bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of endometrial scarless healing using a postpartum mouse model
Authors: Yoshii Asuka , Kitahara Shuji , Ezaki Taichi
Published: June 2021

The repair of endometrium is interesting for the remarkable speed without any scars, the process of which differs from ones in general wound healing. We sought to characterize its healing mechanism using an experimental mouse model. In this model, ICR mice had a Caesarean operation on Day 18 (D18) of gestation. On D1 after operation, a massive leakage of intravenously injected lectin towards the luminal side of the uterus was seen, indicating a continuous bleeding from cut ends of injured vessels. The leakage stopped by D2 may due to the involution by myometrial constraction. The endometrial epithelium completely recovered by D5 without any cicatrization. Surprisingly a large amount of fibrin in the injured endometrium disappeared as arginase1‐positive macrophages increased in and around it. Furthermore, the immunostainings of HIF1a as a marker of factor induced by hypoxia and VEGF revealed that angiogenesis occured in the endometrium before the completion of endometrial repair. We suggest that the change of oxygen concentration due to the rapid myometrial contraction might be involved in triggering the endometrial scarless healing.
Beneficial effects of Inchinkoto, a herbal medicine, against the stress of major liver surgery.
Authors: Yokoyama Yukihiro , Mizutani Tetsushi , Kawai Kiyotaka , Miyake Takashi , Kokuryo Toshio , Nagino Masato
Published: June 2021

Objective

We sought to elucidate whether Inchinkoto (ICKT), a herbal medicine, has a beneficial effect on the outcome of major liver surgery through animal and human studies.

Methods

Wistar rats were divided into three groups: 70% hepatectomy (Hx), hepatic ischemia/reperfusion (IR;15 min) and subsequent 70% Hx with vehicle treatment (IRHxV), IR and Hx with ICKT treatment (IRHxK). Rats were sacrificed 1 h after Hx. In human study, the patients who were planned to undergo major hepatectomy were randomly assigned to the group with or without preoperative ICKT treatment (n=30 in each group). The liver samples were harvested at the laparotomy and after the completion of hepatectomy.

Results

In animal study, the elevation of serum transaminase and the upregulation of inflammatory cytokines in the remnant livers were all significantly attenuated by preoperative administration of ICKT. Moreover, gene expressions of antioxidant enzymes such as glutathione were also higher in the IRHxK group. In human study, the livers with ICKT pretreatment showed a significantly greater upregulation of Nrf2 in response to the surgical stress (i.e., IR and Hx) as compared to the livers without ICKT pretreatment.

Conclusions

Preoperative administration of ICKT may provide beneficial effects through attenuating inflammatory responses and oxidative stress in the liver following IR and subsequent Hx.
Long‐term atorvastatin treatment alters cardiac ultrastructure in healthy mice while preserving systolic cardiac function
Authors: Godoy Joseph C. , Schilling Jan M. , Schwarz Anna , Asfaw Elizabeth K. , Alvarez Erika A. , Dalton Nancy D. , Niesman Ingrid , Patel Hemal H. , Zemljic‐Harpf Alice E.
Published: June 2021

Statins lower low‐density lipoprotein cholesterol (LDL‐C) for prevention of cardiovascular disease. Since statins reduce mevalonate synthesis they also harbor non‐cholesterol dependent effects such as improvement of endothelial function and inhibition of inflammation. Statins are known to cause skeletal muscle side effects, but it is currently unclear if they also affect cardiac muscle. Control mice underwent either atorvastatin (Ator), pravastatin (Prava) (both 5mg/kg/day) or vehicle (Veh) treatment for up to 6.5 months. After two months, Ator and Prava reduced direct non‐fasting LDL‐C compared to Veh. Echocardiography, in two‐week intervals, showed that fractional shortening and wall thicknesses were similar in Ator, Prava, or Veh treated groups. TEM analysis after 6.5 months of treatment showed that only Ator, but not Prava or Veh treated hearts (n=4 each) presented; 1) calcium phosphate granules, either in mitochondria or close proximity, 2) scattered, misorganized, and swollen mitochondria, 3) protein aggregates as well as inclusion bodies, and 4) enlarged, mislocalized T‐tubular structures.

This is the first report showing ultrastructural changes in cardiac muscle after long‐term Ator treatment.
Acute cardiac gene expression changes mediated through beta‐AR‐mediated transactivation of EGFR in vivo
Authors: Carter Rhonda L , Talarico Jennfer A , Grisanti Laurel A , Yu Justine , Tilley Douglas G
Published: June 2021

BetaAR‐mediated transactivation of epidermal growth factor receptor (EGFR) has been shown to promote cardioprotection in a mouse model of heart failure through unknown mechanisms. We hypothesized that this process could impact survival via regulation of gene expression. Thus, acute changes in cardiac gene expression were assessed via RNASeq of C57BL/6 mouse hearts given i.p. injections of isoproterenol in the presence or absence of the EGFR antagonist gefitinib for 1 hour. Total RNA from 4 hearts per treatment group were combined and underwent DNA library generation and SOLiD sequencing analysis, revealing a substantial number of genes regulated by each treatment. Gef alone significantly altered the expression of almost 200 genes compared to control suggesting potential Gef‐dependent alterations in the heart during clinical use. ISO alone and Gef/ISO significantly altered 310 and 481 distinct genes compared to control, respectively. Further statistical analysis was performed, confirming 139 genes significantly altered between the ISO and Gef/ISO groups (ISO‐induced gene up/downregulation antagonized/promoted by Gef), including several genes known to be involved in the regulation of cell death and survival. Thus, BetaAR‐mediated EGFR transactivation induces rapid modulation of cardiac gene expression in response to catecholamine stimulation in vivo.
Glucagon‐like peptide‐1 induced cardiac protection is dependent on caveolin‐3 expression
Authors: Hamaguchi Eisuke , Oshita Shuzo , Tanaka Katsuya , Tsutsumi Rie , Tsutsumi Yasuo M
Published: June 2021

