bluuebunny/crossref_metadata_2025 huggingface dataset:
Eating an avocado a day does not change body composition or blood cholesterol in healthy students
Authors: Johnson Maxwell Bates , Burns‐Whitmore Bonny , Spalding Thomas , Sancho‐Madriz Martin F.
Published: June 2021

Cardiovascular disease is the leading cause of death in the U.S. Primary risk factors includes high serum cholesterol and high percent body fat composition. Diets high in monounsaturated fats (MUFA) have been shown to improve serum lipid profile without negatively altering body composition. Our study sought to determine if daily ingestion of avocados, a food high in MUFA, elicits the same response in healthy people. Free‐living, healthy male and female students (age 18–40) completed a parallel design; a four‐week avocado‐free (AF) run‐in followed by a random assignment to one of two eight‐week treatment periods: AF or avocado‐enriched (AE) treatment. Body composition and serum lipid analyses were measured at the beginning and end of each period. Neither treatment produced a change in total, LDL, HDL cholesterols, triacylglycerides, or body composition. The AE group resisted an increase in the non‐HDL:HDL ratio exhibited by the AF treatment. Though improvement in total serum lipid profile wasn't observed, results suggest that an avocado‐enriched diet might reduce cardiovascular disease risk in a healthy population. Additionally, a diet rich in avocado does not contribute to changes in body composition, suggesting that foods high in MUFA do not alter body composition in free‐living, healthy people.

Grant Funding Source: Agricultural Research Institute and College of Agriculture, Cal Poly Pomona
Glucagon‐like Peptide‐2 Therapy Improves Outcomes of Intestinal Adaptation and Decreases Relative Apoptosis in a Distal Intestinal Resection Neonatal Piglet Model of Short Bowel Syndrome
Authors: Suri Megha , Turner Justine M. , Nation Patrick N. , Wizzard Pamela , Brubaker Patricia L. , Sigalet David L. , Wales Paul W.
Published: June 2021

Glucagon‐like peptide‐2 (GLP‐2) is an intestinotrophic hormone produced in the ileum and colon that has been shown to augment intestinal adaptation in adult short bowel syndrome (SBS). Endogenous GLP‐2 levels and intestinal adaptation are reduced in distal intestinal resection (DIR) animal models of SBS that lack remnant ileum.

We hypothesize that exogenous GLP‐2 will improve clinical, morphological, and histological outcomes of intestinal adaptation in a DIR neonatal piglet model of SBS.

Twelve piglets underwent a 75% DIR with a jejunocolic anastomosis. Parenteral nutrition (PN) started on day 1 and was weaned as enteral nutrition (EN) was advanced. On day 2, piglets were randomized to IV GLP‐2 (11nmol/kg/day) or saline. Comparisons were made using the Student's t‐test, and p<0.05 was considered statistically significant.

GLP‐2‐ vs saline‐treated DIR piglets had fewer days of diarrhea (8.0±0.7 vs 12.3±0.4d), fewer days on PN (10.0±0.6 vs 13.8±0.2d), increased bowel length (19±4 vs −5±3%), and deeper jejunal crypts (248±21 vs 172±12μm). The fold increase in jejunal villus apoptosis was lower for DIR piglets vs surgical controls (1.1±0.1 vs 2.3±0.6 fold) in response to GLP‐2.

GLP‐2 therapy improves clinical, morphological, and histological outcomes of intestinal adaptation in a DIR neonatal piglet model of SBS, and reduces relative apoptosis. These results support a potential role for GLP‐2 therapy in pediatric SBS.
Increasing amylose content of starch stimulates fermentation and is bifidogenic in weaned pigs
Authors: Fouhse Janelle Margaret , Gänzle Michael , Regmi Prajwal , van Kempen Theo , Zijlstra Ruurd
Published: June 2021

Weaning causes digestive disorder in pigs primarily due to unstable gut microbiota. Increasing dietary amylose increases substrate available for microbial fermentation. Weaned pigs (n = 32) were allocated to 1 of 4 diets containing 67% starch with 0, 20, 28 or 63% amylose, respectively, for 21 days. In colonic digesta, Bacteroides increased (P < 0.05) with 63% amylose compared to 20 and 28%. Bifidobacteria spp. increased (P < 0.05) and Clostridium clusters IV and XIVa decreased (P < 0.01) with 63% amylose in cecal and colonic digesta. Lactobacillus decreased (P < 0.05) in cecal and colonic digesta with 28% amylose compared to 0, 20, and 63%. Total VFA concentration in cecal and colonic digesta increased (P < 0.01) with 63% amylose. Propionate concentration in cecal digesta increased (P < 0.01) with 63% amylose. Acetate and propionate concentration in colonic digesta increased (P < 0.01) with 63% amylose. pH decreased (P < 0.01) in ileal, cecal and colonic digesta with 63% amylose. Villus height and crypt depth were not affected by amylose content. Increasing amylose content can modulate gut microbial profiles, VFA concentrations, and pH that may play a protective role against pathogenic bacteria.

Grant Funding Source: Canadian Swine Research and Development Cluster, Danisco, Provimi
Selectins and integrins in ischemia‐reperfusion and sepsis induced murine acute kidney injury
Authors: Herter Jan Michael , Block Helena , Rossiant Jan , Stadtmann Anika , Zarbock Alexander
Published: June 2021

Ischemia‐reperfusion induced kidney injury has been the most frequently used model for acute kidney injury (AKI) in mice. However, sepsis is the leading cause of AKI in critical care patients in a clinical setting. We established a new sepsis induced kidney injury model using cecal ligation and compared it to the established model of ischemia‐reperfusion induced AKI. Furthermore we characterized both models using monoclonal antibodies against the most important selectins and integrins involved in neutrophil recruitment and evaluated leukocyte recruitment using intravital microscopy of the kidney cortex, FACS and histologic analysis of kidney tissue and assessed organ function by measuring serum creatinine. We here report that both models reveal similar characteristics concerning P‐ and E‐selectin as well as the β2‐integrins Mac‐1 and LFA‐1.

