bluuebunny/crossref_metadata_2025 huggingface dataset:
Antioxidant and proteolytic activity of ethanol extract from Crataegi fructus
Authors: Park Sung-Jin , Yoo Seon-Mi , Kim Haeng-Ran
Published: June 2021

Crataegi fructus was an important medicine resource and distributed widely in Korea. In this study, we examined the antioxidant activity of the Crataegi fructus extract in vitro using total phenolic contents, total flavonoids contents, 2,2‐diphenyl‐1‐picrylhydrazyl(DPPH) radical scavenging activity, reducing power assay. The total phenolic and total flavonoids contents of Crataegi fructus extract were 64.1 mg/mL, 6.0 mg/mL, respectively. The DPPH radical scavenging activity of extract varied from 64.85 (200 μg/mL) to 92.36 (1,000 μg/mL). The reducing power of the extract absorbance varied from 2.24 (200 μg/mL) to 2.64 (1,000 μg/mL ), and reducing power of extract presented a concentration‐dependent activity increase. The extract from Crataegi fructus showed the highest activity among all the tested samples including vitamin C, known as antioxidant. Proteolytic activity of Crataegi fructus extract showed strong activity than pancreatin (used as a positive control). However, there is no inhibitory activity against α‐amylase and α‐glucosidase activity. These results indicated that Crataegi fructus extract can be used as a natural resource for development of antioxidant or materials aiding digestion.
Effect of n‐3 polyunsaturated fatty acids on peroxisome proliferator‐activated receptor gamma (PPARγ) expression in adults
Authors: Rajaram Sujatha , Sabaté Joan , Mohan Subburaman
Published: June 2021

Background

N‐3 fatty acids play a role in reducing the risk of cardiovascular disease via lowering blood lipids and suppressing inflammation. Whether or not the anti‐inflammatory effects are mediated through alteration in gene expression is not known. Our objective was to determine the effect of varying dietary ratio of n‐ 6: n‐3 fatty acids on the expression of PPAR‐γ.

Methods

A factorial design approach was used to test the effect of 4 diets on gene expression: 10:1 and 2:1 n‐6: n‐3 fatty acid ratio diets with and without a supplement (S) of eicosapentaenoic and docosahexaenoic acids (EPA/DHA). This study was part of an original randomized cross‐over feeding study (8‐week period each with a washout between periods) on 24 healthy adults. Subjects consumed eucaloric diets [10:1 (Control), 2:1 (α‐linolenic acid, ALA; 6–7g/2400 kcal/d), 10:1+S (0.20/0.72g EPA/DHA per 2400 kcal/d), and 2:1+S. At the end of each diet period, samples of subcutaneous tissue was collected from the abdominal region and used for measurement of PPAR‐γ mRNA levels by real‐time polymerase chain reaction with actin as an internal standard.

Results

Preliminary analyses of the fold changes in PPAR‐γ expression between diets was not statistically significant.

Conclusion

Our initial analyses did not show an effect of dietary ratio of n‐6: n‐3 fatty acids with or without the supplement on the expression of PPAR‐γ for the 8‐week study period.

Funded by the Center for Health and Nutrition Research, University of California, Davis.
Novel sorghum brans containing bioactive compounds alter the production of microbial secondary metabolites in response to a DSS‐induced chronic inflammatory state
Authors: Ritchie Lauren Elizabeth , Carroll R , Weeks B , Rooney L , Turner N D
Published: June 2021

We have shown that black bran containing bioactives reduces colonic injury and NF‐κB activity with chronic inflammation, which may result from altered microbiota and short chain fatty acids (SCFA e.g., butyrate). Rats (n=80) were fed diets with 6% fiber from cellulose (C), or bran from black (B), sumac (S), or high tannin (HT) sorghum (anthocyanins, condensed tannins, or both, respectively). Ten rats in each diet received 3% dextran sulfate sodium (DSS) three times to stimulate inflammation. Feces were collected pre and post DSS to assess SCFA. On day 63, colon tissue was collected to determine injury and gene expression. After DSS, butyrate concentrations were higher (p<0.0001) with C and B (19.7 and 22.6) compared to S and HT (8.5 and 14.1). SCFA transporter (MCT1 and Slc5a8) expression was upregulated in C, S or HT DSS rats (avg 35, 48, and 12%) and downregulated in B DSS rats (avg 18%) compared to controls. In DSS rats, butyrate excretion, as a percent of total SCFA, was elevated with B (23%) compared to C, S or HT (19, 9, 15%). Thus, with chronic inflammation, there was an inverse relationship between the direction of transporter expression change and butyrate excretion. To determine the implications of the diet effects on butyrate and SCFA transporter levels in chronic inflammation, it will be necessary to evaluate proliferation and apoptosis. Funding: United Sorghum Checkoff Board Roo31A‐09.
Tolerance and safety of an avocado‐based ingredient for adult dogs
Authors: Davenport Gary M , Shoveller Anna K , Hayek Michael , Massimino Stefan , Halsey Lori , Minikhiem Debbie , Flickinger Elizabeth , Roth George , Ingram Donald
Published: June 2021

Feeding avocados to dogs is controversial due to a single report of fatal toxicity for two dogs living near a South African orchard that regularly ate whole avocados and other plant parts (Onderstepoort J Vet Res, 1994, 61:107). This 6‐month study evaluated the tolerance and safety of an avocado‐based ingredient fed to healthy adult dogs. The ingredient was a defatted, water‐soluble extract of flesh, skin and pit of Lula (var) avocados (Persea americana). The vacuum‐dried extract contained 61% carbohydrate, 4% protein, 4% fat, 16% ash, 0.1% fiber and 13% moisture. Adult Beagles (30 spayed females; 10 neutered males) averaging 4.4 years of age were assigned to four treatments (n=10/diet): control (Eukanuba Senior Maintenance formula) and three test diets comprised of control diet with 0.22, 0.66 or 1.10% avocado extract. All dogs completed the study and readily consumed all diets. Body weight, normal GI health and stool production were maintained throughout the study. Regular health assessments revealed no abnormalities or inferior health conditions for dogs assigned to any diet group. While transient changes in hematology and serum chemistry parameters were noted, values remained within normal reference ranges for healthy adult dogs indicating no health or safety concerns. Feeding a diet with up to 1.1% avocado extract was well‐tolerated by adult dogs and resulted in no adverse health effects. P&G funded the study.
Validation of a Novel Food/Nutrient Intake Instrument in Georgian Tuberculosis Patients
Authors: Frediani Jennifer K , Tukvadze Nestani , Sanikidze Ekaterine , Kipiani Maia , Hebbar Gautam , Ramakrishnan Usha , Martorell Reynaldo , Ziegler Thomas R
Published: June 2021

