bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of substituted benzazepines as functionally selective ligands of the D1 dopamine receptor
Authors: Conroy Jennie Lynn , Doyle Trevor B , Sibley David R
Published: June 2021

The D1 dopamine receptor has been implicated in numerous neuropsychiatric disorders, and various D1‐selective ligands have shown potential as therapeutic agents. Functionally‐selective, or signaling biased compounds present a unique therapeutic opportunity to target individual pathways while minimizing signaling through others. A series of “atypical” substituted benzazepines, and D1 “typical” agonists, were tested for their functional effects on D1‐mediated cAMP accumulation, D1‐mediated β‐arrestin recruitment, and D1 receptor internalization using live cell functional assays. With respect to β‐arrestin recruitment, some “atypical” benzazepines were found have no agonist efficacy at this response, but instead functioned as antagonists. Other “typical” agonists exhibited a range of efficacies from 20–110%. As with β‐arrestin recruitment, the “atypical” benzazepine agonists exhibited little agonist efficacy in an assay of receptor internalization, yet other “typical” compounds were full agonists. Using two distinct cAMP accumulation assays, “typical” agonists were found to elicit increases in D1 cAMP accumulation similar to that produced by dopamine, while the “atypical” agonists were found to be partial‐full agonists. Taken together, these experiments identify a novel series of substituted benzazepines that are G protein‐biased or functionally‐selective agonists of the D1 receptor.
ageLOC Vitality improves memory and learning ability in memory impairment models
Authors: Tan Ning‐Zhi , Yang Jie‐Ying , Yang Guang , Zhu Jia‐Shi
Published: June 2021

Based on the differential gene expressions found in skeletal muscle and neocortex of Young (5‐mo) and Old (25‐mo) mice, ageLOC Vitality, containing Cordyceps sinensis Cs‐4 and extracts from ginseng and pomegranate, opposes the age‐associated changes in global gene expressions, and improves endurance exercise and sexual functions. We further examined its effect on memory functions at a dose of 200, 400 or 800mg/kg in mice by using both shuttle‐box and Morris water‐maze tests. Three models were used with impairment of memory acquisition (MAI, by scopolamine), memory consolidation (MCI, by sodium nitrite), and memory reproduction (MRI, by ethanol). In shuttle‐box avoidance test, Vitality shortened active avoidance latency (AAL) and increased active avoidance response rate (AARR) in (1) MAI: AAL −47 to −70% (p<0.001) and AARR +109 to +209% (p=0.007); (2) MCI: AAL +9.5 to −44% (p=0.024) and AARR +59 to +149% (p=0.002); and (3) MRI: AAL +10 to −52% (p=0.014) and AARR +17 to +203% (p<0.001). In water‐maze test, Vitality shortened the swim time to platform in MAI: −14 to −57% (p=0.008), and in MRI: −29 to −43% (p=0.019). Our results indicate ageLOC Vitality is effective in improving memory and learning ability in mice with memory impairments, supporting its potential anti‐aging effect.
Effects of M1 and M4 muscarinic acetylcholine receptor positive allosteric modulators on sleep and cognition in rodents
Authors: Gould Robert W , Nedelcovych Michael T , Grannan Michael D , Bubser Michael , Wood Michael R , Lindsley Craig W , Xiang Zixiu , Wess Jurgen , Conn P Jeffrey , Jones Carrie K
Published: June 2021

Clinically available antipsychotics primarily treat the positive symptoms in schizophrenia, but have little or no effect on the negative symptoms or disruptions in cognition or sleep. Muscarinic cholinergic receptor (mAChR) subtypes mediate aspects of learning, memory and sleep/wakefulness. Recently, the M1/M4‐preferring mAChR agonist xanomeline showed efficacy on the positive symptoms and some cognitive impairments in patients with schizophrenia. In the present studies, we evaluated potential cognitive enhancing and wake promoting effects of xanomeline in comparison with the M1 and M4 selective positive allosteric modulators (PAMs) BCQA and VU0152100, respectively, in rodent sleep and cognition models. Electroencephalography data showed that xanomeline increased wakefulness in rats, similar to acetylcholinesterase inhibitors. Ongoing studies are defining the role of M1 and M4 using BCQA and VU0152100. These ligands are also being evaluated in touch screen cognition assays using M1 and M4 KO mice. While M1 and M4 KO mice exhibited no differences on discrimination or reversal learning tasks compared to wildtypes, alterations in cholinergic activity may provide enhanced efficacy in these tasks. Changes in vigilance during wake and sleep efficiency using M1 and/or M4‐preferring mAChR ligands may provide a new approach for the treatment of sleep and cognitive disruptions in schizophrenia.
Metabolomics‐guided amino acid analysis reveals acetaminophen overdose‐induced disruption of arginine metabolism in mouse
Authors: WANG LEI , YAO DAN , CHEN CHI
Published: June 2021

Acetaminophen (APAP) overdose is a major cause of acute liver failure. Significant changes in lipids, glucose, and thiol antioxidants have been observed in APAP‐induced hepatotoxicity (AIH). However, the influences of AIH on amino acid metabolism are not well characterized. In this study, the profiles of free serum amino acids in the wild‐type (WT) and Cyp2e1‐null (KO) mice after APAP (400 mg/kg BW) treatment were defined through metabolomics‐guided biochemical analysis. Consistent to different sensitivity to AIH, serum amino acid levels differed greatly between WT and KO mice. Among observed changes in amino acids, APAP‐induced change in arginine was the most dramatic. After APAP treatment, serum arginine in WT mice became undetectable within 2 hours, while its level in KO mice was reduced at early time points but recovered quickly. This depletion of serum arginine in WT mice was well correlated to the increased arginase activity in serum, suggesting the release of arginase from the liver is the main cause of arginine depletion. Accompanying with rapid degradation of serum arginine, the distribution of arginine in WT mice as well as the levels of arginine metabolites was also dramatically affected. All these observations warrant further investigation on the role of arginine in AIH.
Compartmental distribution and antiviral effect of efavirenz metabolites
Authors: Avery Lindsay B , VanAusdall Jennifer L , Hendrix Craig W , Bumpus Namandje N
Published: June 2021

