bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in Individual and Team Cognition in High Stress Extreme Underwater Saturation Environment Under Intense Workload
Authors: Ari Csilla , D’Agostino Dominic Paul , Bharwani Sahil , Rehsi Anand , Moss Sara , Schmer-Galunder Sonja , Fiore Stephen
Published: April 2020

The objective of this study was to measure potential changes in cognitive function in response to living in an extreme underwater habitat. Chronic high stress and intense workload can affect individual and team cognition. As such, associated variables were monitored during the NASA Extreme Environment Mission Operation (NEEMO) 23 mission at the Aquarius reef Base in June 2019.

Individual cognition was measured on 6 subjects (2 males, 4 females) with 8 tests using a battery of cognitive assessments (Joggle Research platform), 1 day pre‐mission, at mid‐point of the mission and 1 day post‐mission. In addition, 4 cognitive tests were applied on 5 subjects (2 males, 3 females) at the beginning of Extravehicular Activities (EVA) and at the end of EVAs that lasted for ~3 to 5 hours. To monitor changes in stress level, Short Stress State Questionnaire (SSSQ) and Generalized Anxiety Disorder (GAD7) scores were collected. Effects of workload on performance were measured by NASA Task Load Index (TLX). Changes in team cognition was monitored using an assessment of cognitive process measures.

There was a trend of increasing performance in individual cognition in‐habitat and post mission, compared to baseline. The NBACK test, a measure of working memory, significantly improved during the mission, compared to pre‐mission and it further improved post‐mission. The psychomotor vigilance test (PVT) results were non‐significantly lower during the mission. The Motor Praxis Task (MPT) scores decreased at the beginning of EVA and further decreased by the end of EVA. The end of EVA MPT performance was significantly lower, compared to baseline, in habitat, and post mission measurements, suggesting that long duration extravehicular activity had a significant negative effect on sensory motor speed. BART scores were significantly lower at the beginning of EVA, compared to baseline, suggesting that risk taking decreased with the start of the EVA. The PVT score was significantly lower at the end of EVA, compared to baseline, in habitat, and the beginning of EVA, suggesting that the long duration EVAs had negative effect on the ability to quickly respond to the sudden appearance of a stimuli. A trend of increasing GAD7 scores at half time of the mission and post‐mission could be observed. There were differences in NASA TLX and team cognition scores between subjects and at different time points.

The present results suggests that sub‐components individual and team cognitive functions may be differently affected during this extreme environment mission. Further research is needed to isolate which cognitive functions might be altered and how these may be correlated with stress level and workload to affect mission success.

Support or Funding Information

Ketone Technologies LLC provided partial funding for the experiments. SIFT provided partial salary for CA.
Kefir peptides improve gut and blood vessels function by ameliorating intestinal morphology and enriching intestinal microbiota in salt‐induced spontaneous hypertension rat stroke‐prone (SHRSP) animal model
Authors: Chen Yu-Hsuan , Chen Hsiao-Ling , Chen Jen-Kun , Chen Chuan-Mu
Published: April 2020

Kefir peptides are prebiotic products generated by kefir grain fermentation of milk proteins. Effects of kefir peptides for lowered blood pressure, antioxidant and osteoporosis. In this study, we hypothesis that kefir peptides may protect the gut function with microbial dysbiosis changes and decrease the mesenteric blood vessels inflammatory which linked to anti hypertension. The current study found that a high salt diet is considered a cardiovascular risk factor, altered the intestinal bacteria composition and produced gut dysbiosis in rats. Thus, we used the 1% salt (v/w) in drinking water to speed up hypertension of spontaneous hypertension rat stroke‐prone (SHRSP) rats, the rats were assigned to four treatment groups, including normal control group (WKY rat), Untreated group (SHRSP rat fed with normal water), NaCl/Mock group (SHRSP rat fed with 1% salt water) and NaCl/KP group (SHRSP rat fed with 1% salt water+200 mg/kg kefir peptides) for four weeks experiment. Data showed that rats only gave the 1% salt group the gut villus height of small intestine and the diversity of intestinal microbiota were significantly decreased, while supplementation with kefir peptides counteracted the normal control group. In the small intestinal microbiota, the microbiota enriching of 1% salt‐treated Mock group have significantly decreased than the other groups. Oral administrations of kefir peptides could regulate the microenvironment of the intestinal tract for the microbes, provide extra sources of carbon and nitrogen for the microbes to live and grow. Additionally, the genera of Allobaculum, Bifidobacterium and Lactobacillus were found to have a high degree of richness in the kefir peptides group, which are generally considered beneficial taxa with the energy and regulate blood pressure. In conclusion, the results indicated that kefir peptides improve gastrointestinal microenvironment and mesenteric blood vessels function by ameliorating intestinal morphology and enriching intestinal microbiota under the 1% salt‐induced SHRSP rats.
Structure‐Based Design and Evaluation of Peptidomimetic Inhibitors Against the Human Norovirus Protease
Authors: Muzzarelli Kendall Marie , Brunzelle Joseph , Kovari Iulia , Yang Zhe , Kovari Ladislau
Published: April 2020

Norovirus is the leading cause of acute gastroenteritis worldwide and can lead to severe chronic infection in immunocompromised patients. Currently, there are no FDA approved therapeutics or vaccines against Norovirus infection. However, progress has been made in terms of identifying the Norovirus protease as a promising drug target due to its necessity in the production of mature viral proteins. Our study has focused on the development of targeted covalent inhibitors against the human strains of Norovirus protease by employing both computational and enzymatic design and evaluation techniques. Analysis of our recently solved apo and complexed crystal structures of Norovirus GI.1 and GII.4 proteases provided additional insights into the binding pocket and active site composition, which led to improved inhibitor efficacy. Potential compounds were designed and analyzed using Schrodinger (2019‐1) software, allowing for predictive ADME screening, covalent docking of inhibitors to the protein structure, and binding affinity analysis. The lead compounds were synthesized, then enzymatically screened against purified Norovirus protease using a FRET‐based assay, following in silico design and evaluation. To date, our lead compounds exhibit nanomolar activity against the Norovirus GII.4 protease as well as the less infectious GI.1 strain.

