bluuebunny/crossref_metadata_2025 huggingface dataset:
Protective effects of perilla seed meal on tert‐butyl hydroperoxide induced oxidative damage in HepG2 cells
Authors: Sim Ung , Kim In‐Hwan , Lee Junsoo
Published: June 2021

This study aimed to evaluate the protective effect of methanol extract of perilla seed meal (PSE) against oxidative stress induced by tert‐butylhydroperoxide (t‐BHP) in HepG2 cells. The total phenolics and flavonoid contents of PSE were 41.36 ± 1.00 mg gallic acid equivalents/g of residue and 38.20 ± 0.17 mg catechin equivalents/g of residue, respectively. Quantitative analysis of the major constituent phenolic acids and tocopherols was achieved by using high‐performance liquid chromatography. PSE (10–100 μg/mL) pretreatment significantly prevented t‐BHP‐induced cytotoxicity in HepG2 cells. Oxidative damage induced by t‐BHP was accompanied by glutathione depletion, ROS generation, and lipid peroxidation, which were all significantly ameliorated by PSE. The cytoprotective effect of PSE was attributed to the modulation of cellular antioxidant defense enzymes such as glutathione peroxidase, superoxide dismutase, glutathione reductase, and catalase. Collectively, the bioactive compounds of PSE were able to prevent oxidative stress through the enhancement of the antioxidant indices.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Syntaxin 4 (STX4)‐ More than just a SNARE protein: Elevating STX4 content in skeletal muscle to prevent insulin resistance/pre‐diabetes
Authors: Merz Karla Elzein , Olson Erika , Zhang Jing , Veluthakal Raja , Oh Eunjin , Hamilton Angelica , Huss Janice , Thurmond Debbie C.
Published: June 2021

Type 2 diabetes results from a combination of peripheral insulin resistance plus insulin secretion defects. Insulin resistance is characterized by a failure of skeletal muscle to clear excess blood glucose from the circulation, and is central to what is commonly called ‘pre‐diabetes'. While type 2 diabetes afflicts ~10% of the US population, pre‐diabetes is estimated to impact more than a third of our population. Over 80% of glucose clearance occurs via the skeletal muscle, there are no directed therapeutics to restore its function in pre‐diabetics. Several studies have identified correlations between insulin resistance and reduced levels of a SNARE protein named Syntaxin 4 (STX4) in human skeletal muscle. STX4 interacts with other SNARE proteins to facilitate the fusion of glucose transporter 4 (GLUT4) vesicles with the plasma membrane, to then channel extracellular glucose to the interior of the muscle cell, ultimately ‘clearing glucose’ from the circulation. Towards understanding how STX4 content is regulated in muscle, we mimicked the pre/diabetic milieu of high glucose and high lipid content in the L6‐GLUT4myc skeletal muscle cells and demonstrated the same STX4 depletion reported in the human studies. Having established this model system, we next assessed the potential for STX4 overexpression to prevent insulin resistance, and found STX4‐overexpressing cells show less Annexin V and Propidium iodide staining compared with sham‐transfected cells, suggesting that STX4 overexpression exerted a protective effect against diabetogenic stimuli‐induced apoptosis. To next test this in vivo, we have developed an inducible skeletal muscle specific STX4 overexpressing mouse model. Remarkably, under standard non‐diabetogenic conditions, female mice overexpressing STX4 show heightened glucose tolerance and insulin sensitivity compared to control mice. Whether STX4‐overexpressing mice are protected from obesity‐induced insulin resistance, or perhaps whether STX4 upregulation in already‐obese mice might reverse insulin resistance and through which mechanism, are questions currently under pursuit. Taken together, these results suggest that STX4 functions in the skeletal muscle to facilitate glucose uptake function and to protect myocytes from diabetogenic stress induced apoptosis. This suggests that STX4 could be a viable therapeutic target for treatment and prevention of pre‐diabetes.

Support or Funding Information

This research was supported by the National Institutes of Health (DK067912, DK102233) to D.C.T. and also by the H.N. & Frances C. Berger Foundation Fellowship to K.M

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Analyzes of Body Adiposity Index, waist to size ratio, waist to hip ratio, and Heart Score as a better clinical evaluation in postmenopause
Authors: LUGO‐MARTINEZ GABRIELA , Jiménez‐Zamarripa Carlos Alberto , Ocharan‐Hernández Maria Esther , Calzada‐Mendoza Claudia Camelia
Published: June 2021

BACKGROUND

Menopause represents the end of the reproductive stage, and is linked to the increase in obesity and cardiovascular diseases incidence (NAMS, Menopause Glossary).

Body mass Index is positive correlated with major vasomotor symptoms. The classic clinical evaluation of the menopausal women includes variables such as, weight, height, arterial pressure, intensity of the climacteric symptoms and familiar and personal antecedents for oncologic diseases. However, in recent years the reliability of BMI has been questioned; fact linked with the documented body modifications in postmenopause, supports the need to implement a better anthropometric and clinical evaluation.

The heart score allow estimate total cardiovascular risk in apparently healthy individuals, and also can be use to estimate the complications of cardiovascular diseases. This risk estimation is based on the following risk factors: gender, age, smoking, systolic blood pressure and total cholesterol. Total punctuation of heart score greater than 1 indicates the rise of cardiovascular risk.