Glucagon‐like peptide‐1 (GLP‐1) is an incretin hormone secreted by the enteroendocrine cells of the intestine in response to the presence of nutrients. A recent study showed that GLP‐1 directly protects against myocardial infarction via inhibition of apoptosis or the activation of survival factors. Furthermore, caveolin‐3 (Cav‐3) is necessary in cardiac preconditioning. In this study, we focused the effects of Cav‐3 on GLP‐1 induced cardiac preconditioning. Mice were exposed to 30 min ischemia followed by 120 min of reperfusion (control). Some mice were assigned to Exendin‐4, GLP‐1 receptor agonist or GLP‐1 receptor inhibitor, exendin. Exendin‐4 preconditioned significantly reduced infarct size compared to control group (24.7 ± 8.9% [n = 6] vs. 41.6 ± 4.6% [n = 8] respectively, p<0.05) and exendin eliminated the protection produced by exendin‐4 (38.8 ± 7.2% [n = 7]). Additionally, exendin‐4 induced cardiac protection was not observed in Cav‐3 knockout mice (43.2 ± 6.9% [n = 7]). These results reveal that GLP‐1 produces cardiac protection in mice in association with caveolin‐3 expression.
Genetic deletion of the TRPC3 channel blunts the development of angiotensin II‐induced hypertension in mice
Authors: Pathan Asif Rahimkhan , Fields Brandi , Birnbaumer Lutz , Zheng Fang , Rusch Nancy J
Published: June 2021

The Transient Receptor Potential Channel 3 (TRPC3) is a cation – conducting channel in the plasma membrane of vascular smooth muscle cells, which is activated by G‐protein coupled receptors (GPCR) including the α1‐adrenergic receptor. Several reports indicate that TRPC3 are upregulated in arteries of hypertensive animals and humans, but the lack of a selective TRPC3 blocker has thwarted efforts to define its role in blood pressure elevation. Here, we used TRPC3−/− mice to define the contribution of TRPC3 to the pathogenesis of hypertension. TRPC3 mRNA level was similar between mesenteric arteries (MA) of Ang II‐ and saline‐infused WT mice, but TRPC3 protein expression was ~5.7‐fold higher in MA of hypertensive mice. Resting mean arterial pressure (MAP) was not different between TRPC3−/− and WT mice (119±3 vs. 125±3 mm Hg; n=8–10). However, Ang II infusion for 14 days increased MAP in WT mice to 170±3 mm Hg compared to only 146±4 mm Hg in TRPC3−/− mice. PE‐induced constrictions were blunted in pressurized MA of Ang II‐infused TRPC3−/− (53±2%) compared to WT mice (71±5%), and PE‐evoked cation current was lower in patch‐clamped VSMCs of Ang II‐infused TRPC3−/− mice (‐2.5±0.5 pA/pF) compared to WT animals (−5.4±0.6 pA/pF). Thus, TRPC3 appears to importantly contribute to Ang II‐induced hypertension possibly by mediating GPCR cation current in VSMCs. NIH R01 HL064806 (NJR), AHA 12PRE11850002 (ARP) and NIH Z01‐ES‐101684 (LB)
Novel and potent GPR35 agonists aid identification of residues important in ligand interaction
Authors: Mackenzie Amanda Elaine , Neetoo‐Isseljee Zaynab , Southern Craig , Jerman Jeff , McIver Edward G. , Hudson Brian D. , Taylor Debra L. , Milligan Graeme
Published: June 2021

The orphan G protein coupled receptor GPR35 has emerged as a therapeutic target in a number of disease states including metabolic disorder, cardiovascular disease, inflammation and nociception. Efforts to characterize this receptor, however, have been complicated by marked differences in the pharmacology of GPR35 species orthologs. We have identified novel agonist compounds with which to dissect the pharmacology and function of GPR35. Selected compounds were chosen to illustrate the differences in ligand structure activity relationship between human, mouse and rat orthologs of GPR35. Sequence alignment and receptor homology modeling techniques guided site directed mutagenesis, which highlighted a number of residues involved in GPR35 ligand interaction and revealed differences between rodent and human orthologs. These findings will benefit future drug development programs and efforts to generate therapeutically relevant GPR35‐specific pharmacological compounds. This work was funded by a BBSRC CASE studentship with MRC Technology
The analgesic and anticonvulsant effects of Piperine in mice
Authors: Alhumayyd Mohammad Saad , Bukhari Ishfaq Ali , Mahesar Abdul Latif , Gilani Anwar H
Published: June 2021

This study was carried out to investigate the analgesic and anticonvulsant effects of piperine and its possible mechanism of action in mice. The analgesic effect was assessed in acetic acid induced writhing and tail flick tests, while the anticonvulsant effect was studied in pentylenetetrazole (PTZ)‐ and picrotoxin (PIC)‐induced seizure models. The administration of piperine (30, 50 and 70 mg/kg; i.p.) exhibited a significant inhibition (P<0.01) of the acetic acid‐induced writhing. In the tail flick assay, piperine (30 and 50 mg/kg; i.p.) and morphine (5 mg/kg; i.p.) caused a significant (P<0.01) increase in the reaction time of mice. Pre‐treatment of animals with naloxone (5 mg/kg; i.p.), reversed the analgesic effect of both piperine and morphine in the tail flick assay, suggesting the involvement of opioid pathway in the observed analgesic effect of piperine.

On the other hand, piperine (30, 50 and 70 mg/kg; i.p.), significantly (P<0.01) delayed the onset of PTZ and PIC‐induced seizures in mice. Similarly valproic acid and diazepam, used as standard anticonvulsants, suppressed the PTZ and PIC‐induced seizures in mice, suggesting that piperine may produce its anticonvulsant effect by enhancing GABA‐ergic neurotransmission in the brain.