This study was supported by grants from the German Research Foundation (AZ 428/3–1, AZ 428/6–1, SFB 1009/A5 to A.Z and HE‐6810/1–1 to J.H.).
Endothelial apoptosis and glycocalyx morphology in plaque and non‐plaque areas of the mouse atherosclerotic brachiocephalic artery
Authors: Cancel Limary Melissa , Ebong Eno E , Tarbell John M
Published: June 2021

The vascular endothelial cell (EC) layer defends against atherosclerosis. Animal experiments have suggested that both increased EC apoptosis and shedding of the EC surface glycocalyx layer (EGL) play a role in elevated low density lipoprotein (LDL) permeability, a characteristic feature of atherosclerotic‐prone vascular regions. Recently, we confirmed in vitro that LDL permeability correlates strongly with measured apoptosis rates. We have also observed that a compound that enhances the EGL leads to dramatic reductions in LDL permeability in vitro. In the present study, we further elucidate the role of EC apoptosis and the EGL in atherosclerotic plaque formation. Atherosclerotic brachiocephalic (BCA) arteries were dissected from high fat fed Apolipoprotein‐E‐null mice. Fluorescent stained 6‐μm thick BCA cryosections revealed elevated EC apoptosis and degraded EGL on the plaque‐covered vascular wall, compared to non‐plaque areas. This is the first study to visualize the EGL morphology overlying an atherosclerotic lesion, and to compare EGL degradation and EC apoptosis in plaque development. Research support: NIH HL094889.
Interleukin‐17A plays an important role in endothelial cell activation and atherogenesis in ApoE−/− mice
Authors: Mai Jietang , Wang Hong , Yang Xiao‐Feng
Published: June 2021

Interleukin‐17 (IL‐17) cytokines are a family of pro‐inflammatory cytokines. IL‐17A has been implicated in numerous autoimmune and inflammatory diseases including atherosclerosis. However, the role of IL‐17A in early atherogenesis remained to be determined. The objective of our study is to determine the role of IL‐17A in early atherogenesis. Previously, using a bioinformatics approach, we found that IL‐17 receptors A and C, and adaptor protein, ACT1, are expressed in human vascular tissue, suggesting that IL‐17A has a role in chronic vascular inflammation. The data for the expression of these receptors and ACT1 were not available in mouse vascular tissue; however, using immunostaining, we found that IL‐17RA is expressed in mouse aorta. Moreover, plasma levels of IL‐17A are found to be increased in atherogenic ApoE−/− mice compared with wild type mice. IL‐17A treatment was found to induce pro‐inflammatory cytokines production in mouse aortic cells. Furthermore, to elucidate the role of IL‐17A in atherogenesis in ApoE−/− mice, we cross‐bred IL‐17A−/− mice with ApoE−/− mice to generate our double knock‐out mouse model, ApoE−/−IL‐17A−/− mice. Our data suggest that IL‐17A have an important role in early atherogenesis and with our novel mouse model we will be able to elucidate the function as well as the mechanism of IL‐17A in the endothelial cell activation and development of atherosclerosis in ApoE−/− mice.
Role of Caveolin‐1 S‐Nitrosylation, Ubiquitination, and Degradation in Idiopathic Pulmonary Arterial Hypertension
Authors: Bakhshi Farnaz R , Bonini M , Chen Z , NieuwAmerongen GN , Comhair S , Erzurum S , Minshall RD
Published: June 2021

Cav1−/− mice develop pulmonary arterial hypertension (PAH) due to eNOS hyperactivation since inhibition or deletion of eNOS abrogates the PAH phenotype. Here, we tested the hypothesis that oxidative stress upon inflammatory stimuli can induce Cav1 degradation & that this is associated with PAH. We observed TNFα‐induced NO production and S‐nitrosylation (SNO) at Cav1 C156 increased Src activity and destabilized Cav1 oligomeric chains in human pulmonary endothelial cells (PECs), and that this could be blocked by eNOS & Src inhibitors. Cav1 SNO at C156 was associated with reduced Cav1 expression following prolonged (72 hr) stimulation with TNFα or the NO donor, DetaNONOate. Mass spectrometry revealed Cav‐1 ubiquitination (Ub) on K86, supporting the hypothesis that Cav1 SNO at C156 induces Src‐dependent Cav1 Y14 phosphorylation, destabilization of Cav1 oligomers, & Ub of K86 leading to Cav1 degradation. In transduced HEK cells, we observed decreased Cav1 oligomerization with the Cav1 C156S‐mutant, preservation of oligomers with Y14F, and abrogation of Ub & degradation with a K86R mutant compared to WT‐Cav1. We also observed decreased oligomerized & total Cav1 expression as well as eNOS hyperactivation in PECs from 2 patients with IPAH compared to control PECs. Thus, we show oxidative stress‐induced Cav1 SNO, phosphorylation, ubiquitination, & degradation is associated with idiopathic lung vascular disease.
Withapysalin induced cardiac positive inotropism: in vitro and in vivo studies
Authors: Nascimento NIlberto R , Amorim Luciana Siqueira , Gomes Victor M , Santos Claudia F , Fonteles Manasses C
Published: June 2021

The cardiac effects of Withapysalin F, isolated from Acnistus arborescens were evaluated in vivo and in vitro. Guinea pigs were infused with Withapysalin F (25 and 50 μg/Kg/min) and them evaluated parameters such as maximum intraventricular pressure, heart rate, dP/dt maximum, dP/dT minimum and tau, comparing to control and isoproterenol (1 μg bolus). Withaphysalin F increased maximal intraventricular pressure and dP/dt, while there was no significant change in frequency, minimum dP/dT and tau. In vitro assays, concentration‐response curves (0.2 to 145 μM) were made in spontaneous beating right atrium as left stimulated electrically atrium. Withapysalin F increased force in the right atrium by 250 ± 50%, without chronotropic effect. Withapysalin F increased tension by 338.75 ± 21.55% in the left atria. Propranolol (1μM), did not inhibit the positive inotropism. No changes in cAMP levels were detected. Furthermore, pretreatment with H89 (10 μM), a PKA inhibitor, did not decrease Withaphysalin F positive inotropic effect in guinea pig left atrium. However, incubation of ruthenium red (70 μM) abolished the positive inotropism promoted by Withaphysalin F. Withaphysalin F inotropic activity appears to be dependent on the activity of RYR2 receptors regardless of increase of cAMP and activation of PKA. This project was approved by the ethics committee with the protocol number: 12235749–3.