Objective

To validate a novel food/nutrient intake assessment tool in patients with tuberculosis (TB) in Tbilisi, Republic of Georgia.

Methods

The instrument was designed to capture specific foods and meal patterns common in Georgian culture, and involved a 3‐day food recall conducted by an interviewer in a questionnaire format (n= 40). Food intake data was entered into Nutrient Database System for Research (NDS‐R) and mean daily intake of specific nutrients determined. Data were compared with data from three 24‐hour recalls conducted on the same days of the week 7 days apart from the food intake recall interview. Paired t‐tests and Pearson correlations were used.

Results

Pearson correlations were P<0.05 for: calories, total fat, total carbohydrate, total protein, cholesterol, saturated fat, monounsaturated fat, total fiber, thiamin, riboflavin, niacin, pantothenic acid, vitamin B6, total folate, vitamin D, calcium, phosphorus, magnesium, iron, zinc, selenium, sodium and potassium. T‐tests showed similar concordance between methods for most nutrients.

Conclusions

This novel food/nutrient intake tool is valid for nutrient intake assessments in TB patients in Tbilisi, Georgia.

Grant Funding Source : NIH grants D43 TW007124, K24 RR023356, UL1 RR025008 and the Emory Global Health Institute.
Perceptions of diabetic/hypertensive patients and primary healthcare providers regarding disease management (San José, Costa Rica ‐ Chiapas, México)
Authors: Alvarado Nadia , Dengo A. Laura , López Erika , Castro Maricruz , Peña Liz , de Beausset Indira , Martínez Homero , Murillo Sandra
Published: June 2021

Objective

To understand why patients have difficulty managing chronic diseases, we studied the perceptions of type 2 diabetic/hypertensive (DM2/HTN) patients and health personnel regarding services offered at primary healthcare (PHC) centers.

Methods

Focus group discussions were conducted with patients (10 groups, total n=76) and PHC staff (4 groups, total n=25) from 4 urban communities (San José=2; Chiapas=2). We focused on: perceived knowledge and attitudes about disease, self‐care practices, PHC quality, and communication issues.

Results

Patients affirmed that having DM2 and HTN is a physical, financial and emotional burden both to them and their families. Family members were not perceived as supportive, resulting in feelings of isolation. Genetics was considered the main risk factor for DM2, above modifiable lifestyle factors. Most patients stated not knowing how to manage their DM2/HTN. Patients and staff lacked nutritional education; however, they acknowledged the role of nutrition in disease management. PHC staff should encourage more proactive self‐care skills but did not feel prepared to deal with patients’ attitudinal and know‐how issues.

Conclusions

These results will be used to tailor an intervention to strengthen patient‐staff communication, facilitate a bi‐directional learning process, and empower patients to take control of their health.

Funding: NHLBI‐HHSN 268200900028C
Impact of maternal and postnatal nutrition on femoral artery vascular function of offspring
Authors: Taheripour Pardis , Ajuwon Kolapo M. , Donkin Shawn S. , Sheldon Ryan D. , Bahls Martin , Arentson Emily J. , Ragland Darryl , Newcomer Sean C.
Published: June 2021

The genesis of disease development can be linked to the intrauterine environment and maternal nutrition during pregnancy plays a significant role in the future vascular health of the offspring. The purpose of this investigation was to test the hypothesis that a high energy (HE) prenatal diet, HE postnatal diet, or their combination will lead to endothelial dysfunction. Duroc x Landrace gilts (n=16) were assigned to either a HE (11,903 Kcal/day, n = 8) or normal energy (NE: 6975 Kcal/day, n = 8) diet throughout pregnancy. Two male and two female piglets from each sow were randomly assigned within sex to receive either a HE diet or NE diet. At 3 months of age the femoral artery was harvested from male and female offspring, representing each pool of maternal and postnatal diet (n=47). Endothelial‐dependent and ‐ independent vasorelaxation were measured using wire‐myography and increasing concentration of bradykinin (BK) and sodium nitroprusside (SNP) respectively. No significant differences were observed for BK vasorelaxation. However, prenatal HE diet tended (p=0.07) to decrease SNP vasorelaxation. These results suggest that prenatal HE diet alters vascular smooth muscle cell function as opposed to endothelial cell function in swine. Therefore, future studies should elucidate the mechanisms underlying alterations in vascular smooth muscle function induced by a HE maternal diet. Funding: The Showalter Trust Fund
Embryonic caffeine acts via A1 adenosine receptors to induce adverse effects in adulthood
Authors: Wendler Christopher Charles , Buscariollo Daniela , Fang Xiefan , Greenwood Victoria , Rivkees Scott
Published: June 2021

Previous research demonstrated that caffeine exposure during pregnancy had long‐term effects into adulthood, including altered cardiac function and increased body fat. Caffeine is a non‐selective adenosine receptor antagonist and adenosine A1 receptor (A1AR) is the predominant adenosine receptor expressed during early embryogenesis. The purpose of our current study is to determine whether A1ARs mediate the long‐term effects of caffeine.