The aim of this study was to analyze the adiposity index (BAI), waist to size ratio (WSR), waist to hip ratio (WHR), and heart score for a better clinical evaluation in posmenopausal women.

MATERIALS AND METHODS

A descriptive, observational, prospective, transversal, cohort study was carried out in 201 women who attend to climacteric clinic from “Hospital de la Mujer”, anthropometric and clinical data were collected, anthropometric indices was calculated and correlated with BMI, and the heart score was calculated considering the systolic blood pressure, age and smoking history of each patient.

RESULTS

The mean age was 52.3±6.21 years, the average of the heart score was 2, the 25th and 75th percentiles were 2 and 3, respectively, it means that at least 75% of the women included, has elevated cardiovascular risk. Anthropometric indices are resumed in table 1. A positive correlation was found between BMI and anthropometric indices analyzed, the correlation values showed in table 2.

CONCLUSION

Only 25% of the patients has normal BMI. All patients had elevated values for the anthropometric indices. Positive correlation was found between BMI and Antrhopometric indices. All this facts suggest that the evaluation of the waist to hip ratio, waist to size ratio, body adiposity index and the circumferences of waist and hip together with the heart score allows to clinician to perform a better evaluation of cardiovascular risk in postmenopausal women than using BMI alone.

The North American Menopause Society Menopause Glossary. Consulted online https://www.menopause.org/for‐women/menopause‐glossaryon2017.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structural And Functional Characterization Of Ulvan Degrading Polysaccharide Lyase Enzymes
Authors: Ulaganathan ThirumalaiSelvi , Helbert William , Banin Ehud , Cygler Miroslaw
Published: June 2021

Ulvans are the sulfated cell wall polysaccharide present in marine green algae. Ulvan polysaccharides are composed mainly of 3‐sulfated rhamnose, glucuronic acid, iduronic acid and xylose. The sulfation pattern in ulvan resemble glycosaminoglycans in vertebrates. The structural features of ulvan make it a good candidate for variety of industrial applications in agriculture, food, pharmaceutical, chemical, and biomaterials industries. Identification of ulvan‐degrading microorganism and the corresponding enzymes will increase the potential application of this highly abundant naturally occurring polysaccharide.

Bacterial microbiomes associated with the green algae contain enzymes to degrade ulvan by a lytic β‐elimination mechanism. Genome sequencing projects lead to the identification of many such ulvan degrading enzymes from several bacteria Nonlabens ulvanivorans, Pseudoalteromonas sp. strain PLSV, Alteromonas sp. strain LOR and LTR. Using X‐ray crystallography and enzyme activity assays, we solved the structure and biochemically characterized three ulvan lyases, PLSV3936, LOR107 and NLR48. All three enzymes share very low sequence identity and act differentially on substrate.

Despite their low sequence identity, PLSV3936 and LOR107 share the same 7‐bladed β propeller fold. However, the complex structure with the bound tetrasaccharide substrate reveals the difference in the active site and mode of cleavage. Whereas, NLR48 has a β jelly roll scaffold. Complex structure of NLR48 with tetrasaccharide substrate suggest that NLR48 appears to utilize lysine and tyrosine as catalytic residues and the substrate acidic group is neutralized by a glutamine residue. Our results expand the information about ulvan degrading enzymes to potentiate the use of ulvan polysaccharide.

Support or Funding Information

The financial support was provided by a grant from the Natural Science and Engineering Research Council of Canada

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cold active β‐1,3‐glucanase from Antarctic Yeast, Glaciozyma antarctica PI12
Authors: Mohammadi Salimeh , Mahadi Nor Muhammad , Murad Abdul Munir Abdul
Published: June 2021

Glaciozyma antarctica is a psychrophilic yeast that was isolated from the surface of Antarctic sea ice. A key adaptation of psychrophilic microorganisms is to synthesize cold‐active enzymes for survival at low temperatures. In this study, the full‐length cDNA encoding Exo‐β‐glucanase (GaExg55) from G. antarctica PI12 was amplified by reverse‐transcription polymerase chain reaction (RT‐PCR). The cDNA encoded a 480‐residue polypeptide with a putative signal peptide of 20 residues. Subsequently, the novel GaExg55 was expressed in E. coli and purified with nickel affinity chromatography as an approximately 55 kDa protein. The biochemical characterization of purified recombinant GaExg55 (rGaExg55) revealed typical cold‐active enzyme characteristics, such as maximal activity at 20 °C and pH 8.0. The Km and Vmax values of the enzyme using laminarin as the substrate were 10.04 mg mL−1 and 14.12 U mg−1, respectively. An analysis of the three‐dimensional structure of the enzyme was carried out using MODELLER9v16 and compared with homologous mesophilic exo‐β‐1,3‐glucanase. The results of the comparative structural study revealed that the psychrophilic GaExg55 contains longer loops, fewer hydrogen bonds and salt bridges, and a higher total solvent‐accessible surface area which enhanced the protein flexibility for high catalytic efficiency at low temperatures.

