bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel ex‐vivo procedure for monitoring β‐hydroxybutyrate (βOHB) production from isolated whole livers
Authors: Singh Avinash , Chakraborty Saroj , Mell Blair , Wenceslau Camilla , McCarthy Cameron , Joe Bina 
Published: April 2020

When glucose is limiting, such as during fasting, the liver synthesizes and secretes ketone bodies into circulation as an alternate fuel source. Previously, we found that the ketone body beta‐hydroxybutyrate (βOHB) is lowered by salt consumption in hypertensive rats. To identify hepatic‐specific mechanisms that regulate βOHB in vivo is difficult because extra‐hepatic factors impact liver function. Thus, we aimed to develop an ex vivo procedure to continually monitor hepatic βOHB. D‐glucose, but not L‐glucose, is a substrate for ATP generation. We exploited this difference to simulate non‐fasted (D‐glucose) and fasted (L‐glucose) conditions in vivo and hypothesized that livers incubated with L‐glucose will generate more βOHB compared to those incubated with D‐glucose. Portal veins of 27‐week‐old Wistar‐Kyoto rats (n=3/group) were perfused with either D‐glucose or L‐glucose in Krebs buffer. Whole livers were excised and incubated with the same buffers as perfusion. Aliquots were collected from 0–6hrs for quantitating βOHB. A statistically significant time effect was noted in both buffers, with βOHB continually increasing with time [D‐glucose Krebs (mmol/L), 0h: 0.03±0.00 vs. 6h: 0.82±0.02*; L‐glucose Krebs (mmol/L), 0h: 0.04±0.00 vs. 6h: 1.16±0.14*, both *p<0.05]. Confirming the sensitivity and validity of our preparation, there was a significant interaction effect, with livers incubated for 6 h with L‐glucose generating higher levels of βOHB than livers incubated with D‐glucose [D‐glucose: 0.82±0.02 vs. L‐glucose: 1.16±0.14*, *p<0.05). Our novel ex vivo liver preparation simulates fasting and non‐fasting conditions in vivo and can be applied to study liver‐specific regulation of βOHB independent of extra‐hepatic factors.Support or Funding InformationAmerican Heart Association (18POST34060003) and National Institutes of Health (R00GM118885 and R01HL143082).Isolated, whole livers, incubated in Krebs buffer made with L‐glucose, have increased beta‐hydroxybutyrate synthesis compared to livers incubated in Krebs buffer made with D‐glucose. n=3. Two‐way ANOVA: *p<0.05.Figure 1
The Role of the Actin‐Binding Protein Cortactin in the Onset and Development of Sepsis
Authors: Lartey Nathaniel Lartey , Ponce Alexander Garcia , Vargas Hilda , Shibayama Mineko , Schnoor Michael 
Published: April 2020