A1AR knockout mice were mated and injected at E8.5 with 20 mg/kg of caffeine or vehicle. Dams gave birth and offspring were examined at 8–10 weeks of age. Male A1AR+/+ mice treated with caffeine were heavier than controls but there was no difference in percent body fat. Using echocardiography, we revealed that A1AR+/+ caffeine mice had altered cardiac function and morphology as adults. Caffeine caused an increase in % fractional shortening, while also leading to increased thickening of the left ventricle wall concurrent with decreased left ventricle inner diameter. Next, we examined methylation patterns of DNA isolated from adult left ventricles. Methylation arrays from Nimblegen revealed that caffeine increased methylation of DNA in A1AR+/+ hearts compared to A1AR+/+ controls.

Since all parameters examined were only altered when A1AR was expressed and not when A1AR had been deleted, we conclude that long‐term effects of in utero caffeine exposure are mediated by A1ARs.
Despite Pharmaceutical Equivalence, Generic Versions of Enoxaparin May Differ in Their Pharmacodynamic Actions. Potential Clinical Implications
Authors: Jeske Walter , Walenga Jeanine M , Hoppensteadt Debra , McGeehan Elizabeth , Harenberg Job , Ramacciotti Eduardo , Fareed Jawed
Published: June 2021

Introduction

This study compared available generic enoxaparins with the branded version (Sanofi‐Aventis) in various assays and investigated their pharmacokinetic/pharmacodynamic behavior in a primate model.

Methods

Cutenox, Lupenox, enoxaparin Sandoz and Lovenox were compared in terms of molecular weight profiling, NMR profile and in vitro anticoagulant activity. Groups of primates (n=8–10) were administered LMWH (1 mg/kg subcutaneously) and blood samples were drawn over a 24 hour period to measure pharmacodynamic effects. Antithrombotic and bleeding effects were studied in rat models of jugular vein clamping and tail transection, respectively. Complexation studies with PF4 were carried out using a SELDI‐MS method.

Results

Anti‐Xa potency ranged from 86–110 U/mg and anti‐IIa potency ranged from 27–35 U/mg. Mean molecular weight of these LMWHs ranged from 4.2–4.5 kDa. Despite comparable pharmaceutical profiles, some of these products showed differences in the pharmacodynamic effects in terms of TFPI release, TAFI modulation, thrombin generation inhibition and protamine neutralization.

Conclusion

Although the generic enoxaparins are pharmaceutically equivalent, their pharmacodynamic profiles differ from the branded product. Thus, each generic product should be considered as a distinct LMWH and individual safety/efficacy data should be considered prior to its clinical use.
4‐Methylumbelliferone Prevents E‐cadherin Downregulation Induced by Cigarette Smoke in Airway Epithelial Cells
Authors: Medina Maria Elena Monzon , Valencia Monica , Forteza Rosanna Malbran , Casalino-Matsuda Marina
Published: June 2021

Cigarette smoke (CS) exposure induces airway epithelial barrier (EB) disruption that results in structural changes associated to COPD. EB integrity is maintained, in part, by E‐cadherin (E‐cad). We have shown that hyaluronan fragments (SHA) increased epithelial permeability and downregulated E‐cad expression. In the airways, hyaluronan (HA) is present in a high molecular weight form at the epithelial surface and exposure to oxidants induces its degradation. Thus, we hypothesized that SHA are generated in smokers and that inhibition of HA synthesis would prevent EB disruption induced by CS. To test this hypothesis, HA size was determined by agarose electrophoresis in bronchoalveolar samples from smokers and non‐smokers and in apical washes of human bronchial epithelial (HBE) cells exposed to CS or to air. In addition, to confirm the role of HA in EB disruption, HBE cells were pretreated with the HA synthesis inhibitor 4‐ methylumbelliferone (4MU, 1mM) and exposed to CS or to air and E‐cad expression was assessed by qPCR and WB. We found that secretions from smokers and apical washes from CS exposed cells contain SHA (< 100 kDa) and exposure to CS significantly reduced E‐cad mRNA and protein. Treatment with 4MU prevented these effects, confirming that HA fragmentation take place in vivo in the airways of smokers and SHA generation is able to induce EB disruption by regulating, at least in part, E‐cad expression.
Structural determinants of α‐defensin mediated anti‐inflammatory activities
Authors: Rivera Paulina Schmitt , Kamdar Karishma , Schaal Justin , Roberts Kevin , Tran Dat , Selsted Michael E. , Ouellette Andre J.
Published: June 2021

α‐Defensins are cysteine‐rich, cationic host defense peptides that have bactericidal activities and immunomodulatory effects. The peptides are characterized by a canonical tridisulfide array and a 9:1 preference for Arg relative to Lys. Here, we report that mouse enteric α‐defensin cryptdin‐4 (Crp‐4) and rhesus myeloid α‐defensin‐4 (RMAD‐4) exert anti‐inflammatory effects and define the structural features essential for this activity. Crp‐4 and RMAD‐4 inhibit the release of TNF‐α by human leukocytes exposed to E. coli and by THP‐1 cells stimulated with LPS. Disulfide‐null variants of both peptides do not inhibit TNF‐α release, evidence that the anti‐TNF‐α effect requires the β‐sheet fold imposed by the three disulfide bonds. Anti‐TNF‐α activity is dependent on peptide primary structure, because the macaque RMAD‐3 α‐defensin lacks similar activity. Peptide mutagenesis shows that inhibition of TNF‐α release is dependent on electropositive charge contributed by Arg rather than Lys. In vitro, both peptides inhibit the TNF‐α converting enzyme (TACE), the major TNFprecursorconvertase. In vivo, Crp‐4 promotes survival in a mouse model of polymicrobial sepsis, evidence that the molecule induces systemic anti‐inflammatory effects. Thus, Crp‐4 and RMAD‐4 attenuate proinflammatory responses with potential translational applications. Grants DK044632, AI059346, R37AI022931, RO1DE15517.
Upregulation of membrane‐bound metalloendopeptidase neurolysin in a mouse model of focal brain ischemia
Authors: Rashid Mamoon , Yang Li , Arumugam Thiruma V. , Abbruscato Thomas J. , Karamyan Vardan T.
Published: June 2021

