bluuebunny/crossref_metadata_2025 huggingface dataset:
Insulin Resistance and Food Cravings
Authors: Fieselmann Krista N. , Sharma Rashmi S. , Maroney Owen H. , Ebner Rachel E. , McCrory Megan A.
Published: June 2021

BACKGROUND

Individuals with insulin resistance (IR) have been suggested to have more food cravings than those without IR, particularly sweet cravings. However, little research has been done in this area. We tested the hypothesis that individuals with IR would have more frequent and stronger cravings, perceive themselves as less able to ignore cravings, and ‘give in’ to cravings more often than those without IR.

METHODS

We analyzed data from healthy, non‐smoking, weight stable adults (aged 18–58 y; BMI 19–35 kg/m2) who had participated in recent studies in our lab and reported energy intakes (EI) plausibly (within ±25% of energy requirement (Huang et al 2005)). Cravings were assessed by questionnaire (Hill et al 1991). IR was defined as homeostasis model assessment (HOMA) >1.775 (Esteghamati et al 2010).

RESULTS

Individuals with IR (n=28) had higher BMI (p=0.014) and were slightly more active (p=0.08) and younger (p=0.11) than those without IR (n=34). After controlling for confounders, those with IR had 45% less frequent cravings than those without IR (p=0.04), but no other craving variables differed significantly by IR status.

CONCLUSIONS

On average, cravings were not experienced differently in those with IR versus without IR. Nevertheless, perceived cravings may be important if they trigger intake of foods that may exacerbate IR. [Funding: DK075862]
The Relationship between Changes in Serum Leptin Levels and Changes in Bone Mineral Density (BMD) in Diabetes En Balance Program
Authors: Chukwueke Ihuoma Okezie , Fashola Yomi , Beeson Larry , Schulz Eloy , Firek Anthony , DeLeon Marino , Cordero-MacIntyre Zaida
Published: June 2021

Objective

To access the relationship between changes in serum leptin levels and changes in bone mineral density (BMD) in diabetic Hispanics

Methods

Thirty‐four Hispanic diabetic subjects participated in the three‐month study. The fan beam dual energy X‐ray absorptiometry (DXA) hologic 4500 version software was used to measure body composition. Paired t‐test was used to evaluate the mean difference between baseline and 3‐months. Non parametric test was used to evaluate the mean difference at body composition and serum leptin at baseline and three months. Spearman correlation coefficient was used to assess the change in Serum Leptin and Bone Mineral Density.

Results

Significant negative correlations (Spearman's) were observed at three months between transformed values of leptin (adjusted for trunk fat (r = −0.369, p=0.035) and pelvis BMD; leptin (adjusted for total fat), r = −0.372, p=0.033) versus pelvis BMD. The values obtained after three months when compared to baseline were: Total fat (30.41± 10.68 vs 29.50 ± 10.52, p< 0.001), trunk fat (16.54± 5.73 vs 15.93 ± 5.60., p<0.001), serum leptin (19.55±14.99 vs 16.34 ± 12.67 p<0.002).

Conclusion

There is a significant inverse relationship serum leptin and bone mineral density in diabetic Hispanics.

Sponsor: CMS 03‐00335 Health Services Research and NIH award 5P20MD001632
Associations between stable isotope ratios and animal‐derived dietary protein intake in Yup'ik eskimos
Authors: Beasley Jeannette M , Nash Sarah H , O'Brien Diane M
Published: June 2021

Stable isotope ratios, such as δ15N, of tissues reflect the isotopic pattern of food sources. Animal nitrogen has a higher isotope ratio than nitrogen derived from plant sources, because δ15N accumulates up the food chain. The objective of this study was to determine whether the δ15N stable isotope ratio is associated with self‐reported dietary animal protein intake (animal versus vegetable). We hypothesized δ15N would be associated with animal, particularly fish, intake in this population of Yup'ik Eskimos. We examined the relation between red blood cell (RBC) stable isotope ratios and self‐reported animal protein intake derived from four 24‐hour dietary recalls in a community‐based sample of 70 Yup'ik Eskimos. RBC δ15N was strongly correlated with proportion of protein intake from animal sources (r = 0.71). The results support the utility of RBC δ15N as a biomarker of proportion of protein intake from animal sources. Controlled feeding studies to evaluate the validity of stable isotope ratios as a method to assess dietary protein source in epidemiologic studies are warranted. This research was supported by the Institute for Translational Health Sciences Visiting Scholars Program, NIH NCRR (P20 RR16430‐10) and NIH NIDDK R01DK07442.
Parity and Pre‐pregnancy Obesity are Independently Associated with Delayed Lactogenesis in Women with History of Gestational Diabetes: Preliminary Results from the SWIFT Study
Authors: Matias Susana Lilian , Chiang Vicky , Dewey Kathryn G , Gunderson Erica P
Published: June 2021

We used baseline data (n=1014) from SWIFT, a prospective study of women with gestational diabetes mellitus (GDM) within Kaiser Permanente (KP) in California, to determine the incidence of delayed lactogenesis and its association with selected maternal prenatal and delivery characteristics. Onset of lactogenesis II was assessed at baseline (6–9 weeks postpartum) by maternal report as well as pre‐pregnancy and pre‐delivery weights, and height was measured. Delayed lactogenesis was defined as onset of lactogenesis II occurring after 72 hours postpartum. Results from a prenatal 3‐hour 100 g oral glucose tolerance test (OGTT) were obtained from KP electronic records. Severity of GDM was measured by number of abnormal OGTT plasma values. Delayed lactogenesis occurred in 33% of postpartum nondiabetic women. After adjusting for maternal, labor and delivery characteristics, primiparity and pre‐pregnancy obesity were independently associated with delayed lactogenesis in this population. High priority for early postpartum lactation support should be given to GDM women who are obese before pregnancy and primiparous.