Efavirenz (EFV), one of the most widely prescribed drugs used to treat HIV, is extensively metabolized by the cytochromes P450 to a number of oxygenated products; however, the pharmacologic activity and distribution of these metabolites in anatomic compartments relevant to HIV infection beyond blood plasma have yet to be explored. The systemic distribution of EFV oxidative metabolites was examined in blood plasma (BP), seminal plasma (SP), and cerebrospinal fluid (CSF) from subjects on an EFV‐based regimen. The 8‐hydroxy EFV metabolite was detected in BP, SP and CSF, with median concentrations of 314.5ng/ml, 358.5ng/ml, and 3.37ng/ml, respectively. In contrast, 7‐hydroxy and 8,14‐hydroxy EFV were only detected in BP and SP with median concentrations of 8.84ng/ml and 10.23ng/ml, and 5.63ng/ml and 5.43ng/ml, respectively. Interestingly, protein‐free concentrations of metabolites were only detectable in SP, where a novel dihdyroxylated metabolite of EFV was also detected. The metabolite concentrations in 34 paired BP and CSF patient samples were not strongly correlated with viral RNA concentrations indicating that the metabolites of EFV are pharmacologically inactive. These results suggest there may be important compartmental differences in EFV metabolite distribution, and the male genital tract may be a novel compartment in understanding local drug distribution and efficacy of oxidative metabolites.
Identification of morphological and hemodynamic biomarkers for tumor vascular perfusion through mathematical modeling and high‐resolution imaging
Authors: Stamatelos Spyros K , Kim Eugene , Pathak Arvind P , Popel Aleksander S
Published: June 2021

Tumor angiogenesis is a hallmark of cancer, and involves the process of blood vessel formation from pre‐existing host and tumor vascular networks. Tumor vascularity and blood flow are chaotic and heterogeneous, making delivery of therapeutic agents to solid tumors inefficient. Image‐based mechanistic hemodynamic modeling can facilitate the understanding of tumor blood perfusion and the design of drug delivery and anti‐angiogenic therapy strategies.

The experimental protocol involves the development of breast cancer xenografts in mice. Ex‐vivo high resolution imaging is conducted using micro‐computed tomography, and 3‐D image reconstruction and segmentation leads to a “wide‐area” mapping of the vasculature. This process is highly anisotropic involving complex geometries, and it was necessary to develop an image‐bioinformatics algorithm to ensure connectivity among vessels. This methodology includes a 3‐D tracing algorithm which optimizes the geometry based on local cues. The mathematical formulation for blood flow is based on Poiseuille‐like law for pressure drop and nonlinear rheological equations for local apparent viscosity and hematocrit. Comparison of tumor regions across samples and time points, based on a number of morphological and hemodynamic measures, allows the identification of biomarkers for drug delivery.

Supported by NIH grant R01 CA138264 and Komen Foundation Grant KG090640.
Dietary salt intake enhances sympathetic responses to central sodium and sciatic afferent activation
Authors: Stocker Sean D
Published: June 2021

Previous studies indicate that increased dietary salt intake enhances sympathetic nerve activity (SNA) and arterial blood pressure (ABP) responses evoked from the rostral ventrolateral medulla. The present study sought to extend these observations and determine whether SNA and ABP responses to physiological stimuli are also enhanced. Male Sprague‐Dawley rats were fed 0.1% (low) or 4.0% (high) NaCl chow for 14–16 days. Then, animals were anesthetized with Inactin, and various sympathetic reflexes were tested. Electrical stimulation (1–20 Hz, 1 ms pulse, 500 uA) of sciatic afferents produced frequency‐dependent changes in SNA and ABP in both groups. However, rats ingesting 4% versus 0.1% NaCl displayed significantly larger increase in lumbar SNA (5Hz: 213±25 vs 146±15%), renal SNA (5Hz: 187±24 vs 120±11%), splanchnic SNA (5Hz: 203±21 vs 136±9%), heart rate (5Hz: 21±4 vs 13±2 bpm), and mean ABP (5Hz: 28±2 vs 12±2mmHg). Similarly, rats ingesting 4% versus 1% NaCl displayed greater increases in lumbar SNA (128±9 vs 112±2%) and mean ABP (12±1 vs 7±2mmHg) in response to intracerebroventricular infusion of 1M NaCl (5ul/10min). These findings suggest that increased dietary NaCl sensitizes central sympathetic networks to produce larger changes in SNA and ABP.
Median preoptic nucleus (MnPO) neurons activated following chronic intermittent hypoxia project to the paraventricular nucleus (PVN)
Authors: Faulk Katelynn Emily , Little Joel , Cunningham Thomas
Published: June 2021