Support or Funding Information

This research was supported by a research grant from the Ministry of Science Technology and Innovation (MOSTI), Malaysia under the research grants 10‐05‐16‐MB002 and 02‐05‐20‐SF0007.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Physical Resilience as a Determinant of Healthy Aging
Authors: Brown Ashley K. , Mazula Daniel L. , Zhang Bin , Roos Carolyn M. , White Thomas A. , Miller Richard A. , Miller Jordan D. , LeBrasseur Nathan K.
Published: June 2021

Physical resilience, defined as the ability to resist and respond to challenges and return to homeostasis, has been postulated to be a determinant of healthspan and lifespan. To test this premise, we have begun to optimize a standardized, clinically relevant battery of resilience challenges, including anesthesia and cisplatin chemotherapy, and recovery outcome measures in younger (2–3 month) and older (26–29 month) mice. We have also started to test whether interventions shown to increase murine lifespan‐acarbose, rapamycin, and 17a‐estradiol‐impact the resilience of older mice.

Our preliminary data demonstrate that younger mice (n = 10) have faster dorsal to sternal recumbence recovery time compared to older mice (n = 8) (39 ± 1 sec vs. 67 ± 6 sec, p < 0.001) following a 5‐minute 4% isoflurane anesthesia challenge. In response to a 5‐day cisplatin (2 mg/kg/day) challenge, both younger (n=5) and older (n=4) mice lost a significant percent of their baseline body weight (younger mice −10.6 ± 0.8% and older mice −13.2 ± 2.1%, p = 0.26), however, older mice failed to regain body weight 11 days after the challenge (younger mice −0.9 ± 0.8% and older mice −9.8 ± 0.8%, p < 0.01). Changes in body weight in younger and older mice were largely attributable to the loss/regain of fat mass. Interestingly, 8 weeks of either acarbose (1,000 ppm) or 17α‐estradiol (14.4 ppm) administration to older female mice improved body weight recovery after the cisplatin challenge, while rapamycin (14 ppm) had no appreciable effect.

Our preliminary results demonstrate that younger mice, as expected, have greater resilience than older mice to clinically relevant physical challenges. The age at which physical resilience starts to decline requires further investigation. Our early observations suggest interventions that extend health‐ and lifespan may impact resilience in older mice. Ultimately, a standardized battery of resilience challenges may provide a new framework for testing and translating interventions that target the fundamental biology of aging.

Support or Funding Information

This work was supported by grant AG52958 from the National Institutes of Health.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Developing a new polymeric system for release of immunomodulators for treatment of heart failure
Authors: de Jesús Salazar‐Ramírez Felipe , Lozano Omar , Chapoy‐Villanueva Hector , Torre‐Amione Guillermo , García‐Rivas Gerardo
Published: June 2021

During recent years the role of the immune system in the progression of heart failure (HF) has become an area of interest to better understand the physiopathological mechanisms of its progression. Some immunomodulators have been tested for efficacy in reducing inflammatory mediators in patients with HF. However, one of the greatest limitations encountered when trying to bring those treatments to patients is the side effect of these drugs. Keeping this in mind, nanotechnology has been advancing rapidly during the past few years. Nanoparticles can carry medications and have active tissue targeting properties, and such characteristics could increase the drug tolerance, which are known to have a narrow therapeutic window. Using this knowledge we used a well‐known immunomodulator, either free or encapsulated in a polymeric nanoparticle to treat a murine model of HF by inducing an anti‐inflammatory effect, and thus, reduce the progression ventricular dysfuction.

Methods

The immunomodulator was encapsulated in a polymeric nanoparticle by an oil‐in‐water emulsion. The resulting nanovector was analyzed in terms of size, surface charge, drug loading and release profile.

Results

The nanovector was of ~ 100 nm in size, −30 mV in surface charge, 95 % entrapment efficiency, and a release profile consistent with reported data. Mice model of HF shown fibrosis and cardiac remodeling, however this damage was reduced in the animals that received the immunomodulator (1 mg/Kg week). Likewise, a 4‐fold increase in the production of IL‐10 was observed. Moreover, the use of polymeric nanoparticles required a lower dose to reach a similar cardioprotective effect.

Support or Funding Information

Supported by ITESM and CONACyT

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Capsaicin and Its Non‐Pungent Analog Arvanil Sensitize Platinum‐Resistant Small Cell Lung Cancer to Camptothecin‐Induced Apoptosis
Authors: Colclough Kate W. , Friedman Jamie R. , Perry Haley E. , Brown Kathleen C. , Akers Austin T. , Nolan Nicholas A. , Rollyson William D. , Chen Yi Charlie , Denning Krista L. , Brown Linda G. , Dasgupta Piyali
Published: June 2021