This study was funded by the National Institute of Child Health and Human Development (NICHD), R01 HD050625, R01 HD050625‐03S1, and R01 HD050625‐05S. This project was also partially supported by the National Institutes of Health National Center for Research Resources UCSF‐CTSI UL1 RR024131.
Effect of Microfluidization on Bioaccessibility of Carotenoids from Chlorella ellipsoidea during Simulated Digestion
Authors: Kang Kyungsu , Cha Kwang Hyun , Koo Song Yi , Song Dae-Geun , Nho Chu Won , Pan Cheol-Ho
Published: June 2021

This study evaluated the bioaccessibility of carotenoids from marine species Chlorella ellipsoidea using simulated digestion. In order to increase the carotenoids bioaccessibility, C. ellipsoidea was pulverized by microfluidization. The major carotenoid in C. ellipsoidea was zeaxanthin with two minor carotenoids, antheraxanthin and β‐carotene. The mean microalga particle size was significantly reduced from 2463 to 361 nm with the microfluidization treatment. After microfluidization, the content of zeaxanthin and β‐carotene in C. ellipsoidea was not changed compared with untreated group. But, the antheraxanthin content was significantly diminished showing antheraxanthin, a kind of epoxyxanthophylls, was fragile to physical processing. The bioaccessibility of carotenoids in untreated C. ellipsoidea was very low and approximately 97% of total C. ellipsoidea carotenoids was not micellized during simulated digestion. The microfluidized microalga (at 20,000 psi) was ten times more effective for zeaxanthin and β‐carotene micelles production compared with untreated C. ellipsoidea, showing the effect of microfluidization on higher carotenoids bioaccessibility (zeaxanthin: 32.60%, β‐carotene: 18.19%). These results indicate that microfluidization may be useful for improving zeaxanthin and β‐carotene bioaccessibility from C. ellipsoidea during digestion process.
Digital clubbing: have we lost its anatomical form and clinical relevance
Authors: Benninger Brion
Published: June 2021

INTRODUCTION

Hippocrates first described digital clubbing (DC) in a patient with empyema in the 5th century BC. Although the majority of medical trainees and qualified clinicians are aware of DC, few can confirm its diagnosis and are unable to discuss its associations with multiple systems despite its prevalence in the population. DC can be diagnosed with the combination of observation, provocative testing, and palpation. The objective of this study was to investigate the diagnosis of DC, its associations with the body's systems, radiological diagnosis and its fate regarding treatment.

METHODS

Literature search was conducted on journals, specialty texts and websites regarding DC, its medical associations, radiological diagnosis and its role in physical examination.

RESULTS

Literature search revealed that DC could be confirmed using x‐ray. No contemporary articles or studies described its use during clinical examination or its fate.

DISCUSSION

DC is a relatively well‐known term. Few qualified clinicians and fewer trainees can discuss its diagnosis, pathological associations, clinical exam integration or its fate. DC will subside with successful treatment of the associated pathology.

CONCLUSION

This study suggests with anatomical and etiological understanding of DC, health care providers can diagnose and link it to one of its several pathologies and monitor the pathology with the presence of DC.
Development of Integrated Clinical Skills and Musculoskeletal Modules
Authors: Doroudi Majid , Jong Benjamin , Clarkson Paul , Joughin Karen
Published: June 2021

Many students find memorizing MSK materials difficult and daunting, and often find themselves feeling overwhelmed by the number of muscles, bones, innervations, vessels and special tests that they must commit to memory. The overall objective of this study was to encourage students to adopt a more anatomical approach to clinical skills and also to show explicitly the relevance of musculoskeletal anatomy, thus moving away from rote memorization of physical exam skills and anatomical knowledge. In order to accomplish the goal of interactive case‐based modules we used the Articulate Engage software suite to create a module that progresses slide by slide in order to encourage a logical flow from patient presentation through to treatment. Using Articulate we were also able to add interactive labeled diagrams to review MSK anatomy, video and pictures to study clinical skills and integrated quizzes to test student knowledge. The MSK modules were evaluated objectively from the survey questionnaire results and subjectively from the comments from the results. Before the modules 48% of students felt they knew the MSK materials well or very well. After the modules 77% of students felt they knew the MSK materials well or very well. Over 95% of students surveyed strongly or very strongly wanted to see more modules like this.
The effects of consuming a pistachio snack versus a refined carbohydrate snack on blood lipids, blood glucose, body weight, and body composition in young, healthy adults
Authors: Hernandez Lisa M , Meichtry Nicole E , Soto Daniel , Sringkarawat Sopida , Hong Mee Young , Hooshmand Shirin , Kern Mark , Petrisko Yumi
Published: June 2021

A variety of nuts have been well‐established to produce lipid‐lowering effects in hyperlipidemic and/or obese subjects. Fewer studies have investigated the effects of nut consumption in young, healthy adults. The objective of this parallel‐arm study was to determine if pistachio nut snack consumption would favorably affect plasma lipid profiles, food intake, body weight, and body composition in a non‐obese, normolipidemic population compared to a refined carbohydrate snack. Forty‐one participants with a mean age of 23.0 ± 3.4 years, a mean BMI 23.1 ± 3.7 kg/m2, and mean percent body fat of 25.1 ± 7.8% were divided into two groups consuming isoenergetic (150 kcal) snacks of either 26.5 g pistachios (n=20) or 38.5 g pretzels (n=21), twice daily. After 3 weeks, body weight and percent body fat were significantly decreased in the pistachio snack group (P=.014 and P=.026, respectively). No significant differences were detected within or between groups for plasma total cholesterol, LDL‐c, HDL‐c, triglycerides or blood glucose (P>.05), but a trend towards a significant increase in serum triglycerides in the pretzel snack group (P=.056) was detected. These results suggest that short‐term consumption of a pistachio nut snack can decrease body weight and fatness as compared to a refined carbohydrate snack in young, healthy adults. Partial funding support by Paramount Farms, Inc.
Doped Halloysite Sutures for Clinical Medicine
Authors: Vandenberg Herb , Weisman Jeff , Mills David
Published: June 2021

