 months (p<0.001), but no change in daily poly-unsaturated fatty acid (PUFA) intake over time. Compared to baseline, there was a significant increase in serum LDL at 1 month (p<0.001) and 2 months (p<0.01) and an increase in serum TC at 1 month (p<0.01) but not 2 months. DISCUSSION/SIGNIFICANCE OF IMPACT : Despite a significant increase in total fat, saturated fat and mono-unsaturated fat intake as well as an increase in total cholesterol and LDL levels following MAD initiation, dietary fat composition appears to minimally predict serum lipid values in the short term. CONFLICT OF INTEREST DESCRIPTION: Tanya McDonald has received speaking honoraria from Nutricia North America. Bobbie Henry-Barron receives grants from Johns Hopkins Institute for Clinical and Translational Research (ICTR) which is funded in part by Grant Number UL1 TR 001079 from the National Center for Advancing Translational Sciences (NCATS) a component of the National Institutes of Health (NIH), and NIH Roadmap for Medical Research, Nutricia and Vitaflo. Diane Vizthum receives grants from the Johns Hopkins Institute for Clinical and Translational Research (ICTR) which is funded in part by Grant Number UL1 TR 001079 from the National Center for Advancing Translational Sciences (NCATS) a component of the National Institutes of Health (NIH), and NIH Roadmap for Medical Research. Mackenzie C. Cervenka has received grant support from Nutricia North America, Vitaflo, Army Research Laboratory, The William and Ella Owens Medical Research Foundation and BrightFocus Foundation. She receives speaking honoraria from LivaNova, Epigenix, Nutricia North America and the Glut1 Deficiency Foundation and performs consulting with Nutricia North America and Sage Therapeutics and receives Royalties from Demos Health.

Pilot Study on the Effects of a Plant-Strong Diet on Cardiovascular Risk Factors in Veterans (Part 1)

Abstract Objectives A short-term intervention designed to test the feasibility of Veterans to adopt a whole-food, plant-based (WFPB) diet has not been tested. The specific aims of this study were to (1) determine the feasibility of Veterans adopting a WFPB diet within four weeks, following a plant-strong protocol, (2) determine the effectiveness on participant's cardiovascular risk factors and dietary patterns and (3) compare the results to Veterans following the American Heart Association or Diabetic Diets. Methods Prospective, nonrandomized, pilot study of overweight or obese hyperlipidemic Veterans naive to lipid lowering drugs. The first 4 participants were assigned the control group, the next 6 were assigned the intervention. Participants in the control group who met inclusion/exclusion criteria after completion of control intervention could cross-over into the intervention group. The intervention was a four-week program which included individual education on a WFBP diet, (including menu planning and making grocery lists), a grocery store tour, cooking skills, substitutions and food preparation, and challenges dining out. Participants in the control arm received the standard of care. Anthropometrics, biochemical measures, and dietary intake patterns were obtained at baseline and five weeks. Results Eight male Veterans (mean 57 years old) completed the study, (n = 4 control, n = 6 intervention, 2 crossed over). Ability to adopt a WFBP diet was 83%. Data from diet adopters were compared to the control. Both groups had a significant decrease in LDL from baseline (12% control and 19% in intervention, P = 0.04 each). Veterans in the intervention group had a significant decrease in weight (3%, P = 0.04) and reported: gassiness, better digestion, more energy and better sleep. Conclusions Veterans were able to adopt a WFBP diet and experienced a favorable change in LDL cholesterol. Resources needed to conduct the intervention preclude widespread implementation. Further research is needed to determine if less resource intensive programs are feasible and effective for long-term sustainability and determine which Veterans may benefit most. Funding Sources Carl T Hayden Medical Research Foundation.