Membrane‐bound metalloendopeptidase neurolysin was recently identified as the non‐AT1, non‐AT2 angiotensin binding site. Our previous study indicated dramatic over‐expression of this protein in membrane preparations of primary cortical neurons challenged in several in vitro models of cell death suggesting a possible role of neurolysin in neuronal survival. To further confirm the association between upregulation of neurolysin and neuronal cell death, in this work we studied the changes in density of membrane‐bound neurolysin in mouse transient middle cerebral artery occlusion model (60 min occlusion with ~75% local cerebral blood flow drop, followed by 24 h reperfusion). Brain parts were collected after documenting reperfusion injury by neurological scoring and TTC staining. Density of neurolsyin was estimated by radioligand binding assays in cerebral cortical, striatal, hippocampal and cerebellar membranes from ischemic hemispheres. Membrane preparations of the same brain parts from sham operated mice served as controls. Significant up‐regulation of neurolysin in membrane preparations of all forebrain regions from ischemic brains was observed. Density of neurolysin did not differ in cerebellar membranes of ischemic and control brains. These results confirm our in vitro observations further supporting a role of neurolsyin in neuronal cell death. Supported by TTUHSC SOP start‐up funds to VTK.
Rho‐dependent upregulation of endothelial contractility and adhesion disassembly contributes to the enhanced permeability responses to inflammation in diabetic venules
Authors: Yuan Dong , He Pingnian
Published: June 2021

Diabetes mellitus (DM) often results in microvascular complications. Our study demonstrated that DM vessels have moderately increased basal permeability, but manifested augmented responses to platelet activating factor, PAF. The objective of this study is to investigate the mechanisms involved in the increased susceptibility of DM venules to inflammation. Experiments were conducted in individually perfused mesenteric venules of normal and streptozotocin‐induced DM rats. Microvessel permeability was measured by hydraulic conductivity Lp and the changes of VE‐cadherin and F‐actin were examined using confocal imaging with immunostaining. In normal venules, pre‐perfusing Rho kinase inhibitor Y‐27632 (30 μM) did not affect basal Lp but reduced PAF‐induced peak Lp from 10 ± 1.7 to 2 ± 0.4 times the control value. In DM venules, Y‐27632 restored the increased basal Lp to a normal range and markedly reduced PAF‐induced Lp peak from 115 ± 20.0 to 11 ± 2.1 × 10−7 cm.s−1.cmH2O− 1. In Y‐27632 perfused DM vessels, PAF‐induced apparent separation of VE‐cadherin between endothelial cells was restored to the pattern of frequent breaks observed in normal vessels exposed to PAF, and phalloidin staining showed reduced endothelial stress fiber assembly at PAF induced Lp peak. Our results indicated that Rho kinase‐mediated upregulation of endothelial contractility and adhesion disassembly contributes to the augmented PAF response in DM microvessels. Supported by HL56237 and HL084338.
Left ventricular torsion based on 2D speckle tracking echocardiography (STE) is preload insensitive
Authors: Gao Zhaohui , Drew Rachel C. , Momen Afsana , Ndukwu Ifesinachi , Muller Matthew D. , Mast Jessica , Blaha Cheryl , Leuenberger Urs A. , Sinoway Lawrence I.
Published: June 2021

Left ventricular (LV) torsion plays an important role in myocardial performance. 2D special tracking echocardiography (STE) has emerged as a promising tool for LV torsion analysis. It is unknown if LV torsion is affected by alterations in preload. We sought to determine whether acute preload reduction by Lower Body Negative Pressure (LBNP) affects 2D STE derived LV torsion parameters. LV apical and basal rotation and peak early diastolic torsional rate were measured by 2D STE in 12 healthy subjects (26±1 yrs, 6 females) before and during LBNP to −10, −20, −30, −40 and −50 mmHg levels. LV total torsion was calculated as the net difference between systolic apical counterclockwise rotation and basal clockwise rotation. Peak early diastolic torsional rate was also measured. Compared to baseline, LBNP caused significant decreases in LV end‐diastolic volume (P<0.05) and stroke volume (P<0.01) while heart rate increased (P<0.01). Interestingly, multiple indices of LV torsion including LV total torsion, apical and basal rotation and peak early diastolic torsional rate, were unaffected by LBNP (all P>0.05). These data support the concept that LV torsion parameters assessed by 2D STE are preload‐independent indices of myocardial function. Quantification of LV torsion by 2D STE may, therefore, provide a relatively load‐independent measure of myocardial performance.

Supported by P01 HL096570 (LIS); UL1 RR033184 (LIS).
Time course of development of erectile dysfunction and coronary artery endothelial dysfunction in response to a western diet: influence of endothelial nitric oxide synthase uncoupling
Authors: La Favor Justin , Anderson Ethan , Chaaban Miranda , Hickner Robert , Wingard Christopher
Published: June 2021

