bluuebunny/crossref_metadata_2025 huggingface dataset:
Motivating Me: Healthy Minds, Healthy Bodies: Development of a Psychoeducational Program for Obese Children
Authors: Maia Angelina Moore , White Adrienne A
Published: June 2021

The objective of the study was to develop a psychoeducational program for a clinical setting to promote weight maintenance or loss and improve self‐esteem and body image in 9–10 year old obese children. The program, Motivating Me: Healthy Minds, Healthy Bodies, was designed based on a critical review of clinical interventions and informed by the Self Determination Theory. Motivational interviewing counseling techniques were the hallmark of the intervention. The program design included six, hour‐long individual sessions for the child and parent delivered over a 12 week period; telephone calls with child and parent on non‐session weeks to provide encouragement and problem‐solving assistance; and three, 90‐minute group sessions for parents to address and reinforce information covered at the individual sessions. Content areas included eating breakfast, appropriate portions, increasing fruit and vegetable intake, decreasing sweetened beverage intake, improving body image, improving self‐esteem, and individualized goal setting for behavior change. The program was pilot tested with 3 parents and their children (mean age=10.3 years), each at the 97th BMI percentile. Based on preliminary qualitative data, the children were building confidence, relatedness, and making autonomous decisions about food choices. Program refinement will occur before the intervention phase. Funding: Maine Experiment Station.
Innovation in Dietetic Internship Education at Iowa State University
Authors: MacDonald Ruth S , Anderson Jean A , Bergquist Erin , Johnson Janet S , Kruzich Laurie S
Published: June 2021

Iowa State University (ISU) Dietetic Internship (DI) has expanded from 30 to 120 interns – the largest in the US. This was achieved through integration of electronic educational methods and pedagogy combined with a nationwide preceptor network. Cutting edge educational technology including Blackboard Learn, iTunesU, Survey Gizmo, and GoToMeeting are used. Interns are evaluated by preceptors and ISU faculty using online surveys, assignments and projects, exams, simulations and case studies, blogs and discussion postings, guided reflections, and self and peer evaluation. In 2012 ISU adopted the Independent Supervised Practice Pathway (ISPP) option and created 2 international experiences in Ghana (focused on maternal, infant, child nutrition and feeding practices) and Japan (focused on regional school lunch system or Shokuiku). The ISU DI provides a model for expanding opportunities for training Registered Dietitians (RDs) via educational technology. Our outstanding record of accreditation, registration exam pass rate (87% n=298) and employment (94% n= 102) verifies that distance education with appropriate educational technologies and supervision is a format to meet the increasing demand for RDs.
Elevated Gluconeogenesis in Aging and Lung Cancer is Related to Inflammation and Blunted Insulin‐Induced Protein Anabolism
Authors: MacAdams Jacqueline , Winter Aaron , Morais José A. , Burgess Shawn C. , Chevalier Stéphanie
Published: June 2021

Cancer and aging are both accompanied by metabolic alterations leading to muscle loss that may result from an increased use of amino acids for gluconeogenesis (GNG).

Methods

we measured the fractional contributions (%) of glycogen, glycerol and GNG from phosphoenolpyruvate (PEP) to 17h‐fasting endogenous glucose production (EGP), using oral 2H20 and deuterium enrichment of plasma glucose, by NMR spectroscopy. Whole‐body protein and glucose kinetics (1‐13C‐leucine and 3H3‐glucose) were measured fasting and during a subsequent hyperinsulinemic, euglycemic, isoaminoacidemic clamp in: young (Y, n=11; 27±3 y), older (O, n=13; 65±2 y) and older men with lung cancer (CA, n=9; 66±2 y).

Results

Men with CA had greater serum CRP levels and resting energy expenditure (REE) than Y and O, and a lesser clamp glucose disposal, indicating insulin resistance. EGP was elevated with aging and cancer with greater contribution from GNG: Y:37±2, O:43±2 and CA:48±4% (ANOVA p<0.05); % contribution from glycogen and glycerol did not differ. The resulting GNG flux (EGP × %GNG) correlated positively with CRP (r=0.69) and REE (r=0.61), and negatively with fat‐free mass index (r=−0.41), all p<0.05. GNG flux was related to lesser glucose uptake and protein anabolic response (r= −0.54, p=0.001) in response to insulin.

Conclusions

Inflammation may underlie the greater production of glucose from GNG and blunted protein anabolism. (CIHR)
Grain sorghum reduces plasma glucose and insulin levels in healthy men
Authors: Gu Xuan , Poquette Nicole , Lee Sun‐Ok
Published: June 2021

Starch digestion is classified as: rapidly digestible starch (RDS), slowly digestible starch (SDS), and resistant starch (RS). Functional starch fractions, SDS and RS, are attributed to numerous health benefits. Grain sorghum's functional starch content may help in preventing chronic illness, particularly diabetes and obesity. A human study was conducted to investigate the effects of grain sorghum on plasma glucose and insulin levels. In a randomized‐crossover design, 11 male subjects consumed grain sorghum and wheat muffins. Muffins contained 50‐g of total starch (TS) based on flour analysis, and additional ingredients were consistent. Baseline measurement was collected 15 minutes prior, and 0, 15, 30, 45, 60, 75, 90, 120, and 180 minutes after consumption. Muffin RDS, SDS, RS and TS were analyzed, and SDS and RS were 10% higher in sorghum muffins compared to wheat. Plasma glucose incremental area under the curve (iAUC) reduced ~24% and 15–90 min intervals were significantly lower for sorghum (p<0.05). Also, plasma insulin iAUC reduced significantly and 15–60 min intervals were significantly lower with the sorghum muffins (p<0.05). Results indicate grain sorghum would assist in controlling glucose and insulin levels, and may help prevent diabetes and obesity. Funding provided by Arkansas Grain Sorghum Board is greatly appreciated.
Folate deficiency accelerates Alzheimer's disease related pathology in APP/PS1 mice
Authors: Crivello Natalia , Spires‐Jones Tara , Shukitt‐Hale Barbara , Carey Amanda , Roe Annie , Smith Donald , Blusztajn Jan Krzysztof , Hyman Bradley , Rosenberg Irwin
Published: June 2021

