bluuebunny/crossref_metadata_2025 huggingface dataset:
Maturation of Osteoblast‐like cells sensitizes them to Calcium and Phosphate Ion Pair‐Induced Apoptosis
Authors: Genoese Angela Antonia , Saunders Ray , Debo Matthew , Adams Christopher S
Published: June 2021

Our previous work demonstrated the sensitivity of osteoblasts to the presence of microenvironmental apoptogens. As osteoblasts change their functional orientation with maturation, it follows that the sensitivity of these cells to apoptosis must change as well. It is the goal of this study to exam the relationship between the maturation of osteoblasts and the susceptibility of these cells to apoptosis. For this study the Human Fetal Osteoblast (hFOB) cell line was used. These cells exhibit an early osteoblastic phenotype when cultured at 34ºC while above 37ºC, the cells mature. Cells were cultured for 2, 7 and 14 days. Phenotypic changes were confirmed. Cells cultured at both 34ºC and 39.5ºC were exposed to increasing levels of the calcium and phosphate ion pair. Percentages of viable cells were measured using the MTT assay. Apoptosis was confirmed by evaluating Caspase‐3 activation and TUNEL staining. Results indicated that hFOB cells incubated at 39.5ºC exhibited a greater sensitivity to the activation of apoptosis by the calcium and phosphate ion pair. These results suggest that with maturation, osteoblasts become more sensitive to the activation of apoptosis. This work was supported by a grant from the Center for Chronic Disorders of Aging at the Philadelphia College of Osteopathic Medicine.
The role of cAMP/PKA signaling enhancer Protein Phosphatase Inhibitor 1 (I‐1) in the control of Ca2+ cycling and signaling in VSMCs
Authors: Hadri Lahouaria , Regis Bobe , Lopez Jose J , Turnbull Irene C , Kovacic Jason C , Karakikes Ioannis , Kranias Evangelia , Hajjar Roger J , Lipskaia Larissa
Published: June 2021

The Ca2+ cycling in “contractile” vascular smooth muscle cells (VSMC) requires the expression of SERCA2a, whereas in “proliferating” ones is associated with the expression of SERCA2b only. Inhibitor‐1 (I‐1), a highly specific inhibitor of protein phosphatase 1 (PP1), enhances both PKA‐dependent Phospholamban (PLB) phosphorylation and SERCA activity.

The goal of this study was to elucidate the role of I‐1 in the control of VSMC Ca2+ cycling. In humans and rodents, the expression of I‐1 was found to be specific for contractile VSMC, whereas PP1 was highly expressed in synthetic VSMC. Genetic deletion of I‐1 in mice model resulted in lack of PLB phosphorylation in VSMC. Despite the fact that SERCA2a was expressed, VSMCs from I‐1 deficient mice were locked in the proliferating state, with the development of a vascular proliferative disorder and excessive neointimal proliferation after vascular injury. Gene transfer of constitutively active I‐1 (I‐1c) significantly increased PLB phosphorylation and Ca2+ uptake in both synthetic and contractile VSMCs. Therefore, I‐1c prevented proliferation and remodeling of contractile VSMC, but had no effect on synthetic VSMC, which predominantly express SERCA2b.

In conclusion, Gene transfer of I‐1c may be considered as a promising therapeutic strategy for preventing vascular proliferative diseases.
Decreased release of methyl palmitate in perivascular adipose tissue in hypertension
Authors: Chang Hsi-Hsien , Lee Yuan-Chieh , Chiang Chih-Lung , Liu Chin-Hung , Chen Mei-Fang , Chen Po-Yi , Lee Tony J. F.
Published: June 2021

Our recent studies have demonstrated that palmitic acid methyl ester (PAME) is the major perivascular adipose tissue (PVAT)‐derived relaxing factor. We determined if PAME release from the PVAT and/or its vasorelaxing activity decreased in hypertension. PAME release from aortic PVAT preparations of Wistar Kyoto rats (WKY) and spontaneously hypertensive rat (SHR, 20‐week old) was estimated pharmacologically using superfusion bioassay cascade technique and biochemically by GC/MS. Results indicated that the PVAT of WKY and SHR aorta spontaneously released PAME. The release was significantly less but with an increased release of angiotensin II in SHR. Also, PAME‐induced relaxation of SHR aortic smooth muscle diminished drastically, which was reversed by losartan. Incubation of isolated PVAT preparations of SHR in Krebs solution containing losartan (10 £gM) for 30 min enhanced PVAT‐elicited vasorelaxation, with enhanced PAME release. These results suggest a noble role of PVAT & PAME in pathogenesis of hypertension. Antihypertensive effect of losartan is partly attributed to its increasing PAME release and reversing diminished PAME‐induced vasorelaxation (Supported by NSC, Tzu Chi Found & TCU).
Anti‐inflammatory effects of Laminaria japonica in association with the inhibition of nitric oxide and pro‐inflammatory cytokines production
Authors: Kim Young Woo , Cho Je , Kim Sang Chan
Published: June 2021

Laminaria japonica has been frequently used as a drug in traditional oriental medicine, and as food supplements in many of the Asian countries. This research investigated the effects of Laminaria japonica extract (LJE) on the paw edema formation in rats and on the induction of nitric oxide (NO), prostaglandin E2 (PGE2) and proinflammatory cytokines by lipopolysaccharide (LPS) in Raw264.7 cells. The effect of LJE on acute phase inflammation was evaluated in a carrageenan‐induced paw edema model, as assessed by histomorphometric and immunohistochemical analyses. NO, PGE2, TNF‐α, interleukin‐1β and interleukin‐6 contents were assayed by ELISA. iNOS and COX‐2 expressions were determined by western blot analyses. In rats, LJE treatment inhibited carrageenan‐induced paw edema formation, and infiltration of inflammatory cells in H&E staining. LJE treatment prevented the ability of LPS to increase the levels of iNOS and COX‐2 protein in a concentration‐dependent manner. Consistently, LJE suppressed the production of TNF‐α, interleukin‐1β and interleukin‐6. Treatment of the cells with LJE caused inhibition of I‐κBα phosphorylation induced by LPS suggesting LJE repression of NF‐κB activity by LPS. In conclusion, this study showing here may be of help to understand the pharmacology and action mechanism of LJE, and the anti‐inflammatory use of Laminaria japonica.
Desflurane‐Induced Coughing in Conscious Guinea Pigs is Mediated through TRPA1
Authors: Daller Justin , McKee Jeffrey S. , Wong Joseph , Brooks Ben
Published: June 2021

Desflurane is widely used for induction and maintenance of anesthesia, and is associated with a high incidence of perioperative coughing. Desflurane has been shown to activate TRPA1 in vitro and increase total lung resistance in anesthetized guinea pigs that could be mitigated with a selective TRPA1 antagonist.