In the ER, wounds are the third most common injuries treated with sutures that stop the bleeding and close the wound. Subsequent wound management is mainly treatment for postoperative infections. A suture providing sustained release of antibiotics/antiseptics directly into the wound site is highly desired. Halloysite is a biocompatible clay nanotube. It is a double‐layered aluminosilicate, structurally similar to kaolin, with a diameter of 10–50 nm. Halloysite was loaded with antiseptics and antibiotics and mixed with poly‐e‐caprolactone and using the electrospinning technique was spun into various fibrous materials including sutures, gauze and wound dressings. These materials were added to confluent bacterial cultures of S. aureus and E. coli and maintained (via serial culture) over a two‐month experimental period. Control cultures consisted of halloysite/PCL fibrous materials minus antibacterials. Doped halloysite/PCL materials created and maintained a zone of growth inhibition over the two‐month period. Control cultures failed to inhibit bacterial growth. Results suggest that postsurgical infections could be controlled, reducing patient cost, promoting quick recovery, and return to normal health.
Polyphenols‐rich fruits attenuate cell migration in vitro in human umbilical vein endothelial cells (HUVEC) exposed to glucose and free fatty acids
Authors: Chang Claire Wei-Ju , Kangath Archana , Krishnankutty Sandhya , Tadapaneni Ravi Kiran , Edirisinghe Indika , Burton-Freeman Britt , Jackson Lauren
Published: June 2021

Elevated levels of plasma glucose and free fatty acids (FFA) are associated with increased risk of cardiovascular diseases. Previous studies suggested that fruits rich in polyphenolic antioxidants modulate endothelial cell migration via a redox‐sensitive mechanism. We hypothesized that polyphenol‐rich extracts of strawberry (SB) and wild blueberry (WB) would enhance cell migration in vitro in endothelial cells exposed to glucose and FFA. Human umbilical vein endothelial cells (HUVEC) were treated with SB or WB (0.2 mg/mL) for 2 h followed by glucose (5–15 mM) and/or FFA (0.5–2 mM) at concentrations intended to mimic normal or elevated concentrations found in human plasma for 15 h (overnight). Thereafter, HUVEC monolayer was scratched with sterile pipet tip, washed twice (phosphate buffered saline, PBS), and new media containing 2% serum added. Number of cells migrated beyond the scratch line was monitored for 12 h. After 8 h, cells treated with glucose (15 mM) and/or FFA (2 mM) showed significantly less cell migration compared to the PBS control (P<0.05, n=4). However, cells treated with 0.2 mg/mL SB or WB extracts with glucose (15 mM) and/or FFA (2 mM) in the media exhibited significantly (p<0.05, n=4) enhanced migration compared to cells not exposed to fruit extracts. The results suggest a possible protective effect of SB and WB on endothelial cell migration in cells exposed to high concentrations of glucose and/or FFA.
The Effect of Aqueous Mulberry Leaf Extract on Glucose Metabolism in Subjects with Impaired Fasting Glucose
Authors: Ok Hyang Mok , Park Ji Eun , Ryu Na Hee , Kim Joo hee , Kwon Oran
Published: June 2021

Mulberry leaf water extract (Morus alba L.) contains various functional components such as 1‐deoxynojirimycin (DNJ), soluble dietary fibers, resveratrol, quercetin, rutin, astragalin, and ¥ã‐aminobutyric acid. Previously we reported that aqueous mulberry leaf extract (MLE) had effects on elevated expression of insulin receptors as well as alpha glucosidase inhibition activity. Here we report a further study in which the blood glucose‐lowering effects of dose of aqueous mulberry leaf extract (MLE), containing 1‐deoxynojirimycin, was examined in subjects with impaired fasting blood glucose (FBG). Ingestion of MLE for 4 weeks decreased the elevated plasma glucose level in the postprandial blood glucose. The effective daily dose of MLE was observed in the group of 6 g of MLE (corresponding to 25.8mg of DNJ). Similarly, subjects taking MLE showed a decrease in plasma insulin and C‐peptide secretion compared to subjects taking a placebo. * This project was supported by the Ministry of Knowledge & Economy (RITD program, Project No. 70004683) and the Ministry of Education, Science and Technology (Brain Korea 21, Project No. 2006‐0519‐4‐7).
Differentially expressed genes and lincRNAs throughout oral squamous cell carcinoma development
Authors: Oliveira-Costa João Paulo , Carvalho Alex Fiorini , Lourenco Silvia Vanessa , Kowalski Luiz Paulo , Carraro Dirce Maria , Soares Fernando Augusto , Ribeiro-Silva Alfredo
Published: June 2021

Oral squamous cell carcinoma (OSCC) development still poorly understood, in terms of genome wide alterations leading to high invasiveness and aggressive behavior, and also, which long intergenic non‐coding RNAs (lincRNAs) are differentially expressed throughout its progression. Using 36 OSCC samples, we performed cDNA and lincRNA microarrays, comprising whole human genome and more than 7000 lincRNAs, compairing differentially expressed genes and lincRNAs, from T1 staging to T4 TNM staging. We found 94 genes and 12 lincRNAs with expression increasing or decreasing during OSCC development, including ERBB and HOX families. As the first study adressing OSCC developmental changes in either gene and lincRNAs expression, our results point to differentially expressed protein families that might be aims of deeper studies in OSCC, and the characterization of lincRNA mechanisms in OSCC development.
Estrogens inhibit TNF‐alpha induced apoptosis in an autoimmune disease: Sjogren's syndrome
Authors: Chakraborty Rima , Torres-Romero Luis Francisco , Wang Yang , McArthur Carole , Molteni Agostino
Published: June 2021