Previous studies have shown that the APP/PS1, double transgenic mouse model of Alzheimer's disease (AD), develops cognitive dysfunction and Abeta deposits from 5 to 6 months of age. We exposed 1 month old APP/PS1 mice to dietary folate deficiency (FD) for 5 months to test the hypothesis that long term FD accelerates AD‐related pathology and thus contributes to cognitive dysfunction. Results showed a robust disruption in learning and memory performance assessed by the novel object recognition (NOR) test in FD group suggesting abnormalities in the cortex of FD mice. NOR behavior was negatively correlated with the plaque burden in the cortex (r=−.55; P<0.05) but not in the hippocampus (r=−.38; P=.20). The size of the plaques was also significantly larger in the cortex of FD mice as compared to those in the control group; and it was negatively correlated with the cortical choline‐containing lipids (r=−.48; P<.05). There was 30% decrease in the concentration of sphingomyelin in the cortex of FD mice but not in the hippocampus. Our findings suggest a higher sensitivity of the cortex to long term FD and AD related pathology as compared to the hippocampus of APP/PS1 mice.
Inhibition of lysine‐specific histone demethylase LSD1 suppresses melanoma growth
Authors: Lian Christine Guo , Fang Rui , Zhan Qian , Ma Jie , Lee Chung‐Wei , Frank Markus H. , Shi Yujiang Geno , Murphy George F.
Published: June 2021

Lysine‐related histone methylation, a key mechanism of epigenetic regulation, is a reversible event mediated by the lysine‐specific demethylase 1 (LSD1). Here we explored the epigenetic effects of LSD1 inhibition in melanoma. First, the expression of LSD1 was evaluated by RT‐PCR in cultured human melanoma cell lines A2058 and SK‐Mel5 and was found to be significantly higher in the more aggressive line A2058 compared to SK‐Mel5 cells. Next, LSD1 was knocked down in A2058 cells using an adenovirus‐based siRNA delivery system. A cluster of genes including Melan‐A and VEGF, were significantly down‐regulated in LSD1 knockdown A2058 cells. Moreover, melanoma growth in soft agar revealed significant reduction in colony formation by cells treated with LSD1 inhibitors. Finally, LSD1 knockdown A2058 cells demonstrated diminished tumor growth in NOD‐SCID mouse xenografts. Overall, our data reveal that LSD1 inhibition suppressed colony formation in vitro and human‐to‐mouse melanoma xenograft growth in vivo. These findings suggest that repression of LSD1 can suppress melanoma growth, thus providing a rationale for development of targeted LSD1 inhibitors for melanoma therapy. Study funded by NIH‐GM078458 and NIH‐NCI 5P50CA093683–09.
Punishment by intravenous nicotine in rats
Authors: Truong Yen Nhu‐Thi , Woods James
Published: June 2021

Rationale

Although it is known that humans will self‐administer intravenous nicotine, the motivational valences of nicotine are not well understood. In order to address the seeming paradox involving the positive and negative subjective effects of nicotine, we have developed operant choice procedures that allow us to detect when intravenous nicotine results in both increases and decreases in behavior resulting in its self‐administration.

Objective

To examine the effects of contingent and non‐contingent intravenous nicotine on sucrose‐maintained operant responding.

Methods

The effects of intravenous nicotine on sucrose maintained behavior in a two‐lever operant chamber was measured in rats.

Results

Response‐contingent nicotine dose‐dependently decreased sucrose maintained‐behavior in our operant choice procedures, and when available, nicotine increased responding maintained by only sucrose alternative consequence. Delay in response‐contingent nicotine attenuated the punishing effect of nicotine. Non‐contingent nicotine, at total cumulative doses equal to and above those self‐administered did not punish sucrose‐maintained behavior.

Conclusion

Intravenous nicotine may serve as a punisher of sucrose‐maintained behavior in rats. University of Michigan Rackham Merit Fellowship.
Gene expression of NMDA receptor subunits using laser capture microdissection in the rostral ventrolateral medulla (RVLM) of sedentary versus physically active rats.
Authors: Subramanian Madhan , Holt Avril G , Mueller Patrick J
Published: June 2021

The RVLM is important in tonic and reflex control of blood pressure and sympathetic nerve activity (SNA). We have observed enhanced SNA responses in sedentary versus physically active rats after microinjection of glutamate in the RVLM. Changes in glutamate receptor gene expression in the RVLM may contribute to enhanced responses in sedentary rats. In order to test this hypothesis, active (voluntary running for 15–18 wks) and sedentary (no running) rats were injected with Fluorogold (5%) at spinal levels T9‐T10 (n=5 ea). Brainstems were cryosectioned at 8μm for laser capture microdissection of bulbospinal RVLM neurons. RNA was isolated to analyze changes in gene expression of NMDA receptor subunits (NR2A‐D) using RT‐PCR. Gene expression was normalized to a housekeeping gene (HPRT). Preliminary results suggest that expression of NR2A‐D is not different between groups. However, within active rats, total running distance was significantly correlated to NR2C gene expression (r2=0.87, p<0.05). These findings suggest that physical activity contributes to NR2C neuroplasticity in the RVLM. However further studies are needed to address whether NMDA receptor protein expression contributes to enhanced responses in sedentary rats. (R01HL096787; DC007733)
Central modulation of arterial chemo‐baroreflex control by propofol during severe arterial hypoxia in the rabbit
Authors: Quail Anthony W , Cottee David B , Roach Rebecca , White Saxon W
Published: June 2021