Financial Support: FUNCAP
A novel epoxyeicosatrienoic acid analog attenuates hypertension and renal injury in Cyp2c44 KO mice
Authors: Khan Md Abdul Hye , Falck John R , Imig John D
Published: June 2021

Epoxyeicosatrienoic acids (EETs), synthesized by cytochrome P450 monooxygenases (Cyp), have been demonstrated to contribute to vascular tone, endothelial function and blood pressure regulation. Cyp2c44 is the primary enzyme responsible for renal EET generation. Reduced renal expression of Cyp2c44 results in reduced kidney EET levels in salt‐sensitive hypertension. In this study, we investigated anti‐hypertensive, kidney protective and anti‐inflammatory effects of a novel EET analog, EET‐A in Cyp2c44−/− mice receiving high‐salt (8% NaCl) diet for 4 weeks while treated with vehicle (0.05% ethanol and 0.1% PEG‐400) or EET‐A (10mg/kg/day) given in drinking water. Blood pressure was measured using radio‐telemetry throughout the experiment and urinary protein (UPro), albumin (UAlb) and monocyte chemoattractant protein–1 (MCP‐1) were measured on day 28 to assess kidney injury and inflammation. High‐salt diet elevated systolic blood pressure in Cyp2c44−/− (on day 28, 160±9 mmHg) compared to normal SV129 (110±11 mmHg) mice and it was attenuated by EET‐A (121±7 mmHg. EET‐A treatment also reduced UPro, UAlb and MCP‐1 excretion in Cyp2c44−/− mice on high‐salt diet compared to vehicle (UPro, 4±1 vs. 10±1 mg/day; UAlb, 37±10 vs. 112±23 μg/day; MCP‐1, 33±9 vs. 83±33 pg/day. In summary, Cyp2c44−/− mice on high‐salt diet develop hypertension, renal injury, and inflammation that are attenuated by the EET analog.
Functional Interaction between mGluR1 and 5
Authors: Sevastyanova Tatyana , Kammermeier Paul
Published: June 2021

Metabotropic glutamate receptors (mGluRs), class C G‐protein‐coupled receptors, exist and function as dimers. Until recently, mGluRs were thought to mainly form homodimers, but recent evidence is showing that some, including the group I mGluRs (1 and 5), can physically interact. To date, the nature of this interaction has not been addressed, nor have the pharmacological and functional consequences. These questions were addressed by expressing mGluR1 and/or 5 in isolated sympathetic neurons from the rat superior cervical ganglion (SCG), and using modulation of the native calcium currents as an assay for receptor function. Our data suggest that mGluR1 and 5 show a functional interaction we have termed ‘trans‐activation,’ in which ligand binding in one receptor induces G protein activation in the other. Among the data supporting this model is the finding that co‐application of mGluR1‐selective competitive and noncompetitive antagonists prevent signaling in cells expressing both mGluR1 and 5. In these cells, combining allosteric and orthosteric mGluR1 antagonists – BAY 36–7620 (“BAY”) and 3‐MATIDA, completely occluded signaling, while either drug alone only partially blocked the response. These data suggest that while mGluR5 is expressed, it requires mGluR1 (uninhibited by BAY) to signal. These data support the hypothesis that mGluR1 and 5 signal in a mutually dependent manner.
Pharmadynamic (PD) & pharmacokinetic (PK) properties of AF‐219: first in class, selective, clinical P2X3 antagonist in development for chronic pain and related conditions
Authors: Ford Anthony P , Smith Steven A , Dillon Michael P
Published: June 2021

P2X3 receptors are ATP‐gated channels with expression somewhat limited to primary afferent populations suspected of chronic sensitization in pain‐related disorders & offering therapeutic potential for antagonists (Ford, 2012, Purinergic Signal. 8:3–26). We describe preclinical PD & PK of AF‐219 (MW 353), a selective P2X3 antagonist optimized from an HTS lead.

AF‐219 blocked ATP activation of rat & human P2X3 & P2X2/3 trimers in calcium flux & voltage‐clamp assays: IC50 values 25–200 nM; unlike A‐317491 & TNP‐ATP, allosteric inhibition of ATP was displayed. In homogenate binding studies, AF‐219, but not ATP or TNP‐ATP, displaced specific binding of [3H]AF‐353. At 10 μM no activity was seen at other P2X channels & in a battery of receptors, enzymes & channels.

Favorable PK properties were observed for AF‐219 in rat, rabbit, dog & primate: high metabolic stability; oral availability (38–82 %F), elimination half‐life (t½ 1.5–7.6 h) & plasma free‐fraction (55–67% protein‐bound); predominant renal elimination (parent). AF‐219 was dose‐dependently efficacious (1–60 mg/kg, p.o. or s.c.; total plasma range 100–3000 ng/ml) in rodent models of hyperalgesia (neuropathic & inflammatory) or visceral hypersensitivity. The favorable PD, PK, & in vivo activity confirm AF‐219 as an excellent medicinal candidate to establish the role of P2X3 receptors in chronic pain and related conditions.
Delivery of chemotherapeutic paclitaxel by multifunctional nano‐carriers with dual targeting for enhanced anticancer activity
Authors: Dodwadkar Namita Sudhir , Biswas Swati , Torchilin Vladimir
Published: June 2021

Objective

In our previous studies, we demonstrated efficient mitochondrial targeting of nanocarriers by surface modification with triphenylphosphonium (TPP). Our current objective was to develop a dual functionality nanocarrier with transferrin (Tf) for tumor targeting and TPP for mitochondrial targeting.