Sjogrens Syndrome (SS) is a chronic autoimmune disease characterized by keratoconjunctivitis, salivary and lacrimal gland inflammation and rheumatoid arthritis. Tissue destruction in SS is related to cellular apoptosis and it may be clinically treated by glucocorticoids (dexamethasone). Tumor Necrosis Factor alpha (TNF‐α), a proapoptotic cytokine, is increased in SS. Reported here is the the effect of the estrogen E2 administration on TNF‐α induced apoptosis on cultures of human salivary gland (HSG) ductal cells. The cells were harvested after 4 hour culture in presence of a combination of TNF‐α and cycloheximide. Total RNA was extracted by RNA aqueous‐Midi total RNA isolation System (Ambion, Ca) and messenger RNA (MRNA) expression evaluated using selectively targeted gene microarrays on gene known to be involved on human apoptosis (GE arrays, S series) containing 67 genes. Results show a significant reduction of apoptotic activity of HSG cells treated with E2 when compared to controls. Using gene microarray analysis an up and down regulation of different apoptotic associated genes including RIPK2, TNFRF9, TNFRS6, TANK, BIl‐X, BCL2AI was observed. The data suggests that E2 plays a role on the apoptotic cell regulation, opening a potential therapy for this disease.
Neutralization of IL‐13Ra1 receptor on Human CD4+ CD45RO+ Memory T cells Stimulates Release of IL‐17
Authors: Wilkinson Patrick , Farrell Francis
Published: June 2021

Interleukin 13 (IL‐13) has been shown to play a role in airway hyper‐responsiveness and immune cell‐mediated cytokine release in models of allergic airway inflammation. Additionally, neutralization of IL‐13 has been shown to improve lung function in a placebo‐controlled trial of adult asthmatics. Previous studies have shown that Th17 cells express a functional IL‐13 receptor and that IL‐13 attenuates IL‐17A, a cytokine involved in airway inflammation. The existence of a parallel IL‐13/IL‐17 axis in memory T cells, a source of inflammatory cytokines able to respond quickly to extrinsic insult and effect the tissue micro environment, has not been explored. In this study, human CD4+/CD45RO+/CCR7‐ effector memory T cells (TEM) were isolated from human blood and characterized by flow cytometry. Subsequently, cells were activated with anti‐CD3 and anti‐CD28 in the presence or absence of anti IL‐13Ra1 (20ng/ml) and anti IL‐4 (20ng/ml). Blockade of IL‐13Ra1 stimulated release of IL‐17A (8 fold) compared to unstimulated cells on day 5 post‐stimulation. Addition of an IL‐4Ra neutralizing antibody in combination with anti‐IL‐13Ra1 further increased IL‐17‐A expression compared to anti IL‐13Ra1 inhibition alone. These results suggest that increased IL‐17A production as a consequence of IL‐13 neutralization occurs in effector memory T‐cells.
Comparative studies on the effect of newer oral anticoagulants on platelet aggregation. Pharmacological and developmental implications
Authors: Chaudry Talhah , Aranda Christopher , Hoppensteadt Debra , Jeske Walter , Fareed Jawed
Published: June 2021

The newer oral anticoagulants include Dabigatran, Apixiban and Rivaroxiban. These drugs are developed for the DVT and stroke prevention in atrial fibrillation. These drugs are useful as alternate anticoagulants for heparin compromised patients. The purpose of this investigation was to study their effects on agonist induced platelet aggregation. Such agonist as epinephrine, collagen, arachidonic acis, ADP and thrombin were used in whole blood (multiplate) and citrated PRP systems (PAP8). In addition the interaction of these agents with heparin induced thrombocytopenia associated antibodies were also studied. Each of these agents were supplemented in citrated whole blood at a concentration range of 0–10ug/ml, agonists were added and the aggregation profile was recorded in both the whole blood and PRP. Serum from patients with HIT syndrome was also used to compare the aggregation profile of these agents with heparin. None of these newer oral anticoagulants produced any alterations of the aggregation profile in both the whole blood and platelet rich plasma (20–30;p=<0.05) with all of the agonists except thrombin. Dabigatran supplemented systems showed inhibition wheras Apixiban and Rvaroxiban did not produce inhibition of thrombin aggregation. These results suggest that newer oral anticoagulants do not impair the agonist induced platelet aggregation and will be useful in heparin compromised patients.
Acute Oral Bryostatin‐1 Treatment Improves Morris Water Maze Acquisition in a Transgenic Mouse Model of Alzheimer's Disease
Authors: Schrott Lisa M , Yi Ping , Johnson Gabriel S , Basting Tyler , Castor Trevor P , Alexander J. Steven
Published: June 2021

Recent research suggests that Bryostatin‐1 is neuroprotective via activation of protein kinase C isoforms, down regulation of pro‐inflammatory factors and angiogenic processes and the suppression of beta amyloid. Alzheimer's disease (AD) has been associated with these factors and therefore Bryostatin‐1 is an attractive candidate for ameliorating AD cognitive deficits. We evaluated the ability of Bryostatin‐1 to restore learning and memory in the BC3‐Tg(APPswe, PSEN1 dE9) 85Dbo/J mouse model of AD. This strain incorporates the human ‘Swedish’ (swe) amyloid precursor protein and presenilin mutations, which are associated with progressive human AD. A stable oil formulation of Bryostatin‐1 (5 μg/mouse) was administered by oral gavage 3 times the week prior to testing and then daily during 5 days of Morris Water Maze testing (4 trials per day). Treated mice were compared to untreated transgenic controls and wild‐type mice. Tg AD mice treated with Bryostatin‐1 had better performance in the water maze than untreated controls. Treated mice exhibited significantly shorter latencies and swimming distances to find the hidden platform, with no difference in their swimming speeds. The most profound effect was found in the early days of testing. The treated mice had performance nearly identical to that of the wild‐type controls. These data suggest that oral Bryostatin‐1 may be useful in treating cognitive decline associated with AD.