0348 Sleep EEG-Based Brain Age Index is Reduced Under Continuous Positive Airway Pressure Treatment

Abstract Introduction Continuous positive airway pressure (CPAP) is a treatment for apnea. With long-term CPAP, changes in electroencephalogram (EEG) include increased delta power (1 - 4Hz) and sigma power (11 - 15Hz, spindle). However, the short-term EEG response to CPAP in a split-night study is less quantified. We recently developed a “brain age” model using sleep EEG features. The brain age index (BAI) is defined as the difference between chronological age and brain age (BA - CA). Here we first quantify how BAI changes during CPAP in the same patient, and then investigate how much brain age features during the diagnostic part can predict the reduction in apnea-hypopnea index (AHI) during CPAP. Methods The dataset consisted of 160 subjects. The average age was 59 years with 53% male, 24% female and 23% unknown. We extracted 480 features including band powers, and then computed the BAIs for both diagnostic and CPAP parts. To predict the reduction in AHI during CPAP, we fit a Bayesian regression model using the brain age features, demographics, and sleep parameters during the diagnostic part, and assessed the feature importance using dominance analysis. Results The BAI from the diagnostic part is significantly reduced compared to BAI during CPAP for the same subject (paired t-test, p < 0.01). The diagnostic part has an average BAI 2.24 years; and the CPAP part -4.75 years. The brain age features that are increased during CPAP include sigma powers in N2 and N3. The prediction of AHI reduction has Pearson’s correlation 0.85. The features predictive of reduced AHI are the diagnostic AHI (explained variance 69%), followed by high/low waveforms during N2 (e.g. K-complex, measured by kurtosis) (8.6%), delta power during REM (4.5%) and N1 (2%). The feature predictive of increased AHI is frontal alpha power during quiet awake (2.6%). Conclusion The average BAI is reduced during CPAP. BAI provides a novel view of the acute response to CPAP in sleep EEG. Future study with more CPAP failure patients has the potential of predicting CPAP failure. Support MBW is supported by Glenn Foundation for Medical Research. RJT is supported by Category I AASM Foundation.

0035 Resting Metabolism and the Metabolic Response to Exercise Follow Circadian Patterns with Day/Night Differences in Substrate Utilization Between Lean and Obese Adults

Abstract Introduction Resting energy expenditure (EE) follows a circadian rhythm in healthy lean participants, with a nadir in the early morning hours. We determined: (1) whether this pattern persists (or how substrate utilization may change), when challenged with exercise, and; (2) whether obesity affects these responses. Methods Fourteen participants (aged 48.5±12.8y; 6-female; 5-obese, BMI 31.9±1.4kg/m2 [avg±SD]) underwent a 5-day inpatient forced desynchrony protocol, comprised of ten 5h 20min ‘days’ in dim-lighting and free of time cues. Resting EE was measured immediately prior to a 15-minute cycle ergometer exercise bout at 50% of estimated heart rate maximum. Substrate utilization was determined from the respiratory quotient (RQ). Circadian phase was calculated using the salivary dim-light melatonin onset (>3pg/mL threshold). EE data were analyzed using a mixed-effect model with group (lean vs. obese) and circadian phase as fixed factors; subject was a random factor. RQ was analyzed using t-tests to determine day/night differences in groups at rest and in response to exercise. Results Resting and exercising EE both displayed endogenous circadian rhythms (p<0.05) with nadirs in the early morning (~5:30am), without any differences between groups (p>0.22). Resting RQ was similar between the day and night in the lean group (p=0.66), but decreased (suggesting lower carbohydrate utilization) at night within the obese group (-2.5±1.6%, p=0.02). The lean group increased RQ in response to exercise both during the day (+8.9±2.8%) and night (+8.0±2.8%) (both p<0.001), but there was no increase in RQ in the obese group during either day or night exercise (p>0.16). Conclusion These data demonstrate that EE during rest and exercise follows a circadian pattern, with limited influence of obesity. Circadian differences in substrate utilization between lean and obese in the resting state and in response to exercise may play a role in expression and maintenance of unwanted weight gain and impaired metabolic health. Support R01HL125893, R01HL140577, KL2TR002370, K01HL146992, F32HL131308, Medical Research Foundation of Oregon, Ford Foundation, and CTSA grant (UL1TR000128)

Editorial Note for Journal of Chemical Engineering and Process Technology

I am pleased to introduce Journal of Chemical Engineering & Process Technology (JCEPT) a rapid peer reviewed Journal which provides an opportunity to share the information among the medical scientists and researchers. It comes under the various field like physical science, chemistry, physics biochemistry and so on I am pleased to announce that, all issues of volume 10 were published online well within the time and the print issues were also brought out and dispatched within 30 days of publishing the issue online during the year of 2019. The Journals aims to flourish and to publish most complete and reliable source of information on the discoveries and current developments in the mode of original articles, review articles, case reports, short communications, etc. in all areas of the field and making them freely available through online without any restrictions or any other subscriptions to researchers worldwide. The Index Copernicus value of the journal is 97.45. And also the journal is indexed in Google Scholar, CAS Source Index (CASSI), Open J Gate, RefSeek, OCLC- WorldCat, Sherpa Romeo, Genamics JournalSeek, JournalTOCs, Ulrich's Periodicals Directory, Directory of Research Journal Indexing (DRJI), Hamdard University, EBSCO A-Z, Scholarsteer, Publons, Geneva Foundation for Medical Education and Research. During the calendar year of 2019, JCEPT received a total of 40 manuscript, out of which 28 articles (70%) were rejected in the preliminary screening due to plagiarism or being out of the format and peer review process. During 2019 around 10 articles were subjected for publication after they are accepted in the peer review process. In the 5 issues of Volume 10 published during the year 2019, a total of 10 articles were published (at an average of 5 articles per issue) of which, articles were published from authors all around the world. Globally articles have been accessed by the research scientists and also cited. During the calendar year 2019, a total of two Editors, five Reviewers joined the board of JCEPT and contributed their valuable services towards contribution as well as publication of articles, and their valuable reviewer comments will beneficial to publish quality of article in the Journal. I take this opportunity to acknowledge the contribution of Dr. Garry Chen during the final editing of articles published and the support rendered by the editorial assistant, in bringing out issues of JCEPT in time. I would also like to express my gratitude to all the authors, reviewers, the publisher, the advisory and the editorial board of JCEPT, the office bearers for their support in bringing out yet another volume of JCEPT and look forward to their unrelenting support to bring out the Volume 12 of JCEPT in scheduled time.

