bluuebunny/crossref_metadata_2025 huggingface dataset:
Influenza‐induced cardiopulmonary dysfunction and alveolar fluid clearance inhibition are attenuated in F508del CFTRheterozygous mice
Authors: Davis Ian Christopher , Aeffner Famke , Abdulrahman Basant , Hickman‐Davis Judy Marie , Janssen Paul , Hayes Don , Amer Amal , Bedwell David , Sorscher Eric
Published: June 2021

Influenza virus inhibits bronchoalveolar epithelial ENaC‐mediated Na+ absorption and stimulates Cl− secretion via CFTR, resulting in significant impairment of alveolar fluid clearance (AFC). To determine the contribution of CFTR to influenza pathogenesis, adult C57BL/6‐congenic F508del CFTR‐heterozygous mice (HETs) and wild‐type (WT) littermate controls were mock‐infected intranasally with 50 μl virus diluent (0.1% FCS in PBS) or infected with 10,000 FFU/mouse influenza A/WSN/33 (H1N1). Infection‐induced hypoxemia and bradycardia were attenuated in HETs at 2–6 days post‐infection (d.p.i.), although weight loss and viral replication did not differ from WT controls. Pulmonary edema (increasing lung wet:dry weight) developed over the course of infection in WT mice, but was absent in HETs at both 2 and 6 d.p.i. Influenza significantly inhibited AFC in live, mechanically‐ventilated WT mice at 2–6 d.p.i. In HETs, influenza had no effect on AFC at 2 d.p.i. and caused significantly less AFC inhibition at 6 d.p.i. Higher HET AFC rates were associated with greater bronchoalveolar lavage fluid TGF‐β at 2 d.p.i. and a 10‐fold increase in IL‐6 at 6 d.p.i. Systemic antibody‐mediated neutralization of TGF‐β or IL‐6 did not alter outcomes in WT mice, but increased cardiopulmonary dysfunction to WT levels in HETs. Our findings indicate that CFTR activity is an important host determinant of lung dysfunction in influenza.
Effects of NO/cGMP inhibitors in a rat model of anaphylactoid shock
Authors: Fagundes Agnes Afrodite Sumarelli Albuquerque , Carvalho Marco Túlio Menezes , Ferreira Luciana Garros , Silveira Ana Paula Cassiano , de Nadai Tales Rubens , Capellini Verena Kise , Evora Paulo Roberto Barbosa , Celotto Andrea Carla
Published: June 2021

Anaphylaxis is a severe clinical manifestation of allergic diseases. The aim was investigate the effect of NO/cGMP pathway inhibition with L‐NAME and methylene blue (MB) and indigo carmine (IC, α‐adrenergic receptor agonist) in C48/80‐induced shock in rats. Animals were allocated in groups: Control, C48/80, MB, MB+C48/80, C48/80+MB, L‐NAME, L‐NAME+C48/80, C48/80+L‐NAME, IC, IC+C48/80 and C48/80+IC. The drugs were administered 5 min. before or after the C48/80. The systolic blood pressure (SBP) and the survival were analyzed every 10 min. over a period of 60 min. Administration of C48/80 decreased the SBP (89±5 to 27±7 mmHg), led to cyanosis and killed 60% of the animals within 60 min. Plasma NOx levels were increased in C48/80 group compared to Control. The use of MB and L‐NAME prior to shock induction does not prevented the SBP reduction (37±7 and 43±8 mmHg respectively), but increased the survival in 50%. When MB and L‐NAME were administered after C48/80, no improvements in SBP were observed and survival was reduced. The IC before or after C48/80 not inhibited the decrease in SBP and in both groups the survival was lower than C48/80 group. In this rat model of anaphylactoid shock there is an increase of NO production, but the blockade of NO/cGMP pathway did not restores the SBP. However, these blockers increased animal survival. IC not inhibited the decrease in SBP and not prevented animals death by shock. Supported by FAEPA.
Small arteries of gestational rats are less sensitive to relaxation induced by inhibition of rho A‐associated kinase (ROK).
Authors: Chi Mei , Zielnik Barbara , Davidge Sandra , Mitchell Bryan
Published: June 2021

Inhibition of ROK has been proposed as a method to arrest uterine contractions during preterm labor. Because of concern about vascular side effects, we studied the effects of the ROK inhibitor g‐H1152 (g‐H) on relaxation of mesenteric and uterine arteries.

Arteries from non‐pregnant (n=5) and pregnant (gestation day17–21; n=6) rats were studied using wire myography. Concentration response curves (CRC) to calculate IC50 for g‐H were obtained following phenylephrine (PE) or endothelin‐1 (ET‐1) induced constriction. Using vessels from pregnant rats, CRC for PE were performed before and after a 20‐min incubation with g‐H at 300 nM, 1 and 3 μM.

Mesenteric arteries were more sensitive than uterine arteries to g‐H‐induced relaxation in both non‐pregnant and pregnant rats. Both arteries were significantly less sensitive to g‐H‐induced relaxation during pregnancy. Sensitivity to PE was not changed with any dose of g‐H. However, using 2‐factor ANOVA, the maximal PE‐induced tension was significantly reduced by the two highest concentrations of g‐H in both vessels from pregnant rats but to a greater extent in mesenteric vessels (34±5.6%, 18±4.1%) compared to uterine (14±4%, 7±2.7%).