Epidemiological evidence suggests that men experiencing cardiac events may have preexisting erectile dysfunction (ED). Male Sprague‐Dawley rats (n = 5/group) were fed a control or western diet for 4, 8, or 12 weeks. Erectile function was evaluated by calculating the area under the curve (AUC) of intracavernosal pressure/mean arterial pressure in response to electrical field stimulation of the major pelvic ganglion, and again following intracavernosal injection of 10 μM sepiapterin (SA). Coronary artery segments were mounted on a wire myograph, pre‐constricted with serotonin and endothelial function was evaluated by cumulative doses of acetylcholine (ACh) (0.001 – 10 μM), and repeated in one segment following incubation in 10 μM SA. Erectile function was attenuated at 8 and 12 weeks (C: 286 ± 52, 4w: 232 ± 35, 8w: 123 ± 49, 12w: 160 ± 59 (AUC); p = 0.004). SA had no effect on erectile function at C or 4w, but showed improvement at 8w (225 ± 99, p = 0.13) and 12w (211 ± 80, p = 0.24). The western diet increased the coronary ACh EC50 at 12w (C: 0.20 ± 0.18, 4w: 0.25 ± 0.15, 8w: 0.50 ± 0.32, 12w: 0.84 ± 0.55 μM; p = 0.046). SA had no effect on C, 4w or 8w, but improved the EC50 38% at 12w (1.37 ± 2.06 vs 0.85 ± 0.77 μM). ED was manifest prior to coronary dysfunction. There is a trend for improved erectile and coronary function in the dysfunctional state with SA, suggesting that eNOS uncoupling is a common contributor to dysfunction in these vascular beds.
Contribution of prostacyclin regulation in the vascular tone of angiopoietin like‐2 deficient mice
Authors: Yu Carol , Farhat Nada , Thorin Eric
Published: June 2021

Vascular tone, in response to acetylcholine (ACh), is controlled by various endothelium‐derived relaxing factors, which include nitric oxide (NO), prostacyclin, and endothelium‐derived hyperpolarizing factor. In turn, contribution of each factor may be influenced by pathologies. As such, in the insulin resistant state, NO production is impaired. Angiopoietin like‐2 (Angptl2), a member of the angiopoietin‐like protein family, was shown to cause insulin resistance related to obesity (Tabata, 2009).

Objective

An Angptl2 knock‐down (KD) mouse model was generated to study the role of Angptl2 in endothelial function in the femoral artery.

Results

Femoral arteries of 4‐month old Angptl2 KD and wild‐type (WT) mice showed that NO was responsible for the sensitivity of ACh‐mediated vasodilation in both groups, while prostacyclin contributed to maximal dilation in KD but not in WT. Femoral artery wall thickness of KD (20±1μm) at physiological pressure was smaller than that of WT (25±1μm; P<0.01, n=7), while constriction to phenylephrine was similar in both groups. Although KD and WT mice displayed similar metabolic parameters including fasting glucose level, KD mice showed a higher fasting insulin level (P<0.05, n=7–8).

Conclusion

Knock‐down of Angptl2 may lead to differences in the prostacyclin signaling pathway in the femoral arteries between the two genotypes of mice. Supported by CIHR MOP14496 and NSERC.
The Effects of Captopril Treatment and Exercise Training on Blood Pressure and Endothelium Function in the Spontaneously Hypertensive Rat
Authors: Woods Vernikka , Coleman-Shepherd Gwendolynn , Runnels James , Keaton Alphonso
Published: June 2021

Angiotensin converting enzyme inhibitors (ACE‐I) have been show to prevent impairment of endothelial cell function in the Spontaneously Hypertensive Rat (SHR). The purpose of this study was to examine the combined effects of long‐term exercise training and ACE‐I treatment on blood pressure and vascular reactivity in the SHR. Four groups of male SHR were studied: 1) untreated SHR; 2) exercise trained (35 cm/s) for 60 minutes/day, for 5 days/week (SHREX); 3) captopril treated (50 mg/kg/day), treatment was initiated at four weeks of age and continued for 12 weeks (CAP); and 4) captopril treated + exercise (CAPEX). N = 8, for all four groups. All rats were studied at 16–18 weeks of age. Mean arterial pressure (MAP) in the untreated SHR was 130±5 mmHg. Exercise treatment in the SHR significantly decrease MAP in the (SHREX – 114±7 mmHg). Oral administration of captopril was associated with a significant decrease in MAP (97±3 mmHg) compared to untreated SHR and SHREX. Further, captopril treatment + exercise significantly lowered MAP in the CAPEX group (107±6mmHg, vs. SHR). Isolated aortic ring segments were suspended in tissue chambers for measurement of isometric force. Ring segments were exposed to cumulative concentrations of serotonin (3× 10−9 − 3 × 10−5M). Aortic rings from SHR demonstrated increased responsiveness to serotonin induced contraction in comparison to aortic rings from the CAP and CAPEX group. Endothelium‐dependent relaxation to acetylcholine was dramatically impaired in the SHR. Endothelium‐dependent relaxation was markedly greater in both the CAP and CAPEX group. These findings suggest that early long‐term treatment of captopril coupled to exercise training can prevent alterations in endothelial function observed in the SHR….
Maternal Hypoxemia Suppresses Muscarinic Acetylcholine Receptor Dependent Contraction of Pulmonary Arteries from Fetal Sheep
Authors: Paez Ricardo , Nguyen Dan , Papamatheakis Demosthenes , Kim Joon , Blood Quintin , Longo Lawrence D , Wilson Sean M
Published: June 2021

Muscarinic acetylcholine (ACh) receptor (mAChR) activation relaxes arteries through endothelium‐dependent signaling pathways, yet ACh can contract pulmonary arteries (PA). The impact of development and chronic hypoxia (CH) on PA contraction and intimacy between endothelium and myocyte function led to the hypothesis that chronic hypoxia (CH) reduces mAChR‐dependent PA contractility. This was tested by performing wire‐myography on endothelium‐denuded PA rings from term‐fetal or adult sheep that lived at low altitude or 3,200 meters for <100 days. The roles for the three mAChR isoforms were assessed using the mAChR agonist carbachol and various antagonists, including non‐selective (atropine), or M1 (Pirenzepine), M2 (AFDX‐116), and M3 (Dau5884, 4‐DAMP) selective blockers. Carbachol responses as a function of arterial diameter were also examined. Carbachol contracted very few vessels from fetal CH sheep. However, in fetal normoxic and adult CH sheep, smaller arteries contracted more than larger ones, and in adult normoxic medium arteries contracted the most. The M2 receptor predominated in fetal normoxic while M3 was most important in adult normoxic and CH arteries. These results suggest M2 mAChR reactivity contributes to high pulmonary vascular resistance in the fetus and loss of contractility in the CH fetus may influence pulmonary vascular disease of the newborn. NIH P01HD031226, R01HD003807 (LDL).
Caveolin‐1 and BKCa Channel Activity in Pulmonary Arterial Smooth Muscle
Authors: Zhu Shu , Fulton David , Chen Feng , White Richard , Barman Scott
Published: June 2021