Cisplatin or carboplatin‐based chemotherapy is the standard of care for small cell lung cancer patients (SCLC). The challenge with SCLC treatment is that such chemotherapy shows excellent response initially; however, the tumor relapses within a year and is resistant to chemotherapy or radiation. Furthermore, a subset of SCLC is resistant to cisplatin (called platinum‐resistant SCLC) or displays platinum refractory phenotype. There is only one FDA‐approved drug for relapsed SCLC, which is camptothecin (irinotecan). It has been shown to display an objective response rate of about 3%. Therefore, agents which can increase the response rate of camptothecin should be of considerable benefit to SCLC patients. We tested the combinatorial effects of the nutritional compound capsaicin with camptothecin in human SCLC. Caspase‐3 activity assays reveal that the combination of camptothecin and capsaicin displayed greater apoptotic activity than any of the compounds used alone. Chicken chorioallantoic membrane assays confirmed the combinatorial effects of capsaicin and camptothecin. Statistical analysis using the Chou‐Talalay isobologram showed that capsaicin displayed synergistic apoptotic activity with camptothecin. The clinical development of capsaicin as a viable anti‐cancer drug is limited due to its adverse side effect profile. Our laboratory has performed structure‐activity relationship (SAR) studies to identify non‐pungent capsaicin analogs. One of those analogs, Arvanil, has shown synergistic apoptotic activity with SN‐38 (active drug component of irinotecan) in cisplatin‐resistant SCLC cells. Chou‐Talalay isobologram analysis showed that the magnitude of synergy between arvanil and SN‐38 was greater than capsaicin and SN‐38 in cisplatin‐resistant H69 human SCLC cells. The results of our studies will pave the way for novel second‐line treatment regimens for human SCLC.

Support or Funding Information

Funding for our study was supported by an NIH R15‐AREA Grant (2R15CA161491‐02) and a WVU‐MU Health Partnership Grant. This work was supported in part by the West Virginia IDeA Network of Biomedical Research Excellence (WV‐INBRE) grant GM103434 (PI: Dr. G. Rankin).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
An Integrative Approach to Identify Biological and Socioeconomic Contributors to Cancer Incidence and Mortality: The Advancing a Healthier Wisconsin Endowment's Cross‐Cutting Component
Authors: Olson Jessica , Stolley Melinda , Millon‐Underwood Sandra , Cawthra Tobi , Beyer Kirsten , Fraser David , Ignace Lyle , Pinsoneault Laura , Salazar Jose , Walker Alonzo , Willliams Carol , Lucey Paula , Maurana Cheryl
Published: June 2021

Background

Despite advances in breast and lung cancer research, racial and geographic disparities in incidence, late‐stage diagnosis, and mortality persist in Wisconsin. Root causes for cancer‐related health disparities are complex and multifactorial, and require bold, innovative approaches to have a significant impact. The Advancing a Healthier Wisconsin Endowment's Cross‐Cutting Component is dedicated to reducing these breast and lung cancer disparities in Wisconsin by integrating the strengths of research, education, and community and investing up to $20M over a period of 8 years.

Methods

This initiative began with the formation of an interdisciplinary “cells to society” design team comprised of basic scientists, population health experts, clinicians, and community leaders. The team was charged with developing broad approaches to engage scientists and community and achieve the following goals: (1) gain a more contextualized understanding of the molecular, genetic, environmental, cultural, and behavioral factors that contribute to cancer disparities, as well as barriers and facilitators to these discoveries; and (2) facilitate the bringing together of “cross‐cutting teams” to adopt, adapt and/or create innovative strategies to address cancer disparities statewide. To inform the creation of these teams, a series of “listening sessions” with basic and clinical biomedical health professionals, educators, and community members, were created to gather input on needs and opportunities to reduce disparities.

Results

This presentation will review the Cross‐Cutting Component and present results from Goal 1 activities. We have completed: (1) a comprehensive curation of literature detailing strategies to reduce cancer disparities, (2) root cause analyses on breast and lung cancer from a team of multi‐disciplinary experts, (3) maps detailing differences in breast and lung cancer disparities across the state and (4) analyzed themes from discussion sessions held with key stakeholders with perspectives and expertise ranging from T0 – T5 research, education, and the community.

Conclusion

Results highlight common and unique themes among these diverse groups, that will be used to inform the “cross‐cutting teams” that will (1) encourage participants to think beyond their expertise (2) create, adapt or adopt strategies for addressing complex health problems, and (3) develop, test and evaluate initiatives to address breast and/or lung cancer disparities in incidence, late‐stage diagnosis, and mortality.

Support or Funding Information

This work is supported by the Advancing a Healthier Wisconsin Endowment.
Role of HER‐2 in Breast Carcinogenesis
Authors: Link Tina , Jain Supriya , Lu Allison , Nathan Ajay , Shabazaz Tara , Uppalapati Isha
Published: June 2021

Breast cancer has several defining prognostic and predictive markers, one of which is HER‐2. The Human Epidermal Growth Factor Receptor (HER‐2) is a protein which plays a role in cellular proliferation and cell survival. The solved structure revealed that the protein has an extracellular ligand binding domain. However, HER‐2 has no known ligand to bind to its active site. Consequently, its known function is to dimerize with its sister receptor (HER‐3) and initiate a cascade pathway for growth. The upregulation of the HER‐2 neu oncogene can result in the overexpression of HER‐2, causing a change in the ratio of HER‐2 to HER‐3 receptors. Because HER‐2 promotes cellular proliferation and survival, uncontrolled expression of this factor has major implications in carcinogenesis. About twenty percent of all breast cancers are diagnosed as HER‐2 positive. Treatments to inhibit HER‐2 include ligand mimetics which bind to HER‐2, preventing its dimerization with HER‐3 and therefore stopping the uncontrolled cellular proliferation that would otherwise occur downstream. The FDA has approved several anti‐HER2 antibody therapies which have been used in combination with chemotherapy in the adjuvant and neoadjuvant setting. Trastuzumab (Herceptin) and Pertuzumab (Perjeta) are two anti‐HER2 antibodies which, in conjunction with chemotherapy, have resulted in improved outcomes for HER‐2 positive breast cancer patients. The Walton High School MSOE Center for BioMolecular Modeling SMART Team used 3‐D modeling and printing technology to examine the structure‐function relationship between HER‐2 as it relates to carcinogenesis.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Patch Compartment Neurons in Methamphetamine‐Mediated Reward
Authors: Kendrick Troy , Robinson Hannah , Barker Natalie , Patel Birva , Logue Jordan , Horner Ashley
Published: June 2021