This research was supported by a grant from the NIA and NIH (1R44AG034760‐01A1)
Selective Inhibition of HDAC‐6 is Cardioprotective in an Endothelial Cell/Cardiomyocyte Co‐Culture Model
Authors: Procknow Jesse , Liu Yanan , Weihrauch Dorothee , Pratt Phillip , Kersten Judy
Published: June 2021

The acetylation state of α‐tubulin (αt) regulates nitric oxide synthase‐generated nitric oxide in endothelial cells (EC) and this pathway may be important in protecting against cellular injury. αt is a substrate of histone deacetylase‐6 (HDAC‐6); therefore, EC‐mediated protection of cardiomyocytes could be HDAC‐6‐dependent. Human coronary artery EC were co‐cultured with isolated neonatal rat cardiomyocytes at a 1:6 ratio (CC) and subjected to two hours of hypoxia and two hours of reoxygenation. CC were pretreated for one hour prior to hypoxia with a nonselective [trichostatin A (TSA), 750 nM] or a selective HDAC‐6 inhibitor (tubacin, 50 nM). Hypoxia significantly increased HDAC‐6 expression in cardiomyocytes compared to normoxic controls measured by Western Blot (WB) and immunofluorescence intensity. Compared to control CC, lactate dehydrogenase activity, an index of cell injury, was significantly decreased in CC pretreated with tubacin when compared to TSA‐treated cells or vehicle. Tubacin also increased acetylated αt expression compared to control measured by WB. These results suggest that HDAC‐6 may be involved in cardioprotective signaling by modulating the acetylation state of αt. Future studies will examine the importance of HDAC‐6 in regulating other cardioprotective pathways, such as nitric oxide production. This work was supported by NIH Grant 1 T32 GM089586.
Formulation of Compounds by SAR Analysis for HIV‐1 Capsid Core Inhibition
Authors: Brown Joshua D , Summers Michael F
Published: June 2021

In the human immunodeficiency virus replication life cycle, thousands of the viral Gag polyprotein associate at the cell membrane. The virus buds to form a non‐infectious virion. Gag is then cleaved by protease. Capsid protein is released and assembles into the capsid core, creating an infectious virus. The capsid protein is an attractive inhibition site because its formation of the capsid core is crucial for viral infectivity. A compound, CAP‐1, has been identified that binds to the N‐terminal domain of the capsid protein (CAN) and inhibits capsid formation in vitro and in vivo, but the dissociation constant (Kd) is not within an acceptable range for clinical use. We have been testing new compounds for affinity with a Kd in the nanomolar range. NMR HSQC titrations have been used to calculate the Kd. The computer program, PyMOL, has been used to give a visual representation of the binding pocket conformation of the compounds bound to the capsid protein. A correlation has been found between the amount of unoccupied space within the pocket and the Kd. Using this information, several compounds’ pocket conformations and Kd values were screened. Taking a structure activity relationship (SAR) approach, four potential inhibitors were formulated. All four targets are currently in the synthesis phase. The results of the future NMR HSQC titrations will provide the data needed for the next steps in the drug design.
Tissue factor mediated generation of thrombin in prothrombin complexes is inhibited by anti‐Xa based parenteral and oral agents but not by parenteral and oral antithrombin agents. Results from proteomic profiling using protein chip arrays
Authors: Fareed Jawed , Hoppensteadt Debra , Kahn Daniel , Walenga Jeanine M. , Jeske Walter , Lewis Bruce
Published: June 2021

This study investigated the effects of direct factor Xa and thrombin inhibitors on tissue factor (TF) mediated activation of prothrombin complexes. Calcified Thromboplastin C and Innovin were used to activate buffered prothrombin complexes that were supplemented with graded concentrations of Xa inhibitors (Apixiban, Rivaroxiban, Otamaxiban) or thrombin inhibitors (Argatroban, Refludan, Hirudin, Melagatran). These mixtures were analyzed using a surface enhanced laser desorption ionization method over a molecular range of 0–150 kDa. Each of the prothrombin complexes provided a unique molecular spectrum with distinct peaks in 71–72 kDa range representing prothrombin. Upon activation, the prothrombin peak was diminished with the appearance of a 36–37 kDa peak representing thrombin. The factor Xa inhibitors produced a concentration dependent inhibition of the generation of thrombin whereas the thrombin inhibitors did not. As tissue factor plays an important role in the pathogenesis of thrombotic stroke and acute coronary syndrome, this data suggests that the anti‐Xa drugs may be more effective in the management of these syndromes. Furthermore, the observed myocardial infarcation signal with the oral thrombin inhibitors Dabigatran and Ximelagatran may be due to residual tissue factor activation of thrombin in patients treated with these drugs.
Generation of selective ligands for nicotinic acetylcholine receptors
Authors: Yamauchi John , Nemecz Akos , Gomez Kimberly , Ho Kwok-Yiu , Grimster Neil , Fotsing Joseph R , Weide Timo , Stump Bernhard , Talley Todd T , Fokin Valery V , Sharpless K. Barry , Taylor Palmer
Published: June 2021

Nicotinic acetylcholine receptors (nAChRs) are members of the cys‐loop receptor family located in synapses of the CNS, neuromuscular junctions, and autonomic ganglia. The α4β2 and α7 subtypes are the most abundant isoforms in the brain and associated with a range of disorders including; smoking addiction, schizophrenia, and Alzheimer's disease making them attractive pharmaceutical targets. Subtype selective ligands have been difficult to design due to the conserved nature of the binding site. The Aplysia californica and Lymnaea stagnalis acetylcholine binding proteins (AChBPs) are structural surrogates for the extracellular domain of nAChRs. We mutated regions of the binding pocket with the Aplysia AChBP towards α7 to obtain pentameric chimeras. We employed a structure‐guided strategy utilizing wildtype and α7‐chimera AChBPs to catalyze in situ azide‐alkyne triazole cycloaddition reactions generating compounds targeted for the α7 nAChR. Responses for α7 and α4β2 nAChRs were characterized using an in vitro fluorescent functional assay using a genetically encoded Ca2+ indicator. The majority of compounds were α7 agonists with some showing selectivity over α4β2. All compounds with α4β2 activity were identified as antagonists. These results illustrate the utility of in situ click chemistry and its potential expansion into membrane protein templates for generating selective allosteric modulators. Support: R37‐GM18360, UO1‐DA019372, NSF GK12 0742551
Human body‐on‐a‐chip: A tool to reproduce modulations of the methylation pathway
Authors: Prot Jean-Matthieu , Ariza-Nieto Magnolia , Shuler Michael Louis
Published: June 2021