During hypoxia specific deficits in neural control caused by propofol (2,6‐diisopropylphenol) anesthesia (Prop) are unknown. Dose‐related effects were analysed in rabbits with and without carotid body denervation (CBD) to partition arterial chemo‐baroreflex control. Minute ventilation (VE), tidal volume (VT), respiratory frequency (f), heart rate (HR), mean arterial pressure (MAP) were measured in 12 awake rabbits mask‐breathing room air, then 15 minutes of low oxygen air (PaO2 < 35 mm Hg), during Prop alone (0.2 and 0.5 mg kg−1min−1i.v.), and during matched Prop plus hypoxia. Prop alone caused dose‐related depression in f and rise in HR. Prop plus hypoxia abolished bradycardia, while MAP, VE and VT effects remained intact. CBD eliminated all chemoreflex responses to hypoxia and MAP fell while HR rose, effects enhanced by Prop in a dose‐related way. It is concluded that during Prop, carotid body function for respiratory and MAP defense is functionally intact. Central activation of the vagus is inhibited selectively, because electrical stimulation of peripheral vagus during Prop evokes normal bradycardia blocked by atropine. By contrast, Prop inhibits arterial baroreflex‐vagosympathetic defense against widespread hypoxic vasodilatation. We speculate that the data reflect dominant central GABAA receptor agonist effects on the arterial chemo‐baroreflex arc. Funding: ANZCA and HMRI.
Ionotropic but not metabotropic glutamatergic receptors in the Locus Coeruleus modulate the hypercapnic ventilatory response in unanesthetized rats
Authors: Dias Mirela Barros , Taxini Camila L , Puga Cintia C.I. , Gargaglioni Luciane H , Bícego Kênia C.
Published: June 2021

Central chemoreception is an important mechanism by which changes in tissue CO2/H+ regulate breathing, and the Locus coeruleus (LC) is one of many putative central chemoreceptor sites. Here, we studied the contribution of LC glutamatergic receptors on ventilatory, cardiovascular and thermal responses to hypercapnia. To this end, we measured pulmonary ventilation (VE), body temperatures (Tb), mean arterial pressure (MAP) and heart rate (fH) of male Wistar rats before and after microinjection of Kynurenic Acid (KY, an ionotropic glutamate receptors antagonist, 10 mM) or α‐methyl‐4‐carboxyphenylglycine (MCPG, a metabotropic glutamate receptor antagonist, 10 ηM) unilateraly into the LC, followed by 60 minutes of air or hypercapnia exposure (7% CO2). Ventilatory response to hypercapnia was higher in animals treated with KY intra‐LC (1899 ± 83 mL kg−1 min−1) compared to the control group (1141 ± 64 mL kg−1 min−1; P < 0.01). However, the MCPG treatment within the LC had no effect on the hypercapnia‐induced hyperpnea. The cardiovascular and thermal control were not affected by hypercapnia or by the injection of KY and MCPG in the LC. These data suggest that the glutamate acting on ionotropic but not metabotropic receptors in the LC exerts an inibitory modulation of hypercapnia‐induced hyperpnea.

Financial support: FAPESP, CNPq, PROPe.
Inhibition of the receptor of advanced glycation reverses pulmonary arterial hypertension
Authors: Meloche Jolyane , Courchesne Antony , Barrier Marjorie , Tremblay Marie‐Pier , Breuils‐Bonnet Sandra , Tremblay Eve , Provencher Steeve , Bonnet Sebastien
Published: June 2021

Pulmonary arterial (PA) hypertension (PAH) is a vasculopathy characterized by enhanced PA smooth muscle cells (PASMC) proliferation and suppressed apoptosis. A recent proteomic analysis of PAH lungs demonstrated that the receptor of advanced glycation endproducts (RAGE) is one of the most upregulated proteins in PAH patients. We hypothesized that RAGE could play a critical role in PAH etiology and progression. We demonstrated a specific RAGE overexpression in PAH lungs. In human PASMC, we showed that RAGE was implicated in the proliferative and apoptosis‐resistant phenotype. Recent studies demonstrated an altered BMPR2 (Bone Morphogenetic Protein Receptor 2)/PPARγ (Peroxisome Proliferator‐Activated Receptor gamma) axis in PAH. We showed that RAGE is accountable for BMPR2/PPARγ downregulation, thus regulating proliferation and apoptosis. In vivo, RAGE inhibition (siRNA nebulization) in rats with Sugen‐induced PAH showed decreased PA pressure and right ventricular hypertrophy, associated with a significant improvement in lung perfusion and decreased PA wall thickness. To conclude, we demonstrated the implication of RAGE in PAH etiology. Thus, RAGE inhibitors, which are in clinical phase II for Alzheimer's disease, represent potential therapeutic agents for PAH treatment.

A Canadian Thoracic Society scholarship to JM and Canadian Institutes of Health Research grants to SB supported this work.
Bradykinin‐induced pulmonary vasorelaxation is modified by long term hypoxia and postnatal maturation in sheep
Authors: Blum‐Johnston Carla , Blood Quintin , Wee Chelsea , Wilson Rachael , Blood Arlin B , Longo Lawrence D , Wilson Sean
Published: June 2021

Endothelial activation is critical to pulmonary vasorelaxation during the fetal transition. Bradykinin (BK) relaxes vessels through endothelial‐induced nitric oxide (NO) and prostacyclin (PGI2) dependent signaling pathways. Intrauterine long term hypoxia (LTH) can stunt vasorelaxation in the fetal transition and cause pulmonary hypertension of the newborn. We therefore tested the hypothesis that LTH impairs maturation of BK‐mediated vasorelaxation by performing wire‐myography on pulmonary arteries (PA) isolated from fetal, newborn, or adult sheep that lived at low (720 m) or at high altitude (3,801 m) for >;100 days. Blocking eNOS‐dependent NO production with 100 μM LNAME and cGMP generation by soluble guanylate cyclase with 10 μM ODQ reduced 1 μM BK vasorelaxation in fetal and newborn but not adult vessels, effects largely independent of altitude. Inhibition of PGI2 production with 10 μM indomethacin also decreased BK‐vasorelaxation in fetal hypoxic and newborn normoxic PA. Interestingly, LTH enhanced BK‐mediated vasorelaxation in fetal vessels, suggesting a compensatory mechanism. Overall, the data imply that maturation has greater influence than altitude on eNOS ‐ cGMP mediated vasorelaxation. Moreover, cGMP as well as PGI2 pathways are important therapeutic avenues for pulmonary hypertension in newborns and adults. (NSF MRI923559, NIH R03HD69746, R01HD31226, R01HD3807, R01HL95973, P20MD6988)
Activation of calpain in pulmonary arterial smooth muscle cells (PASMCs)
Authors: Kovacs Laszlo , Rafikov Ruslan , Szabo Andras , Bagi Zsolt , Black Stephen M. , Su Yunchao
Published: June 2021