Purpose

To evaluate the mechanism of desflurane‐induced coughing in conscious guinea pigs during anesthetic induction.

Methods

Guinea pigs were placed into a transparent chamber where flow rate, pressure, audio and video were recorded. Animals were induced with 1 MAC (6.4%) desflurane or challenged with a selective agonist for TRPV1 (citric acid, 1 M) or TRPA1 (AITC, 80 mM) with or without pre‐treatment with a selective antagonist for TRPV1 (BCTC, 3 mg/kg IP) or TRPA1 (HC‐030031, 300 mg/kg IP). The number of coughs over a fixed, 5‐min exposure period was recorded.

Results

Desflurane caused 4.9 coughs vs. 0 for the carrier air. AITC caused 10.7 coughs vs. 5.8 when pretreated with HC‐030031 (p<0.05). Citric acid caused 12.4 coughs vs. 3.2 when pre‐treated with BCTC (p<0.002). HC‐030031 markedly attenuated desflurane‐induced coughing (4.3 vs. 1.2, p<0.08) whereas BCTC had no effect (5.4 vs. 4.0, p=0.13).

Conclusion

Desflurane‐induced coughing in conscious guinea pigs appears to be mediated through activation of TRPA1. Supported by Baxter Healthcare Corporation.
Psoriasis – modern and traditional treatment approaches
Authors: Senthilkumar Uma , Sheu Mary
Published: June 2021

Psoriasis is one of the most common immune‐mediated skin disorders. In allopathic medicine the first‐line therapy for patients with moderate to severe psoriasis is the application of topical agents, followed by phototherapy for more extensive disease. Second‐line agents include PUVA, acitretin, methotrexate, cyclosporine or other immunosuppressive agents. New biologics like infliximab, etanercept, alefacept, ustekinumab have revolutionized the management of severe psoriasis. These agents act by blocking the action of T cell, TNF‐alpha or IL‐ 12 and 23. These medical approaches do not offer a complete cure for this disorder especially when initiated long after the occurrence of lesions. These treatment options are associated with moderate to serious adverse drug reactions. With strict dietary and life style modifications Ayurvedic and Siddha medicine treatment approaches, both of which are practiced in India for hundreds of years, offer an alternative approach in the management of psoriasis. Compounds derived from herbs such Wrightia tinctoria, Cynadon dactylon, Melia azadirachta, Aregemone Mexicana, Chirattai thailam, curcumin and almond oil have proved to be beneficial and offer complete cure. Further research to investigate the possible mechanism of action of these drugs and long term case controlled studies are warranted to assess the effectiveness of these treatment methodologies.
Parkin protects dopaminergic terminals against methamphetamine neurotoxicity by inducing inclusion formation
Authors: Liu Bin , Moszczynska Anna
Published: June 2021

Viral vector‐mediated delivery of parkin pervents dopaminergic (DAergic) neurodegeneration in Parkinsonˈs disease models. One of the mechanisms underlying parkin neuroprotection is to promote the formation of á‐synuclein inclusions. It is unclear whether parkin protects against methamphetamine (METH)‐induced degeneration of DAergic terminals. Here we investigated whether parkin overexpression protects against METH neurotoxicity and whether the parkin effect is due to the formation of inclusions. Parkin‐containing adeno‐associated viral 2/6 vector was injected into the left substantia nigra in rats with/without METH treatment. Parkin‐immunoreactivity (IR) in the left substantia nigra and striatum was higher than that of right side. A large portion of parkin‐positive DAergic neurons and terminals expressed tyrosine hydroxylase (TH). Animals overerexpressing parkin showed reductions in METH‐induced decrease in TH‐IR in the striatum. Parkin‐mediated neuroprotection was associated with an increase of punctuated á‐synuclein immunostaining in the striatum. These data suggest that parkin overexpression attenuates METH‐induced damage to striatal DAergic terminals, which may be via increasing á‐synuclein inclusions. It also suggests that future clinical applications to prevent METH neurotoxicity might involve the mediation of parkin function. Support: NIH DA023085
Activating PPARs‐γ alleviates Parkinson disease revealed by behavioral and functional MRI assays
Authors: Chang Chen , Chen Chiao-Chi V , Hsu Yi-Hua , Lin Teng-Nan
Published: June 2021

Neuroinflammation is a suspected cause of dopaminergic neurodegeneration of the substantia nigra (SN) in Parkinson disease (PD). This study aims to understand whether modulating neuroinflammation by activating peroxisome proliferator‐activated receptors‐gamma (PPARs‐γ) produces beneficial effects on PD using in vivo assays including behavioral assessment and functional MRI. A rat PD model induced by lesioning the SN with 6‐hydroxy‐dopamine (6‐OHDA) was used. Upon the lesion, an osmotic minipump filled with 15‐deoxy‐delta12,14‐prostagladin J (PGJ), an endogenous PPAR‐gamma agonist, was implanted in the SN. Saline and three doses were examined. The effects were evaluated by (1) meth‐amphetamine induced rotational behavior and (2) cerebral blood volume (CBV)‐weighted fMRI for the striatum where the dopaminergic fibers terminate. Activating PPARs‐γ by continuous infusion of PGJ into the SN alleviated the behavioral symptoms and fMRI responses specific to PD. Among the three doses, the medium one produced the most beneficial effects on the disease signs. fMRI show that PGJ at the medium dose best preserved the CBV reactions associated with dopaminergic integrity while the saline group exhibited diminished CBV signals due to the significant loss of dopaminergic inputs. The results imply that PD is mitigated by the modulation of neuroinflammatory activity via the activation of PPARs‐γ.
Pharmacokinetic evidence for the long‐lasting effects of nor‐binaltorphimine (nor‐BNI)
Authors: Kishioka Shiroh , Kiguchi Norikazu , Kobayashi Yuka , Ko Mei-Chuan , Woods James
Published: June 2021

