bluuebunny/crossref_metadata_2025 huggingface dataset:
Supplementation of multi‐vitamins, not glutamine alleviate erythrocyte oxidative status in rats with trauma‐hemorrhagic shock and resuscitation
Authors: Lo Hui‐chen , Lin Yu‐Hsin , Lee Chien‐Hsing
Published: June 2021

Resuscitation fluid replacement is the major life saving strategy for patients with hemorrhagic shock; however, resuscitation‐induced oxidative stress may lead to systemic inflammation, multiple organ failure (MOF), and even death. Studies showed that reduction of oxidative stress and preservation of ATP production may improve MOF in animals with trauma‐hemorrhagic shock and resuscitation (THR). In addition, glutamine has been considered as a cellular fuel and is the source of glutathione and multi‐vitamins possess antioxidant activity and play important roles in energy metabolism. Therefore, we investigated the effects of single or simultaneous administration of multi‐vitamins and glutamine on antioxidant defense systems, including enzymatic and non‐enzymatic, in the plasma and erythrocytes of THR. Male SD rats were suffered with THR, including a 5 cm midline laparotomy and mean arterial pressure 30 to 35 mmHg for 60 min followed by resuscitation lactate Ringer's (LR) solution with or without L‐alanyl‐L‐glutamine and/or multi‐vitamins. The sham‐operated, parenterally fed rats were included. After infused with continuous, slow rate of LR solutions for 42 hours, the THR rats had significantly decreased red blood cells and hematocrit and increased erythrocyte glutathione (one‐way ANOVA, p<0.05). Intravenous glutamine significantly increased erythrocyte total and protein‐bound thiol groups and plasma TBARS; intravenous multi‐vitamins significantly decreased erythrocyte TBARS; and combination of glutamine and multi‐vitamins significantly decreased erythrocyte TBARS and the ratio of plasma reduced to oxidized glutathione and increased plasma TBARS. In addition, multi‐vitamins was the main factor to decrease plasma total and protein‐bound thiol groups and plasma and erythrocyte TBARS and to increase the ratio of plasma reduced to oxidized glutathione (two‐way ANOVA, p < 0.05). In conclusion, multi‐vitamins, not glutamine, may alleviate erythrocyte oxidative status in trauma‐hemorrhagic shock and resuscitation.

Support or Funding Information

NSC 102‐2320‐B‐030‐005‐MY3

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Selenium Reduces Lectin‐Like Oxidized LDL Receptor‐1 (LOX‐1)‐ Mediated Vascular Oxidative Stress Induced by Inhalation of Pyrolysis Oil Vapors
Authors: Moustafa Mohamed El‐Sayed , Fawaz Youssef B.
Published: June 2021

Pyrolysis oil (PO) is a renewable fuel produced from municipal solid waste and there is a concern about the adverse effects of its vapors on vascular oxidative stress. In this study, we investigated the effects of exposing mice to PO vapors on lectin‐like oxidized LDL receptor‐1 (LOX‐1)‐mediated vascular oxidative stress. We also examined the effects of selenium supplementation in these mice. Our results showed that the exposure of mice to PO vapors increased aortic levels of malondialdehyde, LOX‐1 and its soluble form sLOX‐1, NADPH oxidase 4 (NOX‐4), NF‐kB (p65) and endothelial NOS (eNOS). The administration of 1.78 μg selenium/kg body weight into these mice 15 minutes before the exposure to PO vapors reduced the increased levels of the above parameters. In conclusion, the exposure to pyrolysis oil vapors contributes to the increased LOX‐1–mediated vascular oxidative stress which can be reduced with selenium supplementation.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In‐vivo evaluation of ethanolic seed extract of Walnut (Tetracarpidium Conophorum) on N‐acetyl‐ 1,4‐benzoquinone imine‐induced oxidative stress in albino rats
Authors: Nosiri Chidi Ijeoma , Anyanwu Chukwuma , Benedict Sandra
Published: June 2021