The results of this study show that piperine possesses analgesic and anticonvulsant properties in mice. Additionally, these effects of piperine may be partly mediated via opioid and GABA‐ergic pathways, respectively. Moreover, these finding may substantiate the folkloric uses of black pepper in the management of pain and epilepsy.
The role of social media in recruiting for clinical trials in pregnancy
Authors: Shere Mahvash , Koren Gideon
Published: June 2021

Objective

Recruitment of pregnant or planning women to clinical studies using traditional advertising is difficult and slow. Given the widespread use of the internet as a source for medical information and research, we analyze the impact of social media as the primary recruitment tool in an ongoing randomized, open‐label clinical trial among pregnant women.

Methods

Recruitment through traditional sources, such as referrals from medical establishments, between April 2007–November 2011 was compared to supplementary online social media recruitment approaches used between December 2011–May 2012. Yearly recruitment and recruitment rates in the two arms were compared using the Mann Whitney U test.

Results

Between 2007–2011, with over 56 months of recruitment using traditional sources, 35 women were enrolled in the study, resulting in a mean rate of ±0.62 recruits per month. In the 6 months implementing recruitment primarily through social media‐based strategies, 45 women were recruited, for a 12‐fold higher rate of ±7.5 recruits per month (p<0.0001). Attrition rates as a function of total recruitment remained constant, suggesting that social media mainly had a positive impact on recruitment.

Conclusions

Clinicians and scientists recruiting for clinical studies should learn how to use online social media platforms to improve recruitment rates, thus increasing efficiency and cost‐effectiveness.
Maternal fat‐rich diet alters vasodilatation response in adult offspring
Authors: Clough Geraldine , Musa Moji , Torrens Christopher , Reynolds Richard , Fraser Paul
Published: June 2021

We have tested the idea that maternal over‐nutrition during pregnancy alters vasodilator signaling pathways in the adult offspring. Female mice were fed either a high fat (HF, 45%) or chow (C) diet for 6 weeks pre‐conception, throughout pregnancy and suckling. After weaning, male offspring were fed the same diet as their dams or switched to the alternative diet to give four dietary groups (CC, HFC, CHF and HFHF), studied at 15 weeks of age. Relaxation to ACh applied topically to cremaster arterioles was attenuated in the CHF and HFC offspring compared to CC. L‐NAME and indomethacin reduced the relaxation in CC offspring by 29% and CHF by 74%, while the HFC and HFHF were unaffected. The specific inhibitors of the SK(Ca) and IK(Ca) channels, apamin and TRAM‐34, abolished the response in the HFC and HFHF groups as well as the residual responses in the CC and CHF groups. These data suggest that the developmental environment results in a shift in ACh‐mediated relaxation from a NO pathway to one that requires opening of Ca‐activated K channels.

Supported by the BHF
Contractile behavior of the uterine lymphatic vessels
Authors: Gasheva Olga , Uddin Mohammad Nasir , Horvat Darjana , Kuehl Thomas , Zawieja David
Published: June 2021

While various lymphatic vessels networks are shown to adapt their contractility to changes in wall stretch and wall shear stress, until now, little is known on physiological characteristics of uterine lymphatic vessels (LV) in both the non‐pregnant and pregnant uterus. In this study we for the first time performed detailed evaluation of the contractile characteristics of the collecting uterine LV from ~250 g female non‐pregnant Sprague‐Dawley rats. Changes in lymphatic diameter were recorded with their subsequent analysis and calculation of the lymphatic pump indices of isolated, cannulated and pressurized rat uterine LV at different ranges of pressure and imposed flow in control conditions and after L‐NAME application. We found, that pressure stimulates contractility of rat uterine LV with contraction amplitude and pumping maximum at 1 cm H2O, while contraction frequency is almost stretch‐insensitive. NO had only a minor involvement in this regulation. These vessels also demonstrated a minor imposed‐flow/ wall shear stress‐dependent inhibition of tone, contraction amplitude and pump flow, but not contraction frequency. L‐NAME did not eliminate this flow/shear dependency, but induced flow‐independent shifts of all parameters of contractility of these vessels. In summary, our preliminary data show that uterine LV possesses unique contractile patterns different from other regional LV. NIH RO1 HL‐070308
Redox‐Dependent Hepatocyte TNFα Secretion Following Reoxygenation Injury
Authors: Spencer Netanya Y. , Zhou Weihong , Li Qiang , Zhang Yulong , Abbott Duane , Banfi Botond , Engelhardt John F.
Published: June 2021

Cell line studies have previously shown that hypoxia/reoxygenation (H/R) leads to the production of NADPH Oxidase 1 and 2 (NOX1 and NOX2)‐dependent reactive oxygen species (ROS) required for the activation of c‐Src and NFκB. We now extend these studies into mouse models to evaluate the contribution of hepatocytes to the NOX‐ and c‐Src‐dependent TNFα production that follows H/R and liver ischemia/reperfusion (I/R). In vitro, c‐Src‐knockout (KO) primary hepatocytes produced less ROS and TNFα following H/R than controls. In vivo, c‐Src KO mice also had impaired TNFα and NFκB responses following partial liver I/R. We found that NOX1 and p47phox are partially required for H/R‐mediated TNFα production by primary hepatocytes. In vivo experiments in KO mice deficient for NOX1, NOX2, p47phox, Rac1, and/or Rac2 showed that compared to controls, all of these KO mice, except NOX1 KO, had significantly decreased serum TNFα after I/R injury. These results demonstrate that NOX2 and its activator subunits p47phox and Rac control liver secretion of TNFα following I/R. Interestingly, NOX1 played a major role in secretion of TNFα by primary hepatocytes following in vitro H/R, but not following in vivo I/R. Thus, NOX2 can compensate for loss of NOX1 in vivo. We conclude that c‐Src and NADPH oxidase components are necessary for redox‐mediated production of TNFα following liver I/R and that hepatocytes play an important role in this process.
Validity, intra‐ and inter‐test reliability of arterial stiffness and wave reflection measured by the new brachial cuff SphygmoCor Xcel
Authors: Hwang Moon‐Hyon , Yoo Jeung‐Ki , Kim Han‐Kyul , Hwang Chueh‐Lung , Mackay Karen , Hemstreet Olivia , Vasconez Estefania , Jacobs Kimberley , Nichols Wilmer W , Christou Demetra D
Published: June 2021