It is well known that a single administration of nor‐binaltorphimine (nor‐BNI), which is a kappa opioid receptor antagonist, has long‐lasting effects in vivo. However, there is no direct pharmacokinetic evidence of prolonged effects of this drug. In this experiment, we quantified both serum concentration and brain content of nor‐BNI in mice, using high‐performance liquid chromatography with the electrochemical detector (HPLC‐ECD) method. After subcutaneous injection of a mixture of nor‐BNI (30 mg/kg) and naloxone (30 mg/kg), serum and brain samples were collected at 1 hr, 1, 3, 7, 14, 21 and 28 days. Serum concentrations of nor‐BNI were detected at 1 hr and 1 day after injection in all mice, at day 3 in three out of seven mice, and at day 7 in one out of six mice. After day 14, serum concentrations of nor‐BNI were below the lower limit of detection in all mice. In comparison, nor‐BNI was detected at in the brains of all mice from 1 hr to day 21, and was detected at day 28 in two out of six mice. On the other hand, serum concentrations and brain content of naloxone were detected only at 1 hr after injection and both serum concentrations and brain content were below the lower detection limit after that in all mice. In conclusion, we detected nor‐BNI and naloxone in both blood and brain using HPLC‐ECD method, and provided the pharmacokinetic evidence for the long‐lasting effects of nor‐BNI in vivo.
CaMKII inhibition in vascular smooth muscle improves angiotensin II–hypertension
Authors: PRASAD ANAND MOHAN , Koval Olha M , Nuno Daniel W , Xie Litao , Li Hui , Li Weiwei , Sanders Philip N , Ketsawatsomkron Pimonrat , Kutschke William , Accaoui Ramzi El , Weiss Robert , Lamping Kathryn G , Sigmund Curt D , Anderson Mark E , GRUMBACH ISABELLA M
Published: June 2021

The multifunctional calcium/calmodulin‐dependent kinase II (CaMKII) is expressed in vascular smooth muscle cells (VSMC) and activated by Angiotensin‐II (Ang‐II). To test the hypothesis that CaMKII in VSMC mediates Ang‐II‐hypertension, we developed transgenic mice that express the specific CaMKII peptide inhibitor CaMKIIN selectively in VSMC (Tg SM CaMKIIN). These mice have a 65 % reduced CaMKII activity in arteries (p<0.01) and identical baseline blood pressure compared to littermate controls. Following Ang‐II infusion at pressor dose, the increase in mean pressure was significantly attenuated (129±4 mmHg vs, 116±3 mmHg, p<0.05). Ang‐II increases Ca2+ influx through L‐type Ca2+ channels (LTCC). Cytosolic Ca2+ ([Ca2+]) rise leads to an increased vascular tone. Compared with littermates, freshly isolated mesenteric VSMC of Ang‐II treated Tg SM CaMKIIN mice had significantly reduced voltage gated L‐type Ca2+ current (ICa), ICa facilitation. Moreover, the [Ca2+] elevation in response to Ang‐II stimulation was reduced by 50 % in VSMC of TgSM CaMKIIN mice (0.045±0.002 vs, 0.025±0.003; ratio340/380; p<0.05) whereas, level of total releasable SR calcium was decreased by 35 %. In conclusion, inhibition of CaMKII in VSMC significantly attenuates Ang‐II induced hypertension through regulation of LTCC and by restoring the calcium homeostasis. CaMKII may be a novel target in treating hypertension.
IMP2 Expression In The Mouse Nervous System
Authors: Chmielewski Sarah Elizabeth , Black Paige E. , Holmes Tiffany , Lau Kelly , Bassell Gary J. , Xu Mei
Published: June 2021

Insulin‐like growth factor‐II (IGF‐II) mRNA‐binding protein‐2 (IMP2) is one of the three homologs (IMP1–3) that play important roles in the posttranscriptional regulation of gene expression in several tissues. IMP1/ZBP1 (zipcode binding protein) has been shown to play important roles in axon guidance and regeneration by regulating the localization and translation of specific mRNAs. However, the function of IMP2 is least understood, largely because an isoform‐specific antibody is not available, which makes the conventional techniques to locate protein expression not feasible. We custom made an IMP2‐specific antibody. We used Western blot and immunocytochemistry to test its specificity on cultured cells following over expression of IMP 1–3 isoforms, respectively. Using this IMP2‐specific antibody, we examined IMP2 expression in the mouse nervous system. We found that IMP2 expression in the nervous system is sustained postnatally, unlike that of IMP1 and IMP3. Ongoing experiments are aimed at further understanding IMP2 expression patterns during injury and assessment of its role to facilitate mRNA localization during axon regeneration in the adult nervous system.