This study evaluated the liver integrity and antioxidant effects of ethanolic seed extract of Walnut (Tetracarpidium Conophorum) on paracetamol (PCM) induced hepatotoxicity via its reactive intermediate metabolite N‐acetyl‐1,4‐benzoquinone imine (NAPQI) in albino rats. PCM is an over the counter drug and commonly abused. Overdose of NAPQI causes oxidative stress to the liver and other tissues as the endogenous enzymes will not be able to handle the toxicity. Tetracarpidium conophorum (TC) nutritionally contains a lot of vitamins C and E. The rats were divided into five groups of five rats each. (Gp1: n‐saline, Gp2: PCM alone, Gp3: PCM + 250mg/kg Extr., Gp4: PCM +500mg/kg Extr. and Gp5: PCM + 750mg/kg Extr.) All the animals were starved for 24hrs before the administration of PCM to groups 2 – 5. After 48hrs, Gps 3 – 5 were given 250, 500 and 750mg/kg of TC extract respectively for 7 days and then the animals were sacrificed. The levels of the transaminase enzymes were used to assess the intergrity of the liver. The antioxidant status in the liver was determined by measuring the activity of Glutathione (GSH), Superoxide dismutase (SOD) and Catalase (CAT). The oral treatment of rats with different doses of the extract showed significant (P≤ 0.05) hepatoprotective activity against NAPQI induced hepatotoxicity by decreasing the activities of transaminases (U/L) (ALT from 53.00 ± 2.10 – 31.40 ± 1.33, AST from 127.20 ± 1.16 – 106.00 ± 2.21, ALP 67.80 ± 0.86 – 57.60 ± 0.93), Total Cholesterol from 87.40 ± 0.61 – 67.68 ± 1.27, Total Bilirubin (g/dl) from 1.50 ± 0.05 – 1.24 ± 0.08 and significantly (P≤ 0.05) increasing the liver homogenate (U/L) of GSH from 28.80 ± 0.73 – 35.80 ±.86, CAT from 17.80 ± 0.28 – 18.5 ± 0.16 and SOD from 15.80 ± 0.37 – 19.60 ± 0.68 activities compared with the negative control. These findings showed that the walnut seed may have cytoprotective and antioxidant properties.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Neuromuscular function during aging is protected in baicalein‐treated C57BL/6 mice
Authors: Bhattacharya Arunabh , Walsh Michael , Hamilton Ryan , Singh Rashmi , Rahman Mizanur , Rodriguez Karl , Chaudhuri Asish , Sharma Ramaswamy
Published: June 2021

The loss of muscle mass and strength during aging is a debilitating condition leading to altered metabolism and the loss of independence in older adults. The causes of this condition, known as sarcopenia, are not known. We previously reported an increase in arachidonic acid and its metabolite 12‐hydroxyeicosatetraenoic acid (12‐HETE), which is generated by 12/15‐lipoxygenase (12/15‐LO), in sciatic nerves of old (28–30 months) C57Bl/6 mice. To determine if 12/15‐LO activity contributes to the age‐related decline in muscle and nerve function, we treated middle‐aged (24 mo old) mice with the 12/15‐LO inhibitor baicalein for 14–16 weeks. Compared to baseline measurements, baicalein treatment improved hindlimb muscle force generated by electrical stimulation. Baicalein treatment partially prevented the decline in fatigue observed in vehicle‐treated old mice compared to baseline. These improvements occurred without any change in muscle mass or sciatic nerve conduction velocity. Baicalein treatment was associated with a reduction in oxidative and ubiquitin modifications to proteins in sciatic nerves from old mice, which may be due to elevated proteasome activity and altered stress response in sciatic nerves from baicalein‐treated mice. Our data suggest that lipid metabolites generated by 12/15‐LO may contribute to the age‐related loss of muscle strength observed during aging.

Support or Funding Information

National Institutes of Health (K01AG038555) and American Federation for Aging Research (AB), K99/R00 Pathway to Independence Award (AG049940‐01A1) (KR), T32 AG021890‐08 (NIA) National Service Research Award to Postdoctoral Fellow Training Grant on the Biology of Aging to Steven N. Austad for trainee (RTH)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Novel Way to Use Vitamin C? Characterizing L‐Ascorbate (Vitamin C) Catabolism in Ralstonia eutropha
Authors: Wille Brendan , Stack Tyler , Carter Michael
Published: June 2021