Pulmonary arterial hypertension (PAH) is a disease characterized by remodeling of vessel walls. Growth factors, inflammatory cytokines, reactive oxygen species (ROS), and calpain contribute to pulmonary vascular remodeling. In the present study, we studied whether calpain is activated by growth factors, inflammatory cytokines, and ROS. Pulmonary arterial smooth muscle cells (PASMCs) were treated with platelet‐derived growth factor‐BB (PDGF‐BB), serotonin (5‐HT), endothelin‐1 (ET‐1), interleukin‐6 (IL‐6) as well as H2O2, and calpain activity was measured using specific fluorescent substrates. Treatment with the stimulators mentioned above caused dramatic increases in calpain activity. The elevated calpain activity correlates with the increases in collagen‐I expression in PASMCs. Incubation of PASMCs with BAPTA/AM, a cell‐permeable intracellular calcium chelator, did not affect the increases in calpain activity induced by PDGF‐BB, 5‐HT, ET‐1, IL‐6, and H2O2.

Conclusion

PDGF‐BB, 5‐HT, ET‐1, IL‐6, and H2O2 induce the activation of calpain. The mechanism is not due to increase in intracellular calcium. Supported by NIH and FAMRI
Loss of PPARγ promotes NF‐κB activation, Nox4 induction, and proliferation of human pulmonary artery smooth muscle cells
Authors: Bijli Kaiser M , Kang Bum‐Yong , Murphy Tamara , Hart C Michael
Published: June 2021

Hypoxia stimulates pulmonary hypertension (PH) in part by increasing the proliferation of pulmonary vascular wall cells. Recent evidence suggests that signaling events involved in hypoxia‐induced cell proliferation include sustained Nuclear Factor‐kappaB (NF‐κB) activation, increased NADPH oxidase4 (Nox4) expression, and downregulation of peroxisome proliferator‐activated receptor gamma (PPARγ) levels. To further understand the role of reduced PPARγ levels in PH pathobiology, siRNA was employed to reduce PPARγ levels in human pulmonary artery smooth muscle cells (HPASMC) in vitro. PPARγ siRNA reduced PPARγ levels to an extent comparable to those observed under hypoxic conditions. PPARγ depletion activated NF‐κB as determined by phosphorylation of RelA/p65 (NF‐κB) at Ser536 and degradation of its inhibitory protein, I‐kappaB alpha. PPARγ depletion also induced the expression of the NF‐κB target gene, Nox4 and increased HPASMC proliferation as determined by MTT assay and elevated levels of proliferating cell nuclear antigen. Furthermore, treatment with PEG‐catalase attenuated HPASMC proliferation caused by PPARγ depletion. Taken together, these findings provide novel evidence that reductions in PPARγ levels are sufficient to promote HPASMC proliferation via NF‐κB activation, Nox4 induction, and H2O2 generation.

Supported by: Atlanta VA Research Service, NIH DK074518 and HL102167, and AHA‐SDG
Mesenchymal stem cells attenuate renal inflammation, microvascular rarefaction and fibrosis in the renovascular hypertension rat model.
Authors: Boim Mirian A , Oliveira‐Sales Elizabeth B , Maquigussa Edgar , Semedo Patricia , Pereira Luciana G , Camara Niels O , Bergamaschi Cassia T , Campos Ruy R
Published: June 2021

Renovascular hypertension induced by 2 Kidney‐1 clip (2K‐1C) results in significant renal functional deterioration in consequence of inflammation, microvascular rarefaction and fibrosis, so we investigated the potential effects of mesenchymal stem cells (MSC) in this model. Three weeks after renal artery occlusion, fluorescently tagged MSC (2×105cells/animal) were weekly injected into the tail vein. Rats were divided in groups control (n=5), control treated with MSC (n=2), 2K‐1C (n=8) and 2K‐1C treated with MSC (n=7). Tracking assay by flow cytometry showed that labeled MSC were present in the cortex and medulla of the clipped kidney. MSC prevented further increase in the arterial pressure in 2K‐1C rats, improved the morphology and decreased fibrosis in the clipped kidney. Renal pro inflammatory cytokines IL‐1β, IL‐6 and TNF‐α mRNA expression levels were significantly decreased after MSC treatment, while the anti‐inflammatory IL‐10 expression was increased. MSC improved microvascular architecture in the clipped kidney and reduced the proteinuria. The present study showed that MSC improved the morphology and attenuated inflammation, microvascular rarefaction, fibrosis and proteinuria in the clipped kidney, suggesting a therapeutic potential of MSC in kidney exposed to chronic hypoxia induced by renal artery occlusion.
Heart rate and metabolic rate of bar‐headed geese flying in hypoxia
Authors: Meir Jessica Ulrika , Jardine Wilhelmina , York Julia , Chua Beverly , Milsom William K.
Published: June 2021