Support or Funding Information

Vera Fay Righthand Fellowship in Virology
Post‐translational modulations of IL‐6 and oxidative stress networks in the hippocampus after epilepsy induced by pilocarpine in rats.
Authors: DA SILVA RUIZ ÉRICO GUSTAVO , ATUM ALLAN LUÍS BARBOZA , MARQUES DANILA , DOS SANTOS FELICIANO REGIANE , GOUVEIA TELMA LUCIANA FURTADO , NATHANSON LUBOV , MORRIS MARIANA , SILVA JOSE ANTÔNIO
Published: April 2020

Epilepsy is a prevalent and serious brain condition, characterized by the recurrence of unprovoked seizures. Disturbances on cellular processes, which are controlled by post‐transcriptional gene expression modulations such as microRNAs (miRNAs) can lower multiple proteins expressions within molecular pathways. In order to elucidate the role miRNAs exert in the hippocampus from rats with epilepsy induced by pilocarpine, expression of miRNAs was quantified by NanoString technology. Furthermore, miRNA targets were identified by in silico analysis. We observed strong changes in miRNA expression profiles in hippocampal cells derived from epileptic rats compared to the control. MiRNAs to be related to inflammation and oxidative stress were found differentially expressed in silico analysis. For instance, in hippocampal tissue, miR‐146 emerged, which are known to interact with interleukin‐6, a major player in inflammatory process. Expression change was observed for let‐7, which targets IL‐6 as well. Further differentially expressed miRNAs were related to oxidative stress and generation of reactive oxygen species, such as miR‐30e. Altogether, the data indicate that miRNAs are an interesting starting point for functional and mechanistic epilepsy research.

Support or Funding Information

1) Universidade Nove de Julho, UNINOVE, São Paulo, Brazil 2) Universidade São Camilo, São Paulo, Brazil 3) Nova Southeastern University, NSU, Florida, USA
ANALYSIS OF RS12976445 VARIANT IN MIR125A AND FEMALE INFERTILITY IN LAGOS, NIGERIA
Authors: Gbenle segun A. , Igwo-Ezikpe Miriam , Taiwo Idowu A. , Salami Fatimah O. , Chukwunonso Chinwedu F. , Ofoma Osita N. , Onawoga Fatimah O.
Published: April 2020

Micro RNAs play very important roles in maintaining normal human body physiology conditions, meanwhile, abnormal miRNA expressions have been found related to many human diseases including male and female infertility. Reports have indicated that disturbed miRNAs expression contributed to female infertility with reference to a polymorphism (rs12976445) within the miRNA125a sequence. This aim of the study was to investigate the effects of MiRNA125a gene disorders on female infertility and how these can be used in enhancing fertility among women seeking in vitro fertilization (IVF) in Lagos, Nigeria. Ethical approval was obtained from the Lagos University Teaching Hospital Health Research Ethics Committee (LUTHHREC) with number ADM/DCST/HREC/APP/2396. Infertile women (n=50) attending various fertility clinics in Lagos, Nigeria were recruited for this study and structured self‐administered questionnaires were distributed to the subjects to obtain their physiological and demographic parameters. DNA were extracted and amplified from blood samples obtained from subjects using primers specific for miR125a sequencing 5′‐TTTTGGTCTTTCTGTCTCTGG‐3′ and 5′‐TGGAGGAAGGGTATGAGGAGT‐3′. PCR products were analyzed using a BaeGI restriction enzyme specific to the rs12976445 variant. The restriction analysis revealed that 6 out of 50 subjects were TT homozygotes (12%) which were normal, 21 (42%) were TC heterozygotes variants and 23 (46%) were CC homozygotes variants. We hypothesized that rs12976445 may be a potential diagnostic marker of miR125a in female infertility among Nigerian women. Hence, mir125a should be considered in female infertility treatment.

Support or Funding Information

NA
Toll‐Like Receptor Expression in the Hypothalamus of Rats Subjected to Chronic Inflammatory Pain Model
Authors: Teel Austin , Winter Lori , Duric Vanja , Clayton Sarah C.
Published: April 2020

Hypertension is the state of having an abnormally and chronically elevated blood pressure. There are many comorbidities of hypertension, which includes cardiovascular disease, stroke, kidney disease, diabetes mellitus, and psychological disorders, such as chronic pain. Clinical observations suggest that approximately 39% of individuals experiencing chronic pain also have hypertension. While inflammation has been documented in areas of the brain in hypertension and chronic pain, the influence of chronic pain on activation of proinflammatory mechanisms within the cardioregulatory nuclei in the brain has not been evaluated. We speculate that chronic pain leads to inflammation in cardioregulatory brain regions, such as the hypothalamus and brainstem, that leads to dysregulation of autonomic tone and increases in blood pressure. Therefore, in the current study, we evaluated mRNA expression of different members of the Toll‐Like Receptors (TLR) family and associated proteins in the hypothalamus of rats exposed to 21 days of chronic inflammatory pain [i.e., 3 weekly injections of Complete Freund’s Adjuvant (CFA) into the hind paw]. Preliminary results show a decrease in the hypothalamic expression of TLR3 and an increase in the expression of TLR5, TLR7, and TRAM as a result of the pain protocol. These data suggest a relationship between chronic pain and hypothalamic inflammation and may suggest a possible target for probing the intersection of chronic pain and the development of hypertension.