Large artery stiffness (AS) and wave reflection are independent predictors of cardiovascular disease. Aortic pulse wave velocity (AoPWV) and augmentation index (AIx) are commonly used as non‐invasive measures of AS and wave reflection, respectively. We sought to examine 1) the validity of the new SphygmoCor Xcel (Xcel; AtCor Medical) compared with the SphygmoCor MM3 (MM3) model in measuring AoPWV and AIx and 2) the intra‐ and inter‐test reliability of Xcel. Twenty‐six adults participated in this study (16 men, 25 to 75 yrs). To assess the validity, AoPWV and AIx were measured using Xcel and MM3. To examine the intra‐test reliability, AoPWV and AIx were measured three times within the same testing session using Xcel. Finally, to assess the inter‐test reliability, AoPWV and AIx measures were repeated on Xcel within 1 week of the first testing session. AoPWV and AIx were not significantly different as measured by Xcel compared to MM3 (mean differences: −0.15±0.16 m/s, P=0.4 and −0.3±2%, P=0.9, respectively) and there were strong correlations between the Xcel and MM3 measures (AoPWV: r=0.84, P<0.001; AIx: r=0.77, P<0.001). Intra‐test (AoPWV: Intra‐class correlation (ICC)=0.996, P<0.0001;AIx: ICC=0.986, P<0.0001) and inter‐test reliability (AoPWV: ICC=0.979, P<0.0001; AIx: ICC=0.936, P<0.0001) were high. In conclusion, assessment of AS with Xcel is valid and highly reliable within and between testing sessions.
Both Kdr and Flt1 play a vital role in hypoxia‐induced Src‐PLD1‐PKCγ‐cPLA2 activation and retinal neovascularization
Authors: Singh Nikhlesh K , Hansen Dale E. , Kundumani‐Sridharan Venkatesh , Rao Gadiparthi N
Published: June 2021

Previously we have reported that Src‐PLD1‐PKCγ‐cPLA2 signaling plays a role in hypoxia induced retinal neovascularization. In the present study we explored the upstream mechanisms of Src‐PLD1‐PKCγ‐cPLA2 activation and retinal neovascularization. VEGFA, while having no effect on Flt1 phosphorylation, induced Kdr phosphorylation in human retinal microvascular endothelial cells (HRMVECs). Depletion of either Kdr or Flt1 attenuated VEGFA‐induced Src‐PLD1‐PKCγ‐cPLA2 activation, DNA synthesis, migration, and tube formation, albeit more robustly with Kdr downregulation. Hypoxia induced tyrosine phosphorylation of Kdr and Flt1 in mouse retina and depletion of Kdr or Flt1 blocked hypoxia‐induced Src‐PLD1‐PKCγ‐cPLA2 activation and retinal neovascularization. Hypoxia induced VEGFA and VEGFB expression in retina, and inhibition of their expression blocked hypoxia‐induced Kdr and Flt1 phosphorylation, respectively. Furthermore, depletion of VEGFA or VEGFB attenuated hypoxia‐induced Src‐PLD1‐PKCγ‐cPLA2 activation and retinal neovascularization. These findings suggest that both VEGFA and VEGFB mediate hypoxia‐induced Src‐PLD1‐PKCγ‐cPLA2 activation and retinal neovascularization via activation of Kdr and Flt1, respectively. Suported by EY014856, National Eye Institute/NIH
Estradiol and Tamoxifen produces acute and chronic neuroprotective effects after spinal cord injury
Authors: Colon Jennifer Marie , Mosquera Laurivette , Santiago Jose M , Torrado Aranza , Melendez Margarita , Segarra Annabell C. , Rodriguez‐Orengo Jose , Miranda Jorge D.
Published: June 2021

Estradiol (E2) is a multi‐active steroid that may offer neuroprotection via diverse mechanisms of action. Data on the neuroprotective role of E2 after spinal cord injury (SCI) is still controversial. We hypothesized that continuous E2 administration will provide beneficial recovery for injured rats at the behavioral, cellular and molecular level. Tamoxifen (TAM) treatment was evaluated in order to reduce possible detrimental effects from E2 administration. Female ovariectomized Sprague Dawley rats were surgically implanted with E2 implants or MPP‐dihydrochloride, an Estrogen Receptor α (ER‐α) antagonist, and then injured at the T10 level. Rats treated with E2 improved locomotor function in three different tests and MPP‐dihydrochloride treatment confirmed that the effects were ER‐α mediated. E2 treated rats also showed a reduction in reactive oxygen species (ROS), a reduced lesion cavity and an upregulated ER‐α. Rats treated with TAM had a reduced ROS levels and recovered some locomotor activity. Results suggest that E2 mediates neuroprotection in SCI by improving locomotor function, reducing the extent of the lesion and ROS while TAM administration suggest a safer, alternative treatment for SCI. Sponsored by the MBRS/RISE (R25GM061838), RCMI (G12RR003051 & G12MD007600), NIH/NINDS (NS39405), and MBRS‐SCORE (2S066M8224) programs.
Anti‐inflammatory action of angiotensin‐(1–7) and the angiotensin type 2 receptor agonist Compound 21 in hypothalamic microglia
Authors: Liu Meng , Shi Peng , Steckelings Ulrike Muscha , Sumners Colin
Published: June 2021