Aberrant DNA methylation patterns are associated with many types of cancer. We are interested in understanding the faith of methyl donors implicated in the remethylation of homocysteine under extreme pathway overload and hepatic insulin resistance. The microenvironments known to affect epigenetic patterns and signaling between organs are being studied using DNA methylation arrays. The body‐on‐a‐chip is a microfabricated device combined with cell cultures. Several cell culture chambers are interconnected by microfluidic channels and present the same order of magnitude as in vivo cellular tissue constructs. This microdevice prevents dilution of important secretory factors with numerous advantages like volume reduction, dynamic cell culture and the possibility to accurately monitor the flow rate inside the microchannels. This human surrogate is being used to create a physiological microenvironment to study modulations of DNA methylation patterns and signaling between organ tissues such as GI/liver/Brain and adipose tissue. Modulations of input and surrogate biomarkers are being developed and plan to be immobilized within the device. This research is funded by The Department of Biomedical Engineering, Cornell University.
Right ventricle dysfunction in heart failure rats following myocardial infarction involves the increment of phospholamban to phosphorilated‐phospholamban ratio
Authors: Fernandes Aurélia Araújo , Júnior Rogério Faustino Ribeiro , Siman Fabiana Dayse Magalhães , Zoghaib Jorge , Vassallo Dalton Valentim , Stefanon Ivanita
Published: June 2021

The aim of this study was to assess right ventricle (RV) contractility and proteins involving in calcium transient early (1 week) and late phase (8 weeks) in rats with or without heart failure (HF) after myocardial infarction (MI). The Increment of left ventricle end diastolic pressure, lung weight and body weight ratio and right ventricle to body weight ratio was used to divide MI rats with (HF) and without HF (Inf). The scar size was not different between groups neither was correlated to signals of HF. Strips from RV were superfused with Krebs in an isometric apparatus. The inotropic response to Ca2+ and Isoproterenol were preserved early and reduced late after MI. Inversely, in the Inf group, the positive intropic response was reduced only in the early phase. The SERCA‐2a, phospholamban (PLB total), PLB phosphorylated and Na+/Ca2+ exchange proteins were not different among groups in the early phase. In the late phase there was a SERCA‐2a overexpression only in the Inf group (Sham=0.79±0.05; Inf=1.81±0.06*; HF=0,88±0.1; *P<0.05). There was an increment on SERCA/PLB ratio also in Inf group (Sham=0.8±0.08; Inf=1.33±0.12*; HF=0.87±0.09; *P<0.05). The PLB/PLBPratio was increased only in the HF animals at 8 weeks (Sham=1.19±0.19; Inf=1.72±0.24; HF=3.51±1.04* P<0.05). These results suggest that increased SERCA‐2a expression contribute to maintain RV contractility in Inf group. The reduced RV contractility in the HF group at 8 weeks could be related to the increment on PLB/PLBP ratio.
Race Affects Arterial and Ventricular Elastance Responses to Endurance Exercise Training
Authors: Lane Abbi Danielle , Yan Huimin , Ranadive Sushant M , Cook Marc D , Kappus Rebecca M , Wilund Kenneth , Woods Jeffrey A , Fernhall Bo
Published: June 2021

The ventricular‐vascular coupling ratio consists of the ratio of arterial load (Ea) to ventricular stiffness (Elv). Blood pressure (BP) and arterial stiffness affect Ea/Elv. African Americans (AA) have higher large artery stiffness and BP compared to Caucasian Americans (CA), but exercise training reduces BP and large artery stiffness. Our purpose was to determine the effects of race on this ratio after 8 weeks of training. We hypothesized a reduction in the ratio due to a reduction in EaI that would be more pronounced in CA versus AA adults. We used applanation tonometry to measure central BP, and central pulse wave velocity (cPWV) to quantify central arterial stiffness and cardiac ultrasonography to determine LV volumes before and after our training period. We used the formulas: Ea=ESP/SV and Elv=ESP/ESV scaled by BSA. AA and CA maintained their pre‐training ratios (p>0.05 for time and interaction effect), but CA did so by augmenting both EaI and ElvI (EaI was 0.76±0.1 and 0.77±0.09 mmHg/ml/m2 pre to post‐training in AA, and went from 0.68±0.09 to 0.75±0.1 mmHg/ml/m2 in CA, p<0.05 for CA; ElvI was unchanged in AA, 1.20±0.11 to 1.22±0.1 mmHg/ml/m2, p>0.05, but increased in CA, 0.99±0.1 to 1.16±0.1 mmHg/ml/m2 p<0.05) despite similar reductions (p<0.05) in SBP in both races. Only AA reduced cPWV after training (p<0.05). We conclude that the training responses of Ea and Elv were different between races.
The effects of acute β‐Adrenergic blockade on aortic wave reflection in post‐menopausal women
Authors: Wolhart Sarah Christine , Hart Emma C. , Barnes Jill N. , Curry Timothy B. , Charkoudian Nisha , Joyner Michael J. , Casey Darren P.
Published: June 2021