Methods

Liposomes were prepared using Tf‐PEG‐PE and TPP‐PEG‐PE or PEG‐PE (8 mole %) for TPP‐PEG‐PE modified liposomes (TPP‐L and Dual‐L) or simply PEG‐PE liposomes (PL). Human cervical carcinoma (HeLa), human ovarian carcinoma (A2780) and normal mouse fibroblasts (NIH3T3) were used for cellular studies. Paclitaxel (PTX) was loaded into 150–200 nm liposomes for cytotoxicity analysis.

Results

Yellow color indicated significant co‐localization of rhodamine labeled TPP‐L and mitotracker green in mitochondria.

Conclusions

TPP‐L were mitochondriotropic. Dual‐L have higher cancer cell‐specific interactions that lead to higher anti‐tumor cell efficacy. This newly developed system suggests a promising treatment strategy for eradication of cancers.

*Supported by NIH grants RO1 CA121838 and RO1 CA128486 to V.P.T.
Role of K‐Channels in Microvascular Flow‐ Induced Dilation of Subcutaneous and Visceral Arterioles in Human Obesity
Authors: Grizelj Ivana , Cavka Ana , Bian Jing‐Tan , Drenjancevic Ines , Phillips Shane A
Published: June 2021

Altered vascular tone, a characteristic of human obesity, has been associated with impaired endothelium‐dependent vasodilation. Prior data suggest that visceral adipose tissue (VAT) is associated to endothelial dysfunction compared to subcutaneous adipose tissue (SAT). The aim of this study was to determine the contribution of vascular potassium channels to flow‐induced dilation (FID) in human VAT and SAT resistance arteries. Microvessels were obtained from VAT and SAT biopsies during bariatric surgery (OB; BMI>;48). Isolated vessels were cannulated for vascular reactivity to flow (pressure gradient 10–100 cmH2O) in the absence and presence of the 4‐aminopyridine (4‐AP; KV channel blocker), BaCl2 (KIR channel blocker), tetraethylammonium (TEA; BKCa channel blocker) and glybenclamide (KATP channel blocker). Dilations to papaverine (10–4M) were determined in each vessel. FID was reduced in VAT compared to SAT (at 60 cmH2O, 56±6% vs. 71±8%, p<0.05). There was no effect of TEA or glybenclamide on FID in SAT of OB subjects, but 4‐AP (p<0.05) and BaCl2 reduced FID in SAT of OB ( at 60 cmH2O and 100 cmH2O; p<0.05) vs. baseline. These data suggest that 1) FID is reduced in visceral compared to subcutaneous fat of obese patients and 2) Kir and Kv channels may be dysfunctional in VAT arterioles of obese subjects since 4‐AP and BaCl2 don't affect vasodilation of them while still reduce it in SAT arterioles.
SEX DIFFERENCE IN BLOOD PRESSURE REGULATION IN AGING SPONTANEOUSLY HYPERTENSIVE RATS
Authors: Maranon Rodrigo Oscar , Carmo Jussara Do , Reckelhoff Jane F
Published: June 2021

Leptin contributes to sympathetic activation via pro‐opiomelanocortin (POMC) neurons and thereby to increased blood pressure (BP). Whether melanacortin (MC3/4R) receptor contributes to hypertension (HT) in aging SHR is not clear. Our hypothesis was that MC3/4R activation contributes to HT in old SHR. Females and males (16–18 mos, n=6/grp) with MC3/4R antagonist SHU‐9119 (1 nmol/h icv for 7 days). MC3/4R antagonism increased the food intake by 56% in females and 71% in males (females: baseline: 17.1±2 g vs treatment: 26.6±1 g, p<0.001; males: baseline: 25±1.7 g, vs treatment: 43±2 g, p<0.001) and body weight by 18% in females and 23% in males (females: baseline: 217±11 g, vs treatment: 256±6 g, p<0.001; males: baseline: 356±27 g, vs treatment: 438±9 g, p<0.05). MC3/4R antagonism decreased BP (telemetry) and heart rate (HR) in males (BP: baseline: 172±5 vs treatment: 152±4 mmHg, p<0.05; HR: baseline: 321±4 vs treatment: 300±7 bpm, p<0.05), but had no effect in old females (MAP, baseline: 175±13 vs treatment: 172±10 mmHg, p=NS; HR: baseline: 345±9 vs Treatment: 332±9 bpm, p=NS). These results show that while MC3/4R activation contributes to the HT in old male SHR, sympathetic activation in old females is independent of the MC3/4R and suggests that sympathetic activation in aging women may have differential causes than in aging men. Supported by NIH PO1 HL51971, RO1 66072.
Early changes on proximal tubule Na+‐ATPase activity precede blood pressure elevation and renal dysfunction induced by intrauterine undernutrition: reprogramming by α‐tocopherol
Authors: Vieira‐Filho Leucio Duarte , Farias Juliane Silva , Cabral Edjair Vicente , Silva Paulo André , Paixão Ana Durce Oliveira , Vieyra Adalberto
Published: June 2021