IntroductionSepsis is a devastating disease, which occurs as a result of an exacerbated immune response to systemic infection. Major hallmarks of sepsis include increased vascular permeability and leukocyte recruitment to secondary organs such as lung and kidney, often leading to organ dysfunction and, in severe cases, organ failure. Cortactin is an actin‐binding protein, expressed ubiquitously with the exception of neutrophils. Endothelial cortactin regulates vascular permeability and neutrophil recruitment by controlling activation of small GTPases. Therefore, endothelial cortactin could play an important role during sepsis pathogenesis by controlling neutrophil influx into affected organs such as lungs.Aim/ObjectiveTo elucidate the effects of cortactin deficiency on the onset and development of sepsis, which is a disease characterized by massive systemic neutrophil extravasation.Experimental StrategySepsis was induced by cecal ligation and puncture (CLP) in age‐matched male cortactin‐deficient (CTTN KO) and littermate wild‐type (WT) mice; and five day survival was monitored. Blood and lung tissue samples were collected 24 h after CLP for protein and gene expression analysis. Lung histology was analyzed after hematoxylin/eosin staining of paraffin‐crossections of lung tissue. Oxidative stress was assessed in lung cross‐sections using dihydroethidium. mRNA and protein levels of inflammatory cytokines were assessed by quantitative real‐time PCR (lung tissue) and bead‐based milliplex assay (peripheral blood) respectively. Neutrophil numbers were determined in blood and peritoneum by flow cytometry.Results and ConclusionsCortactin deficiency improved survival of mice subjected to CLP. Of note, lung tissue of septic KO animals showed better preserved architecture of the alveoli, and absence of oedema, hemorrhage and mucus deposition. Gene expression and secretion of the inflammatory mediators tumor necrosis factor‐α (TNFα) and interleukin‐1β in KO mice were reduced in comparison to septic WT mice. Moreover, we observed less oxidative stress in the lungs of septic KO mice in comparison to septic WT mice. Cortactin deficiency also protected against sepsis‐induced neutropenia. Additionally, in the absence of cortactin, less neutrophils were recruited to the peritoneum without compromising bacterial clearance. In summary, cortactin deficiency attenuates both local and systemic inflammatory responses during sepsis, protects against sepsis‐induced neutropenia and protects from lung tissue damage. Thus, cortactin could be an interesting target as novel treatment strategy for sepsis.Cortactin deficiency protects against sepsis‐induced lung damageFigure 1Expression of inflammatory cytokines in the lung are reduced in the absence of cortactinFigure 2
Effects of 1‐year Aerobic Exercise Training on Cerebral Blood Flow and Arterial Siffness in Amnestic Mild Cognitive Impairment
Authors: Tomoto Tsubasa , Pasha Evan , Sugawara Jun , Tarumi Takashi , Chiles Collin , Curtis Bryon , Cardim Danilo , Zhang Rong 
Published: April 2020

Aerobic exercise training (AET) may improve cerebralperfusion and reduce the risk of dementia. Changes in central arterialstiffness and pulsatile cerebral blood flow (CBF) associated with exercise training may be one of the underlying pathophysiological mechanism. The purpose of this study was to investigate: (1) The effects of AET on central arterialstiffness and CBF, and (2) whether pulsatile CBF mediates the association between changes in arterial stiffness and cerebral perfusion in amnestic mildcognitive impairment (aMCI), a prodromal stage of Alzheimer disease. Seventy aMCI patients were randomized to 12 months of moderate‐intensity AET (n=31) or stretching program (n=39). CBF and carotid β‐stiffness index were assessed by ultrasonography and applanation tonometry. Cerebral perfusion was calculated by normalizing total CBF to brain tissue mass measured by magnetic resonance imaging. Pulsatile CBF was measured from the middle cerebralartery using transcranial Doppler. Cardiorespiratory fitness was assessed by peak oxygen uptake (VO2peak). VO2peak and cerebralperfusion increased and carotid β‐stiffness index decreased in the AET group, while remained unchanged in the stretching group. The change in VO2peak was significantly associated with the change in cerebral perfusion, carotidβ‐stiffness index, and pulsatile CBF (Fig.
1). The decrease in carotidβ‐stiffness was associated with increased in cerebral perfusion (Fig.
2). Mediation analysis showed that changes in pulsatile CBF mediates the association between carotidβ‐stiffness index and cerebral perfusion (Fig.
2). These results suggest that AET decreases arterial stiffness and increases cerebral perfusion, which is mediated by pulsatile CBF in aMCI patients.Support or Funding InformationThis work was funded by the National Institutes of Health ((R01AG033106, R01HL102457, P30AG012300)Associations between the change (Δ) in relative peak oxygen uptake (VO2) and the change in cerebral perfusion, carotid β‐stiffness index and pulsatile cerebral blood flow (CBF).Figure 1Mediation analysis diagram of the effect of the change (Δ) in carotid β‐stiffness index on the change in cerebral perfusion through the change in pulsatile cerebral blood flow (CBF).Figure 2
Epidural electrical stimulation and hemodynamic control after spinal cord injury
Authors: Soriano Jan Elaine , Saleem Saqib , Netoff Theoden I. , Squair Jordan W. , Samadani Uzma , Krassioukov Andrei V. , Darrow David , Phillips Aaron A. 
Published: April 2020