The ROK pathway plays a significant role in agonist‐induced constriction of mesenteric and uterine vessels, though less so during gestation. If ROK inhibitors are used during pregnancy in vivo, maternal blood pressure and uterine blood flow will be assessed.
Increased cerebral oxygen delivery during sodium nitroprusside administration despite reduced blood flow velocity in the middle cerebral artery
Authors: Schwartz Christopher E , Messer Zachary R , Terilli Courtney , Medow Marvin S , Stewart Julian M
Published: June 2021

We investigated whether the nitric oxide donor sodium nitroprusside (SNP) dilates and phenylephrine constricts the MCA in 10 supine young healthy volunteers. We combined transcranial Doppler (TCD) measurements of the MCA with near infrared spectroscopy (NIRS) over the frontal cortex. Cerebral oxyhemoglobin and total hemoglobin increased by 14±1 and 15±1 μM/L with 100μg SNP despite hypotension, and were reduced by 6±1 and 7±1 μM/L with 150μg phenylephrine despite hypertension. SNP increased NIRS derived cerebral blood flow estimates by approximately 40±8% from baseline, while TCD derived CBFv decreased by 15±3%. Phenylephrine decreased NIRS derived cerebral blood flow estimates by approximately 11±5% from baseline, while TCD derived CBFv increased by 5±3%. Studies using upright tilt and lower body negative pressure were consistent with the literature and demonstrated similar relative changes in cerebral blood flow and CBFv as orthostatic stress progressed. We concluded that the MCA dilates to SNP and constricts to phenylephrine, but does not dilate during orthostatic stress in healthy volunteers. Supported by NIH (HL‐074873) and (HL‐087803).
Racial Differences in Blood Pressure Variability, Baroreflex Sensitivity and Heart Rate Variability Following Maximal Exercise
Authors: Kappus Rebecca M , Ranadive Sushant M , Yan Huimin , Lane Abbi D , Cook Marc D , Wilund Kenneth R , Woods Jeffrey A , Fernhall Bo
Published: June 2021

Introduction

Heart rate variability (HRV), blood pressure variability (BPV) and baroreflex sensitivity (BRS), measures of autonomic nervous system function, are associated with cardiovascular (CV) outcome. African Americans (AA) have an increased prevalence of CV disorders in comparison to Caucasians (C).

Purpose

To determine race specific responses in BPV, BRS and HRV before and after maximal exercise.

Methods

37 AA and 37 C (24 years) underwent maximal exercise on a cycle ergometer. BPV, BRS and HRV measurements were assessed using finger plethysmography before and after exercise. Data were log transformed and pre‐and post‐max test values were compared by race.

Results

Acute exercise significantly decreased BRS, LF‐HRV and HF‐HRV and significantly increased LF/HF‐HRV in both AA and C. With exercise, C significantly decreased measures of BRS and HRV more compared to AA; BRS‐SAPRRI transfer mean (C: 2.65±.54 to 1.09±.81 versus AA: 2.70±.40 to 1.70±.92), BRS‐down down (C: 3.21±.55 to 1.74±.49 versus AA: 3.28±1.0 to 2.2±.71), HRV‐LF (C: 6.39±.86 to 3.68±1.42 versus AA: 6.37±1.24 to 5.18±2.14) and HRV‐HF (C: 7.38±1.23 to 3.53±1.73 versus AA: 7.51±1.09 to 5.38±2.16).

Conclusion

C exhibited increased vagal withdrawal and greater reductions in BRS compared to AA in response to acute exercise. The impact of these racial differences in autonomic control on CV risk needs further investigation.
Central sympathoinhibition by moxonidine attenuates brain oxidative stress, prevents cardiac remodeling and improves prognosis associated with reduction of sympathetic hyperactivity in rats with hypertensive heart failure
Authors: Honda Nobuhiro , Hirooka Yoshitaka , Matsukawa Ryuichi , Ito Koji , Sunagawa Kenji
Published: June 2021

Background

Hypertension causes hypertensive heart failure (HHF). In HHF, excessive sympathoexcitation leads to poor prognosis. However, the effect of central sympathoinhibition on survival in HHF remains unclear. Therefore, we examined whether moxonidine, a sympathoinhibitory agent acting on nischarin associated with oxidative stress, improves prognosis in a rat model of HHF and has antioxidant effects in the brain.

Methods and Results

We fed Dahl salt‐sensitive (DS) rats with high salt diet (HS) to create a model of HHF. This model represents a compensated left ventricular (LV) hypertrophic stage, subsequently a HF stage. We used DS rats with HS as HHF and performed intracerebroventricular infusion of moxonidine or vehicle to evaluate the effect of central sympathoinhibition. In comparison with vehicle, moxonidine decreased urine norepinephrine (2.8±0.2 vs. 1.7±0.3 μg/day, p<0.01), improved survival (23 vs. 76 %, p<0.05) as well as LV function (LVDd; 6.9±0.2 vs. 6.4±0.1 mm, p<0.05, %FS; 38±1 vs. 45±1 %, p<0.01, LVEDP; 14.2±2.7 vs. 7.3±1.7 mmHg, p<0.05, LV −dP/dt; −7738±348 vs. −10394±1156 mmHg/s, p<0.05). Moreover, moxonidine decreased LV fibrosis and brain oxidative stress (Signal Decay Rate; 0.12±0.01 vs. 0.10±0.01/min, p<0.05).

Conclusion

Moxonidine‐induced central sympathoinhibition attenuates brain oxidative stress, prevents cardiac remodeling and improves prognosis in rats with HHF.
Autonomic dysregulation in a genetically modified mouse with reduced expression of the vesicular acetylcholine transporter (VAChT KDHOM)
Authors: Durand Marina Toledo , Oliveira Mauro , Becari Christiane , Ananias Alice , Oliveira Aline , Guatimosin Silvia , Prado Marco , Salgado Helio C
Published: June 2021