Recent studies indicate that the hypertensive action of angiotensin II (Ang II) mediated by its type 1 receptors in central cardiovascular control centers includes induction of the resident neuroimmune system, in particular microglia, in the paraventricular nucleus of the hypothalamus (PVN). Our objective here was to investigate the potential counteractive effects of the protective arms of the renin angiotensin system, angiotensin‐(1–7)/Mas and angiotensin type 2 receptors (AT2R), on activation of hypothalamic microglia. Real time PCR analyses revealed the presence of similar levels of Mas and AT2R in the PVN, as well as other cardiovascular control centers, of normotensive (SD; Sprague Dawley) and spontaneously hypertensive rats (SHR). Both Mas and AT2R were maintained in microglia prepared from the hypothalamus these rat strains, according to respective PCR and receptor binding analyses. Treatment of SD or SHR cultures with either Ang‐(1–7) (100–1000nM) or the AT2R agonist Compound 21 (C21; 100–1000nM) for 24 h elicited significant (~30%) decreases in basal levels of mRNAs for CD11b (marker of microglial activation) and the pro‐inflammatory cytokine interleukin‐1β. These data support the idea that both Mas and AT2R have an anti‐inflammatory role, and potentially an antihypertensive role, in the hypothalamus. Supported by HL‐076803
Electrical Restitution Dynamics during Manipulation of Slow Delayed Rectifier Potassium Current in Pig Ventricles
Authors: Jing Linyuan , Brownson Kathleen , Patwardhan Abhijit
Published: June 2021

Blockers and activators of the slow delayed rectifier potassium current (IKs) potentially can impact stability of conduction. However, effects of changes in IKs on restitution and memory in action potential duration (APD), critical factors in electrical stability, are unknown in the pig, which is a widely used model to study arrhythmia. To determine these effects, we recorded transmembrane potentials in right ventricular tissues from 7 farm pigs during reduction and increase of IKs with Chromanol (20 μM) and Mefenamic acid (100 μM), respecitively. Memory in restitution was quantified by using two protocols with sinusoidal changes in diastolic intervals (DI), which ranged between 100 – 700 ms and 10 – 290 ms. Hysteresis in restitution of APD was observed in both protocols; among 5 measures of the hysteresis loop (loop thickness, area, overall tilt, max/min delay), loop thickness and area significantly increased (decreased) after Chromanol 293B (Mefenamic acid), which suggests an increase and decrease of memory, respectively. Standard and dynamic restitution curves obtained from same tissues showed an increase (decrease) in average slope with application of Chromanol 293B (Mefenamic acid). As increase in slope and memory are hypothesized to have opposite effects on electrical stability, these results suggest that reduction and enhancement of IKs likely have offsetting components that effect stability.
Microparticles from PAH animals increase adhesion molecule expression in the pulmonary microcirculation
Authors: Hargett Leslie Anne , McLendon Jared , Gerthoffer William , Bauer Natalie
Published: June 2021

Microparticles (MPs) are submicron vesicles released by cells into the circulation. Once generated, MPs can activate cells downstream. MP counts are increased in many diseases, including pulmonary arterial hypertension (PAH). We investigated whether MPs from animals with PAH stimulate the expression of adhesion molecules in pulmonary endothelium. The Sugen/Hypoxia/Normoxia rat model was used in this study. This model develops vascular lesions which are characteristic of the human PAH pathology. MPs were isolated from the blood of control, Sugen only, and PAH animals. Pulmonary artery endothelial cells (PAECs) and pulmonary microvascular endothelial cells (MVECs) were treated with serum free media with or without MPs for 24 hours. Cells were then lysed. ICAM‐1 and E‐selectin expression was measured by western blotting. No changes were observed in ICAM‐1 expression in PAECs exposed to serum free media, control MPs, Sugen MPs, or PAH MPs. However, preliminary data suggests that both ICAM‐1 and E‐selectin expression are increased in MVECs exposed to MPs from PAH animals. E‐selectin appears to be increased in PAECs in response to control and PAH MPs. Our data suggests that PAECs and MVECs respond differently to MPs from PAH animals. These findings highlight the heterogeneity of endothelial cells in the pulmonary circulation and could indicate that MPs impact the homeostasis of the pulmonary microcirculation in PAH.

This research is funded by AHA 11 (SDG7390037) Grant and NHLBI T32 (HL076125) Training Grant.
Postprandial fatty acid trafficking in obese women after either endurance or resistance exercise
Authors: Davitt Patrick M , Arent Shawn M , Henderson Gregory C
Published: June 2021

We investigated effects of two exercise modalities on postprandial triglyceride (TG) and free fatty acid (FFA) metabolism. Sedentary, obese women were studied on 3 occasions in randomized order: Endurance exercise for 60 min at 65% VO2peak (E), ~60 min high‐intensity resistance exercise (R), and a sedentary control (C). A liquid mixed meal (48% CHO, 36% Fat, 16% Protein) containing [U‐13C]palmitate was consumed 30 min after exercise and subjects were studied over 7 hours. Content and isotopic enrichment of plasma TG and FFA were measured by mass spectrometry. Labeled fatty acids were inferred to be of exogenous (dietary) origin and unlabeled inferred to be endogenous. Total and endogenous plasma TG content were significantly reduced in both E and R compared to C (P < 0.05) with no effect of exercise on circulating exogenous TG content. Exogenous plasma FFA content was significantly increased following both E and R compared with C (P < 0.05), while total and endogenous FFA were elevated only in E (P < 0.05) compared with C. The results reflect integration of TG and FFA metabolism following E and R exercise. The present results indicate that R and E may be equally effective in reducing plasma TG level, and this reduction in postprandial lipemia is not achieved by enhanced clearance of dietary fat, but rather is achieved by reduced circulating abundance of endogenously produced TG.