Spinal cord injury impairs cerebral autoregulation (i.e., the buffering of changes in blood pressure in an effort to maintain cerebral blood flow constant) and baroreflex sensitivity (i.e., the appropriate heart rate responses to changes in blood pressure), due in part to the disruption of supraspinal sympathetic control of circuits below the level of injury. This impairment is almost certainly associated with the greater risk of cerebrovascular and heart disease observed in this population.We have previously shown in people with spinal cord injury that epidural electrical stimulation of the spinal cord below the level of injury (eSTIM) activates disconnected sympathetic circuits. Furthermore, long‐term daily application of eSTIM can in some cases lead to plasticity that restores function, even when eSTIM is off. It is not clear if eSTIM activation of sympathetic circuits restores cerebral autoregulation and baroreflex function after spinal cord injury, or if functionally‐relevant plasticity occurs after long‐term application of eSTIM.Here, we show that activating eSTIM immediately improves cerebral autoregulation and baroreflex sensitivity. These positive immediate effects were repeatable over several months. Long‐term daily eSTIM for at least 12 weeks did not affect cerebral autoregulation or baroreflex function when stimulation was off, indicating that functionally‐relevant plasticity did not occur. In conclusion, immediate application of eSTIM may be a viable therapy for improving hemodynamic control after spinal cord injury.Support or Funding InformationNatural Sciences and Engineering Research Council of Canada, Canadian Institutes of Health Research, Libin Cardiovascular Institute of Alberta, Hotchkiss Brain Institute, Compute Canada.Cerebral autoregulation was consistently improved with the activation of eSTIM as indicated by increased phase lag between blood pressure and cerebral blood flow. Cardiovagal baroreflex function was also improved as indicated by more rapid heart rate responses to changes in blood pressure. **indicates significant difference (p<0.005)Figure 1
Maternal exercise programs skeletal muscle metabolic dysfunction in offspring mice challenged with high fat diet.
Authors: Son Jun Seok , Chen Yanting , Chae Song Ah , Chen Ke , Zhu Mei-Jun , Du Min 
Published: April 2020

Exercise, including maternal exercise becomes increasingly less common. Though improper maternal nutrition and obesity are known to affect fetal muscle development and predisposes offspring to metabolic dysfunction, the effects of maternal exercise (ME) on fetal muscle development and offspring metabolic health are largely undefined.PURPOSETo investigate the effects of maternal exercise on mitochondrial biogenesis during fetal muscle development, and muscle function and metabolic health in offspring following high fat diet (HFD) challenge.METHODSFemale C57BL/6J mice were randomized and assigned into a control (CON) or an exercise (EX) group (n = 6 per group) fed a normal chow. Exercise was performed every morning for 60 min during pregnancy. Then, the animals were mated, and the skeletal muscle samples were collected at embryonic day 18.5, weaning, and after 8 weeks of high fat diet (HFD; 60 kcal% from fat) challenge in both females and males. Statistical analysis were conducted using student’s t‐test and/or one/two way analysis of variance (ANOVA).RESULTSME muscle fiber diameters and enhanced muscle endurance capacity in offspring (P < 0.05), which were positively associated with switching muscle fibers from type IIb/x to type I and IIa and mitochondrial biogenesis (P < 0.05), but negatively correlated with atrophy‐related Atrogin‐1 and Murf1 gene expression (P < 0.05). Mechanistically, ME enhanced DNA demethylation of Pgc‐1a promoter and its expression during fetal muscle development. Importantly, such changes in DNA methylation maintained in offspring muscle before and after HFD challenge, resulting in elevated Pgc‐1a mRNA expression and increased contents of PGC‐1a1 and 4 isoforms, which improved metabolic health.CONCLUSIONSThis study identifies that ME enhances DNA demethylation of the Pgc‐1a promoter during fetal muscle development, which has programming effects on the exercise endurance capacity and metabolic health of muscle in offspring.Support or Funding InformationThis work was supported by National Institutes of Health Grants R01‐HD067449.Maternal exercise (ME) enhances adaptation to endurance training of offspring mice (A,B) and the proportion of muscle type I fiber composition before and following HFD challenge (C). ME decreases muscle atrophy‐related genes following HFD challenge (D,E). Data are mean ± s.e.m. and each dot represents one litter.; two‐sided P values by unpaired Student’s t‐test (D,E) or two‐way repeated measures ANOVA followed by Tukey’s test (B).Figure 1
Quantification of Mitochondrial Membrane Potential in the Isolated Rat Lung Using Rhodamine 6G
Authors: Audi Said H. , Cammarata Anthony , Clough Anne V. , Dash Ranjan K. , Jacobs Elizabeth R. 
Published: April 2020