Support or Funding Information

DMU IOER and MSBS program
Analysis of Undergraduate Retention Outcomes: What do Students Actually Remember?
Authors: Dvoracek Peter , Balta Joy Y. , Osadjan Mark
Published: April 2020

As a basic science, anatomy has commonly drawn criticism as a subject material that can be notoriously difficult for undergraduates to learn. Some common reasons for this scrutiny could be due to students’ perception of anatomy as a course that relies heavily on abstract, rote memorization
[1]. Learning through abstract memorization with high intrinsic cognitive load often produces poor retention, compared to more elaborative and meaningful encoding
[2]. In this study, student’s long‐term retention of practical material from a standard 4‐credit undergraduate human anatomy course was reassessed 5‐month past end of term. This data was compared to their continuous practical assessment throughout the semester. Students who had successfully completed this course in the past semester were invited to take part in this study. Informed consent was obtained and participating students attended a single, one hour post‐assessment during their regularly scheduled lab time. This post‐assessment evaluated their retention of lab course material using free recall type questions. Anonymized data were collected and analyzed using Excel. This study was approved by the Institutional Review Board at University of Minnesota.

Out of the 156 enrolled students, 94 eligible students participated, yielding a 60% response rate. When comparing post‐assessment results to the practical lab exams, a significant linear moderate positive relationship between the two of them was observed (correlation coefficient=.5971, p<.00001). This suggests that practical lab exams are moderately accurate at predicting students’ performance on the post‐assessment. There is no correlation between student performance and memory retention, as indicated by the linear relationship between the data sets. When comparing written to practical exams, written exam scores stayed relatively constant while students’ practical exam scores were found to have steadily and significantly increased across the semester (p<.00001). This suggests that there was significant development of spatial reasoning skills from the lab material itself. More data, using the same metric, will be collected in the coming years from future cohorts to validate the findings of this study.

Support or Funding Information

None
Differential Protein‐Protein Interactions Driven by the Arg/Gly389 Single Nucleotide Polymorphisms in the Cardiac Beta‐1 Adrenergic Receptor
Authors: Bloomfield Christina , Tran Quang-Kim
Published: April 2020

Activation of the β1 adrenergic receptor (β1AR) increases electrically induced calcium signals and accelerates sequestration of calcium in the myocardium, promoting contraction and relaxation. In humans, Arg389 and Gly389 are common single nucleotide polymorphisms (SNPs) in β1AR. The Arg389 SNP is associated with higher responses to β agonists in terms of cAMP production and contractile force development; in living subjects, it also is coupled with faster response to b blockers with respect to reductions in heart rate and myocardial contractile force. Nevertheless, it is unknown if calcium signals following β1AR activation affects the receptor’s activities and changes in Ca2+‐dependent regulation of β1AR can explain the difference between these polymorphisms. Calmodulin is the ubiquitous transducer of calcium signals and is a limiting factor in cardiomyocytes. We tested the hypothesis that β1AR directly interacts with calmodulin and that the Arg389 and Gly389 SNPs are associated with disparate β1AR‐calmodulin interactions. Using novel fluorescent biosensors developed for β1AR, we identified a novel calmodulin‐binding domain located at the juxtamembranous region of β1AR that encompasses the Arg/Gly389 SNP. The Arg389 SNP is associated with a biphasic interaction with Ca2+/calmodulin. However, the Gly389 SNP is associated with a monophasic interaction with CaM and a ~12‐fold lower affinity than the Arg389 SNP. These kinetic differences correspond to significant variances in the response of β1AR to physiological calmodulin concentrations, peaking at ~50 nM. We also identified that β1AR‐calmodulin interactions are calcium‐dependent, with higher Ca2+ sensitivity of the Arg389 β1AR–CaM interaction over the entire physiological Ca2+ range. These results suggest that calmodulin binding to β1AR is important for its functions and our observed different biochemical properties of these interactions may explain the different responses in subjects carrying the Arg389 vs the Gly389 SNPs.

Support or Funding Information

IOER grant 091707 to Q‐KT.
Lack of muscle p62/SQSTM1 protects against high fat diet‐induced muscle wasting
Authors: Yamada Mami , Okutsu Mitsuharu
Published: April 2020