Funding: Busch Biomedical Foundation
Endoperoxide 4 receptors attenuate the exercise pressor reflex in rats with ligated femoral arteries
Authors: Yamauchi Katsuya , Kim Joyce , Ruiz‐Velasco Victor , Kaufman Marc P
Published: June 2021

Ligating the femoral artery for 72 hours in in decerebrated rats exaggerates the exercise pressor response (EPR).The sensory arm of this reflex is comprised of group III and IV afferents, which can be either sensitized or stimulated by PGE2. Endoperoxide (EP) 3 and EP4 (PGE2 subtype) receptors were found to be responsible for the PGE2‐induced sensitization of rat dorsal root ganglion cells. We tested the hypothesis that L798106 (EP3 antagonist) or L161982 (EP4 antagonist) attenuated the exaggerated exercise pressor reflex in rats with ligated femoral arteries. We measured the cardiovascular responses to static hindlimb contraction or tendon stretch before and after iliac arterial injection of L798106 or L161982. The pressor and cardioaccelerator responses to either contraction or tendon stretch were not attenuated by L798106 in either the ligated or freely perfused rats. Likewise in five rats whose hindlimb muscles were freely perfused, the pressor and cardioaccelerator responses to either contraction or tendon stretch were not attenuated by L161982. In the five ligated rats, however, the pressor response to contraction was attenuated by L161982, averaging 37 ± 3 mmHg before, 18 ± 2mmHg afterward (p < 0.05). In addition, western blots revealed that ligation of the femoral artery for 72 hours increased the EP4 receptor protein in the L4 and L5 dorsal root ganglion by 24% (p < 0.05) over their freely perfused counterparts. We conclude that blockade of EP4 receptors, but not blockade of EP3 receptors, attenuated the EPR in rats with ligated femoral arteries.
Development and plasticity of innate immune function in altricial house sparrow nestlings
Authors: Killpack Tess L , Karasov William H
Published: June 2021

Ontogeny of immune function in altricial birds has been understudied. We examined development of the innate immune system in free‐living house sparrows and the flexibility of innate immune response to food restriction (FR) and refeeding in captive house sparrows. Ecological immunology techniques were used to examine components of the innate immune system. Natural antibody agglutination and complement‐mediated lysis increased with age, yet levels in near‐fledgling birds were about 50% lower than in hatch‐year birds and adults. Lysozyme levels declined with age, reflecting catabolism of maternal lysozyme transferred via the egg. In captive house sparrows, FR did not significantly impact lysis, a constitutive component of innate immunity, yet levels of the acute‐phase protein haptoglobin, an inducible component of innate immunity, were reduced in FR birds. Birds refed after early FR showed no reduction in haptoglobin response. These findings indicate that significant maturation of the immune system occurs post‐fledging in altricial house sparrows, and may challenge the notion that juveniles depend primarily on innate immune defenses during the growth period. Also, innate immune function appears to be flexible to resource supply during the nestling period, with no evidence of long‐term effects of early FR on development of the innate immune system. Support: NSF‐GRF, AOU, USDA‐Hatch, Birge Fund, GWIS.
Quantification of hypoxia‐induced blood flow through intrapulmonary arteriovenous anastomoses in healthy humans at rest
Authors: Duke Joseph W , Laurie Steven S , Elliott Jonathan E , Goodman Randall D , Fish Mathews B , Lovering Andrew T
Published: June 2021

Breathing hypoxic gas mixtures has been shown to increase blood flow through intrapulmonary arteriovenous anastomoses (QIPAVA) in healthy humans, as detected by saline contrast echocardiography. Although this method cannot quantify QIPAVA, Technetium‐99m‐labeled macroaggregated albumin (99mTc‐MAA) and quantitative whole body planar scanning has previously been used to quantify QIPAVA at rest and during exercise. We hypothesized that breathing 12% O2 would increase QIPAVA compared to breathing room air. We studied 4 male subjects while they breathed room air for 30 minutes on one day and 12% O2 for 30 minutes on another day in a randomized order, separated by >;6 days. In the final minute, 99mTc‐MAA, filtered through a 20μm filter (median size MAA = 35.3μm), was injected and subjects immediately underwent whole body quantitative planar scanning. A venous blood sample was drawn at the end of the scan to quantify the free 99mTc that became unbound during the scan. QIPAVA significantly increased from 0.1% at rest breathing room air to 1.5% (range: 1.1–1.9%) while breathing 12% O2. These data suggest that breathing 12% O2 increases QIPAVA in healthy humans. If QIPAVA is acting as a source of shunt, as we have previously suggested, then QIPAVA may play a role in the development of arterial hypoxemia that develops at altitude.
STEM CELL FUNCTION IN ATHEROSCLEROTIC APOE −/− MICE
Authors: Porto Marcella Leite , Campagnaro Bianca Prandi , Tonini Clarissa L , Vasquez Elisardo C , Meyrelles Silvana S
Published: June 2021

Background

Stem cell (SC) has been used as a potential therapy for various diseases. However, the beneficial effects of SC are still a subject of controversy. Therefore, the aim of this study was to evaluate how aging and atherosclerosis affect bone marrow SC function in mice.

Methods

Female mice (n= 12) were divided into C57 and apoE−/− (2 and 18‐month‐old) groups. Bone marrow stem cells were isolated and obtained by immunomagnetic depletion. BM‐MNCLin‐ were stained with phycoerythrin (PE)‐conjugated antibody against Sca‐1 (stem cell antigen) or matched isotype control (anti‐rat IgG2a). For apoptosis detection, 105 BM‐MNCLin‐ were incubated Annexin V‐FITC and propidium iodide (PI). A FACSCanto II cytometer was used for the flow cytometric analysis. Data were acquired and analyzed using BD FACSDiva software. Data are mean±SEM. Statistical analysis was performed with two way ANOVA test.