L‐Ascorbate (Vitamin C) is present in the environment in plant tissues, typically at concentrations from 1–10mM, and as high as 300mM. L‐Ascorbate is also an essential radical scavenger (antioxidant) for plants and mammals and is not synthesized by bacteria. Given the ubiquity of L‐ascorbate in the environment, metabolism of L‐ascorbate necessarily contributes to ecological recycling. Ralstonia eutropha H16, a soil bacterium commonly studied for its versatile use of organic and inorganic carbon sources, can use L‐ascorbate as a carbon source via an unknown pathway. Using RNAseq, whole cell transcript levels in R. eutropha were compared between L‐ascorbate‐grown cells and succinate‐or fructose‐grown cells (controls). Putative pathway genes were identified by their increase in expression greater than 50‐fold in R. eutropha grown with L‐ascorbate. Mutant strains (lacking putative pathway genes) were generated by homologous recombination directed allelic exchange. Wild type and mutant strains were grown in liquid cultures with either L‐ascorbate or succinate as their sole carbon source and monitored by optical density to observe the phenotypic result of gene deletion. Growth studies have been performed, and L‐ascorbate no growth phenotype has been confirmed for three mutant strains. Collaborators have characterized putative pathway enzymes. Characterizing the proposed catabolic pathway of L‐ascorbate in R. eutropha will provide information regarding environmental nutrient cycling and contribute novel functions of suspected pathway enzymes to protein databases.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Andrographolide alters metabolism and mitochondrial function in prostate cancer
Authors: Lopez Rivas Andres R , Sanchez Vazquez Maria R , Zuazaga Humberto Ortiz , Torres Ramos Carlos A , Ferrer Magaly Martinez
Published: June 2021

Prostate cancer (PCa) is the second most common diagnosed cancer and the fifth cause of cancer mortality in men worldwide. In the United States, PCa is the most common diagnosed cancer and the third cause of cancer mortality in men. PCa is often treated using radiation, chemotherapy or radical prostatectomy. However, the combination of natural products with the standard of care treatment is an emerging area of cancer therapeutics. Previous studies from our laboratory showed that Andrographolide inhibits prostate cancer progression through a DNA damage response mechanism. In this study, we investigated the role of Andrographolide in metabolic function. Tumor development was performed by using an orthotopic xenograft model in which the prostates of SCID mice were injected with the prostate cancer cell line 22RV1. These mice were treated three times per week with Andrographolide 10 or 25 mg/kg for 4 weeks. Tumor tissue was collected to evaluate the effect of Andrographolide in tumor gene expression. Microarray analysis identified carbohydrate metabolism, connective tissue development and function, and cell cycle as a major molecular and cellular function altered in tumors treated with Andrographolide. Specifically, we found that the expression profiles of SASS6, FBXL4, MYDGF, TKK, MRSP33 and ZNF766 were significantly altered, suggesting that Andrographolide may be altering tumor metabolism. To confirm this observation, bioenergetics profile was determined using an Extracellular Flux Analyzer (Seahorse XF Mito Stress assay). The results of these experiments indicate that Andrographolide treatment decreases the Maximum and Spare Respiratory Capacity of PCa cells. These data suggest that Andrographolide disrupts the bioenergetics profile of PCa cells.

Support or Funding Information

Work funded by Institutional funds from the University of Puerto Rico Comprehensive Cancer Center and the School of Pharmacy. HOZ was supported in part by NIH‐NCI grant U54CA096297. MARC grant 5T34GM007821‐38.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Leucine‐rich diet changes tumor metabolism, reducing glucose consumption and metastasis in Walker 256 tumor‐bearing rats
Authors: Viana Lais ROSA , Tobar Natalia , Busanello Estela , Marques Ana Carolina , Lima Tanes , Machado Gabrielly , Castelucci Bianca , Ramos Celso Dario , Sergio  , Silveira Leonardo Reis , Vercesi Anibal Eugenio , Consonni Silvio Roberto , Gomes‐Marcondes Maria Cristina Cintra
Published: June 2021

Leucine stimulates muscle protein synthesis and is begin used as a potential treatment for muscle wasting conditions, such as cancer cachexia. However, the effects of leucine in the tumor tissue are still unknown. We aimed to describe the effects of a leucine‐rich diet on the Walker 256 tumor tissue. For this purpose, tumor‐bearing Wistar rats were randomly distributed into a control group (normoprotein diet) and leucine group (leucine‐rich diet (18% protein + 3% leucine)). After 20 days of tumor inoculation, the animals underwent to 18F‐FDG PET‐CT and after being euthanized, fresh tumor biopsies were collected to measure the oxygen consumption rate and fixed for electron microscopy analysis. Our results showed that the leucine‐rich diet leads to a lower tumor glucose (18F‐FDG) uptake by tumor and reduced number of metastases sites. Furthermore, leucine‐rich diet changed the mitochondrial morphology and function (from a glycolytic to an oxidative phenotype), associated with a higher gene expression of oxidative phosphorylation components. In addition, tumor tissue from leucine group showed a higher number of apoptotic nuclei with an increased oxidative stress. In conclusion, leucine‐enriched diet was associated with considerable metabolic activity reduction of malignant tissue associated with mitochondrial morphology and function changes in Walker 256 tumor tissue.