Skeletal muscle atrophy is a hallmark of aging and several debilitating diseases (e.g., cancer, congestive heart failure, and type II diabetes) that contribute to morbidity and mortality. Oxidative stress plays an important role in skeletal muscle atrophy in these situations. Therefore, increased antioxidants expression could be a potentially protective measure against oxidative stress and skeletal muscle atrophy. We recently reported that p62/SQSTM1 expression in skeletal muscle contributes to control exercise training‐induced antioxidant enzymes expression with established p62 muscle‐specific knockout mice (p62mKO). However, the role of muscle p62 expression in type II diabetes induced‐muscle wasting was still not understood. Aim of this study was to elucidate the role of muscle p62 in type II diabetes induced‐muscle atrophy. p62mKO and their wild‐type littermates (WT) were fed normal chow (NC) or 45% fat diet (HFD) for 10 weeks. Body weight and food intake were measured every week. Glucose or insulin tolerance test were conducted at the 8 weeks. A grip strength test was performed at the end of HFD feeding. At 10 weeks, mice were sacrificed and harvested soleus (SO), gastrocnemius (GA), tibialis anterior (TA), extensor digitorum longus (EDL), and white vastus lateralis (WV) muscle. These muscles were measured weight and used for evaluation of protein expression. Muscle weight was measured by semi‐micro analytical balances and normalized by tibia length. Carbonylated protein was measures by western blot. Body weight, food intake, glucose and insulin tolerance ability, and grip strength were not significantly different between HFD fed p62mKO (p62mKO‐HFD) mice and HFD fed WT (WT‐HFD) mice. p62mKO‐HFD mice significantly attenuated loss of GA and EDL muscle weight. Carbonylated protein expression in p62mKO‐HFD mice was significantly lower than WT‐HFD mice. These results suggested that lack of muscle p62 protects against high fat diet‐induced muscle wasting via an antioxidants synthesis‐independent pathway.

Support or Funding Information

This study was supported by Grant‐in‐Aid for Research Activity Start‐Up (25882036), Grant‐in‐Aid for Scientific Research (B) (15H03080), Grant‐in‐Aid for Scientific Research (B) (18H03153), The Nakatomi Foundation, and The Uehara Memorial Foundation (to M.O.).
Regulation of the Phenotypic Switch of Vascular Smooth Muscle Cells by Adiponectin
Authors: Cullen Abigail , Centner Ann , Salazar Gloria
Published: April 2020

Vascular smooth muscle cells (VSMCs) play an essential role in the development and progression of atherosclerosis. The phenotypic switch from contractile to synthetic cells induces excess proliferation and migration of VSMCs into the intima contributing to plaque formation. Adiponectin (Adpn) is an adipokine that regulates glucose metabolism through AMPK and plays beneficial roles in atherosclerosis. We hypothesize that Adpn promotes the contractile phenotype of VSMC reducing proliferation and migration of VSMCs, a process that is protective against plaque formation.

Differentiation, proliferation, and signaling through AMPK‐dependent pathways were assessed in VSMCs isolated from aortas of wild type (WT) and Adpn−/− mice. Proliferation markers, including cyclin D1, were increased in Adpn−/− cells, and AMPK activity and differentiation markers were reduced. Cells were analyzed for matrix metalloproteinases (MMPs) which are responsible for extracellular matrix (ECM) remodeling, a contributing factor to the VSMC phenotypic switch. We found that MMP protein expression and activity, measured by zymography, were reduced in Adpn−/− cells. To test whether the phenotypic switch is regulated by AMPK, we treated cells with Metformin, an AMPK activator, or AdipoRon, an activator of Adpn receptors. Metformin and AdipoRon reduced Cyclin D1 and MAPK signaling in Adpn−/− cells, suggesting that AMPK prevents VSMC proliferation; however, none of the treatments yielded an effect on markers of differentiation.

These data illustrated that Adpn regulates proliferation through an AMPK‐dependent signaling pathway, while regulating differentiation by an AMPK‐independent mechanism. The role of MMPs in VSMC differentiation in response to Adpn remains to be elucidated.
A Pipeline for Expression, Functionalization, and Characterization of Human Lectins Involved in Innate Immunity
Authors: Dodge Gregory , Bartlett Helen , Isabella Christine , Diehl Roger , Dugan Amanda , Kiessling Laura , Imperiali Barbara
Published: April 2020

In humans, a collection of diverse proteins known as lectins are responsible for binding to glycan epitopes. Recognition of extracellular glycan epitopes represents a key facet of the innate immune response in eukaryotes. These epitopes can be used to distinguish cell types, self‐from non‐self, and perhaps even pathogenic from commensal bacteria. Understanding human lectin specificity can uncover mechanisms underlying innate immunity. Additionally, lectins represent attractive reagents for the development of glycan‐reading probes. Here, we demonstrate our efforts to develop and deploy a pipeline for recombinant expression, functionalization, and characterization of a subset of human lectins, including C‐type lectins, ficolins, jacalin‐type lectins, galectins, and intelectins. Target lectins can be efficiently expressed and purified, functionalized with either biotin or a fluorophore via sortase‐mediated ligation under flow, and screened for glycan binding. The success of this pipeline has been validated in preliminary studies showing that the recombinant lectins can engage with glycans from mammalian glycan arrays, bacterial glycan arrays, purified mucins, or human biological samples. These experiments highlight the potential of lectins as valuable molecular probes that fill a gap in the current molecular toolkit for glycan analysis.

Support or Funding Information

NIH U01 CA231079‐01
Evaluation of melatonin therapy on improving the metabolic disorders associated to obesity in rats
Authors: filali houda , kadil Youness , Ettaqui Soukaina
Published: April 2020

Obesity is a worldwide epidemic and there are many diseases associated with it. Recent studies suggested that melatonin may be involved in the regulation of body mass and metabolic homeostasis through different mechanisms.

To this purpose, we conducted an experimental study on 24 adults male Wistar rats, separated into 2 control groups and 1 test group. Our experimental protocol took place in three phases, an obesity induction phase in which the rats were subjected to a hyper fat diet “Cafeteria diet”. A therapeutic phase in which the control group received NaCl as a placebo, the test group was treated with melatonin at a dose of 5mg/kg/day. The groups were randomized and received treatment intraperitoneally for 6 weeks. A Biological blinded evaluation for lipidic, glucidic, and inflammatory (CRP) assessments were performed for each group.