Results

Flow cytometric analysis showed increased SC apoptosis in aged compared with young C57 animals (2m: 4,8 ± 0,5% vs. 18m: 39,9 ± 0,8%, p<0,05). This effect was pronounced in apoE−/− group (2m: 38,6 ± 0,3% vs. 18m: 44 ± 0,2%, p<0,05).

Conclusion

The present study demonstrated that aging and atherosclerosis impact on SC function, mainly when these two risk factors are associated. These results provide novel rationales for the optimal use of SC in therapies.
Comparative anatomy and development of the carotid‐basilar arterial trunk in vertebrates
Authors: Rahmat Sulman , Gilland Edwin
Published: June 2021

In order to assess general patterns of vertebrate hindbrain vasculature, we dissected a wide taxonomic range of latex‐injected specimens. Examinations of distantly‐related taxa including cyclostomes, elasmobranchs, actinopterygians and sarcopterygian tetrapods demonstrated a conserved suite of hindbrain vascular features including: 1) the origin and branching of cerebral (internal) carotids; 2) the ontogenetic origins of the basilar artery from paired precursors; and 3) the presence of a quasi‐segmental series of transverse basilar artery branches that extend circumferentially around the brainstem, giving off penetrating vessels. The major exception to this general pattern is found in teleosts, in which a series of midline segmental central arterial stems arise from the basilar artery and branch into intramural vascular trees within the sub‐ependymal zone of the hindbrain. The restriction of penetrating arteries to a series of dominant medial segmental stems in teleosts is derived relative to other vertebrates and may be part of a set of key innovations in this group. Phylogenetic analysis suggests that paired basilar arteries are primitive for jawed vertebrates, with unpaired basilars arising independently in chondrichthyes, teleosts and tetrapods. The hypothesized independent development of unpaired basilar arteries may represent convergence towards similar neurovascular functional designs. Given the highly dissimilar body plans and lifestyles of these groups, the overall conservation of hindbrain macrovasculature likely results from the primitive, shared similarity of embryonic brain development.

Grant Funding Source: None
The Right Gastroepiploic Artery: Anatomical Study on 100 Cadaveric Dissections.
Authors: Settembre Nicla , Malikov Serguei , Champsaur Pierre , Bussani Rossana , Braun Marc , Magnan Pierre Edouard
Published: June 2021

The gastroepiploic artery (GEA) with the greater omentum is known for its high quality as a vascular graft and its resistance to infection when used to perform bypass, free‐flap or flow‐through flap.

Many authors have studied this anatomical unit, but a number of questions remain undiscussed.

The aim of this anatomical study was to evaluate the diameter and length of the GEA graft and its omental branches and to assess the feasibility of the epiploic flow‐through flap.

Methods

We performed 100 cadaveric dissections.

In 70 fresh cadavers we carried out dissections of GEA and of the greater omentum. Transillumination technique was helpful to identify all branches of GEA.

In the other 30 cadavers, prepared with Winkler solution, we injected barium sulfate with colored latex. After four days we harvested the artery with the omentum and a digital radiography was performed in order to report measures of lengths and internal diameters.

The diameter of GEA at its origin was 2.9 mm (2.7–3.4 mm). Its length was 245 mm (230–260 mm).

Conclusion

The anatomical unit based on GEA defines a relatively long arterial graft and a reliable omental flap for vascularization. Our dissection data confirm the anatomical feasibility in all cases and define the limits. This graft can be useful in patients requiring revascularization combined with coverage of tissue loss.
Morphological Plasticity in the Mouse Hallucal Metatarsal
Authors: Byron Craig Daniel , Herrel Anthony , Pauwels Elin , Patel Biren A
Published: June 2021

The objective of this study was to examine if climbing on thin substrates influences mouse hallucal metatarsal (Mt1) morphology. We focused on the Mt1 because mice raised in climbing habitats containing narrow branches frequently utilized secure hallucal grasps while controls did not. Forty male and female, skeletally mature CD‐1/ICR mice (≥4 months) were raised in experimental habitats from weaning (21 days) to adulthood. After euthanasia, Mt1s were isolated and high resolution μCT scans were obtained with a voxel size of 0.003865 mm. Several morphological parameters at bone midshaft were calculated using ImageJ software (BoneJ plugin). These variables included estimates for compressive strength (CA), bending strength (Z), and torsion strength (J), and all were scaled to their biomechanically appropriate scaling variable (bone length and/or body mass). We found that CA (P<0.05) and Zmax (P<0.1) were significantly greater in climbers. These mice had Mt1s that were more robust, with greater resistance to compression and bending moments. We conclude that Mt1 cross‐sectional geometry exhibits significant phenotypic plasticity in response to increased hallucal grasping when locomoting in a fine branch niche. Moreover, our results provide further evidence that post‐natal morphological development is influenced by substrate type and habitual locomotor behavior.
Dux4 target gene expression in mouse muscle transplanted with muscle cells from FSHD patients
Authors: Windelborn James , Rahimov Fedik , Emerson Charles P
Published: June 2021

In order to gain insights into FSHD molecular pathology, we developed a xenograft model by transplanting myogenic cells from patients with FSHD (4qA contractions) as well as from their unaffected relatives into the tibialis anterior muscles of immunodeficient mice. Our findings show that muscle xenografts derived from FSHD myogenic cells express Dux4 target genes, recapitulating the expression of these disease biomarkers in muscle biopsies of FSHD patients. FSHD muscle xenografts provide an animal model for investigations of the molecular pathogenesis of FSHD muscles and for drug development.