We investigated the circadian rhythm, autonomic regulation, baroreflex control and the restrain stress in VAChT KDHOM mice. Wild type (WT, n=9) and VAChT KDHOM (n=8) mice were anesthetized (isoflurane) and telemetry transmitters were implanted into the left carotid artery. Ten days after the recovery, mean arterial pressure (MAP, mmHg) and heart rate (HR, bpm) were recorded for 48 hours (5 min every 1h). Basal MAP and HR were also recorded continuously, followed by methylatropine and propranolol administration. VAChT KDHOM showed an increase in MAP only at night (114±3 vs 107±2), but similar HR as compared to WT mice. VAChT KDHOM exhibited reduced vagal (蜐bpm,144±18 vs 193± 15) and elevated sympathetic tone (−45±7 vs −25±7) evaluated by the autonomic blockers. AP variability was similar between groups, while HR variability (LH/HF: 0.6±0.1 vs 0.9±0.1) and baroreflex sensitivity (3±0.3 vs. 5.1±0.3) were reduced. Restraint stress elicited a greater pressor response (蜐MAP: 30±3 vs 20±3 after 60 min), but similar tachycardia as compared to WT. The results demonstrated that the autonomic cardiovascular regulation of VAChT KDHOM is characterized by reduced parasympathetic and increased sympathetic tone, combined with decrease in baroreflex sensitivity and an increase in MAP at night. Moreover, the greater pressure response to restrain stress suggested vascular abnormalities in VAChT KDHOM.
Psycho Social Stressors in Elderly Assessed with Heart Rate Variability
Authors: Marck Jan Willem , Kallen Victor L. , Issa Amine N. , Kasak Alex J. , Smethana Michael , Johnson Bruce D.
Published: June 2021

BACKGROUND

Psycho‐socio stressors may over time result in dominance of the sympathetic nervous system (SNS) over the parasympathetic nervous system (PNS). Reduced PNS control over cardiovascular processes has been associated with decreased health. Heart rate variability (HRV) has been proposed as a technique to quantify SNS and PNS interactions and thus might offer a noninvasive predictive index.

OBJECTIVE

To investigate the impact of real‐life stressors on HRV fluctuations in independent living elders (EL).

METHODS

For three months every two weeks EL (N=22; Age=75±9; M/F ratio=17/5) performed autonomic tests consisting of quiet breathing, paced breathing and hand grip. All did a questionnaire assessing depression, positive/negative live events, and isolation.

RESULTS

Negative events (r=0.36, p=.01) and isolation (r=0.35, p=0.02) are correlated with HR. Personal changes in HRV, SDNN (r=−0.43, p<0.005) and Poincare Width (r= −0.42, p<0.005), are correlated to negative life events.

CONCULSION

Negative life events reduce HRV which suggests either SNS activation and/or PNS withdrawal.

Research was made possible by Healthy for Life, TNO, and the Center for Innovation, Mayo Clinic.
Blunted cardiovagal arterial baroreflex gain to acute hypertension in young black men
Authors: Keller David M , Holwerda Seth W , Credeur Daniel P , Fadel Paul J
Published: June 2021

Blacks are at an elevated risk for developing hypertension. Although carotid baroreflex control of heart rate in blacks has been investigated, little is known regarding integrated arterial baroreflex (ABR) control of the heart in blacks. Thus, we tested the hypothesis that cardiovagal ABR is reduced in blacks compared to whites. The slope of the relationship (i.e., gain) between changes in systolic blood pressure and r‐r interval were determined in 16 non‐hypertensive men (7 black and 9 white, age 18–27 yr) using 100μg iv bolus of sodium nitroprusside (NTP) followed 1 min later by 150μg bolus of phenylephrine (PE) during rest (3 trials). In addition to analysis of the complete mod‐Ox trials (overall ABR gain), separate linear analyses of the NTP (hypotenion) and the PE (hypertension) phases were also performed. Overall ABR gain was not different between racial groups (black: 14±4 vs white: 13±4 ms/mm Hg, p=0.64). Although the gain for the NTP phase was also not different between groups (p=0.17), the gains for the PE phase were significantly reduced in blacks (10±4) compared to whites (18±10 ms/mm Hg; p=0.04). These findings indicate that blacks have diminished ABR responses to acute hypertensive stimuli and support racial differences in cardiovagal ABR function between black and white men.

Supported by NIH R15HL096103 (DMK) and RO1 HL093167 (PJF)
ACE2 inhibits Endoplasmic Reticulum stress and autophagy associated to neurogenic hypertension
Authors: Xia Huijing , Johnson Tanya , Mungrue Imran , Lazartigues Eric
Published: June 2021

Endoplasmic reticulum (ER) stress has recently been suggested to contribute to the development of hypertension. We hypothesized that ACE2, a counter regulator of the Renin‐Angiotensin System, could oppose the Angiotensin‐II‐mediated activation of ER stress in neurons. In vitro, Neuro 2A cells treated with Ang‐II (100 nM, 24 h) show significant increases (qRT‐PCR) in ER stress and autophagy markers (fold change, P<0.05): IRE1 (2.0 ±0.3), EIF2α (2.0 ±0.1), EIF2α kinase (2.0 ±0.3) and CHOP (1.3 ±0.1). To test whether ACE2 can prevent ER stress and autophagy in neurogenic hypertension, DOCA‐salt hypertension was induced in non‐transgenic (NT, n=6) and transgenic mice overexpressing ACE2 in the brain (SA, n=6). Subfornical organ and PVN punches were collected and ER stress assessed in both NT (136 ±2 mmHg) and SA (126 ± 1 mmHg) DOCA mice. DOCA‐salt hypertension significantly increased (P<0.05 vs. no treatment) ATF4 (1.6 ±0.1; 1.3 ±0.1), spliced XBP (6.5 ±0.9; 1.6 ±0.1) and CHOP (1.5 ±0.02; 1.4 ±0.2) in NT mice SFO and PVN, respectively. However, in SA mice with blunted hypertension, ATF4 (1.2±0.1; 1.0±0.03), spliced XBP (4.5±0.1; 0.9±0.2) and CHOP (1.0±0.1; 1.2±0.1) were reduced (P<0.05 vs. NT+DOCA). Our data suggest that ER stress and autophagy are increased in DOCA‐salt hypertension, through PERK‐eIF2α‐ATF4 and IRE1‐SXBP1 pathways, and ACE2 inhibits both cascades in the brain, thus reducing neurogenic hypertension.