The results of our study showed that the Cafeteria diet significantly increased body weight with a gain of up to 126g (p=0.00). The biological analysis of the blood of rats treated with melatonin showed a decrease in blood glucose levels, triglycerides, LDL, HDL, CRP and total cholesterol compared to the initial values. Plasma analysis results obtained at the end of the therapeutic phase between melatonin and placebo‐treated rats showed a significant decrease in total cholesterol (p=0.002), LDL (p=0.005), HDL (p=0.014) and blood glucose (p=0.013). The difference between the two groups for CRP (p=0.084) and triglycerides (p=0.116) was not significant.

Based on our results melatonin seems to have a beneficial effect against metabolic disorders related to obesity.

Support or Funding Information

N/A
The role of depressive symptoms on brain blood vessel pulsatility in humans
Authors: Sanjana Faria , Richey Rauchelle E. , DeConne Theodore M. , Saunders Erika F. , Brothers R. Matthew , Martens Christopher R. , Greaney Jody L.
Published: April 2020

The prevalence of major depression continues to increase, particularly in young women. Depression is associated with increased risk for cardiovascular disease and cognitive impairment later in life. Peripheral vascular dysfunction, including stiffening of the large elastic arteries, is evident in adults with depression and may mediate the relation between depressive and neurocognitive symptoms by exposing the cerebral vasculature to damaging pulsatile pressure waves. We hypothesized that increased depression symptom severity would be associated with more pulsatile blood flow through the middle cerebral artery (MCA) as assessed by the pulsatility index (PI). Transcranial Doppler ultrasound was used to measure MCA cerebral blood velocity (CBV) in 40 young adults (25 F/15 M, mean age: 24±5 y) having a wide range of depressive symptom severity. MCA PI was calculated as the difference between maximum CBV and minimum CBV normalized to the mean CBV. Self‐reported depressive symptom severity was derived using the Patient Reported Outcome Measurement Information System (PROMIS; T‐score). The association between PI and depressive symptom severity was quantified using linear regression; sex differences were assessed using multiple linear regression. PI (mean±SE: 0.8±0.02; range: 0.6–1.0) was negatively correlated with depressive symptom severity (mean±SE: 51.3±1.6; range: 37.1–71.9) (r2=0.12, P=0.03). There was no difference in the relation between depressive symptom severity and PI between men and women (R2=0.09, P=0.39). Lower pulsatility of the cerebral arteries in adults with more severe depressive symptoms may indicate a possible compensatory role to protect the brain from damaging pressure waves. Future studies should examine the mechanisms by which depression contributes to decreased pulsatility of cerebral blood vessels and stiffening of large elastic arteries in young individuals, including the possible role of augmented cerebral autoregulation, and whether this mechanism changes with advancing age.

Support or Funding Information

NIH Grant P20GM113125
CHRONIC ALCOHOLISM ASSOCIATED WITH DIABETES INDUCED APOPTOSIS IN THE CORPUS CAVERNOSUM IN RATS
Authors: Cirino Mucio Assis , Schimming Bruno , Novais Paulo Cezar , Lizarte Fermino S. , Melo de Carvalho Camila A. , Fazan Valeria P. S. , Tirapelli Daniela P. C. , Molina Carlos A. F. , Junior Silvio Tucci , Tirapelli Luis F.
Published: April 2020

The association between diabetes mellitus and erectile dysfunction has been studied for many years. Erectile dysfunction secondary to angiopathic, neuropathic and myopathic damage is one of the main complications of diabetes in men. Other reproductive complications include ejaculatory dysfunction, hypogonadism, modified semen parameters, and delayed puberty. On the other hand, ethanol has various effects too, on male sexual activity, in small amounts improves erection and increases libido because of its vasodilatory effect; however, large amounts can cause central sedation, decreased libido, impaired the penile tumescence, and transient erectile dysfunction. Chronic alcoholism may cause hypogonadism and polineuropathy that may affect penile nerve function. Therefore, chronic alcoholism induced dysfunction of smooth muscle in the corpus cavernosum in rats. Although, effects of ethanol on the ultrastructure of cavernosal smooth muscle cells, elastic fibers and collagen content; and apoptosis in the erectile tissues of diabetic rats has been showed, no studies have been done to analyse the apoptotic effects of the diabetes associated with ethanol in the corpus cavernosum in rats.

Objectives

To investigate the effects of chronic ethanol consumption and diabetes on apoptotic process in the corpus cavernosum.

Material and Methods

Male Wistar rats were divided into four groups: control, diabetic (alloxan injection), ethanol, and ethanol‐diabetic (12 rats in each group). Fragments of corpus cavernosum were obtained and destined to the immunohistochemistry and real‐time PCR to analyze the protein expression of apoptotic genes CASPASE‐3, and CASPASE‐9.

Results

The immunoreactivity of CASPASE‐3 and CASPASE‐9 was diffuse and higher in the treated groups, however there was no significant difference between the experimental groups, only when compared with the control group. An increase was observed in the gene expression of CASPASE‐3 and CASPASE‐9 in the diabetic and ethanol‐diabetic groups when compared with to control and ethanol groups.