Chronic Intermittent Hypoxia (CIH) is a model for the arterial hypoxemia seen in sleep apnea and is associated with hypertension and increased sympathetic tone. In rats, the MnPO demonstrates increased FosB expression following CIH. A major projection from the MnPO is to the paraventricular nucleus (PVN). This projection may serve as one possible pathway for increased sympathetic tone and sustained hypertension in CIH. The purpose of this study is to further characterize MnPO neurons transcriptionally activated by CIH and determine if they project to the PVN. Isoflurane anesthetized adult male rats were microinjected with 200nl of a retrograde tracer, fluorogold, unilaterally into PVN. Rats were then exposed to CIH for 7 days through 3 minute periods of hypoxia (10% oxygen) and 3 minute periods of normoxia (21% oxygen) for 8 hours per day (0800–1600 h). Controls were exposed to room air. Forebrains were fixed through paraformaldehyde perfusion and later processed for immunohistochemistry. Antibodies used were goat polyclonal FosB (1:2000). The sections were imaged using an Olympus DSU confocal microscope. In the MnPO, CIH exposure increased FosB positive cells (Control 7.75± 1.1; CIH 25.7±4.4; P<0.05) and colocalization of FosB with fluorogold positive cells (Control 0.67±0.4; CIH 7.25±1.2; P<0.05). These results show that MnPO neurons transcriptionally activated by CIH project to the PVN. P01 HL88052
In‐vivo transfection of manganese SOD gene and NFkB shRNA into nodose ganglia improve blunted baroreflex sensitivity in chronic heart failure rats
Authors: Zhang Dongze , Tu Huiyin , Liu Jinxu , Muelleman Robert L. , Yu‐Long Li
Published: June 2021

Blunted baroreflex sensitivity is observed in chronic heart failure (CHF) patients and animals. Our previous study has shown that mitochondria‐derived superoxide‐NFκB signaling reduces the Nav channel activation in baroreceptor neurons. Additionally, reduced Nav channel activiation contributes to the impairment of baroreflex sensitivity in the CHF state. Using in‐vivo gene transfection technique in present study, we examined the effect of Adenoviral Manganese SOD (Ad.MnSOD) and lentiviral NFκB p65 shRNA on the attenuated baroreflex function in CHF rats. CHF were induced by left coronary ligation. CHF significantly elevated mitochondrial superoxide and NFκB p65 expression in nodose neurons, and blunted baroreflex sensitivity. Ad.MnSOD transfection (4×107 PFU) into the nodose ganglia (NG) decreased CHF‐enhanced mitochondrial superoxide and NFκB p65 expression in nodose neurons, and improved baroreflex sensitivity in CHF rats. Similarly local transfection of lentivial NF‐κB p65 shRNA (5×103 IFU) into the NG also improved baroreflex sensitivity in CHF rats. These results suggest that mitochondrial superoxide‐NFκB signaling is involved in the impairment of baroreflex function in CHF rats.
Exaggerated response to acute stress in the spontaneously hypertensive rat (SHR) is linked to dysregulation of the hypothalamic orexin system
Authors: Hayward Linda F. , Castellanos Mabelin
Published: June 2021

Heightened sensitivity to stress is a hallmark of of hypertension. Orexinergic neurons in the hypothalamus are critical for cardiorespiratory arousal and defense responses. This raises the possibility augmented responses to stress observed in hypertension may be linked to dysregulation of the orexin system. Thus, the present study tested the hypothesis that exaggerated cardiovascular responses to acute stress in the SHR are associated with heightened activation of the orexin system. Adult male SHR and Wistar rats (n=4/group) were either sacrificed immediately for mRNA extraction from the hypothalamus or chronically instrumented for blood pressure (BP) monitoring during novel air jet stress. Prepro‐ orexin mRNA was identified to be significantly reduced in the SHR (21±6% of Wistar). Alternatively, 20 min of air jet stress elicited both a heightened BP response in the SHR and a significant increase in the number of c‐fos positive neurons in the lateral hypothalamus (103±10 vs 27±3 neurons, SHR vs Wistar) and an increase in the number of orexin/c‐fos double labeled neurons (25±5 vs 3±1 neurons, SHR vs Wistar). These results demonstrate that the exaggerated autonomic response to acute stress in hypertension is linked to abnormalities in the orexin system, including abnormally low gene expression at baseline and elevated system recruitment in response to acute stress.
Role of mTOR signaling in compensatory hypertrophy following uninephrectomy in the rat
Authors: Myung Choi , Oyekan Adebayo
Published: June 2021

Loss of functional nephrons following kidney injury or uninephrectomy (UNx) often results in functional and structural changes in the kidney such as compensatory hypertrophy. The eNOS/NO pathway has been shown to play a critical role in compensatory renal growth by modulating hemodynamic changes. As NO stimulates the Akt‐mTOR (mammalian target of rapamycin) pathway via the sGC/PKG pathway, this study tested the hypothesis that mTOR is a downstream signaling pathway in compensatory renal hypertrophy. In rats subjected to sham or UNx in the presence or absence of methylene blue (MB), an inhibitor of sGC, with or without rapamycin (Rapa), an inhibitor of mTOR, UNx induced an increase in kidney/body weight ratio (P<0.05) that was not affected by MB or Rapa. However, renal tissue DNA was increased following UNx (34%; P<0.05). MB further increased this (85%; P<0.01) while Rapa abolished the MB effect. Similarly, pAkt expression was increased by UNx (P<0.05) and blunted by MB (P<0.05). These changes were accompanied by UNx‐induced nitrite excretion (UNOxV; P<0.05) which was blunted by MB (P<0.05). Rapa blunted the MB effect (P<0.05). Blood pressure was unchanged by UNx, MB, or Rapa. Proteinuria (UProtV) was also not affected by UNx but MB increased UProtV (P<0.05) and Rapa paradoxically exarcerbated it (P<0.05). Despite this, serum creatinine (SCr) was unchanged in all the groups. However, UNx increased sodium excretion (UNaV; P<0.05) but was unaffected by MB or Rapa. We conclude that the Akt‐MTOR signaling pathway is involved in renal growth and is linked to renal NO production.
Athlete's heart: adaptations in blood flow, oxygen utilization, and efficiency at rest and during exercise
Authors: Heinonen Ilkka , Kudomi Nobu , Kemppainen Jukka , Luotolahti Matti , Sipila Hannu , Duncker Dirk J , Knuuti Juhani , Kalliokoski Kari K
Published: June 2021