Support or Funding Information

The authors are grateful for financial support from CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior), CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnológico #302863/2013‐3; #302425/2016‐9), and FAPESP [grant number #2012/06955‐0; #2014/13334‐7; #2015/21890‐0; #2017/02739‐4].

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of MED28 and Vitamin D on Glycolytic Metabolism and Cell Growth in Human Colorectal Cancer Cells
Authors: Weng Yu‐Ting , Li Chun‐I , Liu Hsiao‐Sheng , Lee Ming‐Fen
Published: June 2021

Mammalian Mediator Complex has been implicated in human diseases, including cancer and metabolic disorder. MED28, a Mediator subunit, was found highly expressed in several types of cancer. Vitamin D, a secosteroid with hormone‐like features, exhibits multiple biological roles, including calcium homeostasis and cell differentiation. Calcitriol, 1,25‐dihydroxyvitamin D3, and its analogues have been suggested to suppress cancer development through vitamin D receptor (VDR). Our laboratory has previously reported that MED28 mediates Wnt/β‐catenin signaling and glucose metabolism in human colorectal cancer (CRC) cells, and we have also reported that calcitriol inhibits MED28‐mediated cellular events; in this study, we further examine their underlying mechanisms. First, by immunohistochemistry, we observed higher expression levels of MED28 in cancer tissues than those of adjacent normal counterparts in a CRC tissue microarray containing 19 paired tissue samples. Secondly, we detected lower protein expression of MED28, glucose transporter 1 (GLUT1), lactate dehydrogenase (LDHA), and cyclin D1, and less lactate production and higher pH value of the medium when we cultured human CRC cells in low glucose medium as compared with high glucose (25 mM) medium. Thirdly, the employment of siMED28 decreased both glucose uptake and lactate production, accompanied by lower expression levels of GLUT1 and LDHA as well as cyclin D1. Calcitriol inhibited the expression of MED28, and the effects of calcitriol on glucose metabolism and cell growth resembled those of MED28 suppression. In contrast, ectopic expression of MED28 increased glucose uptake as well as the protein expression of GLUT1, LDHA, and cyclin D1. Moreover, MED28 appeared to co‐localize and physically interact with VDR. Taken together, our findings indicated that MED28 upregulated Warburg effect and cell growth, including glucose uptake, lactate production, and the expression levels of GLUT1, LDHA, and cyclin D1, and calcitriol inhibited these events, where the interaction of MED28 and VDR may be responsible for the suppressive effect of calcitriol on MED28.

Support or Funding Information

This work was supported by the grant MOST 106‐2320‐B‐039‐062‐MY3 to M‐F Lee.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Loss of the Retinoblastoma Protein Modulates Glucose Metabolism in Lung Cancer
Authors: Clem Brian , Dougherty Susan , Kruer Traci , He Liqing , Yin Xinmin , Zhang Xiang , Reynolds Lindsey
Published: June 2021

Dysregulated metabolism is a hallmark of cancer cells and is driven in part by specific genetic alterations in various oncogenes or tumor suppressors. The retinoblastoma protein (Rb) is a tumor suppressor that was initially characterized in regulating cell cycle progression but recent studies have highlighted a functional role for Rb in controlling cellular metabolism. As such, we aim to characterize the metabolic phenotype caused by genetic loss of Rb in a mouse model of lung cancer. We have employed the Kras+/LSLG12D/Rb1lox/lox and Kras+/LSLG12D/Rb1+/+ transgenic model to study the metabolic effects of Rb1 loss in lung cancer. Cre‐exposed Kras+/LSLG12D/Rb1lox/lox mice exhibit increased tumor burden as well as decreased overall survival compared to Kras+/LSLG12D/Rb1+/+ mice. Our tracer studies using bolus dosing of 13C‐glucose revealed an increase in glucose carbon incorporation into select glycolytic intermediates. Consistent with this result, Rb1 depleted tumors expressed higher hexokinase 2 and glucose transporter 1. Additionally, compared to Rb1‐proficient tumors, Rb1 null tumors exhibit decreased one and three carbon labeled aspartate, which is indicative of pyruvate carboxylase (PC) activity. However, Rb1−/− tumors were found to express higher PC compared to Rb1+/+ tumors, suggesting these tumors may utilize an alternative nutrient source for TCA anaplerosis other than glucose. Taken together, these data have identified several metabolic genes under Rb control, and suggest Rb may regulate differential nutrient utilization in lung tumors.