This work is supported by the Department of Anatomy, PCOM and Center for Chronic Disorders of Aging (CCDA).
On the Anatomy of a Modern Biotelemetry System: A Cell phone Paradigm
Authors: Pitsillides Koullis
Published: June 2021

Totally implantable biotelemetry systems are becoming very useful tools in both academic and pharmaceutical research laboratories. In the past 30 years, implantable biotelemetry systems have made possible a number of studies that required the use remote measurements of parameters such as ECG, blood pressure and temperature. Current and future medical research measurements, however, demand more advanced features and functions from the biotelemetry systems currently available. In the past, biotelemetry implants were very much like the cell phones of that era. Inflexible, unchangeable, preset, analog. Our recently introduced EndoGear modern biotelemetry system, is a powerful, flexible, user configurable, programmable and digital, much like the smartphones of today. The advantages of our biotelemetry implants are the harnessing of demanding applications by incorporating new and advanced sensors, such as blood flow sensors, solid‐state pressure sensors, and modern RF communication protocols. Users can attach and control a variety of add‐on modules that can perform new functions without the need for a completely new implant; i.e., run a new application (app). Commands and data using digital RF communication protocols minimize signal noise and improve signal accuracy. In general, the advancement of modern biotelemetry systems was a direct result of the advancement of modern smartphone technologies. The drive to produce smaller, more advanced electronic parts and more powerful smartphones helped the development of smaller, more advanced and more powerful biotelemetry systems that have the capacity and new capabilities to assist medical researchers to exciting new discoveries.
Myocardial Stunning in the 5/6 Ablation/Infarction Model of Chronic Kidney Disease
Authors: Kuczmarski James M , Martens Christopher R , Lennon-Edwards Shannon L , Edwards David G
Published: June 2021

Myocardial stunning is a form of cardiac ischemia‐reperfusion (I/R) injury characterized by reversible contractile dysfunction following short bouts of ischemia without resulting necrosis. The purpose of this investigation was to assess myocardial stunning in the 5/6 Ablation/Infarction (AI) model of chronic kidney disease (CKD). Further, oxidative stress was explored as a potential mechanism for decreased nitric oxide (NO) and augmented I/R injury in CKD. Cardiac function was measured pre‐ and post 15min ischemia using an isolated working heart model, and left ventricular (LV) tissue was used for biochemical analysis. Cardiac function was impaired during post‐ischemia reperfusion (15–30min) in AI as indicated by cardiac output (66±4 vs. 41±4%PRE‐I p<0.05) and cardiac work (65±2 vs. 41±6%PRE‐I; p<0.05). Urinary NOx was lower in AI (1.3±0.2 vs. 0.6±0.1μM/24hr/100g; p<0.05) and accompanied by reduced LV eNOS (1.0±0.10 vs. 0.62±0.13; relative to β‐actin p<0.05) and NOx (18.9±2.2 vs. 11.3±2.3μM; p<0.05) Altered expression of LV antioxidant enzymes was also observed in AI (SOD1: 1.0±0.12 vs. 0.34±0.08; SOD2: 1.0±0.14 vs. 0.52±0.10; GPX: 1.0±0.1 vs. 1.3±0.08; relative to β‐actin all p<0.05). In conclusion, these findings suggest that myocardial stunning is augmented in CKD and could be, in part, the result of altered antioxidant defense leading to impaired NO‐mediated cardioprotection during I/R injury. Supported by: P20 RR016472
SIRT1 Activation with SRT1720 Reverses Impaired Endothelium‐Dependent Dilation in Old Mice by Augmenting COX‐2 Mediated Vasodilation
Authors: Gano Lindsey B. , Seals Douglas R. , Pasha Hamza M. , Hearon Christopher M. , Donato Anthony J.
Published: June 2021

Endothelial dysfunction develops with aging due to reductions in nitric oxide (NO)‐ and/or cyclooxygenase (COX)‐related vasodilation. Treatment with SRT1720 (Sirtris) increases arterial SIRT1 activity and restores endothelium‐dependent dilation (EDD) independent of NO. We hypothesized that COX‐2 vasodilators mediate the improvements in EDD with SRT1720 treatment. Old mice (29–32 mo) were treated with SRT1720 (100 mg/kg BW; OS) or vehicle (OV) for 4 weeks. Maximal ex vivo carotid artery EDD to acetylcholine was reduced with age (Young, 4–9 mo [Y]: 95±1% vs OV: 83±3%, p<0.01) and rescued with SRT1720 (OS: 95±1%, p<0.01). Inhibition of COX‐1 & −2 (indomethacin) reduced max EDD in Y (88±3%, p<0.01) and OS (82±6%, p<0.01), but not in OV (p=0.55), abolishing all group differences (p=0.64). Inhibition of COX‐2 (NS‐398) reduced max EDD in OS (82±6%, p<0.01), but not in Y or OV (p>0.05), and abolished SRT1720 treatment differences (p=0.69). Aortic COX‐2 protein was reduced with age (Y: 0.9±0.2 AU vs OV: 0.5±0.1 AU, p=0.06) and restored with SRT1720 (OS: 1.1±0.2, p<0.05). COX‐1 protein was not different between groups (p=0.23). These results demonstrate that aging is associated with a reduction in COX mediated EDD. SIRT1 activation reverses vascular endothelial dysfunction with aging via COX‐2 specific vasodilators. These results suggest a novel protective effect of SRT1720 treatment on the aged arterial endothelium.
Role of the Skeletal Muscle Pump in Exercise Onset Hyperemia in the Human Forearm
Authors: Carter Matthew Roderick , Gray Eric J , Rawicki Nathaniel L , Jasperse Jeffrey L
Published: June 2021

Blood flow to active skeletal muscle increases with the first contraction, but the mechanism responsible for this exercise onset hyperemia is unknown. We tested the hypothesis that the muscle pump is necessary for onset hyperemia. Blood pressure and brachial artery blood flow velocity (Doppler ultrasound) were measured in 6 young males in three different trials, each performed with the arm at heart level (HL) and with the arm hanging below heart level (BH) in the dependent position, perpendicular to the floor : 1) a single dynamic handgrip contraction (20% MVC) (HG), 2) following a 10‐minute arm occlusion combined with a 2‐minute isometric handgrip (from 7:30 to 9:30 of the occlusion period) to induce maximal vasodilation in the forearm (OC), and 3) arm occlusion and handgrip followed by a dynamic handgrip performed 5 seconds after cuff release (OC+HG). HG elicited an increase in forearm conductance that was not different between arm positions (247±77 vs 308±35%). OC elicited a large increase in conductance that was not different between arm positions (0.100±0.010 vs. 0.101±0.008 units). OC+HG (.093±0.008 at heart level and 0.086±0.010 units below heart level) did not cause a greater increase in conductance than either the OC trial or the pre‐handgrip value in the OC+HG trial. These data suggest that the muscle pump does not play an important role in immediate exercise hyperemia in the forearm.
Expression of Anti‐angiogenesis protein, Tenomodulin, in Vascular Smooth Muscle Cells
Authors: Xing Enze , Wen Yuan , Franz Michael G
Published: June 2021