The striatum is significantly important in the formation of habitual behaviors as well as reward association. Elicited reward behavior from methamphetamine (METH) is mediated by the striatum. Two distinct striatal sub regions, the patch and matrix, have been shown by previous studies to have key role in the regulation of addiction behavior. The patch compartment, in contrast with the matrix, expresses a high density of mu opioid receptors and receives inputs from limbic regions of the brain. Patch compartment neurons have been shown to contribute to habitual behaviors and it has been hypothesized that these neurons also contribute to reward, although this has yet to be demonstrated. The basis of this experiment was to investigate the role of the patch compartment neurons in METH‐induced reward behavior. This goal was attained by determining if the ablation of the mu opioid receptor‐containing neurons of the patch compartment would alter METH‐mediated condition place reference (CPP), which is an indicator of drug reward, by using Dermorphin‐Saporin (Derm‐Sap) which is a neurotoxin used to accurately target and eliminate mu opiate receptor‐containing neurons. Male and female rats in this study were bilaterally infused in striatum either with Derm‐Sap, or unconjugated Saporin, which was used as a control. After eight days of recovery, subjects were exposed to the CPP paradigm receiving either a moderate dose of METH (2 mg/kg) or saline. After eight days of conditioning, a preference test was conducted on each animal to determine the degree of preference for the drug‐paired chamber. It was found that METH‐mediated CPP was reduced in patch‐lesioned female rats, as compared to control animals, while METH‐mediated CPP was increased in male rats with Derm‐Sap lesions as compared to controls. These results allow us to conclude that patch compartment neurons are necessary for METH‐induced reward behaviors in females, but not males.

Support or Funding Information

Navicent Health Foundation Research Award to AH

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of Eating a High Fat Diet on Sensitivity of Female Rats to the Reinforcing Effects of Methamphetamine
Authors: Galindo Kayla Irene , Beltran Nina , Serafine Katherine
Published: June 2021

Eating a diet that is high in fat contributes to increased risk of chronic diseases and mortality, even in the absence of obesity. Eating a high fat diet can also impact the same brain reward pathways that are targeted by drugs of abuse. Preclinical studies have demonstrated rats eating high fat chow are more sensitive to drugs that act on dopamine systems (e.g., cocaine). However, the majority of evidence demonstrating that diet can impact drug sensitivity has investigated only male subjects, despite potential sex differences. To test the hypothesis that eating high fat chow increases sensitivity of female rats to the positive reinforcing effects of methamphetamine, female Sprague‐Dawley rats (n = 30) were fed either standard chow (17% kcal from fat), high fat chow (60% kcal from fat), or given restricted access to high fat chow (such that their body weight was maintained, without the development of obesity) for 8 weeks. Following catheter implantation, rats were trained to respond for 0.1 mg/kg methamphetamine on a Fixed Ratio 1 schedule. Although there were no differences in acquisition (n = 2–3/group), future work will examine reinforcing effectiveness of methamphetamine across dietary conditions using a progressive ratio schedule of reinforcement. These data expand previous research, which has predominately focused on male rats, and add to the growing literature on the effects of diet on drug sensitivity.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Demand Analysis of the Mu Opioid Receptor Agonist Remifentanil Alone and in Combination with the Cannabinoid Receptor Agonist JWH018
Authors: Maguire David R. , France Charles P.
Published: June 2021

Opioid abuse is a significant public health problem. With continuing emergence of novel psychoactive substances (e.g., synthetic cannabinoids found in “K2”/”spice” preparations), the co‐administration of opioids and other novel drugs is likely to become more prevalent which might increase the risk for abuse and other adverse effects. This study examined the impact of the synthetic cannabinoid receptor agonist JWH018 on the reinforcing effectiveness of the mu opioid receptor agonist remifentanil in rhesus monkeys (n=5) using economic demand analyses. Lever presses delivered iv infusions of a drug or drug mixture according to a fixed ratio schedule. For each condition, the ratio (10–3200) progressively increased across sessions yielding a demand curve: consumption (infusions obtained) plotted as a function of price (fixed ratio value). When available alone, remifentanil (0.00032 mg/kg/infusion) occasioned the highest unconstrained demand (consumption at the lowest cost) and highest essential value, while JWH018 (0.0032 mg/kg/infusion) alone occasioned lower unconstrained demand and essential value. Unconstrained demand for a mixture of remifentanil and JWH018 was lower than that of remifentanil alone but higher than that of JWH018 alone. Essential value of the mixture was not significantly different from that of remifentanil alone. These data indicate that synthetic cannabinoids such as JWH018 might impact some aspects of opioid self‐administration (i.e., decreased consumption at the lowest price) but do not enhance reinforcing effectiveness as measured by sensitivity of consumption to increasing costs. Opioid/cannabinoid mixtures do not appear to have greater abuse potential compared with an opioid alone.