Grant Funding Source: NIH 5U54HD06084805 and FSH Society FSHS‐22012–04
Current Curricular Trends in the Anatomical Sciences: The 2012 Survey
Authors: McBride Jennifer Michelle , Drake Richard L
Published: June 2021

Since the American Association of Anatomists (AAA) published its last survey in 2009, related to the status of courses in Gross Anatomy, Microscopic Anatomy, Neuroscience and Embryology, significant curricular changes have occurred. In this session, data obtained in the 2012 AAA Education Course Survey will be discussed. Survey items queried course directors on several aspects of their course including but not limited to course format (integrated vs stand‐alone), course hours, target audience, textbook usage and assessment style. These data reflect current trends in the teaching of these courses in undergraduate medical education.
Gender Difference In Response to Dietary Genistein‐induced Chromatin Remodeling of Wnt Genes in Preneoplastic Rat Colon
Authors: Bray Julie K , Zhang Yukun , Chen Hong
Published: June 2021

Genistein, a soy‐derived isoflavone, has been reported to reduce the abnormal Wnt signaling that is observed in most colon cancer through epigenetic regulations. In this study, we investigated the effects of dietary genistein on gene expression of the Wnt pathway by histone modifications in the descending colon of rats. Using the azoxymethane‐induced rat colon cancer model, we have identified that dietary genistein reduced the formation of preneoplastic lesions in the colon of female rats more significantly compared to males. Expression of Wnt1, Wnt6, and Wnt8b was upregulated, while c‐Myc was downregulated by dietary genistein in female but not in male. By chromatin immunoprecipitation assay, we identified gender‐related differences in dietary genistein‐induced chromatin remodeling, which includes histone acetylation, methylation and phosphorylation at the above‐mentioned Wnt signaling gene regions. The differences were correlated with the differential response in gene expression to genistein between genders. In conclusion, our results suggest an epigenetic mechanism by which dietary genistein differentially affects the Wnt genes and Wnt signaling in male and female during colon cancer development. This study may facilitate establishment of a gender‐based recommendation on dietary genistein intake in maintaining colon health.

Grant support: Illinois Soybean Association and NIH/NCI
Cutting the cost of crouching: Over‐ground and tunnel locomotion in a tunnel specialist
Authors: Horner Angela , Moritz Sabine , Konow Nicolai
Published: June 2021

The physical constraints of tunnels present a considerable challenge to the animals that must navigate through them and still maintain the ability to efficiently travel over‐ground. Mustelids appear to be adapted for moving in the confined spaces of tunnels and burrows, as they possess elongate, flexible bodies with relatively short limbs. To investigate the relative performance of the mustelid bodyplan in tunnel environments, we studied various performance metrics of locomotion of the domestic ferret (Mustela putorius furo) in a height‐constrained chamber and compared to unrestricted over‐ground conditions. We synthesized these data with separate studies of gait analyses, whole body mechanics, histology, energetic, and electromyography to link tissue level functional morphology to whole‐animal performance. Despite a marked change in posture when ferrets enter tunnels, voluntary gaits and speed are unchanged, and oxygen consumption increases only slightly. Our analysis suggests that, in addition to the more obvious derived features of the musculoskeletal system, mustelids are also physiologically well suited for crouched locomotion.

Grant Funding Source: none
Characterization of PurF‐like proteins from Sulfolobus solfataricus
Authors: Belanger Maura Comeau , Zilinskas Egidijus , Sarisky Catherine A
Published: June 2021

The first step in purine biosynthesis converts PP1‐ribose‐P (PPRP) and glutamine into β‐P‐ribosylamine (PRA) and glutamate. The enzyme that catalyzes this reaction is called phosphoribosylpyrophosphate amidotransferase (GPATase or PurF). This enzyme has been extensively characterized in both prokaryotes (Bacillus subtilis) and in eukaryotes (human) but has never been characterized in an archaeon. In Sulfolobus solfataricus, a well characterized archaeon, the gene most likely to code for the PurF protein is SSO0632. The neighboring gene, SSO0633, while similar to other PurF‐like genes, does not code for the necessary active site cysteine. Both genes have been successfully cloned in pMAL‐c5E vector. Protein expression and experimental characterization of these proteins will be discussed. Funding for this project was received from a Cottrell College Science Award.
Chemical biology analysis of borrelidin identifies an essential translational enzyme as a pro‐angiogenic modulator of tumor progression in multiple human cancers
Authors: Francklyn Christopher S. , Lounsbury Karen , Williams Tamara , Mirando Adam
Published: June 2021

Phenotypic screening is a valuable tool in the discovery of new cancer therapeutics. Borrelidin, an 18‐carbon macrolide antibiotic produced by Streptomyces, inhibits the protein synthesis enzyme threonyl‐tRNA synthetase (TARS), while also blocking angiogenesis in cell culture and metastasis in a mouse model of melanoma. To explain the molecular basis of the effect of borrelidin and its less toxic derivative BC‐194 on angiogenesis, we examined the response of 80 different angiogenesis specific genes in SKOV cells to macrolide. Borrelidin and BC‐194 altered the expression of vascular endothelial growth factor (VEGF) and protein levels of HIF‐1, the hypoxia inducible factor that controls VEGF synthesis. TARS was revealed by the human vein endothelial cell (HUVEC) tube formation and chick chorionic allantoic membrane (CAM) assays to be a pro‐angiogenic factor with a potency rivaling VEGF and basic‐FGF. BC‐194 blocked these pro‐angiogenic functions. TNF‐alpha and VEGF stimulated TARS secretion from HUVECs, and TARS promoted angiogenesis by stimulating cell migration. The clinical significance of these observations is suggested by the involvement of TARS in selected human autoimmune diseases, and by its statistically significant expression increases in tissue samples from ovarian and prostate cancer patients. Supported by NIGMS and the Lake Champlain Regional Cancer Organization