Vascular surgery's identity

Vascular surgery's identity

Structural Disorder and Induced Folding of the WWC Family Proteins

The WWC family consists of three proteins‐ KIBRA/WWC1, WWC2, and WWC3. Each protein contains two WW domains which recognize proline‐rich sequences of the type PPxY (P is proline, Y is tyrosine, x is any amino acid). WW domains are named for two conserved tryptophans spaced 23 residues apart and adopt a three‐stranded antiparallel beta sheet fold. The WWC proteins are “atypical” in that, the second WW domain (WW2) contains an isoleucine rather than a conserved tryptophan. Recent studies of the KIBRA WW domains shows that this substitution leads to unfolding of the WW2 domains. However, the domain undergoes binding‐induced folding when bound to some partners but remains largely unfolded when bound to others. High sequence similarity of the WW domains in the WWC family suggests that unfolding and binding induced folding of the WW2 domain may be conserved in this family, but it is unclear why some partners induce folding of the WW2 domain while others do not. Our working hypothesis is that partners which contain PPxY motifs in close proximity will bind the WWC proteins with relatively high affinity and induce folding of the WW2 domain. In this study, we test this hypothesis with two binding partners, Large Tumor Suppressor 1 (LATS1), a kinase and tumor suppressor, and Synaptopodin (SYNPO), a synaptic and podocyte protein. We demonstrate by isothermal titration calorimetry that wildtype LATS1 and SYNPO which have two PPxY motifs each, separated by 194 and 15 residues respectively, bound KIBRA and WWC3 with modest micromolar binding affinities. However, mutant LATS1 (LATS1‐VA) and SYNPO (SYNPO_AA) constructs in which the PPxY motifs are separated by two amino acids, bound KIBRA and WWC3 with 20 to 180‐fold enhanced affinity. Using solution magnetic resonance spectroscopy, we demonstrated that the WW2 domain of KIBRA and WWC3 remain unfolded when bound to wildtype LATS1 and SYNPO, while the enhanced binding of the mutants LATS1_VA and SYNPO_AA is accompanied by folding of the WW2 domain. The structural heterogeneity of the WW2 domain may be an important for modulating binding to diverse PPxY motif partners.Support or Funding InformationThis work is supported by start‐up funds from Oregon State University (OSU), OSU College of Science Disease Mechanism and Prevention Fund, and a New Investigator Award from the Medical Research Foundation of Oregon. NMR experiments were collected at the Oregon State University NMR Facility funded in part by the National Institutes of Health, HEI Grant 1S10OD018518, and by the M. J. Murdock Charitable Trust grant # 2014162

Sex‐differences in Statin Effects: Long‐term Atorvastatin Administration Reduced LDL‐Cholesterol Levels and Body Weights only in Male Mice, but Decreased Voluntary Cage Activity in Male and Female Mice