Tenomodulin (Tnmd) was expressed predominantly in hypovascular connective tissues with anti‐angiogenesis function. Interaction between endothelial cells and vascular smooth muscle cells (VSMCs) plays important role in maintaining normal vascular structure and function, such as angiogenesis. This study investigated Tnmd expression in rat VSMCs (rVSMCs) and explored regulation of Tnmd expression in response to different stimulations. rVSMCs were isolated from aorta of SD rats and maintained under defined in vitro conditions. Tnmd protein and mRNA were detected by immunohistochemistry (IHC), immunofluorescence staining (IF), Western blot, RT‐PCR, and DNA sequencing, respectively. Cells were treated with proinflammatory cytokines, growth factors or immune modulator to induce changes in Tnmd expression. rVSMCs express Tnmd both at mRNA and protein level. ICH and IF showed that Tnmd was located in smooth cell layer of media artery and rVSMCs. Western blot showed Tnmd specific band of rVSMCs located at the same position as that of fibroblast. DNA sequencing on targeted RT‐PCR product from rVSMCs showed 100% identity with that from rat linea alba fibroblast. Tnmd protein level significantly decreased in rVSMCs after 72‐h treatment of PDGF till 120 h. Real‐time RT‐PCR showed that PDGF treatment decreased Tnmd mRNA expression. Data suggest change of Tnmd expression in VSMCs is involved in local regulation of angiogenesis in certain physiological and pathophysiological conditions. This study was supported by Department of General Surgery, University of Michigan medical School.
Impaired muscular force production in a canine model of muscular dystrophin deficiency
Authors: Yang Hsiao Tung , Shin Jin-Hong , Pan Xiufang , Terjung Ronald L , Duan Dongsheng
Published: June 2021

Loss of muscle force is a salient feature of Duchenne muscular dystrophy (DMD), a fatal disease caused by dystrophin deficiency. Assessment of force production from a single intact muscle has been considered as the gold standard for studying physiological consequences in murine models of DMD. A similar assay has not been established in dystrophic dogs. Therefore, we developed a novel in situ protocol to measure force generated by the extensor carpi ulnaris (ECU) muscle from Normal (n=4) and Affected dog (n=3). Absence of dystrophin resulted in marked histological damage in the ECU muscle. Peak tetanic force was significantly reduced in affected dogs (only 78% of Normal group, p<0.025). The ability to maintain the initial tension during continuous tetanic contractions was similar in both groups. Importantly, eccentric contractions by extending 5% of ECU muscle length while muscle was contracting, induced a much higher magnitude of force loss in the affected dogs (p<0.001). Our data showed that dystrophin‐deficient muscle is sensitive to damage by eccentric contraction. To our knowledge, this is the first convincing demonstration of force deficit in a single intact muscle in the canine DMD model. The method described here will be of great value to study physiological outcome following innovative gene and/or cell therapies.

Supported by NIH AR‐49419, MDA, and Jesse's Journey‐the foundation for gene and cell therapy
Efficiency of a controlled physical reconditioning monitored by maximal cardiovascular and metabolic parameters in patients with chronic low back pain
Authors: Duvallet Alain , duvallet Emilie , Lhuissier François , Beaudry Michele , Duque Ivan
Published: June 2021

Practice physical exercise on a regular basis is admitted to be an important element for health.

In patients untrained during a chronic disease, the return is often difficult. A follow‐up is then necessary for reconditioning which will have to be maintained.

In 101 patients with chronic low back pain, presenting a physical deconditioning, we have done a programmed physical training. They performed a 5 weeks training with 6 hours of exercise per day, 5 days a week. The exercises comprised endurance, muscledevelopment and limbering‐up. Patients were tested when included in the protocol and at the end of the 5 weeks. The exercise test was performed on a bicycle ergometer by progressive graded exercise from rest to maximal effort. The collected parameters comprised heart rate, followed all along the effort, the maximum work rate, the maximum oxygen consumption, the respiratory exchange ratio and the blood lactate level. The statistical comparison was made by paired t test.

Taken together these parameters show a statistically significant improvement after physical reconditioning. These results confirm that physical restoration is a complementary factor of general physical condition as a health factor.
MironRNA‐194 (mir‐194) regulates ROMk channel activity by down‐regulation of intersectin 1 (ITSN1)‐WNK pathway
Authors: Lin Daohong , Yue Peng , Wang Wenhui
Published: June 2021

The aim of the present study is to test whether miR‐194 plays a role in mediating the effect of high K (HK) intake on ROMK channels through inhibiting ITSN1‐dependent endocytosis of ROMK channels. Northern blot showed that a HK intake increased the miR‐194 expression in the mouse kidney. The qPCR further demonstrated that a HK intake increased the miR‐194 transcription. Moreover, a HK intake decreased the expression of ITSN1. The role of miR‐194 in regulating ITSN1 expression is further confirmed by Luciferase Assay. Moreover, down‐regulation of endogenous miR‐194 by the antagomer increased ITSN1 protein expression in HEK cells. The whole‐cell recording and biotin labeling technique further showed that expression of miR‐194 increased ROMK channel activity in HEK cells transfected with GFP‐ROMK1, and effect was abolished by co‐expression of ITSN1. We conclude that miR‐194 regulates ROMK channel activity by modulating ITSN1 translation and that HK‐induced increase in miR‐194 is involved in mediating the effect of a HK intake on ROMK channel activity.
Crosstalk between the Glucocorticoid and G Protein‐Coupled Receptor Signaling Pathways Mediated by Arrestins
Authors: Oakley Robert H. , Cidlowski John A.
Published: June 2021