Support or Funding Information

NCI RO1CA166327, American Cancer Society RSG 13‐139‐01

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TNF ligand related molecule‐1A inhibits atherosclerosis in apoE‐deficient mice
Authors: Zhao Dan , Yang Xiaoxiao , Chen Yuanli , Duan Yajun , Han Jihong
Published: June 2021

TNF ligand related molecule‐1A (TL1A) is one of vascular endothelial growth inhibitors. The regulation of neovascularization by TL1A indicates that it may play a vital role in the development of atherosclerosis. However, the exact effect of TL1A on the development of atherosclerosis and the underlying mechanisms, such as the functions of TL1A on foam cell formation and vascular smooth muscle cell (VSMC) phenotype, remain unknown. In this study, we induced atherosclerosis in apoE deficient (apoE−/−) mice by feeding the animals a high‐fat diet (HFD) while intraperitoneally injecting them recombinant TL1A twice a week, for 12 weeks. After treatment, we collected mouse aorta, serum and liver samples to determine the effect of TL1A on atherosclerotic lesions and fatty liver, and expression of the related molecules. The results of Oil red O, VVG and Alizarin red S staining show that administration of TL1A led to a significant reduction of lesions while enhancing plaque stability. The inhibition of lesions by TL1A was attributed to its effect on formation of foam cells derived from SMCs, but not from macrophages. Corresponding, TL1A activated expression of ABCA1 and ABCG1 in SMCs, but not in macrophages. Both the in vivo and in vitro results showed that TL1A inhibited the transformation of SMCs from contractile phenotype into synthetic one by inducing expression of contractile markers (α smooth muscle actin or αSMA) and inhibiting expression of synthetic marker (osteopontin, OPN). In addition, we determined that TL1A treatment decreased liver triglyceride contents by upregulation of HSL and ATGL expression. Taken together, our work shows that TL1A can inhibit the development of atherosclerosis by regulating SMC/foam cell formation and SMC phenotypes. Our study may also provide a new strategy for atherosclerosis treatment.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
para‐substituted methcathinones as selective and unselective inhibitors of human dopamine and serotonin transporters
Authors: Niello Marco , Cintulova Daniela , Hellsberg Eva , Jäntsch Kathrin , Holy Marion , Kudlacek Oliver , Cozzi Nicholas , Baumann Michael H , Mihovilovic Marko , Ecker Gerhard , Sitte Harald
Published: June 2021

Methcathinone (MCAT) is a compound belonging to the class of cathinones which targets monoamine transporters including DAT and SERT. Despite the importance of DAT and SERT as drug targets in several neurological disorders, the key factors underlying the selectivity profiles of their inhibitors are still poorly understood. Recent findings from rat synaptosomes suggested that increasing the volume of the para‐substituent of MCAT results in a swap of the selectivity between human DAT and SERT (1). Docking studies hint towards Ser149 in DAT and Ala169 in SERT as key residues involved in the difference of activity between DAT and SERT (2). The aims of the present biochemical and pharmacological study are to understand (i) which chemical properties (e.g. volume, polarity, or lipophilicity) of the para‐substituent influence the selectivity profile of MCAT between DAT and SERT, and (ii) whether Ser149 in DAT and Ala169 in SERT can be experimentally verified as key residues. Hence, we combined in silico‐driven synthesis, mutagenesis, and radio‐tracer flux assays in HEK293 cells expressing the human DAT and SERT wild‐type and respective mutants. We found that only MCAT and the para‐CF3‐MCAT showed high selectivity, 200‐fold for DAT/SERT ‐ and 25‐fold for SERT/DAT, respectively. This suggests that the high selectivity achieved is determined rather by specific features of these compounds than by the volume of the para‐substituent. Accordingly, we were not able to find any correlation between the selectivity profile of the tested 4MCATs with either volume, polarity, or lipophilicity parameters. Next, based on our results, we tested the hypothesis that the two amino acids in SERT and DAT do not suffice to explain the selectivity between SERT and DAT. We have tested the para‐substituted methcathinones in the swapping mutations DAT Ser149Ala and SERT Ala169Ser and in line with our hypothesis, these mutations did not revert the selectivity profile found in the wild‐type transporters. Our findings rather suggest that the SERT selectivity achieved by the introduction of the CF3‐group in the para‐position of MCAT is not dependent on the volume of the para‐substituent but on specific chemical features of the fluorine atoms which may influence the on and off rate of the MCAT molecule on DAT and SERT.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Therapeutic effects of melatonergic networks on propionic acid‐mediated autism‐like rat model: Sex difference based study
Authors: Kim Joo‐Heon , Choi Jeonghyun , Hong Yonggeun
Published: June 2021