American Heart Association (12EIA8030004); NIH/NHLBI HL093178.
Effect of repetitive coughing on swallowing in the anesthetized cat
Authors: Spearman Daniel , Pitts Teresa , Rose Melanie J , Mortensen Ashley , Davenport Paul W , Bolser Donald C
Published: June 2021

Aspiration can trigger repetitive coughing, and swallow can be inter‐laced between successive coughs to clear the pharynx of ejected material. We hypothesized that repetitive coughing would increase swallow drive and change swallow phase durations and amplitudes of pharyngeal or laryngeal muscle activities. Cough was induced by mechanical stimulation of the trachea and swallow was elicited by injection of water into the oropharynx in spontaneously breathing cats. Electromyograms (EMG) were recorded from pharyngeal, laryngeal and respiratory muscles. During coughing bouts, EMG amplitudes of the geniohyoid, cricopharyngeus, thyropharyngeus, thyrohyoid significantly increased compared to swallows during eupnea. Swallow‐related phase durations of specific muscles were unchanged during coughing. However, there was a significant reduction in the duration of laryngeal elevation and the total swallow time indicating reduced onset latencies between different upper airway muscle groups. These results support a theory of a highly cooperative airway protective network which responds to aspiration. This network must be present to ensure timely and efficient execution of swallow. Supported by NIH HL89104, HL103415, HL109025.
Ampakines induce a persistent increase in phrenic motor output in rats with cervical spinal cord injury
Authors: Sandhu Milap Singh , Elmallah Mai K , Lane Michael A , Reier Paul J , Greer John J , Fuller David D
Published: June 2021

Cervical spinal cord injury (SCI) typically interrupts descending synaptic inputs to phrenic motoneurons resulting in diaphragm paralysis and respiratory impairment. The primary bulbospinal excitatory inputs to phrenic motoneurons are glutamatergic with AMPA receptors playing a prominent role. Ampakines are a group of compounds that can enhance the function of AMPA receptors, and prior work demonstrates that these compounds can increase respiratory output during conditions associated with blunted respiratory drive. Accordingly, in ongoing studies we are testing the hypothesis that intravenous delivery of an ampakine, CX717, will trigger increases in inspiratory phrenic motor output in rats with chronic cervical contusive SCI. Bilateral phrenic nerve activity was recorded in rats that received cervical (C3) hemicontusion of the spinal cord 8 wks prior. A single dose of CX717 (30mg/kg) triggered a modest increase in phrenic inspiratory burst amplitude ipsilateral but not contralateral to the contusion injury. However, repeated doses (3 × 10mg/kg at 5min intervals) triggered a substantial and persistent increase (lasting at least 60mins) in the inspiratory burst amplitude recorded in both phrenic nerves. These results demonstrate ampakines can increase phrenic motor output after cervical SCI and may provide a new means of enhancing respiratory function after SCI.

Support: Craig H. Neilsen Foundation #220521 (MSS), Parker B. Francis Fellowship (MKE), 1R01NS080180–01A1 (DDF), NIH1R21HL104294–01 (DDF)
Stress‐induced activation of a distinct subpopulation of GABA neurons in the basolateral amygdala of male and female rats
Authors: Bolanos Juan M , Lopez Alejandra E , Middleton Capri B , Del Cid Jiselle J , Hamdan Jameel N , Ruiz Christina C , Gosselink Kristin L
Published: June 2021

Previous studies have shown that a repeated 6‐hr restraint stress causes hippocampal plasticity and a shift in the profile of activated neurons in the basolateral amygdala (BLA), suggesting an alteration in mechanisms that direct learning and memory in repeatedly stressed individuals. The purpose of this study was to determine whether a less severe stress exposure induces similar adaptations and whether this response is sex‐specific. Adult male and female rats were acutely or repeatedly restrained for 30 min/d; brain sections were stained immunohistochemically for Fos and calbindin (CB), a marker of GABAergic neurons, and quantified in the BLA to determine cellular activation along with phenotypic profile. Significance was determined by t‐test, with p<0.05 considered significant. We hypothesized that repeated stress would decrease the number of BLA neurons expressing both Fos and CB, compared to control or acutely stressed rats, but that these effects would not differ between sexes. In support of our hypothesis, the number of CB‐expressing neurons in the BLA did not differ according to sex or stress treatment. However, we did not find any significant differences in the number of activated (Fos‐expressing) CB neurons in either sex in response to stress. Therefore, this specific type of GABAergic influence from the BLA to the hippocampus may not be affected by this level of stress.

Supported by: 8G12MD007592
INSPIRATORY INTERCOSTAL LONG TERM FACILITATION IN UNANESTHETIZED RATS
Authors: Opazo Angela Andrea Navarrete , Mitchell Gordon
Published: June 2021

Moderate acute intermittent hypoxia (AIH) elicits a form of respiratory plasticity known as long‐term facilitation (LTF). AIH‐induced LTF is most frequently observed as a persistent increase in phrenic motor output (phrenic long term facilitation, pLTF), diaphragm activity (diaphragm long term facilitation, diaLTF) or ventilation (ventilatory LTF). Since little is known concerning the potential contributions of accessory inspiratory pump muscles to ventilatory LTF, we investigated the effects of AIH on respiratory activity in the second external intercostal muscle (T2 EIM) in unanesthetized Sprague Dawley rats. Rats were instrumented to enable chronic EMG telemetry from the right T2 EIM and diaphragm. Diaphragm and T2 EIM EMG activities were measured before (60 min), during, and after AIH (10, 5‐min hypoxic episodes, 10.5% O2; 5 min normoxic intervals; n = 8) or time control (continuous normoxia, n = 6). Integrated Dia (22.1 ± 6.6% p < 0.05) and T2 EIM burst amplitude (42.6 ± 3.5 % p <0.05) were significantly increased 60 min post‐AIH; thus EIM LTF is relatively greater than diaLTF in unanesthetized rats. Respiratory frequency (15 ± 2.7%; p < 0.05) and external intercostal minute activity (activity × frequency; 64.1 ± 5.7%, p < 0.05) were also significantly elevated 60 min post‐AIH. TC did not induce diaLTF or EIM LTF. Thus, inspiratory intercostal muscles express greater capacity for AIH‐induced plasticity versus the diaphragm. An understanding of AIH induced respiratory plasticity may inspire development of new therapeutic strategies to treat conditions causing respiratory insufficiency, such as cervical spinal injury. Supported by NIH HL69064 and HL80209.
The Effects of Second‐ and Third‐Hand Cigarette Smoke Exposure on the Adrenal Activity in Swiss Webster Mice
Authors: Shakir Reema , Thao My Lo , Watson Flora
Published: June 2021