Acute ß‐adrenergic blockade (BB) increases aortic wave reflection in young women, however, it is unknown if this effect extends to post‐menopausal women. Thus, we examined systemic BB on aortic wave reflection in 14 post‐menopausal women (58 ± 2 yr). Non‐invasive aortic pressure waveforms were synthesized from high‐fidelity radial pressure waveforms obtained from applanation tonometry before and during systemic BB (0.25mg/kg bolus, with 0.004 mg/kg/min continuous infusion of propranolol). Aortic blood pressure, augmentation index (AIx), AIx adjusted for heart rate (AIx@75), aortic augmented pressure (AP), and wasted left ventricular energy (Ew) were calculated from the aortic pressure waveform. Heart rate (60 ± 2 vs. 51 ± 2 bpm) and mean arterial pressure (95 ± 3 vs. 91 ± 3 mmHg) were reduced with BB (P < 0.05 for both). AIx increased during BB (32 ± 2 vs. 35 ± 2%, P < 0.01), however, AIx@75 was unchanged (25 ± 2 vs. 24 ± 2%, ns). Additionally, AP (20 ± 2 vs. 20 ± 2 mmHg, ns) and Ew (4313 ± 538 vs. 4277 ± 537 dyne*cm2/sec, ns) remained unchanged during BB. When compared to our previously published study in young women, indices of aortic wave reflection were significantly less affected in post‐menopausal women in response to acute systemic BB. Taken together, our data suggest that the negative effects (i.e. increased aortic wave reflection) of non‐selective BB are less pronounced in post‐menopausal women.
Functional Evaluation Of Angiotensin‐Converting Enzyme (ACE) Amino Acid Residues important for the interaction with its inhibitors
Authors: Pereira Larissa Miranda , Affonso Regina , dos Reis Rosana Inácio , Casarini Dulce Elena
Published: June 2021

Hypertension is a worldwide health problem considered to be one of the greatest public health challenges. Since hypertension is a major risk factor for cardiovascular and renal disease, many studies have focused on the Renin‐Angiotensin System (RAS), an endocrine system where renin cleaves angiotensinogen and releases Angiotensin I (AI) which is converted into Angiotensin II (AII) by the action of Angiotensin Converting Enzyme I (ACE). The ACE has a large physiological importance being a target of several therapeutic studies. This work aims to investigate the role of amino acids residues Asp140, Gln259, Ala332, Ser 333, Gln 355, Thr 358, Phe435 and Arg 500, in the interaction with the ACE inhibitor lisinopril as well as other ones.. An analysis of sequence homology to the ACEs of Xenopus laevis, mouse, rat and human, human isoform N‐domain (ND) and isoforms of 65 and 90 kDa of rat showed a high conservation of amino acid residues proposed to be mutated. Primers containing the mutation were synthesized. Following, we used a vector containing the cDNA with the human ACE gene (pACE) to perform PCR reactions; then the products obtained from PCR reactions were digested with the enzyme DpnI and transformed in E. coli DH5α. Positive colonies had the plasmid extracted, sequenced and analyzed concerning the desired mutation. The ACE mutants will be analyzed related to enzyme‐substrate interaction as well as enzyme activity.
Central effects evoked by proline‐rich decapeptide in rats: changes in cardiovascular parameters and neuronal c‐Fos
Authors: Ianzer Danielle Alves , Xavier Carlos Henrique , Madden Christopher J. , Camargo Antonio C. , Morrison Shaun F.
Published: June 2021

Bj‐PRO‐10c (10c) is a proline‐rich oligopeptide from Bothrops jararaca CNP brain precursor and crude venom. 10c evokes antihypertensive and bradycardic effects in SHR. In addition, 10c crosses the blood brain barrier and increases plasma levels of the nitric oxide precursor, L‐arginine. We evaluated whether centrally injected 10c changes blood pressure, heart rate (HR) and the expression of neuronal c‐Fos. 10c (7.1nmol/Kg) or vehicle (NaCl 0.9%) were icv injected in anesthetized male SHR and Wistar rats. Mean arterial pressure (MAP) and HR were recorded for 90min following injection. Brain sections were taken to evaluate c‐Fos and NOS, by double‐labeling immunohistochemistry. Although 10c reduced MAP and HR in both Wistar (ΔMAP: −2±1 saline vs. −6±1mmHg 10c, ΔHR −13±5 vs. −34±5bpm; p<0.01) and SHR (ΔMAP −7±2 saline vs. −16±3mmHg 10c, ΔHR −13±5 vs. −42±7bpm; p<0.01), the changes in MAP were greater (p<0.01) in SHR. At 90min following 10c injection, c‐Fos expression was increased in comparison to saline injections in the nucleus Ambiguus and CVL, areas, which contained neurons double‐labeled for c‐Fos + NOS. We conclude that the central administration of 10c prominently drives a bradycardia, possibly through NO‐related mechanisms.

Support: Fapesp
Association between the Activation of Melanin‐concentrating Hormone (MCH) Producing Neurons and REM Sleep in Rats
Authors: Vas Szilvia , Kostyalik Diana , Katai Zita , Kitka Tamas , Adori Csaba , Molnar Eszter , Petschner Peter , Pap Dorottya , Horvath Beata , Bagdy Gyorgy
Published: June 2021

Introduction

Rapid eye movement sleep (REMS) rebound after REMS deprivation is characterized by increased time spent in REMS and the activation of melanin‐concentrating hormone (MCH) expressing neurons of the hypothalamus, known to have a central role in REMS regulation. The time spent in REMS shows strong correlation with activation of MCH cells of the hypothalamus.

Materials and Methods

Male Wistar rats were selectively REMS deprived (72h, flower‐pot method), followed by a 3h rebound sleep, during which EEG, EMG and motility were recorded. Immediately after rebound, each animal was perfused. MCH/cFos immunohistochemistry (cFos as an indicator of neuronal activation) was performed on hypothalamic setions. Home cage animals were used as controls. Data were evaluated by Spearman‐correlation.

Results

During REMS rebound, both the number of REMS episodes and the average duration of REMS items increased. However, a positive correlation with MCH/cFos double staining was shown only in case of the number of REMS episodes.