Autism spectrum disorder (ASD) is characterized not only by altered neurobehavioral symptoms including repetitive and restricted behaviors, defects in social interactions, intestinal malfunctions such as constipation, and sleep disorders but also pathological brain damages. As the cause of this disorder is not clear, early diagnosis is limited and it may remain permanently disabled. Therefore, this study aims to explain the therapeutic potential of melatonergic network‐related materials in propionic acid‐mediated ASD animal models. Especially, to identify genderdependent changes, male and female 3‐week‐old Sprague‐Dawley rats were randomly divided and used. They were divided into control, propionic acid‐mediated ASD group, ASD with melatonin treatment group, and ASD with fermented milk treatment group. To verify the therapeutic effects of melatonergic‐related materials, intestine length, sleep quality and neurobehavioral tests were performed. ASD animals showed decreased intestine length compared to control. Interestingly, exogenous melatonin and fermented milk treatment to ASD animals restored intestine length in both sexes. However, no difference was found in the intestine length of both genders. Also, melatonin and fermented milk administration to ASD group showed restored sleep quality regardless of sexes. Additionally, ASD animals showed abnormal neurobehaviors compared to control animals, and this abnormality is thought to be recovered by exogenous melatonin and fermented milk in both male and female animals. Accordingly, melatonergic network may exert its potential as a therapeutic agent in ASD.

Support or Funding Information

NRF‐2017R1A2A2A01067169, KGM4611821, 2016 Creative Research Program of Inje University, Republic of Korea

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Biomarkers of Neurodegeneration in Goldfish Exposed to Nitrite
Authors: Valdez‐Oranday Diego Francisco , Martinez Melody , Huertas Mar
Published: June 2021

Goldfish have been used in numerous studies involving behavior and toxicology, however, their viability as an animal model to study neuropathies remains to be elucidated. We have previously discovered that the nitrite ion (NO2−) acts as an olfactory disruptor in goldfish. We hypothesize that this disruption is due to neurotoxic effects of NO2− in olfactory receptors and the brain. In this study, goldfish were subjected to a subacute, sublethal 1.0 mM sodium nitrite treatment, and had their brains and olfactory tract dissected. The tissues were mounted in paraffin blocks for sectioning and visualized with hematoxylin and eosin (H&E), Alcian blue, terminal deoxynucleotidyl transferase dUTP nick‐end labeling (TUNEL), caspase immunohistochemistry, and Congo red β‐amyloid staining. We discovered that olfactory tissue from treatment fish displayed signs of hyperplasia and hypertrophy when compared to the olfactory tissue from control fish. Furthermore, signs of increased apoptotic processes, in the form of blebbing, were observed in treatment fish. Alcian blue staining for mucosal cells demonstrated that the microvilli lining the epithelia had lost their brush‐like morphology and were covered by a thick layer of mucus in treatment fish. TUNEL, caspase immunohistochemistry, and β‐amyloid stains are currently being analyzed. Our results indicate that NO2− acts as a neurotoxin in subacute, sublethal doses. Because of the similarities in olfactory processing between goldfish and other vertebrates, it is possible that NO2− may also play a similar role in other aquatic vertebrates. The general availability and easy handling of goldfish make it an ideal candidate to study neuropathies.