Cigarette smoke is a mixture of chemicals, the majority of which have detrimental effect on the body at the physiological and cellular levels. The effects of chronic cigarette smoke exposure on hormone secretions are mediated by the pharmacological action of nicotine and other toxins found in cigarette smoke. Exposure to cigarette smoke can be first‐, second‐, or third‐hand smoke. Firsthand cigarette smoke (FHS) is the inhaled smoke from a burning cigarette. Second‐hand cigarette smoke (SHS) is exhaled smoke from a burning cigarette and smoke from the filter or mouthpiece end of a cigarette. Third‐hand cigarette smoke (THS) is often smelled but not seen after tobacco has been smoked. THS encompasses the environment in which smoking has occurred as well as the chemical residues that coat all indoor surfaces such as walls, carpet, and furniture. The specific aim of this study is to investigate the effects of SHS and THS exposure on cortisol levels. Cortisol is a type of corticosteroid that is secreted in response to stress. Long term increased cortisol levels have been shown to have a large impact on glucose metabolism (Targher, 2005) and may also contribute to diabetes. We hypothesize that SHS and THS exposure will have an increase in cortisol levels. Mice were randomly divided into experimental (SHS and THS) and control groups. The experimental mice were exposed to either second‐ or third‐ hand cigarette smoke once a day, five days a week, for 37 weeks. Fecal pellets were collected for cortisol extraction. The levels of cortisol were determined using ELISA (Enzyme‐linked immunosorbent assay). The resting cortisol levels were significantly lower after seven months of cigarette smoke exposure. However, immediately following smoke exposure, there was an increase in cortisol levels. This result parallels the effects in humans (Winternitz & Quillen, 1977; Wilkins et al., 1982; Seyler et al., 1984).
Selective antagonism of mitogen‐activated protein (MAP) kinase ‐, but not phospholipase C (PLC)‐, dependent angiotensin II (Ang II) actions in brain
Authors: Yosten Gina L.C. , Liu Jun , Sandberg Kathryn , Samson Willis K.
Published: June 2021

Hallmark central nervous system actions of Ang II include stimulation of ingestive behaviors and activation of the autonomic nervous system. We identified a 7 amino acid peptide (PEP7) encoded within a short open reading frame in exon 2 of the rat angiotensin type 1a receptor (AT1aR) gene that inhibits Ang II‐induced activation of extracellular–regulated kinase 1&2 (ERK1/2), suggesting that PEP7 inhibits AT1aR action. We investigated the potential for PEP7 to antagonize the central actions of Ang II. Central administration of 1 nmole PEP7 significantly abrogated ingestion of 1.5% NaCl, but not water, in response to a stimulatory dose of Ang II (25 pmole) in male rats. Since saline drinking in response to Ang II is dependent upon the MAP‐kinase pathway, while water ingestion is activated by a protein kinase C (PKC)‐dependent pathway (Daniels et al. Exp Physiol 94:130,2008), these results suggest that PEP7 is a selective antagonist of MAP‐kinase‐dependent actions of Ang II. PEP7 pretreatment did not alter water drinking in response to overnight water restriction or the central autonomic actions of Ang II, suggesting that in those models, the actions of Ang II are more dependent upon PKC signaling than the ERK1/2 pathway. Thus PEP7 may be an important tool with which the solute‐dependent actions of Ang II can be isolated from those related to volume homeostasis. [NIH 066023 (WKS), NIH HL‐57502 (KS)]
Differential Expression of Toll‐like Receptors in the Cecum of Hibernating Ground Squirrels
Authors: Sprenger Ryan J. , Carey Hannah V. , Kurtz Courtney C.
Published: June 2021

Mammalian hibernation is a response to periods of low temperature and limited food availability. The GI tract of hibernators undergoes significant physiological changes. Recent studies have found shifts in the microbial populations inhabiting the cecum during hibernation. We used immunoblotting to examine the expression of Toll‐like receptors (TLRs) in the cecum of summer, hibernating and spring ground squirrels. TLR4 expression decreased during the hibernation season with the lowest levels found in squirrels that had been hibernating at least 3 months (late‐season hibernators). TLR5 expression showed the opposite trend with increased expression in late‐season hibernators compared to summer. Levels of TLR5 in spring squirrels were similar to summer, but TLR4 was similar to late‐season hibernators. Expression of both TLRs did not differ between torpid and aroused hibernators at the same point in the season (early versus late). Together, these results suggest that hibernation induces changes in microbial sensing, perhaps as a result of increased permeability of the GI epithelium and shifts in the composition of the microbiota. Supported by UW‐Oshkosh Student‐Faculty Collaborative Grant Program.
MicroRNA‐182 targets FoxO3 and attenuates glucocorticoidinduced atrophic signaling
Authors: Hudson Matthew B , Woodworth‐Hobbs Myra E , Zoromsky Sara A , Zheng Bin , Franch Harold A , Price S. Russ
Published: June 2021