We investigated whether intrauterine undernutrition that induces hypertension and renal dysfunction were preceded by early changes in proximal tubule ouabain‐resistant Na+‐ATPase activity, as well as, whether maternal α‐tocopherol treatment prevents these changes. Male Wistar rats obtained from undernourished dams during gestation presented increased Na+‐ATPase activity in proximal tubule membranes at age of 30 and 60 days. At 30 days of age, PKA activity was lowered, while at 60 days‐old PKC activity was elevated and AT2R expression was reduced. Systolic arterial pressure was increased from age of 105 days and lowered creatinine clearance, increased plasma volume, increased tubulointerstitial ANGII‐positive cells and increased renal p47‐ phox expression was seen at age of 150 days. Maternal α‐tocopherol treatment during lactation recovered these changes. These findings indicate a key role of Na+‐ATPase on blood pressure and renal function modulation, besides to support a non‐antioxidant effect of α‐tocopherol. CAPES, CNPq, FAPERJ, FACEPE.
Acute Hypovolemia Does Not Affect Dynamic Cerebral Autoregulation in Humans
Authors: Serrador Jorge M , Falvo Michael J , Maracaja Luiz , Blatt Melissa M , Caine Timothy L , Stenger Michael B , Platts Steven , Knapp Charles F , Evans Joyce M
Published: June 2021

While it is well known that cerebral perfusion pressure is the primary determinant of cerebral blood flow, it has been suggested that cardiac output and stroke volume may also play an important role. To test the role of stroke volume and cardiac output on dynamic cerebral autoregulation we performed sit to stand tests on a group of volunteers prior to and following administration of furosemide to reduce blood volume. These subjects were participating in a larger artificial gravity study. As expected furosemide resulted in a significant reduction in stroke volume while seated (63±5 vs. 51±4 mL, P=0.000). In contrast cardiac output was unchanged due to increases in heart rate (72±3 vs 94±3 bpm, P=0.000). Upon standing subjects demonstrated similar drops in SV, cardiac output and increases in HR from pre to post infusion. Blood pressure was unchanged by infusion while sitting and had similar drops upon standing (−20±4 vs −21±12 mmHg). Similarly cerebral flow velocity decreases were unchanged (−15±7 vs −17±8 mmHg). Examination of the Autoregulatory Index found no change from pre to post infusion (4.7±0.6 vs 4.8±0). These data demonstrate that hypovolemia and reduced stroke volume do not affect dynamic cerebral autoregulation or the cerebrovascular response to standing. Further work is needed to determine the role of cardiac output. Supported by NASA and VA.
Neural signature of cerebral activity of the fetal cholinergic anti‐inflammatory pathway derived from heart rate variability
Authors: Durosier Lucien Daniel , Xu Alex , Matushewski Brad , Cao Mingju , Herry Christophe , Batkin Izmail , Seely Andrew J.E. , Ross Michael G , Richardson Bryan S , Frasch Martin G
Published: June 2021

Fetal cholinergic anti‐inflammatory pathway (CAP) activity parallels changes in fetal heart rate variability (fHRV) in response to acidemia or inflammation. We hypothesized that 1) a multi‐domain fHRV measures approach will detect CAP activation early and 2) the degree of CAP activation will reflect the level of cerebral inflammation independent of the type of stimulus ‐ spontaneous hypoxia (n=5, arterial O2sat<50%), LPS (n=6) or control (n=9).

Chronically‐instrumented near term fetal sheep underwent increasingly intense umbilical cord occlusions until fetal pH<7.00 (~3–4 h). FHRV measures Skewness, Assymetry Index (AsymI), RMSSD, Sample Entropy (SampEn) and Fuzzy Entropy (FuzEn) were calculated continuously using the automated and standardized Continuous Individualized Multiorgan Variability Analysis, and correlated to microglia (MG) counts in specific brain regions. RMSSD and FuzEn increased and SampEn decreased early. Measures of fHRV symmetry did not change with worsening acidemia but correlated to MG counts, with CA1 MG counts correlating only to AsymI. SampEn correlated most often to MG counts in all regions except the DG, which correlated to FuzEn, Skewness and RMSSD. The spatiotemporal pattern of correlation to fHRV measures of different property domains provides a neural signature of cerebral CAP activity.

Funding: Women's Development Council, FRSQ, CIHR, MITACS/NeuroDevNet
Central neural mechanisms underpinning amplified respiratory‐sympathetic coupling in the spontaneously hypertensive rat.
Authors: MORAES DAVI J.A. , MACHADO BENEDITO H. , PATON JULIAN F.R.
Published: June 2021

Sympathetic over activity in the spontaneously hypertensive rat (SHR) is in part due to coupling with the central respiratory pattern generator. Using recordings of sympathetic and respiratory nerves with intracellular recordings of pre‐sympathetic and respiratory neurons in in situ preparation of SHR (4–6 week old), we evaluated whether sympathetic over activity is due to changes in the electrophysiological properties of pre‐sympathetic neurons and/or synaptic drive from respiratory neurons. In SHR, we found that respiratory modulation of the internal (iCSN), external (eCSN) and lumbar sympathetic outflows (lSN) was larger than in the Wistar rats. The iCSN also had an additional discharge in the late‐expiratory (late‐E) phase. Despite an increase in the firing frequency of RVLM pre‐sympathetic neurons during late‐E, there was no difference in their intrinsic electrophysiological properties compared to neurons in Wistar rats. SHR also had a greater late‐E discharge in hypoglossal and laryngeal motor nerves and a reduction in glottal resistance during the late‐E. These changes are due to increased firing frequency of pre‐BötC pre‐inspiratory neurons and not altered late‐E retrotrapezoid neurons activities. We conclude that the respiratory network of the SHR is reconfigured to a pattern dominated by pre‐I activity, in which provides additional excitatory drive to RVLM pre‐sympathetic neurons.
Lactational exposure to DEHP induced adrenocortical hypertrophy and anxiety‐like behavior in rats
Authors: Lin Ming‐Lu , Wang Dean‐Chuan , Chen Shih‐Chieh
Published: June 2021