Pulmonary arterial hypertension (PAH) is characterized by an elevated, sustained increase in pulmonary artery pressure and eventual right heart failure. Caveolins (cavs) are the primary structural proteins of caveolae that function as chaperones and scaffolding proteins to caveolae for temporal and spatial regulation of signal transduction. Evidence suggests that caveolin‐1 (cav‐1) gene and protein expression is altered in severe pulmonary hypertension. BKCa channels are important regulators of pulmonary arterial smooth muscle cell (PASMC) function, and evidence suggests a functional interaction between cav‐1 and BKCa channels. The present study determined the effect of cav‐1 signaling on BKCa channel activity in hypertensive PASMC of the Fawn‐Hooded rat, and in normotensive PASMC of the Sprague‐Dawley rat. Cav‐1 gene and protein expression was decreased in hypertensive PASMC and caveolin‐1 disruption decreased pulmonary vasodilation by BKCa channel activation in normotensive pulmonary arteries to a greater extent than in hypertensive vessels. These results suggest that cav‐1 positively regulates BKCa channel activity in the pulmonary vasculature. Supported by NIH and AHA.
High fat diet increases adipose tissue mass, but not lean tissue mass in age‐relevant postmenopausal mice in association with induction of VEGF expression and angiogenesis in adipose tissue
Authors: Gu Jian-Wei , Makey Kristina L. , Shareet Mohammad A , Chinchar Edmund , Pei Ivy , Miele Lucio
Published: June 2021

The role of angiogenesis in obesity has not been clearly characterized. In the present study, ovariectomy (OVX) was performed in 8 eighty week‐old C57BL/6 mice, then followed by a low‐fat (4%, LF, n=4) or a high‐fat (60%, HF, n=4) diet for 4 wks. The lean mass (LM) or fat mass (FM) composition was monitored by EchoMRI weekly. At the end of the experiment, blood samples, visceral fat, and the heart and limb muscles were collected for measuring VEGF expression using ELISA and vascular density (VD) using CD31 immunochemistry. Increased body weight (47.6±3.5 vs. 34.6±2.8 g; P<0.01) was due to increased FM (23.3±2.7 vs. 11.6±2.1 g; P<0.01) in OVX/HF vs. OVX/LF mice. But, a high fat diet didn't change LM (22.8±2.1 vs. 21.5±1.9 g) in OVX/HF mice, compared to OVX/LF mice. There was a two‐fold increase in the ratio of visceral fat/BW in OVX/HF mice, compared to those in OVX/LF group (0.084±0.005 vs. 0.043±0.003; P<0.01). Plasma VEGF levels (70±7 vs. 49±3 pg/ml) and visceral fat VEGF levels (431±39 vs. 210±22 pg/mg protein) were significantly increased in OVX/HF mice, compared to OVX/LF group, respectively (4=6; P<0.01). There was an increased VD in visceral fat but not in the heart and limb muscles in OVX/HF mice, compared to OVX/LF group. The results support the hypothesis that adipose tissue responds more sensitively to a high fat diet due to increased VEGF expression and angiogenesis.
Specific Endothelin A (ETA) Receptor Blockade Results In Reduced Expression of Endoplasmic Reticulum (ER) Stress Proteins in Renal Medulla of Type‐1 Diabetic (T1D) Rats
Authors: De Miguel Carmen , Pollock David M. , Pollock Jennifer S.
Published: June 2021

Renal levels of endothelin‐1 (ET‐1) are upregulated in diabetic kidney injury; however, the molecular mechanisms by which ET‐1 leads to renal damage are unclear. We hypothesized that ET‐1, through activation of ETA receptor, increases renal ER stress, thus, ETA blockade diminishes renal medullary ER stress. ER stress is caused by misfolded protein accumulation in the ER, leading to increased transcription of ER chaperones and proteins, eventually leading to cellular death. T1D was induced in Sprague‐Dawley rats by i.v. injection of streptozotocin (65mg/kg); control animals received an injection of saline. Initial immunohistochemical studies demonstrated elevated presence of ER stress markers 78KDa glucose regulated protein (GRP78) and C/EBP homologous protein (CHOP) in renal medulla 10 weeks post‐T1D induction. Immunoblotting analysis confirmed that CHOP expression in isolated medullary thick ascending limbs (mTAL) was 10 times greater in diabetic rats than in controls (0.018±0.004 R.D.U. vs. 0.002±0.002 R.D.U., respectively). T1D rats received either ABT‐627 (specific ETA receptor blocker; 5 mg/kg/day) in drinking water for 10 weeks or untreated water (n=8–10/group). ABT‐627 treatment resulted in reduced outer medullary expression of GRP78 and CHOP by immunohistochemistry. These results implicate the direct participation of ET‐1/ETA pathway in ER stress development in the diabetic kidney.
Losartan prevents body weight gain in diet induced obese rats
Authors: Smith Pauline M , Hindmarch Charles C.T. , Murphy David , Ferguson Alastair V.
Published: June 2021

The renin angiotensin system (RAS), best known for its involvement in the regulation of cardiovascular function and salt/water balance, has recently been suggested to play a role in the regulation of energy homeostasis as interference with systemic RAS activity has been shown to decrease body weight and fat. The present study was undertaken to determine whether losartan, a non‐peptide ATII Type 1 receptor blocker, would influence daily body weight gain in diet induced obese (DIO) rats.

Male Sprague Dawley rats (125–150g) were exposed to either a HFD or fed standard rat chow. After 10 weeks, rats on HFD were divided into DIO or diet resistant (DR) and at 18 weeks, body weight, and food and water intake were measured daily for 5 weeks. After a 14 day control period, rats were administered losartan (30mg/kg) in drinking water for 14 days, followed by return to tap water for an additional 7 days and effects on body weight and food and water intake assessed.