Skeletal muscle atrophy occurs in response to glucocorticoid administration, disuse, chronic kidney disease, diabetes, and other systemic diseases and chronic conditions. Previous studies demonstrate that upregulation of Forkhead box O (FoxO) transcription factors induces skeletal muscle atrophy by activating the ubiquitin‐proteasome and autophagy‐lysosome systems. The purpose of the present study was to identify a microRNA that targets and inhibits FoxO transcription factors. In silico target scan analysis revealed miR‐182 as a potential regulator of the FoxO proteins. To test whether miR‐182 targets the FoxOs, C2C12 myotubes were transfected with miR‐182 and levels of FoxO1 and FoxO3 were evaluated by western blot analysis. Overexpression of miR‐182 suppressed FoxO3, but not FoxO1. Utilizing glucocorticoids to induce muscle atrophy, we tested whether dexamethasone (1uM, 6 h) altered the level of mir‐182 and found it was suppressed by 44% (P<0.05). Additionally, overexpression of miR‐182 prevented the glucocorticoid‐induced upregulation of FoxO3 target genes including MAFbx/Atrogin‐1, ATG12, Cathepsin L, and LC3. These data indicate that miR‐182 is a key regulator of FoxO3‐mediated signaling in muscle cells, and a potential therapeutic agent to prevent FoxO3 mediated muscle wasting.
DELETION OF MBTPS1 IN BONE LEADS TO ENHANCEMENT OF MUSCLE MASS AND FUNCTION IN MATURE MICE
Authors: Gorski Jeffrey , Huffman NT , Breggia Anne , Rosen Cliff J. , Chittur Sridar V. , Stern Amber , Dallas Mark , Seidah N G , Brotto Leticia , Bonewald Lynda F. , Brotto Marco
Published: June 2021

Cis‐Golgi protease Mbtps1, a proprotein convertase, is responsible for activating a small family of transmembrane spanning b‐ZIP transcription factors. We previously demonstrated Mbtps1 is required for mineralization of osteoblastic cultures. To understand its role in skeletal development, we conditionally deleted MBTPS1 using osteocyte restricted DMP1 Cre recombinase. Despite the absence of an early bone phenotype, KO male mice abruptly begin to gain weight at skeletal maturity. At 40 weeks, knockout mice were 7.3g larger (p=0.034) than littermate controls. MicroCT and piximus analysis of bone and fat content cannot account for this difference in mass. Since MBTPS1 was specifically ablated in osteocytes, we initiated systematic muscle physiological studies to test our recently proposed hypothesis of a bone‐muscle crosstalk axis in health and disease. We found that the extensor digitorum longus and soleus muscles were significantly heavier and stronger in cKO mice as compared to controls. We also observed a leftward shift in their force vs. frequency relationships, suggesting that cKO muscles could be more sensitive to calcium. Interestingly, leptin was found to be 2‐fold higher in serum from cKO mice than controls (p=0.018). We postulate that MBTPS1 ablation in osteocytes leads to augmented serum levels of leptin that acts systemically to alter muscle function. Support: NIH‐NIA P01 AG039355 and MOLSRB.
Tadalafil‐sensitive impairment in muscle blood flow during exercise in Duchenne Muscular Dystrophy
Authors: Nelson Michael , Martin Elizabeth A. , Tang Xiu , Johannes Jimmy , Lewis Joshua , Gurudevan Swaminatha V. , Victor Ronald G.
Published: June 2021

Out‐of‐frame mutations in the dystrophin gene cause Duchenne muscular dystrophy (DMD)—a devastating X‐linked muscle wasting disease for which there is no treatment other than corticosteroids. In DMD, loss of the cytoskeletal protein dystrophin impairs sarcoelmmal targeting of nNOS, which is the main source of skeletal muscle‐derived nitric oxide (NO). We previously showed that loss of nNOS impairs the normal exercise‐induced attenuation of reflex vasoconstriction in dystrophic skeletal muscle, thus implicating a putative vascular component to the pathogenesis of DMD. Here we present data on a second phenotype, that muscle blood flow (BF, measured by Doppler ultrasound of the brachial artery) fails to increase normally during mild rhythmic handgrip exercise in 6 boys with DMD (7–13 years of age) compared with 8 age‐matched male controls (Ctrls): ΔBF:+13±5% vs. +81±10%, respectively (p<.05). Moreover, we show that the phosphodiesterase 5 inhibitor Tadalafil, restores active hyperemia in boys with DMD in a dose‐dependent manner: 0.5 mg/kg, +56±13%; 1.0 mg/kg, +72+18% These data significantly advance the vascular hypothesis of DMD and implicate PDE5 inhibition as a putative therapeutic treatment option.

Support: Parent Project Muscular Dystrophy, Heart and Stroke Foundation of Canada (MN)
Sonic Hedgehog Acts Via A Smoothened‐Dependent Pathway As A Macrophage Chemoattractant
Authors: Schumacher Michael , Aihara Eitaro , Herbert DeBroski , Zavros Yana
Published: June 2021

Helicobacter pylori (H. pylori) infection elicits recruitment of macrophages (Mø) to the stomach. In mice expressing a deletion of the Sonic Hedgehog (Shh) signaling protein Smoothened (Smo) within the myeloid lineage, Mø are not recruited to the stomach in response to H. pylori infection. Therefore, we hypothesize that Shh acts as a Mø chemoattractant via a Smo‐dependent mechanism. Bone marrow‐derived macrophages (BM‐Mø) were collected from control mice and transgenic mice lacking the Hedgehog signaling protein Smo (LysMCre/SmoKO) in cells of myeloid lineage. Migration of BM‐Mø was performed using ibidi μ‐Slide Chemotaxis chambers following addition of Shh or chemokine ligand CCL2. A stable chemotaxis gradient was confirmed by diffusion of lucifer yellow. Forward migration index, a measure of directed movement, was significantly elevated in control Mø migrating towards Shh compared to media. In contrast, Smo‐deficient Mø did not migrate towards Shh. Control, but not Smo‐deficient, Mø migrated toward chemokine CCL2 indicating involvement of Smo in macrophage migration. In conclusion, Shh acts as a Mø chemoattractant via a Smo‐dependent mechanism. Such findings are consistent with in vivo studies showing that Smo is required in macrophages for their recruitment to the stomach following H. pylori infection. Funding: NIH 1R01DK083402–01 (Y Zavros), Albert J. Ryan Foundation Fellowship (MA Schumacher)
Ursodeoxycholic acid inhibits colonic mucosal cytokine release and prevents colitis in a mouse model of disease
Authors: Ward Joseph Bernard Jude , Kelly Orlaith B , Jefferies Caroline , Murray Frank E , Fallon Padraic G , Keely Stephen J
Published: June 2021

Toll‐like receptors (TLR) promote cytokine release from intestinal epithelia, thereby driving inflammation. Although ursodeoxycholic acid (UDCA) has anti‐inflammatory effects in the liver, its potential for treating intestinal inflammation has not been well‐studied. Here, we investigated UDCA effects on TLR‐driven colonic epithelial cytokine release and induction of inflammation in a mouse model of colitis.