IntroductionStatins inhibit HMG‐CoA reductase and are used to lower low‐density lipoprotein‐cholesterol (LDL‐C) for cardiovascular disease (CVD) prevention. Besides the benefits of reducing CVD‐related morbidity and mortality, adverse events have been reported by the FDA. Statin‐induced skeletal myopathy is the most reported statin adverse event. While the precise mechanism is undefined, there is considerable evidence that a statin‐induced impairment of oxidative phosphorylation within mitochondria is involved. We and others have previously shown that Atorvastatin (Ator) administration induces mitochondria alterations.RationalBecause observational studies report a high incidence of statin‐myopathy and females seem to be more susceptible to statin adverse events, we aimed to generate a mouse model mimicking statin‐induced muscular side effects.Methods8‐week‐old male (M) and female (F) C57BL6J mice received either Ator (5mg/kg/day) or vehicle (Veh, 10% EtOH in water,) by daily oral gavage (4 groups, M+Ator, F+Ator, M+Veh, and F+Veh, n= 9–11 each). Body weights (BW) were recorded in two‐week intervals. 3‐month(m) post statin administration a lipid panel, liver enzymes, and creatine kinase (CK) was analyzed (n=8 each). Voluntary home cage activity was assessed 4‐m post statin treatment.ResultsA) M+Veh mice presented higher total‐C and high‐density lipoprotein (HDL)‐C serum levels than F+ Veh mice (total‐C mg/dl, Mean ± SEM; M+Veh 131.4 ± 7.4, F+Veh 90.5 ± 4.8, M+Veh vs F+Veh, * p<0.0004, and HDL‐C mg/dl, Mean ± SEM; M+Veh 82.8 ± 3.3, F+Veh 58.6 ± 3.4, M+Veh vs F+Veh, * p<0.0002). Long‐term Ator administration; B) induced lower BW (g) in male mice, but not in females starting at 1m post Ator administration (6m post body weight, Mean ± SEM; M+Veh 32.9g ± 0.8, M+Ator 28.4g ± 0.7, M+Veh versus M+Ator * p<0.0005; F+Veh 23.2g ± 0.6, F+Ator 22.7g ± 0.4, F+Veh vs F+Ator, not significant (ns)), C) reduced LDL‐C serum levels in M+Ator, but not in F+Ator mice (LDL‐C direct, Mean ± SEM; M+Veh 8.25 ± 0.41, M+Ator 7.12 ± 0.29, M+Veh versus M+Ator * p<0.04; F+Veh 9.25 ± 0.45, F+Ator 8.38 ± 0.56, F+Veh vs F+Ator, ns), D) did not alter total‐C or HDL‐C levels, E) elevated alkaline phosphatase levels in M+Ator but not in F+Ator mice, 5) did not alter CK serum levels, F) induced the development of gray hair only in M+Ator, and G) reduced voluntary cage activity in M+Ator and F+Ator mice, when compared to M+Veh and F+Veh.ConclusionLong‐term Ator administration reduced LDL‐C serum levels and BW in male mice but not in female mice. Although CK‐serum levels, a marker for skeletal muscle damage, were not significantly changed, voluntary cage activity was reduced in male and female Ator treated mice when compared to Veh treated controls. We suggest for the first time that long‐term Ator administration can induce a mouse‐model of statin‐induced myopathy.Support or Funding InformationVeterans Medical Research Foundation Pilot Project Award: Alice Zemljic‐Harpf (PI), University of California, San Diego, ANES Research Seed Award 2019: Alice Zemljic‐Harpf (PI)

The inflammatory cytokine, interleukin‐1‐beta, drives central immune cell infiltration, changes in sympathetic nerve activity and blood pressure

IntroductionThe area postrema (AP) is a brain region that lacks a blood brain barrier allowing blood‐borne factors to access the brain. Since the AP is connected to numerous brain regions that play a key role blood pressure regulation, we hypothesised that blood‐borne inflammatory factors such as interleukin‐1 beta (IL‐1β) might play a role in the development of hypertension. In support of this hypothesis, plasma Interleukin‐1 beta is increased in hypertensive patients (Dalekos et al., 1997, J Lab Clin Med,129: 300–8). Furthermore, the receptors for IL‐1β are abundantly expressed in the AP. Here we test whether IL‐1β can act on the AP to cause an increase in blood pressure and renal sympathetic nerve activity (RSNA) in rats.MethodsBlood pressure and renal sympathetic nerve activity (RSNA) were recorded and analyzed after microinjection of different doses of IL‐1β into the AP (10ng and 25ng) of anaesthetised rats. In order to show that IL‐1β, specifically causes an increase in blood pressure, specific IL‐1 receptor blocking antibodies (IL‐1Ra) were injected into the AP prior to a 25ng IL‐1β injection. The AP was extracted for fluorescent activated cell sorting (FACS).ResultsExperiments showed that microinjection of 25ng IL‐1β caused a significant (p<0.01; Mann‐Whitney test) increase in both systolic blood pressure (19.6 ± 3.6 mmHg) and heart rate (55 ±10 bpm) compared with control injections of saline (SBP: 5.47 ± 1.58; HR 12 ± 6). Pre‐treatment with IL‐1Ra attenuated the response to IL‐1β, and was comparable with saline controls. IL‐1β also caused an increase in RSNA (137% ± 43%) and FACS revealed an increase in immune cell infiltration. Immunohistochemistry also revealed that IL‐1β injection results in Fos activation in the AP.ConclusionsIL‐1β injection appears to activate neurons in the area postrema which leads to an increase in blood pressure. By providing new insight into the actions of IL‐1β in the brain, we show that IL‐1β might be a potential target for future treatments for hypertension.Support or Funding InformationAustralian Research Council (FT170100363)National Health and Medical Research Council (GNT 1079680)Rebecca L Cooper Medical Research Foundation