There was no difference in daily body weight gain during in the control period between chow fed (2.6±0.5g, n=8), DR (2.1±0.5g, n=8) or DIO rats (3.4±0.4g, n=8, p=.18, one way ANOVA). Although losartan treatment attenuated daily body weight gain in chow fed rats (1.25±0.5g, n=8), it completely abolished daily weight gain in DR (−0.65±0.3g, n=8) and DIO (−0.43±0.7g, n=8) rats. Upon return to normal tap water, all rats gained weight as they had prior to losartan administration (chow 3.3±0.2g, DR 2.9±0.5g, DIO 3.8±0.5g).

Our data demonstrate that losartan attenuates body weight gain in rats fed either a HFD (DR and DIO) or chow diet, an effect which was more pronounced in animals fed the HFD. These data suggest that the RAS plays a role in body weight homeostasis and that pharmacological agents that interfere with the functioning of the RAS may have be useful in the treatment for obesity.
Functional reassembly of NBCe1‐A from co‐expressed cytosolic and transmembrane domains
Authors: Parker Mark D , Wass Amanda B , Lee Seong-Ki , Rahman Farah , Grant Corey , Boron Walter F
Published: June 2021

The renal electrogenic sodium bicarbonate cotransporter NBCe1‐A is composed of three major domains: a ~45 kDa cytosolic amino‐terminal domain (Nt), a ~60 kDa transmembrane domain (TMD), and a ~10 kDa cytosolic carboxy‐terminal domain (Ct). Expressed in isolation in Xenopus oocytes, neither the Nt nor an NBCe1 construct that lacks the Nt (TMD/Ct) mediates detectable electrogenic NBC activity. However, oocytes co‐expressing Nt and TMD/Ct from two separate cRNAs exhibit substantial NBCe1 activity. Furthermore, in oocytes that are expressing TMD/Ct from cRNA, NBCe1 activity can be elicited by injection of recombinant Nt protein. The reconstitution of NBCe1 activity by Nt and TMD/Ct does not require the NBCe1‐Ct, as evidenced by reconstitution of NBCe1‐like activity in oocytes that are co‐expressing Nt and the chimeric TMD/Ct construct “NBCe1‐TMD/AE1‐Ct” in which the Ct of NBCe1 is replaced by the Ct of AE1. These data are consistent with the hypotheses that the Nt and the intracellular face of the TMD of NBCe1 contain reciprocal binding sites and that binding of the Nt to the TMD is required to produce functional NBCe1. Control of Nt occupancy at its binding site on the TMD could represent a novel means by which NBCe1 activity, and thereby HCO3− reabsorption, is regulated.
Non‐invasive integrative analysis of cardiac energetics, translation to human heart in vivo
Authors: Diolez Philippe Henri , Deschodt-Arsac Veronique , Chapolard Mathilde , Hocini Meleze , Jais Pierre , Haissaguerre Michel
Published: June 2021

We propose MOCA, an inovative non‐invasive clinical analysis of heart energetics, based on the combination of non‐invasive MRS & MRI techniques and the analytical background of metabolic control analysis. MoCA brings new informations on the links between ATP demand and supply and delivers the description of heart function through integrated internal regulations, currently not accessible clinically. We anticipate that these informations may be of great help in the future for diagnosis and therapeutics follow‐up in human heart pathologies.

By combining 31P‐NMR spectroscopy with direct measurement of heart contractile activity, we applied MoCA approach to beating Langendorf perfused rat heart and demonstrated the parallel activation of mitochondrial activity and myofibrils ATPases by Ca2+, which explains the energetic homeostasis of the heart. Further, we could describe quantitatively the global action of modulators of heart contraction (adrenaline, levosimendan) or of a pathology (hypoxia), and the relative importance of the different routes within the heart by which the effects are transmitted.

Application to the study of contraction energetics in healthy and pathological heart in human may be a key to the understanding of the links between molecular events in pathologies and whole organ function/dysfunction.
Enantiomeric separation of doxapram reveals a superior respiratory stimulant, GAL‐054
Authors: Golder Francis John , Gruber Ryan B , Baby Santhosh M , Puskovic Veljko , Ideo Courtney M , Peng Sean , Dax Scott L , MacIntyre D Euan , Mannion James C
Published: June 2021

Doxapram is a clinical respiratory stimulant but is limited by analeptic and cardiovascular adverse events (AEs). We hypothesized that the enantiomers of doxapram may diverge with respect to efficacy and tolerability. In the 1st series of experiments, we administered vehicle, doxapram (4, 20, & 40 mg/kg, IV) and equivalent doses of GAL‐054 and GAL‐053 (each at 2, 10, & 20 mg/kg, IV) to conscious rats. Doxapram and GAL‐ 054 dose‐dependently increased tidal volume, respiratory frequency, and minute ventilation (Vmin) compared to vehicle, whereas GAL‐053 did not. The magnitudes of the effect of 20 mg/kg doxapram and 10 mg/kg GAL‐054 on Vmin were equivalent. All rats that received 40 mg/kg doxapram seized and died, whereas 2 of 6 rats that received 20 mg/kg GAL‐053 seized and one died. In the 2nd series of experiments, all 3 compounds (each at 1 – 30 mg/kg, IV) were administered to anesthetized rats. The ED50 values for doxapram and GAL‐054 for increasing Vmin were 6.4 and 1.8 mg/kg, respectively, but GAL‐053 was essentially inactive. Dysrhythmias were observed with doxapram and GAL‐ 053. These data indicate that the (+) enantiomer, GAL‐054, retains all of the respiratory stimulant properties of doxapram, whereas the AEs (dysrhythmias, seizures, and death) are restricted to the (−) enantiomer, GAL‐053. Thus, of the enantiomers of doxapram, GAL‐054, is the eutomer, and displays a markedly higher therapeutic index than GAL‐053.
Inhibitory Mechanisms of Ethanolic Extract of Fruiting Body of Antrodia Cinnamomea in Cell Growth, Migration and Drug Resistance in Human Lung Cancer‐Derived A549 Cells
Authors: Wu Chieh-Hsi , Liu Fon-Chang , Pan Chun-Hsu
Published: June 2021