Conclusions

There was an increase in apoptosis in ethanol and ethanol‐diabetic rats. The association of these factors (ethanol and diabetes), jointly with neuropathy and arteriopathy could lead to development of erectile dysfunction.
Cotinine enhances the nicotine‐mediated decrease in respiratory burst amplitude recorded from the hypoglossal nerve in the brainstem spinal cord preparation from nicotine‐exposed neonatal rats
Authors: Flanigan Emily Gayle , Fregosi Ralph Frank , Wollman Lila Buls
Published: April 2020

Cotinine, the primary metabolite of nicotine, can be detected in the blood of infants born to mothers who used nicotine products during pregnancy and breast feeding, and is used as a marker of nicotine exposure. Previous research shows that prenatal nicotine exposure alters the regions of the brain that control breathing and results in impaired protective breathing reflexes in exposed human and animal neonates. These abnormalities are proposed as a mechanism to explain Sudden Infant Death Syndrome (SIDS), and maternal smoking is the number one risk factor for SIDS. While it has long been known that nicotine has direct effects on brain function and development, it has only recently been established that cotinine may act on the brain as well. Both nicotine and cotinine bind to nicotinic acetylcholine receptors (nAChRs) which are expressed on neurons throughout the nervous system. Preliminary data using the in vitro brainstem spinal‐cord preparation from nicotine‐exposed neonatal rats shows that activation of nAChRs produces a decrease in the amplitude of respiratory bursts recorded from hypoglossal nerve roots (−30.74±18.2 % baseline, n=4), and that this effect is enhanced by cotinine (−60.34±16.1 % baseline, n=4). This supports the hypothesis that cotinine modulates the function of nAChRs in regions of the brain that control breathing (P<0.0001).

Support or Funding Information

5R01HD071302‐07
Effects of the G Protein‐coupled Estrogen Receptor on Cardiovascular Remodeling Induced by Chronic Sympathetic Activation
Authors: Tan Derick , Twohey Stephanie , Allen Patrick , Giles Jennifer , Winter Lori , Clayton Sarah , Tran Quang-Kim , Eric Wauson
Published: April 2020

Estrogens modulate cardiovascular function via the classical nuclear hormone receptors ERα and ERβ, and the more recently discovered G protein‐coupled estrogen receptor (GPER). While the activation of GPER elicits many cardioprotective effects, the mechanisms behind these effects are incompletely understood. Sympathetic nervous activities are essential for normal cardiac functions; however, chronic sympathetic activation is an important contributor to maladaptive cardiovascular remodeling that leads to cardiovascular diseases. Our recent data demonstrate that GPER activity moderates the increases in myocardial contraction and calcium dynamics stimulated by acute activation of the cardiac beta1 adrenergic receptor. However, the effects of GPER activation on the cardiac manifestations in response to chronic sympathetic activation are unknown. We hypothesized that GPER activation would reduce cardiovascular remodeling induced by chronic norepinephrine (NE) administration. In adult female ovariectomized mice, administration of NE (10 mg/kg/day) via osmotic pumps for 8 weeks increased heart weight, kidney weight, cardiomyocyte size as assessed by wheat germ agglutinin staining, left ventricular wall thickness, and aorta wall thickness. These changes occurred without an apparent increase in blood pressure assessed by tail cuff measurements. These effects were reduced by the co‐administration of GPER agonist G‐1 (120 mg/kg/day), while administration of G‐1 alone at the same dose had no effect. These initial data indicate that the GPER agonist G‐1 can reduce cardiovascular remodeling caused by chronic sympathetic activation.

Support or Funding Information

DMU IOER 05‐19‐06
Evaluation of FDA Drugs to Identify Modulators of Multidrug Resistance Associated Protein 1 (MRP1) Expression Levels in HEK293 MRP1 Overexpressing Cells.
Authors: Loecker Josh , Poku Vivian Osei , Iram Surtaj
Published: April 2020

The Multidrug Resistance Associated Protein 1 is a key feature in drug resistance within cancerous cells. This protein is involved in efflux of a broad spectrum of substrates, including organic anions and several chemical therapeutic agents, like doxorubicin. In addition, MRP1 has implications in separate physiologies such as cardiovascular disease and age‐related macular degradation. As such, its overexpression has been implicated in multidrug resistance of several cancers.

This project is aimed at evaluating the effect of chemotherapeutic agents on the expression levels of MRP1 in HEK293/MRP1 cells. A unique library of FDA drugs were screened using a high throughput In‐Cell ELISA Assay. A total of 88 drugs were screened. Drugs that showed modulation of MRP1 protein expression above 30% were considered as Hit compounds. A significant number of hits were identified, from which some of the drugs increased MRP1 protein expression and decreased its expression.

This protein enhances the cells ability to efflux toxic compounds, such as anticancer drugs, out of the cell. A cell’s ability to increase expression of this protein results from a genetic mutation. As a result, higher concentrations of traditional anticancer drug compounds are required to demonstrate similar results as cells that do not over express MRP1. However, discovery of drugs that can decrease the expression of this protein would play a valuable role in assisting the fight against cancer.

Support or Funding Information

I would like to thank Jack and Mary East for their support and funding for this project
Cyclooxygenase Inhibition Increases the Sympathetic Response to Hypercapnia
Authors: Eisenmann Nicole A. , Howery Anna J. , Miller Kathleen B. , Eldridge Marlowe W. , Barnes Jill N.
Published: April 2020

Cyclooxygenase (COX) catalyzes the formation of prostaglandins and influences blood flow regulation. Previous studies have shown that COX inhibition reduces cerebral blood flow velocity and cerebrovascular reactivity to CO2 (CVR), however, the effects of COX inhibition on neurovascular regulation is unknown. Sympathetic nerve activity is an important modulator of blood pressure and blood flow. The purpose of this study was to determine the effect of COX inhibition on the muscle sympathetic nerve activity (MSNA) response to hypercapnia during CVR testing.