The nonvisual arrestins, βarrestin1 and βarrestin2, are multifunctional adaptor proteins that play a central role in G protein‐coupled receptor (GPCR) signaling. Alterations in the cellular complement of arrestins occur in various human diseases and their genetic ablation in mice has severe consequences. Surprisingly, the factors that directly control arrestin gene expression remain unidentified. We now show that glucocorticoids differentially regulate βarrestin1 and βarrestin2 gene expression in human lung adenocarcinoma cells (A549). Glucocorticoids act through the glucocorticoid receptor (GR) to induce the expression of βarrestin1 and repress the expression of βarrestin2. The glucocorticoid‐dependent regulation involves the recruitment of ligand‐activated GR to a conserved and functional glucocorticoid response element (GRE) in intron‐1 of the βarrestin1 gene and to a conserved and functional negative GRE in intron‐11 of the βarrestin2 gene. The marked rise in βarrestin1 following glucocorticoid treatment leads to enhanced activation of the MAP kinase pathway by the protease activated receptor‐1 (PAR1). These studies demonstrate that glucocorticoids can modulate the signaling mode of GPCRs through alterations in arrestin gene expression, revealing a new paradigm for cross‐talk between nuclear and cell surface receptor signaling pathways.
Delineation of the Intestinal Glucose Sensing Pathway in Obese and Diabetic Humans Undergoing RYGB
Authors: BHUTTA HINA YASEEN , DEELMAN TARA , TAVAKKOLIZADEH ALI
Published: June 2021

Roux‐en‐Y Gastric Bypass (RYGB) surgery for obesity achieves weight loss with additional myriad metabolic benefits. Mechanisms explaining these changes are unknown. We hypothesized that RYGB improves glucose homeostasis by excluding glucose sensors and/or transporters from the enteral pathway.

Whilst undergoing RYGB surgery for obesity, jejunal biopsies were taken 40cm (proximal) and 140cm (distal) from the ligament of treitz (Figure 1) of 23 patients (Table 1). mRNA expression of SGLT1, GLUT2, and T1R2 (part of the sweet taste receptor heterodimer) was measured with RT‐PCR. Statistical analysis used paired students’ t‐test.

Intestinal expression of transporters SGLT1 and GLUT2 was higher in proximal jejunum forming the biliopancreatic limb (p<0.001 and p<0.05 respectively). Conversely, expression of glucose sensor T1R2 was higher distally towards the common limb (p<0.01) (Figure 2).

Glucose transporters and sensors show topographic variation in human jejunum. SGLT1, which accounts for active transport of glucose across the intestinal apical border, and GLUT2, responsible for glucose transport at the basolateral border, are concentrated in jejunum which is bypassed in RYGB surgery, rendering upregulation in response to T1R2 activation redundant post‐RYGB. Replicating exclusion of intestinal glucose transporters in a less invasive manner could provide a valuable way to treat metabolic disease.
Effect of estrogen on molecular and functional characteristics of the rodent vaginal muscularis
Authors: Basha Maureen Elizabeth , Chang Shaohua , Burrows Lara J , Lassmann Jenny , Wein Alan J , Moreland Robert S , Chacko Samuel K
Published: June 2021

Vaginal atrophy is a consequence of menopause, however little is known concerning the effect of estrogen on vaginal smooth muscle function. The goal of this study was to determine the effect of estrogen on molecular and functional characteristics of the vaginal muscularis using a rodent model of surgical menopause. 3–4 month old Sprague Dawley rats underwent sham laparotomy (Sham) or ovariectomy (Ovx). Two weeks following surgery, animals received a subcutaneous osmotic pump containing vehicle (Sham, Ovx) or 17‐ β estradiol (Ovx). Animals were euthanized one week later and the proximal vagina was collected for molecular and in vitro contractility studies. Measurements were analyzed using one way ANOVA followed by post hoc analysis. Ovariectomy decreased 1) mRNA transcript (P<0.05) and protein (P < 0.001) expression of carboxyl‐terminal myosin heavy chain isoform (MHC), SM1, 2) protein expression of h‐caldesmon (P < 0.01) and 3) peak force of vaginal strips in response to KCl (P < 0.001). Estradiol replacement reversed these changes. No differences were detected in the % vaginal muscularis, amino terminal MHC isoform expression and maximal velocity of shortening. Our results suggest that menopause is associated with changes in the vaginal muscularis which may contribute to the increased incidence of pelvic floor disorders with age.

Funding: AUA Research Scholar Award, Maureen Basha
BiP deficiency and ER stress in skeletal muscle of a mouse model of amyotrophic lateral sclerosis
Authors: Chen Dapeng , Mazala Davi A.G. , English Samuel A , Chin Eva R
Published: June 2021

Mutations in Cu/Zn superoxide dismutase (SOD1) is a cause of familial amyotrophic lateral sclerosis (ALS). Mutant SOD1 protein induces cell death in motor neurons and skeletal muscle, although the molecular mechanism of mutant SOD1‐induced cell death remains controversial. The purpose of this study was to determine whether the endoplasmic reticulum (ER) stress pathway and apoptosis are activated in skeletal muscle with ALS. Gastrocnemius muscle (GAS) was removed from 120–140d old wild‐type control (CON) and SOD1G93A transgenic (ALS) mice and analyzed for ER stress signaling proteins: protein kinase RNA‐activated like ER kinase (PERK), inositol‐requiring kinase 1‐alpha (IRE1α), protein disulfide isomerase (PDI), immunoglobulin binding protein (BiP) and ER stress‐specific induced apoptotic mediator C/EBP homologous protein (CHOP). PERK and IRE1α were up‐regulated 5.2 ± 0.9 and 4.9 ± 0.1‐fold (p<0.05) in ALS vs. CON GAS, respectively, while PDI was up‐regulated 2.3± 0.1‐fold (p<0.05) in ALS mice. Unexpectedly, BiP was found to be deficient in all GAS muscles examined for ALS mice. CHOP was dramatically up‐regulated 13.3 ± 1.7‐fold (p<0.05) in ALS vs. CON mice, in conjunction with caspase‐12 activation. This study shows that ER stress is induced in skeletal muscle of transgenic SOD1G93A mice. Further, findings of a deficiency in ER chaperone BiP, up‐regulation of CHOP and activation of caspase‐12 cleavage suggest that defects in the ER stress response induce apoptosis and may contribute to the muscle atrophy process in ALS.
Modulation of Vagus Nerve Stimulation via Ganglionic Plexus activation and Ventricular Ischaemia
Authors: Brack Kieran Eugene , Coote John Haven , Ng G. Andre
Published: June 2021