Non‐small cell lung cancer (NSCLC) is generally insensitive to chemotherapy, and Antrodia cinnamomea (AC) has a long history used as a natural remedy for health improvement. The present study attempted to explore the anti‐cancer activity and mechanisms of the ethanolic extract of AC (EEAC) in human lung epithelial carcinoma A549 cell lines, a NSCLC cells. The results showed that EEAC treatment markedly decreased cell viability, arrested cell cycle in G0/G1 phase, and increased the activation of AMPK as well as downregulated the phosphorylation of Akt, mTOR, RB and ERK 1/2 proteins. Additionally, EEAC inhibited cell migration and reduced the expression level of matrix metalloproteinase (MMP)‐2 and 9 proteins. It has also noticed that EEAC can suppress the expression of Cav‐1 protein to enhance the chemotherapy sensitivity of A549 cells to paclitaxel. Our study suggested that EEAC might have potential activities to be developed as adjuvant remedy for lung cancer chemotherapy.
TNF‐α inhibits breast cancer cells adhesion via HIF‐1α mediated VASP suppression
Authors: Su Ke , Tian Yihao , Wang Jing , Shi Wentao , Zhang Jingwei , Wei Lei
Published: June 2021

TNF‐α is implicated in tumor invasion and metastasis, but its mechanism remains elusive. Herein we provide a study underlying the role of TNF‐α in inhibiting MCF‐7 breast cancer cells adhesion which initiate breast cancer metastasis process. In this research, to evaluate the effect of TNF‐α (100 ng/ml) on MCF‐7 cells adhesion ability, MTT assay were performed. We observed that TNF‐α administration led to decreased adhesion ability of MCF‐7 cells in a dose dependent manner. Secondly, TNF‐α attenuates MCF‐7 cells adhesion may via suppressing VASP (vasodilator‐stimulated phosphoprotein) protein expression levels analysing by westernblot. Additionally, TNF‐α had no obvious effect on HIF‐1α mRNA expression levels analysing by Real‐time PCR. In contrast, TNF‐α upregulated HIF‐1α protein expression levels. Furthermore, using promoter assay, EMSA and ChIP approaches we identified that VASP as a direct target gene of HIF‐1α and HIF‐1α repress VASP at transcription level. Consistently, knockdown of HIF‐1α abrogated TNF‐α mediated VASP suppression. Collectively, we provided evidence that HIF‐1α may acts downstream of TNF‐α to inhibit VASP expression and modulate the adhesion of breast cancer cells. (This work was supported by National Natural Sciences Foundation of China under Grant No.30971132, 81172043 and the Hubei Science Foundation under Grant No. 2009CDA074)
Atomic Structure of Chimeric Hepatitis E Virus Designed for Targeted Nano‐Carrier and Delivery System
Authors: Jariyapong Pitchanee , Xing Li , Weerachatyanukul Wattana , Cheng Holland
Published: June 2021

The development of either oral vaccines against pathogens or biological container to carry materials for local targeting in digestive mucosa relies on the chief principle in tolerance of vehicle to harsh environment of digestive system to efficiently carry and deliver the cargo to its destination. Delivery system such as enteric‐transmitted virus‐like particles (VLPs) is advantageous because of the inherited acid‐ and proteolysis‐resistance of the parental viral capsids. In the present study, we engineered hepatitis E (HEV) VLP by inserting an antigenic V3 loop of human immunodeficiency virus (HIV) derived peptide, p18, into the recombinant HEV capsid protein (p18‐VLP) after residue tyrosine 485 on the protruding domain (P domain). Using cryo‐transmission electron microscopy (cryo‐TEM) and three dimensional (3D) image reconstruction, quaternary protein structure of the chimeric p18‐VLP reassembled to that of wild type HEV‐VLP with a slight difference at the protruding spike. This engineered particle also reacted to a specific HIV monoclonal antibody, but not HEV antibody at P‐domain suggesting that the inserted p18 was exposed on the surface of VLPs. Even though p18‐VLP was vulnerable to enzymatic cleavage, presumably occurred at the C‐terminal end of the inserted p18 peptide, the p18‐VLP remained structurally intact and maintained particle integrity. The results thus indicate the further application of this HEV‐VLP system as a candidate of oral delivery vector.

This research is supported by Commission on Higher Education of Thailand

Grant Funding Source: Commission on Higher Education of Thailand
Telomere Homeostasis in Budding Yeast is Regulated by a Single Mechanism Involving Replication Fork Collapse
Authors: Lundblad Vicki , Paschini Margherita
Published: June 2021

In budding yeast, telomere homeostasis relies on a telomere‐dedicated RPA‐like (t‐RPA) complex composed of the single‐strand DNA binding protein Cdc13 in association with Stn1 and Ten1. While bound to telomeres, the t‐RPA complex recruits telomerase to its site of action, and it performs an essential function which is widely assumed to be protection from unregulated resection. A current model assumes that these two events occur following completion of replication. We present here a substantially different model. Specifically, we propose that the t‐RPA complex is a replication factor which associates with telomeres as the replisome proceeds into the G‐rich telomeric duplex tract. Once associated with the replisome, the t‐RPA complex facilitates replication of duplex telomeric DNA by (i) preventing replication fork collapse, which we propose is the essential role of the t‐RPA complex and (ii) resolving collapsed forks by recruiting telomerase to the site of fork collapse. This model is supported by analysis of the sequence of duplex telomeric DNA in mutant strains, which reveals a role for both t‐RPA and other telomere proteins in progression of the replisome through duplex telomeric DNA.