Bar‐headed geese accomplish the extraordinary feat of migrating over the Himalayas, where oxygen (O2) levels are only ½ ‐ 1/3 those at sea‐level. Although physiological responses and adaptations relevant to this species’ success at high altitude have been previously documented, only one study has measured physiological variables in this bird during flight, and only under conditions of normoxia. In order to assess the roles of the cardiovascular and respiratory systems in maintaining oxygen delivery during flight in hypoxia, we trained bar‐headed geese to fly in a wind tunnel while wearing our physiological data‐logger and a mask system. We were successful in measuring heart rate and metabolic rate in flying geese under conditions of normoxia and hypoxia (10.5% O2 and 7% O2, equivalent to altitudes of ~5,500 and ~8,500 meters respectively). Surprisingly, bar‐headed geese exhibited a remarkably wide range of heart rates and metabolic rates while flying at their preferred flight speed, even on an individual bird basis. Based on preliminary data, mean heart rate during flight changed very little with increasing levels of hypoxia, though mean metabolic rate was reduced. This suggests that the birds settled into more efficient flight patterns in the more challenging hypoxic environment. Bar‐headed geese sustained the same flight durations at equivalent flight speeds under conditions of normoxia and severe hypoxia.
Effects of acetylcholine and cholinergic antagonists on the activity of nucleus of the solitary tract (NTS) neurons
Authors: Furuya Werner Issao , Colombari Eduardo , Ferguson Alastair Victor , de Almeida Colombari Debora Simoes
Published: June 2021

It has been shown that acetylcholine (ACh) can depolarize neurons in the NTS (Shihara et al., 1999). Thus, in the present study, we tested the electrophysiological effects of ACh, alone or combined with cholinergic antagonists, on the commissural NTS (cNTS) or intermediate NTS (iNTS). Coronal slices of the brainstem containing either cNTS or iNTS subnuclei, 600 μm apart, were obtained from male Sprague‐Dawley rats (p21–28 days) and used in whole cell patch clamp – current clamp recordings. ACh (10 mM, 1 min) was applied to slices 10 min before and 5 min after application of atropine (ATR, non‐selective muscarinic antagonist, 10 μM, 5 min) or mecamylamine (MEC, non‐selective nicotinic antagonists, 10 μM, 5 min). In cNTS, 7/12 neurons (58%) were depolarized by ACh (6.9 ± 1.5 mV), effects which were maintained in the presence of ATR (Control 5.1 ± 1.4, ATR 4.8 ± 0.7 mV, p>;0.05, n=6). However, MEC inhibited the ACh actions on cNTS (Control 10.4 ± 2.7, MEC 1.3 ± 0.9 mV, p<0.05, n=5). ACh also depolarized (9.8 ± 2.4 mV) 13/17 neurons (76%) of the iNTS. ATR reduced the depolarization induced by ACh (Control 12.5 ± 2.8, ATR 7.8 ± 2.4 mV, p<0.05, n=8), as well as MEC (Control 9.5 ± 2.0, MEC 2.9 ± 1.0 mV, p<0.05, n=8). These data demonstrate that ACh depolarizes cNTS neurons through actions on nicotinic receptors, while depolarizing effects in iNTS are apparently mediated by both receptors.

Supported by: FAPESP, Canadian Inst. for Health Research
Chronic Rapamycin Treatment Confers Partially Reversible Diabetes Mellitus
Authors: Schindler Christine Elyse , Swoap Steven J
Published: June 2021

Chronic rapamycin treatment, initially shown to promote longevity in mice, is now known to cause impaired glucose homeostasis and insulin resistance. It was therefore hypothesized that chronic rapamycin treatment would cause type II diabetes in mice. Outbred mice were treated with oral rapamycin for a period of one year and were assessed with glucose, insulin and pyruvate tolerance tests (GTT, ITT, PTT). Area under the curve analysis (AUC) of GTTs showed glucose intolerance in rapamycin‐treated males as compared to controls (AUC 10418±1839 vs. 2975±318 min‐mg/dL respectively) and in rapamycin‐treated females vs. controls (AUC 4744±867 vs. 2936±261). In a PTT, rapamycin‐treated males showed pyruvate intolerance as compared to controls (AUC 7614 ±1044 vs. 3006 ± 987), suggesting excess gluconeogenesis. The ITT showed normal insulin sensitivity in both rapamycin‐treated males and females suggesting diabetes complications are a result of damaged islet cells. When rapamycin was removed from the diet, fasting blood glucose in males fell from 294±28 mg/dL to 128±11mg/dL (ad lib cohorts: 84±5 mg/dL) and had increased glucose tolerance (AUC 4877±742). Females also exhibited increased glucose tolerance (AUC 2595±527) after removal of rapamycin treatment. These data suggest impairment of islet cell function in rapamycin treated mice that is partially reversible in males and fully reversible in females.
The role of β1 adrenergic receptor in Non‐Alcoholic Fat Liver Disease development
Authors: Fernandes Gustavo Werpel , Marcelino Cícera Pimenta , Brum Patricia Chakur , Passarelli Marisa , da Costa Bocco Bárbara Miranda Leite , Ribeiro Miriam Oliveira
Published: June 2021

Non‐Alcoholic Fat Liver Disease (NAFLD) is characterized by the development of macrovesicular steatosis in the absence of significant consumption of alcohol, which can progress to nonalcoholic steato‐hepatitis (NASH). Knockout mice for the β1 adrenergic receptor (β1KO) are obese when placed on high fat diet (HFD) and develop NASH with severe steatosis and fibrosis. The aim of this study was to determine if β1 adrenergic receptors have a direct role in NASH development or if this abnormality is due to the severe obesity observed in these mice. For that we analyzed genes related to lipid metabolism in the liver of obese β1KO. Fatty acids, triglycerides and cholesterol synthesis were increased in β1KO HFD (ChREBP: 5.55±0.2 vs. 2.28±0.04 WT, p<0.05; SREBP1c: 2.52±0.4 vs. 1.32±0.02 WT, p<0.05; DGAT2: 2±0.3 vs. 1.3±0.1 WT, p<0.05; SREBP2: 3.22±0.5 vs. 0.67±0.06 WT, p<0.05) resulting in hepatic triglycerides accumulation whereas hepatic secretion of lipoproteins were decreased (MTTP: 0.32±0.01 vs. 0.6±0.03 WT, p<0.05). These data showed an imbalance in lipoproteins synthesis and export, resulting in NASH. In conclusion, β1 adrenergic receptor has a direct influence on expression of genes related to lipid metabolism, and its absence leads to NASH when mice are treated with HFD.