Support or Funding Information

Supported by the NIH (R01DA005018) and the Welch Foundation (AQ‐0039).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of Heat Shock Protein 90 in the Spinal Cord Strongly Promotes Morphine Anti Nociception by Increasing ERK MAPK Activation
Authors: Duron David Isaiah , Lei Wei , Blagg Brian , Streicher John
Published: June 2021

Hundreds of millions of individuals suffer from acute and chronic pain in the United States alone. The current gold standard of treatment for patients with moderate to severe pain is the use of opioid analgesics. Although the efficacy of these medications is often superior to other options in the clinic, their use is significantly limited due to their side effects and potential for abuse. Recent progress in the research of opioid receptor signaling has shown that manipulation of opioid signaling by biased agonists or other means can improve the therapeutic profile of opioids; however, the field is limited by a lack of understanding of the opioid signaling complex. We have previously shown that inhibition of the chaperone protein Heat shock protein 90 (Hsp90) in the brain strongly blocks opioid anti‐nociception by blocking ERK activation. Here we demonstrate a reversed effect in the spinal cord in that intrathecal (i.t.) injections of the Hsp90 inhibitors 17‐AAG or KU‐32 in CD‐1 mice strongly increase morphine anti‐nociception in paw incision and tail flick pain. Seeking a mechanism for these changes, we found that i.t. Hsp90 inhibition strongly increased ERK activation in the spinal cord in response to opioids, which is opposite of what is observed in the brain, and also implies that Hsp90 acts to repress opioid‐stimulated ERK signaling in the spinal cord. By combining Hsp90 inhibition with the ERK inhibitor U0126 in the spinal cord, we found that ERK inhibition fully reversed the effects of Hsp90 inhibition while having no effect on its own, supporting the hypothesis that increased ERK signaling in spinal cord is responsible for the effects of Hsp90 inhibition. We also performed experiments with systemic and combined i.c.v. and i.t. Hsp90 inhibitor to demonstrate that Hsp90 inhibition in the brain dominates over other circuits when all are inhibited. This data implies that signaling localization within the pain circuit could be key for the targeting of proteins to modify the therapeutic index of opioid analgesics like morphine. These results also suggest that spinal Hsp90 inhibitors could be used as an opioid dose‐reduction strategy which could preserve analgesic efficacy while reducing side effects.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endosomal GPCR signaling: Tyrosine Phosphorylation of a Peptide Linker in NEDD4‐2 Increases Ligase Activity to Promote p38 Proinflammatory Signaling
Authors: Grimsey Neil J. , Rada Cara C. , Narala Rachan , Stephens Bryan S. , Mehta Sohum J. , Lapek John J. , Handel Tracy M. , Zhang Jin J. , Gonzalez David J. , Trejo JoAnn
Published: June 2021

The regulation of G protein‐coupled receptor (GPCR) signaling is essential for vascular homeostasis and inflammatory responses during disease and injury. MAPK p38 is a key mediator of vascular inflammation, but how GPCRs regulate MAPK p38 signaling is relatively understudied. Post‐translational modifications play a key role in regulating spatiotemporal GPCR signaling and our studies have shown that ubiquitination of GPCRs is not just a cue for trafficking and degradation. Instead, GPCR ubiquitination plays a critical role in endosomal activation of an atypical MAPK p38 signaling pathway. Ubiquitinated receptors recruit the adaptor proteins TAB1 and TAB2 to endosomes and enable auto‐activation of p38, to induce proinflammatory signaling in the vasculature. We have shown that this pathway is conserved for GPCR activation by multiple proinflammatory agents in the vasculature including, thrombin, histamine, ADP and PGE2. Defining how GPCRs activate E3 ubiquitin ligases to initiate endosomal signaling is the focus of our current studies. Using FRET biosensors and phosphoproteomics studies we have discovered that GPCRs activate the E3 ubiquitin ligase NEDD4‐2 by c‐Src dependent tyrosine phosphorylation at a critical tyrosine residue (Tyr485). Tyr485 resides within a predicted auto‐inhibitory linker peptide between the protein binding domains WW II and WW III. Phosphorylation at Tyr485 is required for NEDD4‐2 activation, recruitment to GPCRs and the induction of ubiquitin dependent endosomal p38 signaling. Mutation of Tyr485 blocks thrombin and ADP induced p38 activity and significantly reduces thrombin mediated endothelial permeability. These studies are the first to define how NEDD4‐2 is activated by c‐Src tyrosine phosphorylation, and provides greater clarity into how GPCRs induce proinflammatory p38 signaling from endosomes. Future studies will critical for defining how to selectively block GPCR induced atypical p38 signaling.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of Novel Therapeutic Targets Against Organophosphate‐Induced Axonal Transport Deficits
Authors: Naughton Sean X.
Published: June 2021