Resected human colonic mucosa and T84 colonic epithelial cells were treated with specific TLR agonists, in the presence or absence of UDCA. Released cytokines were measured by ELISA. Dextran sodium sulphate (DSS) colitis was induced in C57BL/6 mice along with daily intraperitoneal injections of UDCA (30 mg/kg) for 5 days. Colitis severity was recorded as disease activity index (DAI). In T84 cells, UDCA reduced TLR2 and 4‐stimulated RANTES release and TLR3‐stimulated TNFα release, compared to controls. TLR4‐driven IL‐8 release was abolished by UDCA in human colonic mucosa. Inhibitors of the transcription factors, NFκB and AP1, mimicked UDCA effects suggesting these pathways may be involved. Furthermore, UDCA treatment significantly reduced DAI in DSS‐treated mice.

In conclusion, our data suggest that, by virtue of its effects in preventing TLR‐induced proinflammatory cytokine release, UDCA may be useful for treating intestinal inflammation.

This work was funded by Science Foundation Ireland.
DGAC is a Novel Mouse Model of EMT‐mediated Renal Fibrosis.
Authors: Chen lihe , Wu Hongyu , Zhang Xi , Zhou Qiaoling , Zhang Wenzheng
Published: June 2021

The role of the epithelial‐mesenchymal transition (EMT) in the renal fibrosis is not clear, partially due to a lack of reliable animal models mimicking chronic kidney disease (CKD) in humans. Here we report DGAC mice to fill this gap. DGAC is a triple transgenic line in which Dot1l loss and GFP expression occur upon Aqp2‐ promoter‐driven Cre‐mediated recombination. DGAC is novel and unique since 1) Cre is exclusively expressed in Aqp2+ principal cells (PC). 2) Dot1l is solely responsible for histone H3 K79 methylation (m2K79). 3) Loss of m2K79 not only causes cell reprogramming as evidenced by derivation of intercalated cells from PC, but also can be used as a tracing marker for fate mapping. 4) Fibrogenesis in adult DGAC mice rarely occurs in the normal condition, but can be extensively induced by chronic Na+ loading and Streptozotocin‐induced diabetes, or during aging. 5) EMT occurs in fibrotic DGAC kidney, as evidenced by GFP+FSP1+ and m2K79−FSP1+ cells in the collecting duct and interstitial space. And 6) AQP2+FSP1+ and m2K79−FSP1+ cells were also observed in kidney biopsies from patients with CKD, indicating the relevance of DGAC. Therefore, DGAC is a novel model of EMT‐mediated renal fibrosis.
Anatomy at West Virginia University: 1867–2012.
Authors: Culberson James L.
Published: June 2021

As the AAA celebrates 125 years, West Virginia University completes 145 years teaching anatomy. Since WVU's founding in 1867, MD‐trained profs who offered courses in anatomy and medicine worked through administrative high points and headaches to grow into a sound four‐year medical program by the early 1960s. This presentation offers a review of the growth and development of anatomy at West Virginia University. Those 1870s‐era courses in anatomy, physiology and hygiene were replaced by a medical “department” ten years later, and a real two‐year College of Medicine emerged by 1912. Nearly 50 more years of erratic “growth” climaxed in a four‐year medical program (1957) and the growth of a modern academic anatomy department (expanded to 15–20 faculty) offering professional anatomical instruction. Since 1960 the department has matured into a balanced academic unit with increased participation in and support for research, education and professional service. About 50 PhD graduates now staff biomedical programs and activities in the US and abroad, and a substantial number of past and present faculty members have served the discipline well beyond the local level. Five of these have chaired anatomy departments; others have assumed leadership roles in the AAA (Sec.‐Treas, two Presidents, board members) and other professional organizations.
Relationship between spatial abilities and cross‐sectional drawings
Authors: Langlois Jean , Dagenais Yvan , Bellemare Christian , Lemieux Renald , Martin Marcel , Lecourtois Marc , Yetisir Elizabeth , Bergeron Germain , Hamstra Stanley , Wells George A.
Published: June 2021

Objective

Spatial abilities have been related to cross‐sections of anatomical structures. The objective was to assess the relationship of spatial abilities to positioning and assessing area of structures in cross‐sectional drawings.

Methods

Fourth‐year medical students (n = 49) were recruited in a prospective study. Spatial abilities were measured with a redrawn Vandenberg and Kuse Mental Rotations Test in two (MRTA) and three (MRTC) dimensions and the Surface Development Test (SDT). Participants had to draw by observation the four lateral sides of objects made of several structures. After the objects were removed from view, cross‐sections of the objects were drawn. Position and area of structures within drawn objects were compared to standard. Results were expressed as means ± standard deviation and Spearman's correlation coefficient was used to assess the relationship of positional and area errors of drawn structures to MRTA, MRTC and SDT scores.

Results

Positional error (9805 ± 6699 pixels) was inversely related to MRTA (13.5 ± 5.2), MRTC (9.7 ± 4.5) and SDT (43.4 ± 10.0) scores; with a correlation of − 0.3718 (p = 0.0085), − 0.4682 (p = 0.0007) and − 0.3461 (p = 0.0149), respectively. Similarly, area error (440416 ± 131870 pixels) was not related to MRTA, MRTC and SDT scores.

Conclusion

Spatial abilities were related to the position, but not to the area, of structures in cross‐sectional drawings of objects.