Discreet regions in the heart, called ganglionic plexuses (GPs) are a network of integration centres that process afferent and efferent nerve activity to modulate the heart. We aimed to investigate the role of the right atrial GP (RAGP) in controlling heart rate (HR).

Methods

The decentralised innervated Langendorff perfused rabbit heart model was used (n=6). 1) RAGP was stimulated using 0.1–0.4mg nicotine (0.1ml) in 2) the presence of 0.1μm Atropine (At) or 1.8μm Metoprolol (Met), 3) The right vagus nerve was stimulated (VNS, 7H10V) before and after RAGP stimulation and 4) VNS before and following coronary artery occlusion. Data are mean±SEM, repeated measures ANOVA or Students t‐test.

Results

i) RAGP stimulation elicited an initial bradycardia (145±9 to 109±3bpm, P<0.05) and secondary tachycardia (226±12bpm, P<0.001, n=5) ii) At abolished the bradycardia (139±8 to 138±10bpm, P=NS), but not the tachycardia (247±9bpm, P<0.001). Met did not affect the bradycardia (135±7 to 106±4, P<0.05) but abolished the tachycardia (128±6bpm, P=NS, N=5. iii) VNS‐bradycardia was augmented after RAGP stimulation (−81±8 vs 142±9bpm, P<0.01, n=3). iv) Coronary artery occlusion increased VNS‐bradycardia (−45±6 vs −68±8bpm, P<0.05, n‐4).

Conclusion

This is the first data representing GP modification in the rabbit showing that RAGP activation elicits a mixed autonomic HR response, via vagal and sympathetic activation that enhanced VNS‐bradycardia. In addition, novel data shows that acute ventricular ischemia augments the HR effect of cervical VNS confirming that the RAGP is important in controlling HR in this species.

Research support from the University of Leicester
Activation by Oligomerization of an Allosteric Sequence Specific Endonuclease
Authors: Horton Nancy C , Park Chad K , Stewart Andrew M , Shah Santosh , Talley Heather , Ma Xin , Wysocki Vicki , Piperakis Michael , Cosstick Richard , Jacovetty Erica
Published: June 2021

SgrAI is a type II restriction endonuclease that cleaves an unusually long recognition sequence and exhibits allosteric self‐modulation of DNA cleavage and sequence specificity. We have shown that SgrAI forms very large aggregates (High Molecular Weight Species, HMWS) in the presence of sufficient concentrations of SgrAI and primary site DNA, that are dependent on the concentration of the DNA bound SgrAI dimer. We propose that the oligomers of DNA bound SgrAI (ie. HMWS) are the activated form of the enzyme. The unusual biochemical activity of SgrAI may have evolved to sequester activated SgrAI endonuclease on invading phage DNA and away from genomic DNA. We present new data showing that an uncleavable (3′S modified) primary site DNA activates SgrAI, therefore providing conclusive evidence that the primary site need not be cleaved by SgrAI in order to induce the activated conformation. Finally, while we have solved several crystal structures of SgrAI bound to DNA, including one showing two DNA bound SgrAI dimers interacting in a domain swapped tetramer, the structure of the activated oligomeric state has remained elusive. We present here new structural information gleaned from electron microscopy as well as ion mobility mass spectrometry (IM‐MS). The new data show that SgrAI/DNA oligomers form regular, rod shaped structures, with the DNA bound SgrAI dimer as the basic building block. IM‐MS collision crosssections are consistent with the rod‐like morphology of the oligomer seen in the electron microscopy, and a model of the oligomer based on SgrAI/DNA crystal structures has been proposed.
The efficacy of tart cherry supplementation in the prevention of age‐related bone loss in C57BL/6 mice
Authors: Chongwatpol Pitipa , Rendina Elizabeth , Graef Jennifer L. , Clarke Stephen L. , Lucas Edralin A. , Smith Brenda J.
Published: June 2021

Osteoporosis continues to be a major health problem and new prevention strategies are needed. This study examined the efficacy of dietary supplementation with tart cherry on age‐related bone loss in female (5 mo) C57BL/6 mice (n=44) assigned to a baseline group or groups receiving 0, 1, 5, or 10% cherry for 90 d. Mice were weighed weekly and bone density and trabecular and cortical bone microarchitecture were assessed at the end of the study. No difference in body weight due to treatment was observed. The groups receiving the 5 and 10% cherry diet had higher (P<0.05) whole body and tibia bone density than the control and baseline groups. The age‐related decrease in trabecular bone volume of the distal femur metaphysis was prevented with cherry supplementation (5 and 10%). In these two treatment groups vertebral trabecular volume and cortical area and thickness of the femoral mid‐diaphysis was increased (P<0.05) compared to the control and baseline cohorts. These findings suggest that tart cherry not only prevented age‐related bone loss, but increased cortical and vertebral trabecular bone above that of baseline which is consistent with an anabolic response.