Supported: FAPESP/CAPES
Chronic Gastric Inflammation Drives the Malignant Transformation of Bone Marrow‐Derived Mesenchymal Stem Cells
Authors: Donnelly Jessica , Engevik Melinda , Xiao Chang , Worrell Roger T. , Zavros Yana
Published: June 2021

Bone marrow‐derived mesenchymal stem cells (BM‐MSCs) promote gastric cancer progression with chronic inflammation. In culture, BM‐MSCs are prone to mutation with age but whether this occurs in vivo in response to gastritis is unknown. Sonic Hedgehog (Shh) is often secreted by tumors and may contribute to BM‐MSC transformation. We tested the hypothesis that malignant transformation of BM‐MSCs in response to chronic gastric inflammation is mediated by Shh signaling. GKO mice displayed severe atrophic gastritis accompanied by elevated gastric tissue and circulating transforming growth factor‐beta (TGFβ) by 3 months of age. BM‐MSCs isolated from GKO mice displayed an increased proliferative rate and elevated phosphorylated‐Smad3 indicative of active TGFβ signaling when compared to those cells isolated from BL/6 bone marrow. In xenograft assays, mice injected with BM‐MSCs collected from 6 month old GKO animals displayed tumor growth. BM‐MSCs collected from age matched BL/6 mice injected into inflamed GKO animals also developed tumor growth that was mediated by TGFβ or Shh signaling. Shh pathway activity within BM‐MSCs was downstream of the TGFβ signaling. This data establishes chronic gastric inflammation induces the malignant transformation of BM‐MSCs. Funded by American Cancer Society Research Scholar Award 119072‐RSG‐10–167‐01‐MPC (Y. Zavros) and the Albert J. Ryan Foundation Fellowship (J. Donnelly)
Intranasal administration of IL‐37 attenuates bleomycin induced experimental lung injury/fibrosis
Authors: Park sung woo , Kim Cho Rong , Lee Eun Hee , Kim Jung Hyun , Park Jong Sook , Lee June Hyuk , Jang An Soo , Kim Do Jin , Uh Soo‐taek , Kim Yong Hoon , Park Choon‐sik
Published: June 2021

Lung fibrosis can occur as a result of various inflammatory and oxidative conditions. Idiopathic pulmonary fibrosis(IPF) is the most common entity with a poor prognosis resulting high mortality rate. Currently, there are no proven effective treatment for inhibitor of innate inflammation and immunity. To evaluate intranasal treatment of IL‐37 have therapeutic effect on bleomycin‐induced lung injury/fibrosis in mice.

Histological examinations, collagen assay, ROS assay and cytokine measurements including IL‐1b, TNF‐a and TGF‐b1 were performed in bleomycin‐treated mouse lung tissues with or without the administration of IL‐37. Lung IL‐37 expression from the patient with IPF and controls were measured by immunohistochemistry and immunoblot. H2O2 induced apoptosis that were measured by Annexin V/PI stain in A549 cells with or without presence of IL‐37.

Compared with control mice, IL‐37 treated mice had significantly less lung inflammation, collagen deposition, oxidative damage as well as decreased levels of BAL fluid IL‐1b, TNF‐a and TGF‐b1. Treatment of IL‐37 significantly inhibit cell apoptosis dose dependent manner. Lung IL‐37 protein expression was significantly decreased in patients with IPF than controls.

Local treatment with IL‐37 is effective against the development of experimental lung injury/fibrosis. Anti‐apoptotic and anti‐oxidative activity of IL‐37 contribute at least partly its anti‐fibrotic effect on the experimental fibrosis.
Development of biased agonists at the kappa opioid receptor.
Authors: Phillips Angela , Zhou Lei , Lovell Kimberly , Frankowski Kevin , Slausen Stephen , Prisinzano Thomas E , Aube Jeff , Bohn Laura M
Published: June 2021

Kappa opioid receptors (KOR) are the primary neuronal targets of the stress peptide, dynorphin, and regulation of the KOR may present a means to modulate responses to psychological stressors. Moreover, the development of therapeutics antagonizing the KOR may be useful for treating stress‐induced relapse in those suffering from substance addiction. βArrestins are key regulatory proteins of G protein‐coupled receptors (GPCR), such as the KOR. Given that there is some evidence to suggest that βarrestin‐mediated signaling may promote the negative aspects associated with dynorphin signaling, there is considerable interest in selectively dampening transduction via the KOR‐βarrestin signaling node, while potentially maintaining the G protein coupling of the receptor. Here we describe a number of newly synthesized KOR ligands that possess varying potency and efficacy profiles across multiple signaling assays. Our early efforts hold promise for biasing drug actions toward KOR‐mediated G protein coupling and ERK activation and away from inducing βarrestin recruitment in cell based assays. NIH/NIDA grant: 1R01DA031927–01
Decreased fatty acid oxidation impairs contractile recovery of the insulin resistant heart post‐ischemia
Authors: Harmancey Romain , Vasquez Hernan G. , Guthrie Patrick H. , Taegtmeyer Heinrich
Published: June 2021

We investigated the effects of insulin resistance on contractile recovery of the heart post‐ischemia. Insulin resistance was induced by feeding male Sprague Dawley rats a high‐sucrose diet. Hearts were then perfused ex vivo in the working mode with glucose (25 mM), oleate (0.8 mM), and insulin (5 ng/ml). Before ischemia, myocardial insulin resistance was confirmed by a 17% decrease in the rates of glucose uptake compared to controls (p<0.05). After 15 minutes of global ischemia, hearts were reperfused for 30 minutes. During reperfusion, control hearts recovered to 93% of their pre‐ischemic contractile performance, whereas cardiac power fell to 80% of its initial value for the insulin resistant hearts (p<0.001). Markers of ROS generation, endoplasmic reticulum stress, necrosis, and apoptosis were similar in both groups, suggesting that decreased contraction of the insulin resistant heart was due to a deficit in ATP production, rather than to an increase in cell death. In insulin resistant hearts, decreased AMPK activity (decreased pAMPK(Thr172)/AMPK ratio) correlated to a 35% reduction in acetyl‐CoA carboxylase inhibition (p<0.01). However, rates of glucose uptake were similarly decreased in both groups during reperfusion, and the rates of glucose oxidation remained unchanged. We conclude that impaired fatty acid oxidation may be the reason for impaired post‐ischemic recovery of the insulin resistant heart.
Differences in the Biosynthesis of Long Chain Omega‐3 and Omega‐6 Polyunsaturated fatty acids (PUFAs) between African and European American patients with Coronary Artery Disease
Authors: Wilson Bryan Anthony , Ivester Priscilla , Sergeant Susan , Murphy Karin , Hundley William G , Chilton Floyd H
Published: June 2021