Sex Differences in the Oxidative Stress and Inflammation Response During and After Simulated Hemorrhage in Humans

IntroductionHemorrhage (i.e., massive blood loss) induces an oxidative stress and inflammatory response that can persist even following hemostasis and resuscitation. These responses can result in tissue and organ damage if therapeutic inventions are delayed. Pre‐menopausal females exhibit a survival advantage following hemorrhage compared to males of a similar age. The role of oxidative stress and inflammation on increased survival from blood loss in young females is under investigation. Examining the potential mechanisms underpinning this survival advantage following hemorrhage may facilitate the development of sex‐specific therapies. In our laboratory, we utilize lower body negative pressure (LBNP) to simulate blood loss in conscious human subjects. In this study, we hypothesized that young males would elicit a greater oxidative stress and inflammatory response compared to young females, both during and after a simulated hemorrhage via LBNP.MethodsYoung, healthy human subjects (10F; 10M) participated in a stepwise‐LBNP protocol to presyncope. Stroke volume was estimated via finger photoplethysmography as a marker of central hypovolemia (indexed to body surface area). Venous blood samples were collected at baseline, at the onset of presyncope, and 60‐min into recovery (i.e., following “resuscitation”). The oxidative stress response was assessed via measurement of circulating F2‐Isoprostanes (F2‐IsoP) using gas chromatography‐negative ion chemical ionization‐mass spectrometry. The inflammatory response was assessed via measurement of circulating interleukin (IL)‐6 and IL‐10 using a MSD® Multiplex assay. Unpaired t‐tests were used to compare LBNP tolerance and stroke volume responses between male and female subjects. Two factor (time and sex) linear mixed model analyses with repeated measures were performed for all other comparisons, followed by Holm‐corrected post‐hoc tests. All data are represented as mean ± SE.ResultsLBNP tolerance time was similar between male and female subjects (Males, 1592 ± 124 s vs. Females, 1437 ± 113 s; P=0.37), and stroke volume index decreased by a similar magnitude at presyncope (Males, −50.2 ± 6.3% vs. Females, −49.4 ± 3.2%; P = 0.87). There was no effect of time or sex on [F2‐IsoP] or on the %Δ [F2‐IsoP] during or after LBNP (P ≥ 0.12). However, male subjects exhibited a greater increase in both the %Δ [IL‐6] and %Δ [IL‐10] compared to female subjects at the 60‐min recovery time point (IL‐6: Males, 101.4 ± 138.9% vs. Females, 12.3 ± 34.0%; P = 0.06. IL‐10: Males, 71.1 ± 133.3% vs. Females, −2.2 ± 11.8%; P = 0.06).ConclusionThese data suggest that there may be a sex difference in the inflammatory response to blood loss and subsequent fluid resuscitation. Future clinical studies should continue to explore sex differences in inflammatory responses to actual blood loss, and tailor therapeutic interventions accordingly.Support or Funding InformationWilliam and Ella Owens Medical Research Foundation

Suppression of histamine H1 receptor gene expression by antihistamines with inverse agonist activities

Allergic rhinitis is an incurable disease whose prevalence rate is over 30% in Japan. Antihistamines are first‐choice drugs for the therapy of the disease. However, sole treatment of antihistamine is not enough to obtain satisfactory control. We elucidated histamine‐induced histamine H1 receptor gene expression as a pathological mechanism of allergic rhinitis. Treatment of epinastine, an antihistamine, to allergic rhinitis patients showed alleviation of nasal symptoms and correlated suppression of histamine H1 receptor mRNA elevation in nasal mucosa. We observed inverse agonist activities in some antihistamines. Suppression of inositol phosphates accumulation and suppression of histamine H1 receptor gene expression showed good correlation. These results suggested that antihistamines with inverse agonist activities have advantage for the therapy of allergic rhinitis.Support or Funding InformationResearch Grant from The Osaka Medical Research Foundation for Intractable Diseases

Strategies for Autonomic Nervous System Mapping of Fiber Composition and Neural Circuitry Using Transmission Electron Microscopy in Support of SPARC Projects