Support or Funding Information

Texas State University start‐up funding for Dr. Mar Huertas

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Natural Rubber Latex as Stem Cell Scaffold for Muscle Regeneration
Authors: Floriano Juliana Ferreira , de Oliveira Graeff Carlos Frederico , Barbosa Angélica Mércia Pascon , Marcondes João Paulo , Pinto Taís Silva , Zambuzzi Willian Fernando , Rudge Marilza Vieira Cunha
Published: June 2021

Natural rubber latex (NRL) is a product extracted from rubber trees. The most common source of NRL is Hevea brasiliensis, NRL is a mixing colloidal rubber particle stabilized by a thin layer of phospholipids and proteins. The NRL has several features that make it an excellent scaffold and biomaterial to promote the growth and repair of tissues, due to their characteristics, like biocompatibility, angiogenic potential, flexibility, mechanical stability, surface porosity, permeability and low cost. The present study aimed at assessing the potential NRL use as stem cell scaffold, for the future use in muscle regeneration and cell therapy. The NRL membranes were produced with latex collected from RRIM 600 clone, the NRL was stored at a low temperature to prevent spontaneous coagulation and was centrifuged for 5 min at 1,500 rpm to remove impurities gathered in the collection process. The membranes were prepared by depositing a thin layer of NRL covering the bottom of a polystyrene petri dish measuring 60 mm in diameter and carried to a oven, where it was dried at 50 °C. The study was approved by Botucatu Medical School ethics committee on the use of animals (CEUA number 1234/2017). The mesenchymal stem cells were isolated from the bone marrow of the femur of young rats and cultured until the fourth passage. After which the cells were characterized by immunocytochemistry and immunophenotyping techniques, cell adhesion on the surface of the NRL membrane after 7 days of sowing the cells was assessed by scanning electron microscopy (SEM), confocal microscopy, zymography techniques for the evaluation of metalloproteinases 2 and 9, electrophoresis and protein expression for integrins. Our partial results showed that 98% of the cells expressed CD90, besides adhesion to plastic and the ability to differentiate and formation of fibroblast colonies, according to the methods indicated by the International Society for Stem Cell. After being seeding the surface of the NRL membranes it was verified high cell viability, excellent cell adhesion high cell proliferation rate, demonstrating the great potential NRL as stem cell scaffold.

Support or Funding Information

This project is funded by São Paulo Research Foundation – FAPESP (number grant 2016/01743‐5)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The effects of hyperglycemia on functionality of Rectus abdominis muscle of pregnant women
Authors: REYES DAVID RAFAEL ABREU , Barbosa Angélica Mércia Pascon , Floriano Juliana , Quiroz Sofia Mércia Vega , Enriquez Eusebio Mércia , Marcondes Joao Paulo , Okoshi Marina Mércia Politi , Rudge Marilza Mércia Vieira Cunha , Calderon Iracema Mércia , Raghavendra Hallur Mércia
Published: June 2021

Introduction

Gestational diabetes mellitus (GDM) is an important risk factor for urinary incontinence (UI) two‐year post caesarean section (CS). Previous studies using rectus abdominis muscle (RAM) of pregnant rats with severe and moderate diabetes confirmed diabetic myopathy as a risk factor to UI. But the pathophysiological mechanisms of UI are not completely understood. Indeed, hyperglycemia can damage skeletal muscle with atrophy due to neuropathy or direct effect on muscle cell metabolism. In this context, this study evaluated for the first time the effects of hyperglycemia on the biomechanics of the RAM of pregnant women using Miography − an innovative technique, ex‐vivo, sensitive, that evaluates the function of the myocardial fiber through a controlled electrical stimulation.

Objective

To identify the possible deleterious effects of hyperglycemia on RAM functionality (ex vivo contractility) in pregnant women with and without GDM.

Methodology

RAM of four primipara women with and without IU (three normoglycemic and one hyperglycemic) were collected during CS. The surgeon collected 1 cm in length of RAM and placed it in Krebs solution at 4°C. The changes in isometric tensile strength were recorded using LabChart software (LabChart 7 for Windows, AD Instruments) coupled to PowerLab (The PowerLab Data Acquisition System, AD Instruments) software for capturing and storing muscular responses. Muscle mechanical responses were expressed as a percentage of maximal contraction induced by electrical stimulation (Grass Model S48). We performed a multiple regression analysis adjusted to confounders using SPSS v21.o software. p < 0.05 was considered as statistically significant.

Initial results

We obtained results of the RAM function after electrical stimulation during one hour of the four women. We noticed that the RAM of pregnant women does not sustain the electrical response during one hour of the experiment. We have not enough data to compare both groups, but I confirmed that the myography is effective to evaluate the RAM functionality.