Di‐(2‐ethylhexyl)‐phthalate (DEHP), a toxic agent leached from plastic products, may act as an endocrine disruptor affecting the development and function of reproductive system. However, there is no study revealing the effect of DEHP exposure on the function of adrenal gland. Stress coping is the main function of adrenal gland, therefore, the purpose of this study is to explore whether adrenocortical development and stress coping ability may be impaired by neonatal exposure to DEHP. Sprague‐Dawley female rats were orally fed with DEHP (10 mg/kg/day) or corn oil everyday during lactational period. After weaning (postnatal day 22), pups were divided into 2 groups: control and DEHP. At the age of 3 and 8 weeks, rats were exposed to the elevated plus maze for assessment of anxiety‐like behaviors. After behavioral assessment, rats were sacrificed and their adrenal glands were dissected for histological observation. The results from histological observation show that the thicknesses of zona fasciculata were increased significantly in DEHP group, indicating that neonatal DEHP exposure may induce adrenocortical hypertrophy. In addition, decreased open‐arm navigations in elevated plus maze in DEHP group indicates the expression of anxiety‐like behaviors. In summary, the results demonstrate that neonatal exposure to DEHP may disrupt the development of adrenal glands and stress coping ability in puberty rats.
Effects of Age and Exercise on Fiber Type and Total Fiber Number in Skeletal Muscle
Authors: Hoffman Marie , Akins Tiffany , Barton Gregory , McKiernan Susan , Aiken Judd , Diffee Gary
Published: June 2021

There have been varied reports of the effects of age and exercise on fiber type composition in skeletal muscle. To elucidate these discrepancies, we conducted a one year endurance exercise study in male FBN rats, starting with rats age 24 mo and continuing daily until age 36 mo. Rats were divided into 3 groups; high intensity (H), and moderate intensity (M) treadmill exercise, and sedentary (S). M and H animals trained at 13 m/min for 30 min/day, with the H group at 5% incline. At the end of the training protocol the plantaris muscle was removed, frozen and sectioned. Fiber type was determined by IHC using an anti‐type I MHC antibody (BA‐D5s). Total fiber number and number of type I v. type II fibers were determined from high resolution images of muscle cross sections. Total plantaris fiber number declined with age (decrease of 34% in 36 mo compared to 24 mo) but fiber number was not significantly different between the 36 mo exercise groups and the sedentary animals. The proportion of fibers that were type I fibers was 15% in 24 mo, and then 18%, 20% and 18% in S, M, and H respectively. None of these percentages were significantly different from one another, indicating no shift in fiber type with either aging or exercise. These results indicate that total fiber number decreases with age, but this fiber loss may not be preferential to specific fiber types, and also suggest that exercise is unable to minimize fiber loss.

Supported by: NIH AG030423
Occludin limits epithelial survival by inducing caspase‐3 expression
Authors: Shen Le , Lingaraju Amulya , Wang Yitang , Edelblum Karen L. , Khramtsova Galina F. , Turner Jerrold R.
Published: June 2021

The tight junction protein occludin regulates the paracellular barrier, but is also implicated in other epithelial functions. To determine if occludin regulates survival, responses of occludin knockdown (KD) Caco‐2 and enterocytes in occludin knockout (KO) mice to pro‐apoptotic stimuli was assessed. Occludin‐deficiency reduced extrinsic pathway apoptosis (TNF) and intrinsic pathway apoptosis (5FU, staurosporine, camptothecin), both in vitro and in vivo. Caspase‐8 and ‐9 were activated normally in both occludin‐deficient models, but caspase‐3 activation was reduced. Consistent with this, cytochrome C and dATP failed to efficiently activate caspase‐3 in cytosolic extracts of occludin‐deficient cells. Further, occludin‐deficient intestinal epithelia demonstrated decreased caspase‐3 promoter activity as well as mRNA and protein expression. Exogenous caspase‐3 expression reversed apoptotic resistance in vitro. Thus, occludin‐dependent caspase‐3 expression is required for intestinal epithelial apoptosis. The biologic relevance of this observation is emphasized by the resistance of occludin KO mice to DSS‐induced epithelial apoptosis, colonic damage, and systemic disease. Thus, disease‐associated occludin downregulation may limit epithelial apoptosis. While acutely beneficial, we speculate that this may allow damaged cells to survive and increase neoplastic risk in chronic disease.
Statistical analysis of Anatomy final examination grades of Hungarian, German and English speaking medical students at the Semmelweis University
Authors: Székely Andrea Dorottya , Attardi Stefanie M , Rogers Kem A
Published: June 2021

Semmelweis University teaches an identical medical curriculum to students in three different languages; Hungarian, English or German. In the Hungarian group, both the students and the lecturers are native speakers, while in the German group, the students are native speakers and the lecturers use German as a foreign language. In the English speaking international group, both the students and lecturers use English as foreign language. We have recorded the results of 3550 Anatomy final examinations held between 2002 and 2011 to determine if there are differences in performance among the groups as measured by the number of students who have passed the practical (dissection) part of the exam followed by the theory exam. Statistical analysis has shown evidence of a relationship between a student's language and their performance on the practical exam. A greater proportion of German students (84.3%) passed the practical exam and then took the theory exam compared to the English (73.8%) or Hungarian students (72.0%) (p < 0.0001, Chi‐square = 46.805, df = 2, Cramer's V = 0.118 ). However, there were no differences among the languages in relation to the grades the students achieved on the theory part of the examination (p = 0.389, Kruskal‐Wallis H = 1.886, df = 2), thus suggesting that the level of tuition was the same for all three groups.
Your voice sounds fine: Making podcasts to promote learning
Authors: Dewar Eric Walter
Published: June 2021