The optimization of neuromodulation strategies is important for a development and refinement of therapies. The design of customized tools, such as electrical stimulating devices, depends on the detailed knowledge of nerve fiber composition and inter‐relationships between target organs, nerve populations, and synaptic circuitry. The autonomic nervous system gives support to organs and are composed by myelinated and unmyelinated fibers as well as pre‐ and post‐ganglionic neurons. Mapping an autonomic nerve such as the pelvic or vagus nerve that innervates peripheral tissues and understanding the synaptology of neurons in the cardiac and stellate ganglia may provide advanced knowledge for improved neuromodulation strategies. Transmission electron microscopy (TEM) is a high‐level technology that helps to identify and characterize axonal myelination and ultrastructure of unmyelinated fibers. TEM also allows for fine structural analysis of neural circuitry and innervation of autonomic targets. Immunogold detection techniques using TEM permits the identification and characterization of neuronal tracers in the nervous system. With the support from the Stimulating Peripheral Activity to Relieve Conditions (SPARC) program at NIH, we developed customized ultrastructural studies of the autonomic nervous system to facilitate refinement of neuromodulation strategies related to the autonomic control of cardiac, gastrointestinal, and lower urinary tract systems. Morphometric analysis is performed by automatic segmentation, and the development of a new strategy to calculate measurements of axon diameter, fiber diameter, myelin thickness, and G‐ratio provides a better understanding of fiber size for modelling studies. Both males and females from different species, including humans, are analyzed and compared. Implementation of common sampling techniques for nerves and ganglia across species provides increased rigor and reproducibility. TEM studies promote an improved understanding of functional studies and provide critical ultrastructural information for the development of new neuromodulation strategies.Support or Funding InformationSPARC/NIH 1OT2OD026585‐01 and Dr Miriam and Sheldon G. Adelson Medical Research Foundation (AMRF)

Parallel TXNIP‐ and TLR2‐ Dependent Pathways Synergistically Mediate Saturated Fatty Acid‐Induced Apoptosis in Cardiomyocytes

BACKGROUNDSaturated fatty acids (SFA) induce apoptosis in many cell types in vitro, and high SFA diet increases apoptotic cardiomyocytes, but mechanism remains unclear. We had previously demonstrated that p38α mitogen activated protein kinase (MAPK)–dependent signaling mediates cardiomyocyte apoptosis induced by palmitic acid (PA), a SFA. More recently, direct binding of thioredoxin interacting protein (TXNIP) and p38 MAPK has been demonstrated. SFA has been shown to bind to and activate Toll‐like receptor‐2 (TLR2), as well as the TLR4 accessory protein, MD2, triggering inflammation, oxidative stress, and cell death. We hypothesized that parallel TXNIP‐, and TLR‐ dependent pathways, both triggered by SFA, synergistically mediate apoptosis in cardiomyocytes.METHODS AND RESULTSA human cardiomyocyte line, AC16, was used in the study. TXNIP was inhibited by pre‐treating (24hours) cells with SBI‐477, a chemical inhibitor of MondoA/ChREBP. TXNIP knockdown was achieved with siRNA. TXNIP was overexpressed by transfecting with TXNIP plasmid, then 48 h later treated with TLR2 ligand, peptidoglycan (PGN) or TLR4 ligand, lipopolysaccharide (LPS), for an additional 2–3 days. TXNIP expression was determined by Western blotting of whole cell lysates. Apoptosis was measured by flow cytometry using Annexin V/PI staining.PA increased TXNIP protein levels dose‐dependently (1.5‐ and 2.3‐fold increase at 150 uM and 300 uM PA respectively) (ANOVA p<0.01, n=3). PA did not affect MondoA and ChREBP protein expression but increased nuclear translocation 2‐fold. TXNIP inhibition reduced PA‐induced apoptosis by 79% (p<0.05; n=3). TXNIP knockdown reduced PA‐induced cardiomyocyte apoptosis by 61% (p<0.01; n=7).TXNIP overexpression for 24 h increased apoptosis 1.8‐fold compared to control plasmid (PCDNA), but this was attenuated by siRNA knockdown of p38α or c‐fos, 59% and 84% respectively. TXNIP overexpression for 4–5 days markedly increased apoptosis rate to 4.0‐fold. The combined stressors of transfection with control plasmid and TLR ligand increased cell death rate 3.2 and 4.6‐fold for PGN and LPS respectively, compared to PCDNA + Vehicle. TXNIP overexpression followed by PGN increased apoptosis rate 8‐fold, whereas LPS reduced it to 2.7‐fold.CONCLUSIONSThis study in a human cardiomyocyte line demonstrated that (1) PA increased TXNIP levels, (2) TXNIP inhibition or knockdown attenuated PA‐induced apoptosis, (3) TXNIP overexpression increased apoptosis, (4) TXNIP overexpression combined with a TLR2 ligand, PGN, synergistically amplified the apoptosis rate, whereas LPS, a TLR4 ligand did not. These findings suggest that TXNIP‐dependent pathway synergistically combine with TLR2‐dependent signaling, but not TLR4‐, to amplify apoptosis, and suggest a mechanism by which elevated plasma SFA levels may lead to development of lipotoxic cardiomyopathy.Support or Funding InformationThis work has been funded by VA VISN 18 New Investigator Grant (to C.O.), CSR&D 5I01CX000598 grant (to P.R.), and VA Merit Grant BLRD I01BX007080 (to R.M.). The study was supported by VA Employment and Carl T. Hayden Medical Research Foundation. K.T. is supported by the M. Lowell Edwards Endowment.