We investigated here the characteristics of athlete's heart by applying [15O]‐labelled radiotracers and positron emission tomography. We measured myocardial blood flow (MBF), oxygen extraction fraction (OEF) and consumption (MVO2), and efficiency in 13 highly trained male endurance athletes (30 ± 5 yrs) and 13 untrained males (30 ± 5 yrs) at rest and during supine cycling exercise (100 watts). Echocardiography was also performed at rest and during exercise and the radial artery was cannulated for measurements of 2,3‐Diphosphoglycerate (DPG) and oxygen content. Many of the athletes had participated in Olympics and various world cups in running, XC‐skiing, cycling and walking, while untrained subjects did not exercise regularly. MBF was lower in athletes both at rest and during exercise. Arterial DPG was unchanged, but OEF increased in response to exercise in both groups, being higher in athletes both at rest (70 ± 21 vs. 63 ± 11 %) and during exercise (86 ± 13 vs. 73 ± 10 %). MVO2 per gram of myocardium trended (p=0.08) lower in athletes both at rest and during exercise. Myocardial efficiency was similar between the groups, although it trended higher in controls and lower in athletes in response to exercise. In conclusion, elite endurance athletes have lower than normal myocardial blood flow, higher oxygen extraction, and their cardiac oxygen consumption per gram of tissue trends lower both at rest and during exercise.
Vitamin D receptor and Estrogen receptor alpha Genotype affects Exercise related Bone mass in Japanese healthy women
Authors: Kondo Hiroyo , Fujino Hidemi , Murakami Shinichiro , Fujita Naoto , Nagatomo Fumiko , Ishihara Akihiko
Published: June 2021

PURPOSE

Osteoporosis is a systemic skeletal disease characterized by low bone mass and/or fragile bone. The vitamin D receptor (VDR) and estrogen receptor alpha (ESR1) are a good genetic candidate for a prime regulator of bone metabolism. The purpose of this study was to assess the interactive effects of exercise, food intake, VDR and/or ESR1 polymorphism on bone mass.

METHODS

240 healthy young women (20–23 yr) were recruited in this study. Bone mass and body composition were measured. Food frequency and exercise‐related questionnaires were used to estimate calcium intake and exercise intensity. The VDR (rs7975232) and ESR1 (rs2234693) polymorphisms were genotyped using the TaqMan‐based SNP method.

RESULTS

Exercise and milk‐intake were significantly associated with bone mass in the whole subjects. The bone mass in non‐exercised participants with CC homozygote in ESR1 was lower than that in TT homozygote. The bone mass in high‐intensity exercised participants carrying CC heterozygotes in ESR1 and VDR was significantly higher than that in low‐intensity.

These results suggest that exercise attenuates are the increase of peak bone mass in young women carrying C allele in VDR and ESR1.

Supported by Grants‐in‐Aid for Science Research from the Japanese Ministry of Education, Culture, Sports, Science and Technology.
Does Arhgef7, a guanine nucleotide exchange factor, regulate epithelial sodium channel activity and lung aeration?
Authors: Trac David , Helms My N.
Published: June 2021

Preterm infants have higher incidence of respiratory failure than term infants. These respiratory disorders are caused in part by delayed clearance of lung fluid. At birth, lung fluid clearance and aeration are mediated by active salt and water transport by epithelial sodium channel (ENaC) activity. Our lab has shown that oxidative environments can enhance ENaC activity, most likely via Nadph oxidase (Nox)‐mediated O2− release. In this highly regulated process, we have shown that the small G‐protein Rac1 plays an important role upstream of Nox signaling. However, the Rac1‐guanine nucleotide exchange factors (GEFs) involved in Nox‐mediated ENaC activity are unknown. Herein, we utilized quantitative real‐time PCR analysis to examine the expression profile of Rac1‐GEFs that may play an important role in ENaC regulation in the neonatal lung. Our major findings thus far are that transcript expression of Arhgef7>;>;>;Sos1>;Rasgrf2>;Tiam1>;Sos2, and that Arhgef7 transcript is expressed 1.5‐fold higher in neonatal (post‐partum day 7) mouse lung than adult mouse lung (n=8). This evidence suggests that Arhgef7 may play an integral role in neonatal lung aeration. Funding support: R00HL09226 awarded to MNH; Children's Center for Development in Lung Biology Grant awarded to MNH; and Emory University Summer Undergraduate Stipend awarded to DT.
ZnT2‐null mice have distinct morphological defects in the mammary gland that impair development and function
Authors: Lee Sooyeon , Hennigar Stephen R , Alam Samina , Kelleher Shannon L
Published: June 2021