Mitochondrial membrane potential (ΔΨm) plays a key role in vital mitochondrial functions, and its dissipation is a hallmark of mitochondrial dysfunction. The objective of this study was to develop an experimental and computational approach for estimating ΔΨm in intact rat lungs using the lipophilic fluorescent cationic dye rhodamine 6G (R6G). Rat lungs were excised and connected to a ventilation‐perfusion system. The experimental protocol consisted of three single‐pass phases: loading, wash, and uncoupling, in which the lungs were perfused with R6G‐containing perfusate, fresh R6G‐free perfusate, or R6G‐free perfusate containing the mitochondrial uncoupler FCCP, respectively. This protocol was carried out with lung perfusate containing verapamil vehicle or verapamil, an inhibitor of the multi‐drug efflux pump P‐glycoprotein (Pgp). Results show that the addition of FCCP resulted in an increase in R6G venous effluent concentration (Figure
1), and that this increase was larger in the presence of verapamil than in its absence. A physiologically‐based pharmacokinetic (PBPK) model for the pulmonary disposition of R6G was developed and used for quantitative interpretation of the kinetic data, including estimating ΔΨm. The estimated value of ΔΨm (−139 ± 21 (SD) mV) was not significantly altered by inhibiting Pgp with verapamil, and is consistent with that estimated previously in cultured pulmonary endothelial cells. These results demonstrate the utility of the proposed approach for quantifying ΔΨm in intact functioning lungs. This approach has potential to provide quantitative assessment of the effect of injurious conditions on lung mitochondrial function, and to evaluate the impact of therapies that target mitochondria.Support or Funding InformationSupported by NIH 2R15HL129209‐02 and VA Merit Review Award BX001681.Solid symbols: R6G venous effluent concentrations during the loading phase, wash phase, and uncoupling phase using FCCP (67 μM). Values are mean ± SE (n = 7 for loading and wash phases, and n = 5 for uncoupling phase). Solid line is PBPK model fit to data.Figure 1
Synthetic cadavers improved laboratory test grades in an undergraduate human anatomy course
Authors: Gregory Diane , Zwambag Derek , McFall Kristen , Andrews David , Richardson Nicolette 
Published: April 2020

This study explored the use of an intriguing instrument which may provide a very similar, yet more sustainable, experience to human cadaver dissection in anatomy courses. The synthetic cadaver is structurally similar to a human cadaver but does not carry the same ethical and sustainability challenges. Students in an undergraduate human anatomy course completed two laboratory tests: Test 1 was completed prior to the introduction of two synthetic cadavers to the laboratory where the primary laboratory resources were plastic models. Test 2 was completed after the synthetic cadavers were introduced. Comparison of student lab test grades both pre and post synthetic cadaver introduction (“Experimental”) as well as to “Historical” cohorts of the course were used to assess student success. In addition, surveys comprised of 5‐point Likert items were used to assess student perceptions. Test 1 grades in the Experimental and Historical cohorts were compared to ensure that no academic differences existed between groups (p = 0.7653; z‐statistic = 0.2985). Students in the Experimental group performed better on Test 2 (median = 73.8% (95% CI: 72.0–75.0%) compared to the Historical group (70.1% (95% CI: 68.4%–70.1%), p < 0.0001). Students perceived laboratory resources to be necessary (95%), helpful (94%) and effective (87%), yet there was no change in these perceptions following the introduction of the synthetic cadavers. This investigation indicates that the synthetic cadavers were an effective learning tool in undergraduate human anatomy, and that student success was improved after using the synthetic cadavers, compared to when only plastic models were available.Support or Funding InformationNo funding was received for this work.Probability density estimates for laboratory test grades in the Historical (light blue) and Experimental (dark blue) cohorts. Grey areas represent the 95% confidence interval for the median grade. There was no difference in median Test 1 grades; however, the median Test 2 grade in the Experimental cohort was significantly higher than in the Historical cohort.Figure 1
Assessing and improving the performance of consensus docking strategies using the DockBox package
Authors: Preto Jordane , Gentile Francesco 
Published: April 2020