In vitro Metabolism studies of Catechol‐Containing Natural Products and Analogues

Many biologically active natural products contain phenol groups that are prone to rapid phase‐II metabolism. In order to better understand the structural basis for these metabolic transformations, we have investigated the in vitro metabolism of rooperol, a bis‐catechol with anti‐cancer activity, and caffeic acid phenethyl amide (CAPA), a synthetic analog of a natural product with antioxidant and cytoprotective activities. In the presence of pig liver microsomes supplemented with UDP‐glucuronic acid, both of these catechols undergo rapid phase‐II metabolism to form mono‐ (CAPA and rooperol) and bis‐ (rooperol) glucuronide metabolites. The identity of these metabolites was determined by HPLC‐MS. We followed the kinetics of these in vitro metabolism reactions using HPLC. In both cases, the disappearance of the compound over time corresponded to a first‐order process with half‐life of 4.0 ± 0.5 min for rooperol and 5.1 ± 0.8 min for CAPA. In the case of rooperol, this data is in agreement with previously published pharmacokinetic (PK) studies in humans, in which only phase‐II metabolites were detected after oral administration, as well as PK studies in baboons which also report rapid phase‐II metabolism following IV administration. While human or non‐human primate PK data for CAPA have not been reported, our data suggest that this compound will also suffer from rapid phase‐II metabolism, and thus more metabolically stable analogs of both rooperol and CAPA are required.Support or Funding InformationThis research was supported by the U.S. Department of Education HSI STEM program (841.031c), Award #P021C160036The Robert and Ella Owens Medical Research Foundation

The ICAReB Platform: A Human Biobank for the Institut Pasteur and Beyond

The ICAReB platform of Institut Pasteur provides access to human bio-resources for academic and private research teams worldwide, essentially in the fields of infection, immunity and neurosciences. More than 134,000 human quality controlled, duly annotated samples (mainly whole-blood derived products, but also stool, urine, saliva, swabs….), from both healthy and diseased cohorts with open, regulated access, are available upon request. Both clinical investigation and biobanking activities are certified following ISO 9001:2015 and NF S96-900:2011 standards, respectively. ICAReB is a member of PIBnet (Pasteur International Biobanking network), BIOBANQUES and BBMRI, the French and pan-european biobanking networks, respectively. Funding statement: Funds come from IP (facilities and staff), and from various funding organisms or agencies (for specific projects): ANR (French National Research funding Agency): OH!Ticks project (see https://www.ohticks.fr/ 2017–2020) AP/HP (Assistance Publique/Hopitaux de Paris): MonaLisa (Multicentric Observational National Analysis on Listeriosis and Listeria) and ListeriaGEN projects, 2009–2022 and 2015–2024, respectively AVIESAN (French National alliance for Life Sciences and Health). Study of the innate immunity and the microbial flora during aplasia (PAMPA project, 2013–2016) Bioaster (French Investissements d’Avenir funding, Infectiology and Microbiology platform, Lyon-Paris, 2010–2020) Biomerieux (Lyon, France; cardio-vascular biomarker, 2013–2015) FRM (Medical Research Foundation): Hidradenitis Suppurativa project (2011–2015) IBiSA (Biology, Health and Agronomics infrastructure) selection of ICAReB platform in 2009 INCA (National Institute for Cancer research): INECOC project, 2009–2011 TOTAL foundation (Afribiota project on Environmental Pediatric Enteropathy, 2016-2020) WHO (for The WHO Human African Trypanosomiasis specimen Bank, 2008–2018, 2019–). Since three years, public-private relationships were proposed and research collaborations accepted with industrial partners, for example the Lyon-Paris ‘Technological Research Institute’ (BioAster) following a national Investissements d’Avenir program launched in 2012.