We have previously shown racial differences in Omega‐6 (ω‐6) long chain polyunsaturated fatty acid (LC‐PUFA) levels and the frequency of genetic variants in genes that enhance ω‐6 LC‐PUFA synthesis. In this study we examine if racial differences also exist in Omega‐3 (ω3) and ω6 LC‐PUFA metabolism in patients with coronary artery disease (CAD). We used Gas Chromatography to measure the LC‐PUFA precursors α‐linolenic (18:3 ALA) and linoleic (18:2 LA) and the major ω3/ω6 LC‐PUFA products, eicosapentanoic (22:5, EPA), docosapentaenoic (24:5 DPA) docosahexanoic (24:6 DHA), gamma‐linoleic (18:3 GLA), dihomo‐gamma linoleic (20:3 DGLA) and arachidonic (20:4 AA) in fasting serum from 74 African Americans (AfAm) and 334 European Americans (Eam) with CAD. Compared to EAm, AfAm had lower ALA (0.71 ± 0.22 vs 0.79 ± 0.26 p=0.01), whereas DHA levels were higher (2.12 ± 0.69 vs 1.71 ± 0.68 p<0.001). AfAm also had higher AA levels (10.00 ± 2.07 vs 7.95 ± 2.11 p<0.0001). Compared to EAm, PUFA product:precursor ratios were higher in AfAm for DHA:ALA (3.33 ± 1.6 vs 2.38 ± 1.20 p<0.001), DHA:EPA (3.28 ± 1.56 vs 2.47 ± 1.04 p<0.001), DHA:DPA (3.77 ± 1.13 vs 2.85 ± 0.96 P<0.001), AA/LA (0.33 ± 0.08 vs 0.27 ± 0.08 p<0.001) and AA/DGLA (6.93 ± 2.17 vs 5.35 ± 1.99 p<0.001). Product:precursor ratios of PUFAs provide a surrogate measure of activity in LC‐PUFA synthesis and suggests enhanced precursor conversion in AfAm which may be genetically determined.
In vivo imaging of intracellular Ca2+ after muscle contractions and direct Ca2+ injection in rat skeletal muscle in diabetes
Authors: Eshima Hiroaki , Tanaka Yoshinori , Sonobe Takashi , Inagaki Tadakatsu , Poole David C , Kano Yutaka
Published: June 2021

The effects of muscle contractions on intracellular Ca2+ ([Ca2+]i) accumulation kinetics in Type I diabetes are unclear. To test the hypothesis that, following repeated bouts of muscle contractions, the rise in [Ca2+]i evident in healthy rats would be increased in diabetes rats and that these changes would be associated with a decreased cytoplasmic Ca2+ buffering capacity. Adult male Wistar rats were divided randomly into diabetic (DIA: Streptozotocin i.p.) and healthy (CONT) groups. Four weeks later animals were anesthetized and spinotrapezius muscle contractions (10 sets of 50 contractions) were elicited by electrical stimulation (100 Hz). Ca2+ imaging was achieved using fura2‐AM in the spinotrapezius muscle in vivo (i.e., circulation intact). The ratio (340/380 nm) was determined from fluorescence images following each contractions set, for estimation of [Ca2+]i. Also, muscle Ca2+ buffering was studied in individual myocytes microinjected with 2 mM Ca2+ solution. During muscle contractions, [Ca2+]i in DIA increased earlier and more rapidly than in CONT (P < 0.05 vs. pre‐contraction). Peak [Ca2+]i in response to the Ca2+ injection was significantly higher in CONT (25.8%) than DIA (10.0%). Diabetes markedly impairs Ca2+ homeostasis following muscle contractions. Markedly different responses to Ca2+ injection suggest fundamentally deranged Ca2+ handling in spinotrapezius muscles of diabetic rats.
Augmented sympathetic activity does not attenuate cerebral vasomotor reactivity to hypercapnia during heat stress
Authors: Brothers R. Matthew , Lee Joshua F. , Christmas Kevin M. , Hurr Chansol , Harrison Michelle L. , Kim Kiyoung
Published: June 2021

Recent work identified that augmented sympathetic activity, via lower body negative pressure (LBNP) induced central hypovolemia, attenuates cerebral vascodilation to hypercapnia during normothermic conditions. If a similar response occurs in individuals with elevated internal temperature remains unknown. This study tested the hypothesis that, relative to normothermic conditions, heat stress would attenuate cerebral vasodilation during hypercapnia and that this response would be further attenuated when LBNP is superimposed upon the heat stress. In 10 healthy individuals (9 men) cerebral vasodilation was assessed as the increase in cerebral vascular conductance (CVC) during 3, 6, 9, 12, and 15 Torr elevations in end‐tidal carbon dioxide (PETCO2) during the following conditions: normothermia, control heat stress (Δ 1.5 ± 0.2°C), and heat stress + 20 mm Hg (Δ 1.6 ± 0.2°C). Due to logistical reasons the normothermic trial was always performed first followed by control heat stress, then heat stress + LBNP. In all thermal conditions CVC increased in a hypercapnic dependent fashion (main effect of PETCO2: P < 0.01). However, contrary to our hypothesis the CVC response to elevations in PETCO2 were similar during the three thermal conditions (main effect of thermal condition: P >; 0.45). These data suggest that CVMR is not influenced be elevated sympathetic activity induced by control heat stress and heat stress + LBNP.
Direct calorimetry exposes inadequacies of respirometry in the measurement of resting metabolic rate
Authors: Burnett Colin M.L. , Grobe Justin L.
Published: June 2021