The chemicals known as the organophosphates (OPs) have a wide array of uses in industrial and agricultural settings, most notably as pesticides, but they have also been used on occasion as chemical warfare (nerve) agents. OPs are highly toxic and they have been associated with a variety of acute and long‐term neurologic deficits including impairments in cognition in humans and animal models. The mechanism of the acute toxicity of OPs, cholinesterase inhibition is well documented; however, the neurobiological basis for the long‐term effects of OPs, especially when the exposure levels were below the threshold for acute toxicity is less clear. It is imperative that the mechanism of these deleterious effects of OPs be elucidated so that effective therapeutic strategies can be developed. Previous research in our laboratory conducted both in vivo and in vitro indicated that deficits in axonal transport may represent one mechanism of the long‐term neurologic deficits associated with OPs. In the current series of experiments, we are analyzing post mortem brain tissues from OP‐exposed rats to identify non‐cholinesterase targets that might contribute to the OP‐related deficits in axonal transport. As the axonal transport‐related targets of OPs are identified post mortem, subsequent pharmacological experiments are being conducted in vitro to exploit these targets as a first step toward developing novel therapeutic strategies. In the first series of experiments in tissues collected from rats treated with the OP‐nerve agent, diisopropylfluorophosphate (DFP), 1.0 mg/kg every other day for 30 days, we have identified (using standard immunoblotting techniques) alterations in the phosphorylation of several signaling kinases (e.g., ERK, Akt, and GSKIIβ). We have also identified DFP‐related changes in post‐translational modifications of structural proteins that affect axonal transport through the regulation of microtubule dynamics and stability (e.g., Tau phosphorylation, Tubulin Acetylation). Using a live imaging technique in rat cortical neurons for evaluating fast axonal transport of fluorescent, (APPGFP‐labeled) membrane bound organelles, we have determined that compounds that target signaling kinases (e.g., GSKIIIβ), microtubule stability, and mitochondrial function, can attenuate the DFP‐related impairments in axonal transport. These data not only further elucidate potential non‐cholinesterase targets of OPs, but also provide an initial step toward the design of new therapeutic strategies against OP toxicity.

Support or Funding Information

This work was supported by the Congressionally Directed Medical Research Programs (CDMRP), specifically, the Gulf War Illness Research Program (GWIRP), grant number W81XWH‐12‐1‐0536.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Discovery of MOR Selective, Reversible Opioid Antagonists for Potential Use in Treatment of Drug Dependence
Authors: Jali Abdulmajeed , Obeng Samuel , Zhang Yan , Selley Dana E.
Published: June 2021

Opioid dependence/addiction is a major public health problem that is associated with multiple health and social costs. Opioids were responsible for the majority of deaths resulting from a drug overdose in 2015 (63% of 52,404). The FDA has approved methadone and buprenorphine for long‐term agonist maintenance therapy. However, a large portion of patients relapse to heroin or illicit use of prescribed opioids. One approach to minimize drug‐induced relapse risk is switching patients to long‐term antagonist therapy after weaning off with an agonist. Naltrexone (NTX) is the only opioid antagonist that is approved for opioid dependence treatment. Nevertheless, NTX is associated with side effects including dysphoria and depression, which contribute to poor patient compliance. Some side effects of NTX could be due in part to its lack of selectivity, as NTX has moderate affinity for the kappa opioid receptors (KOR). Currently, there are no highly‐selective, reversible, non‐peptide μ opioid receptor (MOR) antagonists. Therefore, 18 novel indole‐substituted epoxymorphinan ligands were designed based on the message‐address concept and molecular modeling studies, and synthesized. These novel ligands were hypothesized to bind selectively to the MOR based on interaction with a key aromatic residue in extracellular loop 3 of the MOR (Trp318), such that the added indole substituent in the structure might favor interaction with the indole ring in Trp318 in MOR over KOR and delta opioid receptors (DOR). The purpose of this study was to identify potential MOR‐selective antagonists from this series of compounds using radioligand and [35S]GTPγS binding assays in Chinese Hamster Ovary cells that were stably expressing mouse opioid receptors of interest (MOR, KOR or DOR) and ICR male mouse thalamus. Results showed that all 18 compounds bind to the MOR with high affinity (Ki ≤1 nM) and exhibit a range of efficacies for MOR‐mediated G‐protein activation. Two compounds were discovered to possess most of the desired features. Both compounds have high MOR‐affinity (<1 nM), high MOR selectivity over DOR (>50‐fold) and moderate selectivity over KOR (~8‐fold,) as well as low efficacy to activate the MOR (<20% of the full MOR agonist, [D‐Ala2, N‐MePhe4, Gly‐ol]‐enkephalin (DAMGO)). They also competitively right‐shifted DAMGO‐concentration effect curves for G‐protein activation similarly to naltrexone, indicating they act as competitive MOR antagonists. Our results are encouraging to advance these two compounds for further in‐vitro and in‐vivo pharmacological characterization, and provide important pharmacological information to aid future ligand design.