Women with genetic variation in ZnT2 have ~75% lower milk zinc (Zn) levels during lactation. ZnT2 imports Zn into mitochondria and secretory vesicles in mammary epithelial cells (MECs), which suggests that ZnT2 plays critical roles in mammary gland (MG) function and Zn secretion into milk. We used female ZnT2‐null (KO) mice to explore effects on MG function. While plasma Zn was similar, MG Zn content was significantly lower in adult KO compared with wild‐type (WT) mice (5.6 ± 0.5 vs 11.4 ± 0.9 μg/g; p<0.05). KO MGs had irregular ductal branching, tightly clustered alveoli and greater collagen deposition around collapsed ducts. MG Zn content increased in both genotypes during lactation, but metallothionein expression was ~7 fold higher (p<0.05) in KO compared with WT mice, suggesting Zn sequestration. Lipid accumulation was noted in MECs and the alveolar lumen of KO mice, suggesting secretion defects. Mice were bred and although pregnancy rate, litter size and weight were similar, KO mice had fewer litters surviving past d1 compared with WT mice (41% ± 46 vs 96% ± 6.1; p<0.05). Interestingly, pups from KO dams had greater total body Zn content compared with WT pups (77.7 ± 3.2 vs 56.9 ± 7.4 μg/g; p<0.05), suggesting Zn transfer in utero was enhanced. Our data implicate ZnT2 as a critical mediator of MG development and function and suggest that women with genetic variation in ZnT2 may have substantial lactational defects beyond the inability to transfer sufficient Zn into milk.

Supported by NIH #HD058614 to SLK.
Distribution of cyclooxygenase (COX) isoforms in the developing ovine kidney
Authors: Lewis Megan Leigh , Qi Wei , Smith Francine Gabriel
Published: June 2021

COX is a rate limiting enzyme in the generation of prostaglandins. COX‐1, the predominant isoform, regulates physiological processes, whereas COX‐2 is typically low and upregulated during inflammation. In the adult kidney, both isoforms are present and localized to various structures implicating their roles in regulating renal haemodynamics and function. When administered during pregnancy or after birth, however, COX inhibitors can decrease renal perfusion and ultrafiltration potentially leading to acute kidney injury in the newborn period. To date, which COX isoform plays a more important role during kidney development is not known. In this study, we assessed the intrarenal distribution of COX‐1 and COX‐2 mRNA and protein expression in the developing ovine kidney, by examining cortical and medullary kidney tissue from lambs aged one day, one‐, three‐, six‐, and twelve weeks (N=3 for each age group). In all age groups, both isoforms were expressed at the level of mRNA and protein. Abundance of COX‐1 mRNA varied between cortex and medulla with an increased expression in the medulla at six and twelve weeks. COX‐2 mRNA was increased in the cortex at day one and week one and in the medulla at three and twelve weeks. Both COX‐1 and COX‐2 protein were greater in medulla than cortex and showed increased expression with postnatal age. These data support a role for both COX isoforms in the developing kidney.
Overexpression of Toll‐like receptor‐2 in human carotid artery plaques reflects clinical severity of atherosclerosis
Authors: Cherian George , Tan Xiaoyu , Cherian Rachel , Dileepan Kottarappat N
Published: June 2021

Toll‐like receptors (TLR) and cyclooxygenases (COX) are increasingly implicated in the pathogenesis of atherosclerosis. In this study, carotid endarterectomy plaques collected during surgery from nine patients with significant occlusive disease were quantitatively analyzed for the expression of TLR2, TLR4, COX1 and COX2 by qRT‐PCR, and for the presence of mast cells, T‐lymphocytes and macrophages by immunohistochemistry. Atherosclerotic plaques predominantly occurred at the carotid bifurcation, and had a 2–16 fold higher expression of TLR2 mRNA compared to the shoulder area. Localized plaques occurring away from the carotid bifurcation were predominantly fibrotic and had the lowest levels of TLR2 expression. The gene expression and inflammatory profile of each specimen were subsequently compared with the corresponding patient's clinical severity of atherosclerotic disease. The patient with the highest expression of TLR2 in the plaque also had the highest levels of TLR4 and COX2 mRNA, and the highest number of T‐lymphocytes and macrophages. This patient developed atherosclerotic disease at an earlier age with severe involvement of carotid, coronary and peripheral arteries, and had significant cutaneous S. aureus infections. These results suggest that the magnitude of TLR2 expression in the plaque has a direct association with the severity of widespread atherosclerotic disease. (Supported by NIH R01‐ HL070101 and 3R01‐HL070101–04S1, the Carey Arthritis Fund, the Audrey E Smith Fund, and the Dept. of Medicine Research Office, University of Kansas Med Ctr.)
Positive student response to Camtasia video recordings that supplemented classroom presentations
Authors: Sekar M. Chandra
Published: June 2021

Based on last year's student survey, I modified my cardiovascular pharmacology course for fall of 2012. This course was taught at the University of Findlay, College of Pharmacy. I recorded the first ten of my presentations using Camtasia software, and they were available for students to preview before coming to listen to live presentations. Since presentations were available before class, only 30 minutes were required for lecturing during class. This left time to ask students questions about important points and concepts using an “audience response system”. At the completion of this project, students were provided with a “google docs” survey that they completed anonymously. Results of the survey were extremely positive and are reported here. Fifty‐nine of the sixty‐nine students enrolled in the course completed the survey. Fifty eight percent of students had seen all 10 posted videos; twenty percent – 7 to 9 videos; nineteen percent – 4 to 6 videos; two percent – 1 to 3 videos and two percent did not see any of the posted videos.