Science students have before them the task of learning both novel and enduring ideas, particularly in a courses about anatomy and physiology. I have implemented changes in my undergraduate anatomy and physiology course sequence, often with the use of podcasts. Podcasts are audio or video files that can be accessed by the user at a time of their choosing, either with a computer or a mobile device. I have tracked the effectiveness of the following uses of podcasts on my students’ learning: (1) stand‐alone lecture modules of primary course content; (2) interviews with scientists; (3) small study groups involving students that model effective learning; and (4) simple lecture recordings of traditional class meetings. These do require extra time on the part of the instructor and the student, but frees face‐to‐face time to focus on the more difficult content or discussion of case studies and the primary scientific literature. Use of this relatively simple and usable technology increases the overall student time spent on task for the course, generates reusable documentation for the instructor, and even frees us to consider what the purpose of the lecture is.
Movement guided learning© as a novel means of musculoskeletal anatomy instruction
Authors: Bentley Danielle Christine
Published: June 2021

Our bodies can be physically stretched, palpated, and manipulated. These movement opportunities can be used in anatomical instruction, essentially turning a student's body into their own educational tool. This research details the initial development and use of a novel teaching approach, Movement Guided Learning© (MGL). Following prior instruction on both the skeletal system and the muscular system, students participated in a tutorial‐styled learning activity that relied on the MGL teaching approach. The learning activity was exploratory, guiding students through physical movements/stretches, directing students through surface palpation/visualization, and challenging students to collectively brainstorm on the application concepts. Musculoskeletal anatomy knowledge was assessed before and after the MGL activity, with improvements on open‐ended concept questions (t(9) = −3.65, p=.005). The MGL activity was well received by students, with all students indicating they believe it should be used in future classes. MGL as a means of musculoskeletal anatomy instruction provides an opportunity for students to independently construct deeper, more integrated, concept understanding.

Grant Funding Source: none
The role of BCL11 transcription factors in mouse craniofacial suture development
Authors: Kyrylkova Kateryna , Iwaniec Urszula T. , Leid Mark
Published: June 2021

Craniofacial sutures are the fibrous joints that unite the bones of the skull and are the major sites of bone growth during craniofacial development. These joints normally close postnatally, allowing the brain and the skull to reach the proper size and shape. The premature fusion of one or more sutures leads to a condition called craniosynostosis, which represents a congenital defect affecting approximately one in 2000 live births. Craniosynostosis results in craniofacial morphologic abnormalities and increased intracranial pressure, which can lead to severe neurological and cognitive deficits in children. Surgery, with its inherent risks, remains the only therapeutic modality. Therefore, it is important to understand the molecular mechanisms underlying craniosynostosis in order to develop targeted biologically‐based treatment.

The transcription factors BCL11A and BCL11B play essential roles in the development of immune, central nervous, and cutaneous systems. We show that ablation of BCL11 transcription factors in mice leads to craniosynostosis, which results in severe midfacial hypoplasia and malocclusion. We conclude that BCL11A and BCL11B are essential for normal development of craniofacial bones and restrict premature suture fusion. The results suggest the possibility of modulation of BCL11 expression for the treatment of craniosynostosis.

Grant Funding Source: DE021879 from NIDCR to ML
Characterization of the avian Talpid2 mutant
Authors: Brugmann Samantha A , Chang Ching‐Fang , Schock Elizabeth N. , Snyder Jonathan , Delany Mary E
Published: June 2021

Talpids are naturally occurring avian mutants characterized by limb and craniofacial anomalies. Our objective was to explore the genetic, cellular and molecular etiology for the craniofacial phenotypes of the Talpid2 mutant. Using a 60kb SNP array and whole genome sequencing, we identified the causative genetic element for the Talpid2 mutation. We found that this locus was distinct from the causative locus of the Talpid3, the sister strain of Talpid2. Similar to Talpid3, the Talpid2 mutation also resulted in disrupted primary cilia, as indicated by glutamylated‐tubulin staining. Cellularly, we found aberrant levels of both proliferation and apoptosis within the facial prominences of Talpid2 mutants. Molecularly, we detected disruptions within the Hedgehog pathway of Talpid2 mutants. We found that post‐translational processing of both Gli2 and Gli3 was disrupted in the facial prominences of Talpid2 mutants. We detected a significant accumulation of full‐length (FL) Gli2 and Gli3 in Talpid2 mutants as compared to controls. Accumulation of Gli3FL was at the expense of the Gli3 repressor, however accumulation of Gli2FL was not at the expense of the Gli2 repressor. In sum, these results address the genetic, cellular and molecular etiology of the Talpid2 mutant and present a novel avian ciliopathic model that can be used to understand the role of primary cilia during development.
On the use of 3D anatomical software as a study aid in gross anatomy courses
Authors: Daniel Joseph C
Published: June 2021

A multiyear study investigated usage and efficacy of three‐dimensional (3D) anatomy software as a supplement to standard atlases. In year one, all first‐year medical students in the gross anatomy (GA) course at the University of Arkansas for Medical Sciences (UAMS) were offered use of the program “Interactive Functional Anatomy” (Primal Pictures). Due to limited rotational axes and perceived slowness of the software, few students used it and none extensively. In year two, students were given access to www.anatomy.tv (Primal Pictures), an online version with substantial performance improvement in line with student responses. Student interest and perception of the program greatly increased. No significant differences were seen in test scores, either in class or on the National Board of Medical Examiners standardized test by students using the website. Even so, more students are aware of and expect 3D anatomy software during their education (0% of GA students in 2010, 45% in 2013). Successful software should include tablet portability, fast response, easy search and labeling, 3D axes rotation, ability to hide or make transparent specific structures or groups, pruning, and quizzes. Funding by UAMS.
Incorporation of a Blended Learning Format in a Neuroanatomy Course
Authors: Franklin Samuel R.
Published: June 2021