Methods

Fifteen healthy adults were evaluated (M/F = 7/8; age = 40 ± 21 years; BMI = 24 ± 2 kg/m2) before (Pre) and 90 minutes after oral administration of the COX inhibitor indomethacin (Indo), given at 1.2mg/kg. Mean arterial pressure (MAP), heart rate (HR) and MSNA were measured continuously throughout the protocol. After baseline measurements were recorded, participants were evaluated breathing 6% CO2. Results are reported as mean ± SE.

Results

At baseline, HR was 54 ± 2 bpm, MAP was 100 ± 4 mmHg, MSNA burst frequency was 24 ± 2 bursts/min and MSNA burst incidence was 45 ± 4 bursts/100hb. The change in HR when exposed to hypercapnia was not different after Indo (Pre: 6 ± 2 bpm vs. Indo: 4 ± 1 bpm; p>0.05). The change in MAP (Pre: 0 ± 2 mmHg vs. Indo: 6 ± 1 mmHg; p<0.05), MSNA burst frequency (Pre: 1 ± 1 bursts/min vs. Indo: 4 ± 1 bursts/min; p<0.05), and MSNA burst incidence (Pre: −3 ± 2 bursts/100hb vs. Indo: 4 ± 2 bursts/100hb; p<0.05) was greater after Indo administration.

Conclusion

The sympathetic response to hypercapnia was greater following the administration of Indo. Taken together, these results suggest that hypercapnia‐induced sympathoexcitation may be altered following COX inhibition. Future studies could evaluate if the sympathetic responses to hypercapnia vary with age or sex.

Support or Funding Information

NIH HL118154
Precisely Targeted Reporters Reveal ABCG2 Regulation in Live Lung Cancer Cells
Authors: Kovacsics Daniella , Brózik Anna , Tihanyi Borbála , Matula Zsolt , Borsy Adrienn , Mészáros Nikolett , Szabó Edit , Németh Eszter , Fóthi Ábel , Zámbó Boglárka , Szüts Dávid , Várady György , Orbán Tamás I. , Apáti Ágota , Sarkadi Balázs
Published: April 2020

Expression of the ABCG2 multidrug transporter is a marker of cancer stem cells and an indicator of poor disease outcome. Understanding the effect of pharmacotherapy on ABCG2 expression is critical in guiding therapy recommendations and may help rational drug development. Engineered reporter cell lines are useful in monitoring gene regulation and protein activity in live cells but rely on precise targeting to preserve native regulatory functions. Here, we generated a number of precision‐engineered, fluorescent human cell lines that interrogate ABCG2 regulation in live lung cancer cells. Using homology‐based CRISPR engineering, we targeted a promoterless eGFP to the translational start site of the ABCG2 gene, generating ABCG2 knockout and in situ tagged ABCG2 reporter cells. Using the engineered cell lines, we show that anti‐cancer medications, HDAC inhibitors and hypoxia‐mimicking agents induce ABCG2 expression. In addition, we provide evidence that the glucocorticoid receptor is a specific, positive regulator of the ABCG2 gene. To our knowledge, this is the first fluorescent reporter assay designed to follow the endogenous regulation of a human ABC transporter in live cells. The information gained may direct therapy recommendations and assist rational drug design.

Support or Funding Information

This research has been supported mainly by OTKA‐NKFIH grant NK 115375, but also by NKFIH K104903 and the NVKP_16‐1‐2016‐0005 program from the National Research, Development and Innovation Office. This work has also been supported by research grant OTKA‐NKFIH K128011, awarded to GV. A Borsy was supported by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences (HA: BO/00579/17/5), through the New National Excellence Program of the Ministry of Human Resources (HA: ÚNKP‐18‐4‐SE‐11).
The Development of an Educational e‐Module to Teach the Pathogenesis of Cancer Utilizing an Anatomical Donor Case Study
Authors: Fortney Lane , Kowalewska Jolanta , Kolheffer Kerwin , Elzie Carrie , Wellman Laurie
Published: April 2020

Introduction

There is a significant need for understanding the complexity and gravity of cancer pathogenesis as it continues to be a leading cause of death worldwide. During a gross anatomy lab, students encountered a rare malignancy, Uterine Leiomyosarcoma, in one of the donors. After further investigation and confirmation of the diagnosis, this experience was used to develop an educational e‐module to teach cancer pathogenesis.

Methodology

Relevant measurements and photographs were obtained, as well as representative sections of the uterine tumor specimen and distant metastases. The Eastern Virginia Medical School Biorepository performed histological preparations from these tissues, including standard hematoxylin and eosin preparations, as well as staining for smooth muscle actin (SMA). A faculty pathologist confirmed the diagnosis of leiomyosarcoma, applying current inclusion criteria. The Storyline Articulate 360 software platform was used to generate a case‐based interactive e‐module with pre‐ and post‐test questions addressing the concepts of cancer pathogenesis, tumor heterogeneity, cancer nomenclature, lineage of differentiation, and cancer classification, relating these topics to the findings and likely clinical course of this case.