Statement
# Student
Statement
# Student

Video good quality
55 (95%)
Helped me learn better
57 (98%)

Watched video only once
39 (66%)
Watched video more than once
15 (26%)

Watched before class
37 (64%
Watched after class
14 (24%)

Watched before and after class
21 (36%)
Continue future postings
57 (98%)

Replayed portion of video
54 (93%)
Plan to see videos not seen
37 (64%)
When a cadaver presents a multi‐systemic dilemma
Authors: Locke Ben Charles , Dillard Attallah Siedah , Nusse Gloria
Published: June 2021

Factors affecting the actual death of an individual can be convoluted, ambiguous, and tangled. When cadavers arrive in the San Francisco State University anatomy lab they are accompanied by a cause of death report. Such was the case with an 83‐year‐old adult female cadaver. Her cause of death was respiratory failure with contributing causes listed as degenerative arthritis, depression, dementia, and reflux esophagitis. Although respiratory failure was the primary cause of death, upon dissection her body revealed several abnormalities that were not mentioned in the death certificate. Foremost of which was a 2.5 cm depression in the right frontotemporal lobe as well an unusual digestive tract, which was almost completely avascular in appearance. This poster presents these unusual findings.

Grant Funding Source: none
The star of venous revealed
Authors: Tugbenyoh Mawuli Kofi‐Doe
Published: June 2021

The “star of venous” is a structure that is grossly overlooked in undergraduate anatomy classes as well as in many textbooks. The more recognizable name for this structure is the saphenofemoral junction. In an average of 37% of cases, a patient presents with a true “star of venous”. It is a venous junction, not duplicated in arteries, between the greater saphenous, great circumflex, epigastric, femoral, external iliac and one or several of the external pudenal, veins. The “star of venous” is located within the femoral triangle deep and inferior to Allan Burns’ ligament.

The “star of venous” carries great significance when preforming surgeries such as inguinal hernia repair, or on an ectasia of the saphenofemoral junction, or during greater saphenous stripping for coronary artery revascularization.

Dissection of the “star of venous” is relatively easy to do but also easy to remove if you don't know it is there. It is these types of amazing and elegant structures that are small but carry a large significance in the human body. If students are aware of the “star of venous” when learning anatomy they may gain a real relevance and perspective to its importance.

Grant Funding Source: Biology Department
The flexor digitorum breves manus and contrahentes muscles in Afrotheria and other mammals
Authors: Voegele Georgina
Published: June 2021

The mammalian clade Afrotheria was created as a result of Stanhope et al, 1998. Paleontologists and comparative anatomists have struggled to find unique skeletal and dental features that characterize the clade, but there has been very little research on the soft tissue anatomy of afrotherian mammals. My dissertation research has focused on dissecting the forelimbs of members of Afrotheria: three sengis (Order Macroscelidea), two hyraxes (Order Hyracoidea), one aardvark (Order Tubulidentata), two tenrecs and one golden mole (Order Afrosoricida), and comparing their anatomy with published descriptions of other mammals. Features of particular interest within the afrotherian hand are the robust and unusual flexor breves digitorum manus and the pattern of the contrahentes. Also of note are the sesamoid bones in the flexor retinaculum and over the carpals, the relative unimportance of the flexor digitorum superficialis, and the distribution of the median and ulnar nerves.

Grant Funding Source: none
Undescribed variation of the sciatic nerve
Authors: Jacomo Alfredo Luiz , Saleh Samir Omar , De Ataíde Gamaliel Silva Alves , Andrade Mauro , Akamatsu Flávia Emi
Published: June 2021

Piriformis muscle syndrome has been increasingly recognized as a cause of leg pain. Overuse, strain, or anatomical variations of the relationship between the nerve and the piriformis muscle are thought to be the underlying causes of the entrapment of the sciatic nerve. We report a previously undescribed variation found during a routine dissection

During a routine dissection of the left gluteal region of an adult male cadaver we observed a high division of the sciatic nerve and the presence of an accessory piriformis muscle. The sciatic nerve divided beneath the piriformis muscle and the common fibular nerve passed over the accessory piriformis muscle, whereas the tibial nerve reflected anteriorly to pass between the accessory piriformis and the superior gemellus muscle. Additionally, both nerves communicated with a side branch under the inferior border of the accessory piriformis muscle and the inferior gluteal nerve originated from the fibular nerve.

Anatomical variations in the relationship between the piriformis muscle and the sciatic nerve may be present in up to 17% of the population. Six different variations have been described and none of them is similar to our description. Though complete understanding of the physiopathology of the piriformis muscle syndrome remains to be elucidated, knowledge of the possible anatomical variations may be useful for its adequate diagnosis and treatment.
Histological and immunohistochemical comparison of major salivary glands in young and old mice
Authors: Nonaka Naoko , Nakamura Masanori
Published: June 2021

Pituitary adenylate cyclase‐activating polypeptide (PACAP) is now recognized as the multi‐functional neuropeptide in various organs. We have examined the effect of PACAP for saliva secretion. In this study, we compared the parenchymal structure and the distribution of PACAP receptor, PAC1R, in three major salivary glands; parotid (PG), submandibular (SMG), sublingual (SLG) glands of young (8 weeks old) and old (8 months old) male C57BL/6 mouse. In PG, PAC1R was localized in the striated duct. In SMG, PAC1R was detected in the tall columnar epithelial cells in granular ducts and some of the cells in the striated ducts. In SLG, PAC1R was localized in the striated duct. No apparent difference of PAC1R was detected between young and old mice glands. In SLG of old mice, granular epithelial cells are detected in the striated duct. Furthermore, massive infiltrations of CD3‐ positive or B220‐positive lymphocytes were detected in old PG and SMG. These results suggested that PACAP regulated the saliva secretion by modulating the activity of striated duct and that lymphoid infiltration in PG and SMG but not in SLM might give the useful information for the etiology of autoimmune disease of salivary gland.
Aortic depressor nerve morphometry in male and female adult spontaneously hypertensive rats.
Authors: Amorim Milena Menezes , Castania Jaci Airton , Salgado Helio Cesar , Fazan Valéria Paula Sassoli
Published: June 2021