Novel Compounds Identified by Structure-Based Prion Disease Drug Discovery Using <i>In Silico</i> Screening Delay the Progression of an Illness

The accumulation of abnormal prion protein (PrPSc) produced by the structure conversion of PrP (PrPC) in the brain induces prion disease. Although the conversion process of the protein is still not fully elucidated, the intramolecular chemical bridging in the most fragile pocket of PrP, known as the “Hot Spot”, can stabilize the structure of PrPC and inhibit the conversion process. Using our original structure-based drug discovery algorithm, we identified the low molecular weight compounds that can bind to the hot spot. Several candidates were identified as anti-prion drugs with strong binding to recombinant PrP in vitro that significantly reduced the levels of PrPSc. These compounds were able to decrease the number of aggresomes in persistently prion-infected cells. Remarkably, treatment with the predominant candidate compounds significantly prolonged the survival period of prion-infected mice and dominantly suppressed disease-specific pathological damage, such as vacuole degeneration, PrPSc accumulation, microgliosis and astrogliosis in the brain, suggesting their possible clinical use. Fragment molecular orbital (FMO) analysis indicated that the high affinity of the candidates to the protein is largely dependent on non-polar interactions, such as van der Waals interactions. Our results suggest that in silico drug discovery using NUDE/DEGIMA may be useful to identify candidate compounds that effectively stabilize the protein. Funding Statement: This work was supported in part by the a grant from a Grant-in-Aid of the Research Committee of Prion Disease and Slow Virus Infection from the Ministry of Health, Labour and Welfare of Japan (N.N.); a Grant-in-Aid of the Research Committee of Molecular Pathogenesis and Therapies for Prion Disease and Slow Virus Infection, the Practical Research Project for Rare and Intractable Disease from the Japan Agency for Medical Research and Development, AMED (D.I.); a grant from the Takeda Science Foundation (N.N. and D.I.); a grant from the Japan Intractable Disease Research Foundation (D.I.); a Grant-in-Aid from the Tokyo Biochemical Research Foundation (D.I.); a grant provided by the YOKOYAMA Foundation for Clinical Pharmacology (Grant No. YRY1502) (D.I.); the grant provided by the Ichiro Kanehara Foundation (D.I.); a grant provided by the Mochida Memorial Foundation for Medical and Pharmaceutical Research (D.I.); a grant provided by the Waksman Foundation of Japan Inc. (D.I.); and a grant provided by the Center for Clinical and Translational Research of Kyushu University Hospital (D.I.). Declaration of Interests: The authors state that they have no conflicts of interest. Ethics Approval Statement: Animal experiments were approved by the Animal Care and Use Committee of the Nagasaki University. All animals were treated and cared for in accordance with the Guidelines for Animal Experimentation of Nagasaki University.

Compensatory Changes in Parenchymal Mass and Function after Radical Nephrectomy.

Loss of renal function remains a major limitation of radical nephrectomy. The extent of renal functional compensation by the preserved kidney after radical nephrectomy has not been adequately studied in this elderly population with comorbidities. A total of 273 patients treated with radical nephrectomy without end stage renal disease with available preoperative nuclear renal scans were included in the analysis. Renal functional compensation was defined as percent change in estimated glomerular filtration rate of the preserved kidney after radical nephrectomy. Estimated glomerular filtration rate was calculated by the Chronic Kidney Disease-Epidemiology Collaboration formula up to 5 years postoperatively. Preoperative/postoperative parenchymal volumes of the preserved kidney were measured from cross-sectional imaging. Multiple regression was used to identify predictive factors for renal functional compensation. Median age was 67 years and 67% of the patients were male. Overall 70% had hypertension, 26% diabetes and 37% preexisting chronic kidney disease. Locally advanced (T3a or greater) tumors were found in 53% of cases. Renal functional compensation was observed at 2 weeks (median 10%) and increased during the first 3 months (median 26%) after radical nephrectomy. Functional stability was then observed to 5 years. Renal parenchymal volume increased a median of 10% at 3 to 12 months but in addition, the functional efficiency per unit of parenchymal volume also increased 8% (estimated glomerular filtration rate units/cm3 of parenchyma was 0.236 postoperatively vs 0.208 preoperatively, p=0.004). Age (-0.85, p <0.01), global preoperative estimated glomerular filtration rate (-0.28, p <0.01) and split renal function of the removed kidney (0.61, p <0.01) were independent predictors of renal functional compensation. Percent renal functional compensation after radical nephrectomy is greater in younger patients, when preoperative estimated glomerular filtration rate is lower and when the removed kidney has more robust function. Increases in measurable parenchymal mass and functional efficiency contribute substantially to renal functional compensation.