This study was supported by an internal grant from the Department of Surgery, University of Sherbrooke, Sherbrooke, QC, Canada.
Medical Student Feedback on Participating in a Combined Anatomy Curriculum, 2010–2012
Authors: Guzman Carlos , Young Stephanie , Wimmers Paul , Byus Craig , Wisco Jonathan
Published: June 2021

First‐year medical students at the David Geffen School of Medicine at UCLA and the Thomas Haider Program in Biomedical Sciences at UCLA/UCR were asked about their experiences and their perception of the value anatomy has in the medical curriculum. Annual responses were compared to determine whether the qualitative responses changed from year to year. We conducted three annual surveys of 2nd – 4th year medical students. Students were asked how the anatomy curriculum affected their confidence in physical examination skills, preparation for USMLE board exams, and preparation for 3rd and 4th year clerkships. Most students viewed the anatomy curriculum highly in helping them prepare for USMLE board exams (89% positive response), 3rd year clerkships (95%), and developing physical exam skills (65%). When asked about how the anatomy curriculum helped prepare students for 4th year clerkships, there was a downward trend with a 92% positive response rate for 2010, 90% for 2011, and 83% for 2012. These results appear to indicate a perceived diminishing value of either anatomy pedagogy as the medical experience becomes more clinical.

Grant Funding Source: AAA
Peer teaching in an online histology classroom
Authors: Barbeau Michele , Rogers Kem A
Published: June 2021

The impact of a peer teaching exercise for an online histology course has been assessed using a mixed method approach. Students signed up for a time they would be available to meet live in the virtual classroom. Each week, students were given specific virtual slidebox assignments which they uploaded into the classroom with full annotations. At their group meeting time, members would log into the classroom and take turns presenting their slides; presentations were archived for future reference. Other students were free to ask questions and the course instructor was either present live or viewed the presentation archive at a later time to ensure accuracy or provide clarification. Laboratory grades improved significantly compared to the previous year when peer teaching was not included (+9.4% (p<0.05)). Student surveys indicated that students believed that teaching others was “very helpful” (67%) or “helpful” (33%) for enhancing their understanding of course material compared to 100% who found attending presentations “helpful” for enhancing their understanding. In summary, we have shown that peer teaching in the online environment is an effective means to enhance student learning.

Grant Funding Source: Social Sciences and Humanities Reseach Council of Canada
Anatomy Academy promotes Systems‐Based Practice among medical and undergraduate students
Authors: Ojukwu Kenechukwu , Padilla Jessica , Diaz Molly , Wisco Jonathan
Published: June 2021

Systems‐Based Practice (SBP) is difficult to define and successfully incorporate into medical education. Nonetheless, it is one of the six Accreditation Council for Graduate Medical Education (ACGME) core competencies that physicians are to attain. Competency in SBP includes understanding of the health system to allow adequate assessment and improvement of the quality of patient care and patient safety. Anatomy Academy (AA) provides a setting where UCLA medical and undergraduate students enter the community to teach fifth graders complex health concepts. Students are encouraged to compliment their pathophysiology and diagnostic training with active practice in communication, program assessment, conflict resolution, and problem solving as they teach complex medical concepts such as problematic cardiac blood flow, lung disease, and gastrointestinal function. Written reflections, program‐ and self‐evaluations suggest that AA provided opportunities to gain basic SBP skills such as improved conflict management and problem solving. Community programs such as AA can contribute to medical education curriculum by allowing early medical student exposure to SBP.

Grant Funding Source: Albert Schweitzer Fellowship, UCLA Monica Salinas Fellowship
Our success in translating Anatomy Academy, an intervention program for 5th and 6th grade students, from Los Angeles, CA to Salt Lake City, UT.
Authors: Steed Kevin S , Diaz Molly , Ojukwu Kene , Padilla Jessica , Jenkins Katherine , Lassetter Jane , Wisco Jonathan J.
Published: June 2021

Anatomy Academy (AA) is an educational intervention program to fight obesity and promote both healthy lifestyles and interest in science in children, through the teaching of basic anatomy and physiology to 5th and 6th grade students in didactic small group and physical activities. AA's potential to cultivate lifestyle change children at the community, state, and national levels is only as effective as its scalability. We report the successful adaption of AA from Stella Middle Charter Academy (Los Angeles, CA) to Salt Lake Arts Academy (Salt Lake City, UT). Volunteer undergraduate and graduate students from UCLA, BYU, and University of Utah Medical School provide great service to their communities while gaining invaluable mentoring experience. AA provides a uniting curriculum and structure, with the flexibility for adaptation to individual site's needs, allowing for creativity to flourish from both students and mentors. Communication via social media has allowed sharing of ideas and experiences across sites. With 5 additional schools in Salt Lake and Utah Counties adopting AA in 2013, including several Title 1 schools, AA is making significant progress to fight obesity one community at a time.

Grant Funding Source: The Albert Schweitzer Fellowship and the UCLA Monica Salinas Fellowship
Identification of nicotinic acetylcholine receptor subunit expression in early avian embryos
Authors: Haroon Zoha , Stathos Joseph , Higgins Daniel , Olley John T. , Brauer Philip R. , Reedy Mark V.
Published: June 2021

Most previous studies on nicotine used relatively high concentrations of nicotine, administered later in development. Using chick embryos as our model system, we have found a single low dose of nicotine (100 nM) administered during early development significantly retarded embryo growth, reduced heart rate, and increased the incidence of defects four days later. It is often assumed that nicotine's teratogenic effects are mediated by nicotinic acetylcholine receptors (nAChRs), whose expression at very early stages of embryonic development has not been thoroughly investigated. The objective was to identify which nAChR subunits are expressed during early avian embryogenesis. We performed RT‐PCR expression analysis for eleven nAChR subunits, including alpha1‐alpha7, alpha9, alpha10, beta3, and beta4, at 24‐, 48‐, 72‐, and 96‐hrs of development. Surprisingly, we find nAChR expression is widespread during early development, which suggests the teratogenic effects of nicotine were mediated by the early activation of nAChRs.

Nebraska NIH‐INBRE Program; Nebraska Health and Human Services (LB595); Ferlic Undergraduate Summer Research Program, Creighton University College of Arts and Sciences

Grant Funding Source: none current