Grant Funding Source: Cherry Marketing Institute
Anthropometric differences and body composition among ethnic minorities in northern Vietnam
Authors: Nguyen P H , Katz B , Lowe A E , Nguyen H , Truong T , Pham H , Nguyen S , Reinhart G A , Harding K B , Neufeld L M , Martorell R , Ramakrishnan U
Published: June 2021

Little is known about ethnic differences in body size and composition among minority groups in Vietnam. We used data from an ongoing study (PRECONCEPT) of women of reproductive age (WRA) in Thái Nguyên province, Vietnam to 1) compare selected anthropometric measures and body composition indices across ethnic groups; and 2) evaluate cutoff points for mid‐upper‐arm (MUAC) and calf circumference (CC) for screening undernutrition. A battery of anthropometric measurements (weight, height, MUAC, CC, triceps and subscapular skinfold thickness) was obtained for 4918 WRA. Body mass index (BMI), mid upper arm muscle (MUAMA) and fat area were calculated. The prevalence of underweight (BMI < 18.5 kg/m) and overweight (BMI >; 23 kg/m) were 32% and 5%, respectively. Mean (± SD) MUAC for the largest ethnic group, the Kinh (24.3 ± 2.1) was significantly lower than both Tay (24.6 ± 2.2) and San Chi (24.8 ± 2.2). Similar differences were seen for BMI (Kinh: 19.4 ±1.9; Tay :19.6 ±1.9, and San Chi:19.9 ± 2.0). The differences in BMI and MUAMA remained significant after adjusting for age, socioeconomic status, energy intakes, education and food insecurity. ROC analysis showed high predictive strength of MUAC and calf circumference in screening underweight women. Further studies are needed to explain differences in body composition among ethnic minority subpopulations.

Funding: The Mathile Inst. and the Micronutrient Initiative
Low calcium intake is associated with increased blood pressure among Filipino women
Authors: Lee Nanette R , Bas Isabelita , Largado Fe , Borja Judith , Duazo Paulita , Adair Linda
Published: June 2021

The role of dietary calcium (Ca) as a determinant of blood pressure (BP) has not been firmly established, especially in developing countries like the Philippines where Ca intake is low and hypertension (Htn) prevalence is high. We examined how Ca intake relates to BP among Filipino women using data from 1,997 women (ages 35–68y) in the 2005 Cebu Longitudinal Health and Nutrition Survey. Dietary Ca intake was measured using two 24‐hr recalls (mean: 309 mg/d). Major dietary sources of Ca were fish (37%), creamers/milk/milk products (23%), and vegetables (11%). About 27% of women had Htn (BP>;=140/90 mmHg). Regression analyses adjusted for age, smoking, energy intake, and other confounders showed that low Ca intake (lowest quartile) is associated with higher BP and odds of having Htn. Women with low Ca intake had 3.8 mmHg (95% CI: 1.6–6.1) higher systolic BP and 46% higher odds of having Htn [OR 1.46 (1.11–1.92)]. For diastolic BP, the effect of low Ca intake increased as WC increased (p for interaction <0.1). For example, for women with 75cm WC, the effect of low Ca intake was 1.6 (.1–3.1) while for those with 95cm WC, the effect is 3.8 (1.5–6.0) mmHg. Although Ca intake has been associated with fat metabolism and deposition, the biological mechanism of this interaction is yet to be elucidated. Results suggest that increasing Ca intake may help decrease BP in this population and preventing high WC may enhance this benefit. R01TW008288
Past International Residence and Current Gut Microbiome
Authors: Frankenfeld Cara , Poudrier Jill K , Sikaroodi Masoumeh , Waters Nigel , Shoemaker Sarah , Gillevet Patrick M
Published: June 2021

Intestinal bacteria contribute to the overall host (human) metabolism, and the microbiome (profile of microorganisms that inhabit the human body) is a multidimensional system. Several factors have been identified in the development of the microbiome, and evidence suggests that exposures to environmental factors (including diet) at various time points in life may play a pivotal role in the development of the adult human microbiome. The objective of this study was to evaluate the current microbiome in adults in relation to their residential history. Thirty‐one adults completed an in‐person interview to ascertain, at all points in life, residence locations in which they lived at least three months. In addition, participants completed a 4‐day food record, and provide a fecal sample. The gut microbiome was characterized from the fecal samples using 454 pyrosequencing. RDP10 was used for taxonomic classification of bacteria. In individuals who had never lived outside of the United States, abundance of Bacteroides ranged from 7 to 68% and of Prevotella ranged from 0 to 49%. In individuals who had lived outside of the United States for at least three months at any time in their life, abundance of Bacteroides ranged from 0 to 17% and of Prevotella from 0 to 63%. These results suggest which bacteria have dominant abundance is related to residential history, particularly having lived outside of the United States.
Efficacy of an Evidence‐based Food Guide for Vegetarians and Vegans
Authors: Knurick Jessica R , Fladell Lauren , Johnston Carol S
Published: June 2021

The 2002 modified food guide pyramid for vegetarians was recently updated to incorporate scientific findings from the past decade. The new guide promotes novel food groups (green leafy vegetables, nuts/seeds, beans/quinoa, and soy) to assure nutrient adequacy. In addition, the guide recommends that vegetarians increase protein intake to 125% of the DRI and increase zinc intake to about 250% of the RDA to account for low bioavailability. Additional recommendations address nutritional adequacy for calcium, vitamin D, selenium, iron, vitamin B12, and the omega‐3 fatty acids, including maintaining the omega‐6/omega‐3 ratio between two and four. To examine the efficacy of these new guidelines, 5 vegetarian and 5 vegan menu plans were developed based upon the serving recommendations of the updated food groups. Nutrient recommendations were exceeded for protein (+20%), vitamin B12 (+60%), selenium (+210%), calcium (+40%), and omega‐3 (+230%). The omega‐6/omega‐3 ratio averaged 2.8. Inadequate intakes were noted for iron (−40% for females), zinc (− 60%), and vitamin D (−70%). Menus developed utilizing the new food groups provided for adequate nutrient intakes, with the exception of iron, zinc, and vitamin D. To meet these recommendations, an individual following this food guide would be encouraged to specifically select cereals fortified with these micronutrients.