Discussion

The activation of MCHergic hypothalamic neurons seems to be in a strong connection with the number of REMS, suggesting that REMS deficiency may be compensated via the enhancement of the number during the rebound rather than the average duration of REMS items.

This work was supported by TAMOP‐2.2.1. B‐09/1/KMR‐2010‐0001
Aquaglyceroporin expression and erythrocyte omoregulation in cultures from the freeze tolerant anuran, Cope's gray tree frog, Hyla chrysoscelis
Authors: Mutyam Venkateshwar , Frisbie James , Goldstein David L , Krane Carissa M
Published: June 2021

Cope's Gray Tree Frog, Hyla chrysoscelis is a freeze tolerant anuran that accumulates glycerol during cold acclimation. We hypothesize that the osmoregulatory capabilities of erythrocytes‐‐ to tolerate changes in cell volume and to affect rapid water and solute fluxes‐‐are likely to be most important during the events of freezing and thawing, and are dependent upon the expression and function of HC‐3, an ortholog of the aquaglyceroporin AQP3. Erythrocytes of H. chrysoscelis cultured for 48 hrs in media made hyperosmotic (400 mOsM) through the addition of 150 mM glycerol or urea showed enhanced HC‐3 glycosylation and membrane localization compared to those cultured in control or hypertonic media (250 mOsM culture media + 150 mM sorbitol or 75 mM NaCl). Cultured erythrocytes transferred from hyperosmotic culture media (glycerol or urea) to a hypotonic solution containing 250 mM glycerol or urea underwent a series of shape changes as they swelled, initially elongating, then becoming swollen and round. Cells cultured in hypertonic media (sorbitol or NaCl) responded with significantly less osmotically‐induced swelling (i.e. cell shape change). When aquaporins were blocked by HgCl2, cells retained normal dimensions, indicating the importance of aquaporins in conferring water and solute permeability in these cells. This research was supported by NSF Research Grant IOS‐1121457.
The Nematode C. elegans as a model for heat acclimation research‐molecular characterization of the response to heat stress
Authors: Abbas Atallah , Treinin Millet , Horowitz Michal
Published: June 2021

This study is aimed to reveal essential gene/molecular pathways in heat acclimation (AC). Ceanorhabditis elegans wild type (N2) and HIF‐1 loss of function mutant (KO‐hif1) were used. The N2 worm, acclimated to heat at 25oC enhances survival when exposed to heat stress at the extreme temperature of 35oC. Concomitantly, the HIF‐1 loss of function mutant is unable to acclimate. Hence, comparing the wild type N2 to the KO‐hif1mutant enabled us to study the importance of HIF‐1 targets in AC. To gain a broader perspective on the molecular pathways involved in the acclimation process, we compared the trascriptome profile of control and AC worms of both the N2 and the mutants, using a two‐color array and bioinformatic tools. Survivorship curves of N2 worms with RNAi blockade of the identified genes confirmed their importance in AC. hsp70, ƒÑ£] crystaline, gst20, his71 which encodes Histone 3 and flp1 encoding the FLP1 protein of the worm nervous system were among these genes.
IL‐6 Indirectly Modulates The Induction Of Glyceroneogenic Enzymes In Adipose Tissue During Exercise
Authors: WAN ZHONGXIAO , Ritchie Ian , Chan Catherine B. , Wright David
Published: June 2021

Glyceroneogenesis is an important step in the control of fatty acid re‐esterification with PEPCK and PDK4 being key enzymes in this process. It has been suggested that IL‐6 regulates adipose tissue metabolism and gene expression during exercise. The purpose of this investigation was to determine the role of IL‐6 in modulating the effects of exercise on the expression of glyceroneogenic enzymes in mouse adipose tissue. To achieve these goals, we utilized IL‐6 ex vivo treatment in cultured adipose tissue in combination with IL‐6−/−mice model. Treatment of cultured epididymal adipose tissue (eWAT), increased the phosphorylation of AMPK, ACC and STAT3 and induced SOCS3 mRNA levels while decreasing PEPCK mRNA. The activation of AMPK was independent of increases in lipolysis. An acute bout of treadmill running did not result in the induction of SOCS3 or increases in the phosphorylation of STAT3 in eWAT from WT mice. Exercise‐induced increases in PEPCK and PDK4 mRNA expression were attenuated in eWAT from IL‐6−/− mice in parallel with a greater relative increase in AMPK phosphorylation compared to exercised WT mice. These changes occurred independent of alterations in beta‐adrenergic signalling in adipose tissue from IL‐6−/− mice. Our findings question the role of IL‐6 signalling in adipose tissue during exercise and demonstrate an indirect effect of this cytokine in the regulation of adipose tissue gene expression.

This work was supported by a Bridge Funding Operating Grant from the Canadian Institute of Health Research, Institute of Nutrition, Metabolism and Diabetes to David C. Wright.
Selenoprotein mRNA control
Authors: Seyedali Ali
Published: June 2021

Selenoproteins are defined by the presence of the unique non‐canonical amino acid selenocysteine (sec). The stop codon UGA serves as the triplet which facilitates Selenocysteine's incorporation into the nascent polypeptide. All selenoprotein mRNA's (selenotranscripts) therefore have two stop codons, the bona‐fide and the one providing the code for sec. The sec‐specific UGA can be considered a premature termination codon (ptc) and any regular transcript with a ptc is subject to degradation by the cells quality control mechanism called the nonsense‐mediated decay pathway (NMD). NMD protects the cell from translating aberrant transcripts that harbor ptc's which may have been acquired by mutation or a number of other possibilities. The NMD pathway has specific limitations and exceptions that have been observed with recent research. Our aim is to study factors involved in selenotranscript susceptibilty to NMD. Our primary hypothesis is that under limiting selenium conditions certain selenotranscripts that are predicted to be degraded by NMD will be downregulated while others will remain unchanged. We see that in HEK293t cells and HEPG2 cells almost all selenotranscripts predicted to remain unchanged do not vary while those expected to change vary in their response.