Previous studies form our laboratory [1,2] in spontaneously hypertensive rats (SHR) have demonstrated morphological alterations on the aortic depressor nerve (ADN), known to contain mainly barorreceptor fibers. Nevertheless, those studies used male and female rats indistinctly. We are investigating if there are morphological differences related to gender in the ADN of adult rats. Male and female SHR aged 20‐week were anesthetized for arterial pressure (AP) and heart rate (HR) recordings, together with the ADN spontaneous activity. After the recordings, the left ADN were prepared for light microscopy and morphometry. Comparisons were made between proximal and distal segments in the same gender and between genders. Males are significantly heavier than females but no differences in AP and HR between genders were found. There was a tendency towards larger fascicle values on males compared to females. Interestingly, this tendency was inverted on the myelinated fibers and respective axons sizes. Nevertheless, no significant differences were observed between genders on the morphometric parameters. We conclude that adult males and females can be used indistinctly for morphometric studies of the ADN if physiological parameters are similar between genders.
Structure and function of Holliday junctions complexed with ions and HU protein
Authors: Nocka Laura Margaret , Mukerji Ishita , Beveridge David L. , Wheatley Liz , Vitoc C. Iulia
Published: June 2021

DNA four‐way junctions (4WJ) are central intermediates in processes such as replication and recombination. This project studies how different conformations of the junction relate to function, and how proteins interact with the junction to enhance or repress function. Previously, we have examined the 4WJ structure in the presence of different ions, and used Förster Resonance Energy Transfer (FRET) to map the binding site of the protein HU onto the junction. These FRET measurements were used to build four different models for the binding of HU protein to the Holliday Junction, an interaction with a stoichiometry of 2 to 1. Molecular dynamics simulations ensured that the models were energetically stable. The three stable models included HU bound to the major groove, minor groove, or face‐to‐face. The fourth model, which contained an open junction, though stable, did not agree with the FRET data acquired for the system. Further FRET measurements are needed to narrow the possible orientations for HU binding the junction. To address the function of the protein‐bound junction, a new, migrating 4WJ is being created and characterized. This 4WJ will be analyzed by FRET in the presence of ions and proteins to compare to the characteristics of the stationary junction. The free energy of both the mobile and immobile junctions will be analyzed by molecular dynamics to predict stability of each construct alone and when complexed with HU.

This research is supported by an the NSF (MCB‐0316625; MCB‐0843656 to I.M.), the NIH (GM‐076490 to D.L.B.), and by NIGMS Training grant T32GM08271.
Intracellular Trafficking of an Antibody Bipolar Bridged Complex of HSV‐1 gE‐gI, IgG, and a Viral Antigen
Authors: Ndjamen Blaise , Farley Alexander , Fraser Scott E. , Bjorkman Pamela J
Published: June 2021

Herpes Simplex Virus 1 (HSV‐1) glycoproteins, gE and gI, mimic a human Fc‐receptor to hinder host immune responses. gE‐gI binds the Fc region of human IgG (hIgG), which can be internalized and released into endocytic organelles. gE‐gI also mediates antibody bipolar bridging (ABB), a process whereby IgG Fabs bind an antigen while its Fc region binds to gE‐gI. The cellular fate of the ABB complex is unknown. Here, we engineered an in vitro system to determine whether ABB complexes are internalized and degraded in host cells. Human cell lines transiently expressing gE‐gI and an HSV‐1 antigen, gD, normally localized at the cell surface of HSV‐1 virions and infected cells, were incubated with two forms of a monoclonal IgG anti‐gD antibody: one with human Fc to allow ABB and one with mouse Fc, a control which binds gD but not gE‐gI and cannot enable ABB. A hIgG against an irrelevant antigen was used as another non ABB control. 4D time‐lapse confocal fluorescence data indicate that gD is internalized in an ABB specific fashion and internalization is dependent upon gE‐gI. Internalized hIgGs bound to a surface viral antigen were targeted to lysosomes and biochemical analysis of enriched lysosomal compartments revealed degraded gD. These results suggest that gE‐gI plays an active role in clearing the infected cell surface of both host IgG and viral antigens, providing HSV‐1 with a mechanism to evade immune responses.
MeCP2's Role in Chromatin Structure and Stability
Authors: Riedmann Caitlyn , Fondufe‐Mittendorf Yvonne
Published: June 2021

Autism spectrum disorders (ASDs) affect 1 in 110 children. Rett Syndrome (RTT) is one of the few ASDs that have a direct genetic link, which make it great model to understand some underlying causes of ASD. RTT is caused by mutation in the X‐linked Methyl CpG Binding Protein 2 (MeCP2). MeCP2 is associated with transcriptional repression and can act on naked DNA or, if found in high enough quantities, alters chromatin structure. Therefore understanding the functional implications on both specific genes and chromatin structure is critical in developing therapeutic strategies. After neural development MeCP2 is found in levels equivalent to H1 in neuronal cells and is believed to compete with H1. Like H1, MeCP2 binds at the entry/exit site of nucleosomal DNA, and could regulate the rate of wrapping and unwrapping around the nucleosome, thus being critical in target gene accessibility. Our goal is to understand how MeCP2 alters gene accessibility through dynamic changes to chromatin structure. We will use FRET measurements to determine the influence of MeCP2 on nucleosome stability, site accessibility, and higher order compaction; providing insight to the mechanistic functions of MeCP2 on gene expression.
Electrochemical Characterization of Superoxide Radical Chemistry and Scavenging by Natural Products
Authors: Lee Grace , Belli Stuart , Caruso Francesco , Rossi Miriam
Published: June 2021