Support or Funding Information

NIH grant R01‐DA024022; Jazan University

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
3,3′‐Dichlorobiphenyl (PCB 11) Promotes Dendritic Arborization in Primary Neurons via CREB‐Dependent Mechanisms
Authors: Sethi Sunjay , Keil Kimberly , Wei Feng , Dong Yao , He Guochun , Li Xueshu , Pessah Isaac , Denison Michael , Lehmler Hans‐Joachim , Lein Pamela
Published: June 2021

PCB 11 is a byproduct of contemporary paint pigment production that has been detected in environmental media around the world, including milk produced for human consumption. We previously reported that PCB 11 enhances dendritic arborization in primary rat neuronal cell cultures. Since increased dendritic arborization, or hyperconnectivity, is a shared characteristic of many neurodevelopmental disorders (NDDs), our findings suggest this chemical poses a risk to the developing brain. The goal of this study was to understand how PCB 11 promotes dendritic growth. We tested canonical mechanisms of PCB developmental neurotoxicity, including signaling mediated by the ryanodine receptor (RyR), aryl hydrocarbon receptor (AhR), thyroid hormone receptor (THR), and cAMP response element binding protein (CREB). [3H]Ryanodine binding assays were used to assess effects of PCB 11 on RyR activity; luciferase‐reporter cell lines, to assess effects on AhR or THR activation. To assess a functional role for these signaling molecules in PCB 11‐mediated dendritic growth, primary neuron‐glia co‐cultures dissociated from neocortices of neonatal Sprague Dawley rats were transfected at 6 days in vitro (DIV) with a MAP2B‐FusRed construct to label the dendritic arbors of individual neurons. At 7 DIV, cultures were pretreated with pharmacological antagonists and then co‐treated for 48 h with PCB 11 or vehicle (0.1% DMSO). Dendritic morphology was quantified in FusRed‐labeled neurons by quantifying the number of dendritic tips and primary dendrites. PCB 11 had no activity at RyR, AhR or THR, and pharmacological antagonism of these receptors had no significant effect on PCB 11‐induced dendritic growth. However, the dendritic promoting effect of PCB 11 was blocked by the selective inhibitor of CREB, 666‐15 (3‐(3‐Aminopropoxy)‐N‐[2‐[[3‐[[(4‐chloro‐2‐hydroxyphenyl)amino]carbonyl]‐2‐naphthalenyl]oxy]ethyl]‐2‐naphthalenecarboxamide), or by co‐transfection with a short hairpin RNA construct specific for CREB. These results indicate that PCB 11 promotes dendritic growth via interaction with CREB or CREB‐dependent mechanism(s), but not directly interfering with known molecular targets of the higher chlorinated PCBs. In light of emerging evidence implicating perturbations of CREB signaling in the etiology of many NDDs, these findings add to the weight of evidence suggesting that PCB 11 poses a threat to the developing human brain.

Support or Funding Information

Supported by NIEHS (R01 ES014901 to PJL & INP; T32 ES007059 to SS; P01 ES013661 to HJL) & NICHD (F32 HD088016 to KPK and U54 HD079125).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Retrospective Study of the Relationship Between Testicular Damage and Urinary Creatine Excretion: Possible Markers of Testicular Toxicity Induced by Drugs
Authors: Yono Makoto , Kimura Megumi , Inoue Yumi , Furukawa Kohichiro , Fujiyama Mai , Mugitani Ayumi , Hori Masaharu , Tsuji Shigeki , Tanaka Takanori , Irie Shin , Latifpour Jamshid
Published: June 2021

Objectives

Creatine and creatine phosphate act as a buffer system for the regeneration of ATP in tissues with high and fluctuating energy demands. Total creatine content of the rat testis is high compared to all other tissues except skeletal muscle and creatine excretion in rat urine has been shown to increase following testicular damage caused by toxicants. Because the use of biomarkers in toxicology is becoming increasingly important, we investigated whether urinary creatine might be a useful marker for testicular damage in human.

Methods

In a retrospective study, the records of 22 male patients with urological malignancies treated with cisplatin, leuprorelin or goserelin at 4‐week intervals between October 2005 and October 2015 were evaluated. Urinary creatine and serum follicle‐stimulating hormone (FSH), luteinizing hormone (LH) and testosterone levels were assessed in all patients. Single‐voided urine samples were collected to determine urinary creatine‐to‐creatinine ratios. Blood sample collection was done in the morning when serum testosterone levels are around their peak: such morning time sampling for testosterone measurements is recommended in the clinical setting and routinely used in epidemiological studies.

Results

Seven patients with urothelial carcinoma treated with cisplatin based chemotherapy (mean age, 64.1 years) showed no significant changes in urine creatine/creatinine ratio during treatment and had a significant increase in FSH and LH levels and a significant decrease in testosterone levels compared to those at baseline. Similarly, no significant changes in urine creatine/creatinine ratio occurred in patients with prostatic carcinoma treated with leuprorelin (n=8; mean age, 70.0 years) or goserelin (n=7; mean age, 70.4 years), both of whom had castrate levels of testosterone (≤0.5 ng/mL) and suppressed levels of FSH and LH during treatment.

Conclusions

Our data demonstrate that testicular damage could not be detected by urinary creatine in patients treated with drugs showing testicular toxicity. Two major testicular functions, i.e., the production of testosterone and formation of haploid germ cells, are generally considered to be regulated by pituitary gonadotrophins, with LH acting on the testosterone‐producing Leydig cells located in the interstitium and FSH affecting Sertoli cells in the seminiferous tubules. Thus, our hormonal data may indicate the presence of cisplatin‐induced Sertoli cell dysfunction and inhibition of Leydig cell function by LH‐RH analogues.

Support or Funding Information

None

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.