Resting metabolic rate (RMR) in animals is frequently estimated by respirometry, but the dependence of this method upon untested or untestable assumptions (such as negligible anaerobic metabolism, simultaneously measured or negligible protein oxidation, and no substrate inter‐conversion) render respirometry inappropriate or inadequate for examining novel genetically‐manipulated animal models. A Seebeck‐style direct calorimeter apparatus was constructed to allow simultaneous measurement of total heat dissipation and respirometry gasses from unrestrained mice. Double‐transgenic “sRA” mice exhibit elevated brain renin‐angiotensin system activity by expression of human renin via the synapsin promoter and human angiotensinogen via its own promoter. Respirometry underestimated the RMR in sRA mice by 16% (n=6, male, 11.0±0.6 wks, 20.97±0.68 g; 0.889±0.036 vs 1.057±0.034 kcal/100g lean/hr, P=0.008), and by 14% in control littermates and C57Bl/6J mice (n=10, male, 11.6±0.5 wks, 27.02±0.60 g; 0.712±0.025 vs 0.828±0.018 kcal/100g lean/hr, P=0.005). While respirometry faithfully detected the 26% increase in sRA RMR vs control (P<0.001 by both methods), the magnitude of RMR estimation error by respirometry was similar (~15%). Reanalysis of various other genetically‐manipulated models by direct calorimetry may therefore uncover previously unappreciated physiological regulators of RMR.
Altered skeletal muscle respiration in patients with COPD: mitochondrial density or dysfunction?
Authors: Gifford Jayson Reed , Garten Ryan S , Trinity Joel , Park Songyoung , Layec Gwenael , Larsen Steen , Richardson Russell S
Published: June 2021

INTRODUCTION

We sought to determine if the blunted mitochondrial respiration typically observed in the skeletal muscle of patients with chronic obstructive pulmonary disease (COPD) is solely related to changes in mitochondrial density or if qualitative changes in respiration occur.

METHODS

Combinations of substrates (malate, glutamate, succinate, and ADP) and inhibitors (oligomycin) were used to measure complex I (CI) and complex II (CII) driven state 3 respiration separately and in combination in permeabilized muscle fibers from the vastus lateralis of 9 healthy controls (67±2 years) and 11 COPD patients (FEV1= 56±6%; 63±3 years). State 4 respiration and citrate synthase (CS) activity were measured using these same fibers.

RESULTS

Compared to controls COPD patients had blunted CI driven state 3 respiration (37.2±4.1, 23.7±2.7 nM/min/mg, respectively) and blunted state 3 respiration with CI and CII (47.2±5.1, 33.7±3.8 nM/min/mg, respectively) (P<0.05) while state 3 driven by CII alone was similar between groups. CS activity and state 4 respiration were not different between groups.

CONCLUSIONS

Given that CS activity, an indicator of density, was similar between groups, the blunted respiratory capacity in these COPD patients appears to be related to an attenuation of CI driven respiration. These data indicate that COPD results in qualitative changes in mitochondrial respiration of skeletal muscle.
Structure and function of the patellar tendon in patients with classic Ehlers‐Danlos syndrome
Authors: Nielsen Rie Harboe , Couppé Christian , Olsen Morten Raun , Jensen Jacob Kildevang , Svensson René , Heinemeier Katja Maria , Magnusson Peter , Remvig Lars
Published: June 2021

Classic Ehlers‐Danlos Syndrome (cEDS) is mainly caused by collagen V mutations and the phenotype is associated with joint hypermobility, pain and dislocations. Eight patients clinically diagnosed with cEDS (age 39 ± 11 years, BMI 24 ± 4 kg/m2, mean ± SD) were matched with eight healthy controls (age 41 ± 14 years, BMI 25 ± 5 kg/m2). Patellar tendon tissue was imaged with transmission electron microscopy (TEM), tendon size was obtained with magnetic resonance imaging (MRI) and measures of tendon biomechanical properties were done by ultrasonography‐based tendon elongation during isometric knee contractions. TEM images revealed large irregular collagen fibrils (cauliflower‐like) in cEDS patients. MRI showed no differences in tendon size between cEDS and controls. The biomechanical properties of the patellar tendon were severely altered in cEDS patients; showing >;60% reduction in stiffness (1486 ± 698 vs. 4029 ± 1645 N/mm, p<0.01) and Young's modulus (0.56 ± 0.29 vs. 1.68 ± 0.75 GPa, p<0.05) compared to controls. In conclusion, the altered tendon ultrastructure and stiffness in cEDS patients indicate that tendon tissue can, at least partly, explain the observed joint hypermobility in cEDS.
Mathematical Model of Aortic Coarctation to Elucidate Mechanisms of Vascular Remodeling
Authors: Higareda Deidre J. , Tran Nga V. , Bustillos Jacob L. , Nguyen Phuc H. , Quick Christopher M.
Published: June 2021

Aortic coarctation is of interest to both clinicians and mechanobiologists, because it induces high upstream pulse pressures (PP) and vascular remodeling. Interpreting either clinical or experimental data to infer adaptive mechanisms is challenging: vascular remodeling induces changes in PP, and changes in PP induce vascular remodeling. Further complicating analysis, PP depends on the architecture of the systemic arterial network as well as the properties of each component artery—radius (r), wall thickness (h), and stiffness (E). The resulting complexity has made it difficult to test one of the most enduring hypotheses: high pulsatile wall stress (σ) causes arterial stiffening. Therefore, we used a classical human arterial system model with realistic architecture (Westerhof, 1968) to predict local PP from the r, h, and E of each of its 121 arterial segments. Assuming Laplace's Law, σ was calculated from local PP, r, and h. We then made vessels adaptive by assuming E increases linearly with σ. The result was a fundamental instability leading to positive feedback between changes in E and PP. Assuming instead that E decreases with σ not only led to realistic values of local vessel E and PP, it correctly predicted an increase in compliance upstream of a coarctation and a partial restoration of normal pulse pressure.