This study describes how a blended learning format has been incorporated into the design of the Neuroanatomy for Physical Therapy Students course at the University of Kentucky. In the modern biomedical course a wide array of teaching tools and strategies are available for use including face‐to‐face lectures, video lectures, wet laboratories, text books, selected videos, web applications, kinetic learning, and team based learning exercises. A balance between face‐to‐face exercises and the use of technology for guided independent study must be achieved in order for all types of student learners to be as successful as possible with assimilating the learning objectives of the course. The type of student feedback and assessment for the course, as well as for each exercise, is an important part of the learning process and must be considered when implementing a blended learning curriculum. This study outlines faculty and student reflections about the successes, and failures, during the course and how they will effect the future direction of the neuroanatomy course.
Cyrtophora citricola as a potential component of an integrated pest management program for citrus in Puerto Rico
Authors: Olivera Stefany Yanira , Arroyo Nancy , Asencio Carmen
Published: June 2021

The trend in agriculture towards reducing pesticide use and fostering ecological sustainability has led to a growing interest in spiders as biological control agents of insect pests due to their ability as intensive predators. Previous studies show that spiders may decrease the pest densities and may level out the densities along the way. Cyrtophora citricola, a spider from the Araneidae family, known for its tent shaped dense web, enables the spider to capture a high number of small to medium sized arthropods, water bugs, flies, wasps, thrips, small moths, small butterflies and mosquitoes. C. citricola is considered a voracious feeder of many of these insect pests. Due to recent reports of C. citricola appearance in Puerto Rico in 2008 this work explores this organism's beneficial impact in agriculture without disregarding its deleterious effects. This study has focused primarily on citrus plants, known to be host plants for C. citricola, and analyze its biological control potential for damaging pests like Diaphorina citri. The latter is a well‐known vector of the severe citrus disease, Huanglongbing or greening disease. The first part of this study has focused on plants/pathogen/antagonists interactions. The availability of D. citri genome sequence and our molecular studies on C. citricola have shed light on these interactions. This understanding will provide an important contribution in the management of HLB on citrus plants.
Calcium triggers assembly of ESCRT proteins at the cell membrane
Authors: Scheffer Luana Lea , Sharma Nimisha , Medikayala Sushma , Brown Kristy , Jaiswal Jyoti K
Published: June 2021

Intracellular calcium increase regulates variety of cellular processes, including trafficking of molecules to the plasma membrane. However, the compartments and the mechanisms involved in this trafficking are not well understood in non‐secretory cells. We have recently developed a proteomic approach that allows monitoring transient and rapid changes in the cell membrane proteome. Using this approach we have examined calcium‐triggered changes in plasma membrane proteome of muscle cells. We find that calcium increase triggers cell surface translocation of several proteins in the endosomal pathway. Notable among them are the endosomal sorting complex required for transport (ESCRT) proteins. Western blot analysis validated these proteomic findings. Using total‐internal‐reflection fluorescence microscopy of live cells we visualized calcium‐triggered cell surface translocation of fluorescently‐tagged ESCRT proteins. This showed that calcium increase triggers movement of ESCRTs directly from the cytosol to the cell membrane resulting in assembly of the late stage ESCRTs at the cell membrane. The kinetics of calcium‐induced cell membrane assembly of these ESCRTs is similar to what has been reported for virus‐induced ESCRT assembly. Our studies reveal calcium as a novel regulator of the assembly of late stage ESCRTs at the cell membrane.
Dynamic changes of the Escherichia coli transcriptome and proteome exerted by glucose repression
Authors: Borirak Orawan , Bekker Martijn , Rolfe Matthew D , Dekker Henk L , de Jong Luitzen , de Koster Chris G , Hoefsloot Huup C J , de Koning Leo J , Hellingwerf Klaas J
Published: June 2021

A system biology approach, including multi‐omics analyses (i.e. time‐resolved monitoring of mRNA and protein levels) with an advanced culturing method, was applied in order to identify posttranscriptionally regulated genes in E. coli under Carbon catabolite repression control. Approximately 800 proteins were quantified using 15N‐metabolic labelling, in 3 parallel chemostat cultures. Of these, about 400 proteins were found to change significantly (p<0.05) in expression level, upon a glucose pulse, as compared to the reference conditions. Of almost half of the proteins that are significantly up‐ or down‐regulated, the levels of mRNA and protein are not correspondingly altered. These genes are classified as potential post‐transcriptionally regulated (PPTR) genes. Most of the PPTR genes are involved in transcription, nucleotide‐ and amino acid biosynthesis, membrane transporter expression, and central metabolism. Interestingly, only 30% of the PPTR genes has been reported to be regulated at the post‐transcriptional level by small regulatory RNAs or by a riboswitch, while the regulator(s) and/or the mechanism of post‐transcriptional regulation of the remaining PPTR genes (70%) is still unknown.
Pharmacological evaluation of a new series of benzofuran‐2‐ carboxamides as antihyperlipidemic agents in rats
Authors: Al‐Qirim Tariq Musbah , Shattat Ghassan , Sweidan Kamal , Sheikha Ghassan Abu
Published: June 2021

The aim of this study is to investigate the pharmacological activity of a new series of benzofuran‐2‐carboxamides (3a‐3d and 4a′‐4c′). Compounds (3a, 3b, and 4a′—4c′) were tested in vivo using Triton‐WR‐1339‐induced hyperlipidemic rats as an experimental model. The tested animals were divided into eight groups: control, hyperlipidemic, compounds 3a, 3b, 4a′, 4b′, 4c′, and beza□brate. At a dose of 15 mg/kg, the elevated plasma triglyceride (TG) levels were signi□cantly reduced in compounds 3b (p<0.0001) and 4c′ (p <0.05) after 12 and 24 h compared to the normal control group. Furthermore, high‐density lipoprotein‐cholesterol levels were remarkably increased in compounds 3b (p<0.001) and 4c′ (p <0.05). Meanwhile, compound 4b′ slightly reduced the TG levels after 12 and 24 h. The present study demonstrated new properties of the novel series benzofuran‐2‐carboxamides 3b and 4c′ as potent lipid‐lowering agents. It is, therefore, reasonable to assume that compounds 3b and 4c′ may have a promising potential in the treatment of hyperlipidemia and coronary heart diseases.

This research is financially supported by Al‐Zaytoonah University of Jordan