Results

The e‐module was beta tested for usability and accuracy with a focus group of faculty members who provided qualitative and quantitative feedback. The resulting e‐module will be implemented into a general pathology course taken by medical and graduate students in the spring semester of 2020. Data on usability, knowledge, and application of an anatomical donor case for understanding cancer pathogenesis will be collected. The outcomes of this process will be evaluated for future curriculum implementation.

Conclusion

The interactive e‐module developed is an effective pedagogical tool to reinforce students’ knowledge of cancer pathogenesis using an anatomical donor case study. Inspired by a pathological finding in a gross anatomy lab, this interdisciplinary educational opportunity integrates and enhances a pathology course from a dissection experience. The integration of this interactive e‐module into medical school curriculum has the potential to be a powerful resource for understanding the fundamental principles of neoplasia through a novel approach of utilizing findings from an anatomical donor.

Support or Funding Information

Support and Funding provided by the Contemporary Human Anatomy Program at Eastern Virginia Medical School
Pilot Evaluation of Telemetered Cardiac Hemodynamics in the Setting of Fully‐Implanted Programmable Drug Delivery in Rats: A New Tool for Cardiovascular Drug Discovery?
Authors: Bennett Nikki Luis , Zambataro Christopher L. , Velie Troy , Bogie Heather , Shahidi-Latham Sheerin Luis , Ferguson Brian , del Rio Carlos Luis
Published: April 2020

Introduction

The mechanistic understanding of pharmacokinetics–pharmacodynamics (PK/PD) relationships is a key component of modern cardiovascular drug discovery. However, given that both PK properties and/or experimental techniques (such as anesthesia) can alter auto‐regulatory/autonomic control in vivo (thus masking true PD responses), tools permitting the determination of translationally‐relevant PK/PD relationships for cardio‐active agents are limited. This pilot study established the feasibility of combining implantable programable infusion pumps and telemetered cardiac hemodynamic monitoring for the assessment PK‐independent PD responses in untethered conscious rats.

Methods

Healthy Sprague‐Dawley rats (n = 7) were chronically instrumented with both a digital telemetered micro‐manometer catheter (HD‐S21, Data Sciences International) in the LV, and with a fully‐implanted programmable/refillable infusion pump in a femoral vein (iPRECIO® SMP‐200). The pumps were programmed, a priori, to perform (after a 21‐day recovery period) a two‐arm drug delivery protocol, consisting (each arm) of a 72h active period (ON, @12.5μL/hr) followed by a 96h washout (OFF period, on patency mode @1μL/hr); prior to each active period the pumps were loaded with either vehicle (CTRL) or a novel pure negative inotrope (MYK‐581), while normal saline was loaded prior to OFF periods. Left‐ventricular pressures (LVP) and derived functional indices were evaluated continuously (via telemetry) before/during active periods, while cardiac function/geometry were assessed (via echocardiography) prior to the end of the active period.

Results

Fully implanted conscious rats had normal cardiac hemodynamics (HR: 339 ± 8 bpm, LVPmax: 119±3 mmHg) and cardiac function (EF: 76 ± 2%, FS: 47± 1%, and dLVP/dtmax: 9275 ± 369 mmHg/s), showing normal circadian oscillations. Similar observations were made during vehicle administration (e.g., EF: 77 ± 1%, and dLVP/dtmax: 9585 ± 891 mmHg/s), while marked reductions in both echocardiographic (EF: 45 ± 8.7%, FS: 24 ± 6%, P < 0.05 vs. VEH) and LVP‐derived (dLVP/dtmax: −21 ± 6%, or 7313 ± 819 mmHg/s, P < 0.05 vs. VEH) functional indices were observed during the MYK‐treatment period.

Conclusions

Taken together, these data demonstrate that telemetered hemodynamics and fully‐implanted programmable/refillable infusion pump technologies can be successfully integrated in vivo in order to generate translationally‐relevant PK‐independent pharmacodynamics data in untethered conscious rats.

Support or Funding Information

These studies were supported by MyoKardia
Pyruvate Kinase M2 Deficiency Promotes Brown Fat Adipogenesis in Vitro and Enhances Adaptive Thermogenesis in Vivo
Authors: Alquraishi Mohammed , Puckett Dexter L. , Chahed Samah L. , BAETTAIEB AHMED
Published: April 2020

The prevalence of obesity and its comorbidities has sparked a worldwide concern to address rates of adipose tissue accrual. Pharmacological approaches to combat obesity have met with limited success despite intensive investments in their development. Although many differing strategies are being explored in cellular and molecular studies, many have proven ineffective at achieving long‐term positive outcomes. Thus, the search for novel approaches and new target must continue. Brown adipocytes are specialized to dissipate chemical energy as heat and are associated with improved metabolic phenotypes. Herein we demonstrate that altered expression of the glycolytic enzyme pyruvate kinase M2 (PKM2) promotes adipogenesis of mouse and human brown adipocytes. Additionally, modulation of PKM2 expression results in increased lipolysis through modulation of the activity of protein kinase A and hormone‐sensitive lipase (HSL). Importantly, PKM2 deficiency in brown adipocytes increases ATP turnover, and enhances basal and maximal mitochondrial respiration. Furthermore, mice with specific deletion of PKM2 in brown adipose tissue exhibited increased energy expenditure and resistance to cold‐induced alteration in core body temperature. Collectively, these findings identify PKM2 as a novel component of the molecular circuit that contributes to brown fat adipogenesis and adaptive thermogenesis, which may have potential therapeutic implications.

Support or Funding Information

This work was supported by the National Institute of Diabetes and Digestive and Kidney Diseases (R00DK100736) to A.B.