Molecular docking is a well‐established computational method to predict how a ligand binds to a specific protein target as well as to assess the strength of the binding. Although docking programs are used worldwide for drug discovery, it is not always simple to identify which program or combination of programs provides the best results for a target of interest. Here we present DockBox, a computational package designed to facilitate the use and comparison of multiple docking and scoring programs at a time, and to combine them using different consensus strategies. A new consensus docking method called scoring‐based consensus docking (SBCD) is introduced, which significantly improves success rates in pose prediction as well as virtual‐screening (VS) hit rates compared to standard consensus docking. Several test cases are presented to illustrate the success of the SBCD method in docking and VS scenarios. SBCD requires the same computational cost as standard consensus methods in terms of number of required programs, computational power and time, making it a promising new approach for the screening of large chemical libraries.Presentation of the DockBox packageFigure 1
Is Naloxone use in Southeast Alabama Rising? Evaluation of Naloxone Administration in an Emergency Department Setting
Authors: Parker Robert Wayne , Thorn Morgan , Patel Divyesha 
Published: April 2020

BackgroundFatalities caused by drug‐overdose have increased greatly in recent years across the US and is now the leading cause of injury‐related death in the United States. Drug related deaths with cointoxicants are being seen far too often and the increase in prevalence of opioid overdoses with possible concomitant medication use is a public health concern that needs to be addressed. Naloxone (Narcan ®) is an opioid antagonist approved in November 2015 by the FDA to help combat the effects of opioid overdose. Awareness of naloxone’s availability over the counter and education on opioid related overdoses for communities in the United States has been shown to improve opioid overdose deaths. In this study, we document naloxone administration in the Southeast Health Emergency Department to help identify high risk sub‐populations in this region that present with signs of potential opioid overdose and the connection between overdoses for opioids or a cointoxicant.Study DesignA retrospective study was conducted evaluating the administration of naloxone in individuals arriving in the emergency department at Southeast Health from August 01, 2011 to August 31, 2018. Patient cases were collected based on time, route, and dose of naloxone given, and data was then selected based on ICD 10 codes and documented positive reaction to naloxone. Patients who were not between ages 18–89 or were incarcerated or pregnant prior to their hospital were excluded from this study. Further data was collected on patients’ socioeconomic status, medical history, and the course of their encounter.ResultsOf the 721 charts reviewed, 261 were deemed true overdoses and 51 were possible overdoses. While there were more overdoses in individuals with a history of chronic pain or mental illness, this was not statistically significant. The data showed most patients who overdosed were taking both opioids and benzodiazepines.ConclusionsIt is apparent that disabled/retired females over 65 with Medicare or Medicaid were the most likely to be a true opioid overdose.Due to the patient’s mental status upon arrival, not all charts had complete histories or medication records. There were no clear documentation standards, so this varied from by provider and year, making it difficult to determine if there was a true overdose. Data was also dependent on patient honesty, relying on them to admit a history of abuse or what substances they had taken. The number of cases of overdose is likely under reported. Southeast Health is not the only hospital in the area. Emergency responders or home naloxone may have been administered decreasing the number of emergency consults. When considering patients insurance status, only the primary insurance listed was recorded. A patient could have also had a Medicaid plan indicating low income, but this was not noted.Age DistributionFigure 1Naloxone and Past Medical HistoryFigure 2