The relationship between biological antioxidant molecules and superoxide free radical is a major curiosity. Superoxide radical is the leading contributor to ailments such as cardiovascular disease and cancer, and may play a role in the aging process. While it has been observed that antioxidants sequester these harmful radicals, the exact mechanism is still a mystery. In an effort to better understand the antioxidative properties that attribute to the sequestering of superoxide radical, the radical was studied in vitro in an electrochemical cell using cyclic voltammetry with a gold working electrode. Oxygen gas was converted into superoxide radical and stabilized in an aprotic solvent (DMSO) whereupon the resultant radical was exposed to various molecules thought to be beneficial to health. The effectiveness of said biomolecules was evaluated by quantifying the change in concentration of superoxide radical before and after exposure to antioxidants, as determined by calculating the change in the integral of the resultant currents produced within a set interval of voltage potential. Tocopherol (vitamin E), olive oil, grape seed oil, quercetin, 22–0 docosenoate, and nitro blue tetrazonium were tested for their antioxidant qualities. Spectrophotometry was used in conjunction with cyclic voltammetry to evaluate the effectiveness of certain antioxidants. Supported by HHMI 52006322 and VC URSI 2012
Exercise on a moving platform elicits different gene expression of neurotrophins and receptors in the spinal cord of aged rats
Authors: Kanemura Naohiko , Imagita Hidetaka , Maejima Hiroshi , Kokubun Takanori , Murata Kenji , Takayanagi Kiyomi
Published: June 2021

Proprioceptive training is highly common in rehabilitation of aged people to prevent injures or to enhance performance. This study aimed at determining whether the exercise on a moving platform induces changes in neurotrophins expression patterns in the lumber spinal cord of aged rats. Twelve aged male rats (2 years old) and ten adult male rats (10 weeks old) were randomly assigned to each two groups: exercised and non‐exercised groups. Animals were trained on a moving platform for 5 days a week for 4 weeks. We measure the relative of levels of neurotrophins (BDNF,NGF,NT3) and their receptors (TrkA, TrkB, TrkC ) using quantitative real‐time PCR. The expression of each gene was compared between depots using comparative Ct method. Statistical analysis was undertaken using the one way ANOVA with the Tukey post hoc test.No statistically significant differences of the level of NT3, TrkC, and TrkA mRNAs were found among the groups. The expression of BDNF mRNA was significantly higher in the adult exercised group compared with the aged exercised rats. Level of NGF and TrkB mRNAs in the both old and adult non‐exercised groups were significantly higher than the adult exercised groups. TrkB level of expression in adult non‐exercised was significantly higher than aged non‐exercised rats. The changes in neurotrophins and receptors may have important consequences for neuroal plasticity and regeneration in the neuromuscular system.
Variations in body iron status determine variations in body energy expenditure and brain dynamics as a function of perceptual and cognitive workload
Authors: Wenger Michael J , Murray‐Kolb Laura E. , Haas Jere D.
Published: June 2021

Iron deficiency (ID), with and without anemia, has been shown to have deleterious effects on physical performance. Animal models have demonstrated that physical endurance and the concentrations of oxidative enzymes and respiratory proteins all decrease in ID, even at levels that do not reach the point of anemia. Human studies have documented reductions in endurance in ID without anemia, with effects being seen in measures of energy expenditure and work efficiency. The present effort extends the examination of the effect of ID to consideration of its effects on energy use in the performance of mental work. ID women(ferritin < 16 g/dl, n = 20) and age‐, education‐ and activity‐level‐matched controls (n = 17) provided blood samples for measures of iron status; completed a set of questionnaires probing general health and activity levels, menstrual status, and consumption of iron‐rich foods; completed a working memory task (as a general measure of cognitive efficiency); and performed a visual Sternberg task with a concurrent mental math task. While performing the visual Sternberg task, participants’ brain and metabolic states were measured using electroencephelography (EEG) and measures of the volume of O2 and CO2 in expired air, respiratory rate, and heart rate. We show statistically significant relationships among behavioral, EEG, and metabolic measures, all as a function of variations in task difficulty and iron status. The data suggest the potential for using EEG for indirect estimation of the effects of ID on brain energy.
Household food insecurity during the pre‐harvest period is associated with respiratory infections, but not stunting among 6–11 month old infants in rural Ghana
Authors: Cofie Agartha , Marquis Grace S , Lartey Anna
Published: June 2021

Food insecurity is prevalent in rural Ghana, particularly during the months prior to harvest. We examined the association between pre‐harvest food insecurity and infants’ health and nutritional status. The cross‐sectional survey of 333 mothers and their infants aged 6–11 mo living in the Upper Manya Krobo district, included reported household food insecurity (HHFI), anthropometric measurements of infants and mothers, and mothers’ recall of symptoms of infants’ illnesses during the previous seven days. Multiple logistic regressions were conducted to examine how HHFI was associated with stunting and morbidity. Over one‐fifth of households experienced food insecurity in the previous month. Compared to infants in food secure homes, infants living in food insecure homes were twice as likely to experience cough (aOR = 2.26, 95% CI: 1.24 to 4.13), and tended to experience a runny nose (aOR = 1.82, 95% CI: 0.98 to 3.39). HHFI was not associated with diarrhea, fever or stunting. Efforts to improve infant's health status may need to include strategies that improve household food security, particularly during the pre‐harvest period. Funded by IDRC Doctoral Research Award 105938–99906075‐038 and McGill University.