Conclusion

These initial results confirmed myography as a viable technique to analyze the functionally of the RAM in pregnant women, in a controlled way. The effects of hyperglycemia on the functionality of the RAM were identified.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Topical Application of Recombinant Human MG53 Protein Facilitates Healing of Chronic Wounds in Diabetic Mice
Authors: Zhou Xin , Li Haichang , Zhou Xinyu , Tan Tao , Xu Xuehong , Ma Jianjie
Published: June 2021

Wound care is a major challenge to public health and the burden for wound treatment is growing due to the aging population and the sharp rise in the incidence of diabetes and obesity, diseases that contribute to poor wound healing. While multiple factors may contribute to complications associated with wound healing, compromised tissue repair represents an underlying cause for non‐healing wounds in diabetes. We previously identified a novel TRIM family protein, named MG53, as an essential component of the cell membrane repair machinery. Here we tested the therapeutic value of using recombinant human MG53 (rhMG53) protein to treat chronic wounds. We developed a prototype formulation of rhMG53 in a topical cream that remains stable for long‐term storage at 4°C and is suitable for wound healing applications. We developed a mouse model of chronic excisional dermal wound using the db/db mice. Under control conditions, dermal wounds in the db/db mice would not heal until day 21. Treatment with topical cream containing 1.5 and 10 mg/oz of rhMG53 accelerated wound healing. Application of the topical cream without rhMG53 was ineffective. Wounds treated with 10 mg/oz rhMG53 cream healed nearly completely by day 12. Histological analyses revealed remarkable improvement of hair follicle structure at the dermal wound site in db/db mice that received treatment with rhMG53; the healing site that received cream alone showed defective hair follicles. We have previously demonstrated that genetic ablation of MG53 led to defective hair follicle structure in the mg53−/− mice. Our findings suggest that MG53 could improve chronic wound healing in diabetes by enhancing the hair follicle structure and the regenerative capacity of the skin.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Blockade of the AT1 receptor and silencing of CD44 gene expression are associated with inhibition of angiotensin II‐activated cardiac fibroblasts
Authors: Bai Feng , Yang Guang‐Zhao , Wang Ning‐Ping , Zhao Zhi‐Qing
Published: June 2021

Objective

We have previously demonstrated that the stimulation of fibroblasts by angiotensin II (Ang II) contributes to cardiac fibrosis via transforming growth factor beta (TGF‐β1)‐mediated collagen production. The cellular/molecular mechanism responsible for fibroblast activation, however, is not well understood. CD44, a cell‐surface glycoprotein, is associated with cell‐cell interactions, cell adhesion and migration. This study examines the roles of Ang II AT1 receptor and CD44 gene expression in the activation of cardiac fibroblasts.

Methods

Cardiac fibroblasts were isolated from the neonatal rat hearts, the assays of cell viability and migration were selected to demonstrate the activation of fibroblasts, and silencing of CD44 gene expression on fibroblasts was conducted with small interfering RNA (siRNA).

Results

Immunofluorescence staining with vimentin and discoid in domain receptor‐2 revealed the purity of isolated fibroblasts. Ang □ significantly reduced the cell viability and increased the cell migration in a dose‐dependent manner with a maximal stimulation as seen at 50 nM. In coincidence with fibroblast activation, mRNA and protein level of CD44 were up‐regulated and the protein levels of TGF‐β1, Smad2/4 and collagen I were significantly increased (all p<0.05 vs. unstimulated cells). Addition of Ang II AT1 receptor blocker, telmisartan into the cells abolished Ang II‐elicited stimulating effects on these parameters measured (all p<0.05 vs. Ang II activated cells). Consistent with data showing that there is a linear correlation between CD44 and TGF‐β1 as demonstrated by Pearson correlation analysis (r=0.955, p<0.01), gene transfection of fibroblasts with Ad‐CD44 siRNA as identified by mRNA and protein levels of CD44 demonstrated a significant attenuation in the production of collagen I, suggesting a role of CD44 in the activation of fibroblasts.

Conclusions

These results indicate that cardiac fibroblasts, in response to Ang II stimulation, express high levels of the AT1 receptor and CD44, which reduce the cell viability and facilitate cell migration. Such a signaling mechanism could be crucial for leading to the deposition of collagen and the production of cardiac fibrosis.

Support or Funding Information

This study was supported in part by grants from the Mercer University School of Medicine, the Medcen Community Health Foundation, Georgia and the National Natural Science Foundation of China (81470436).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Preventative Compositions of Azadirachta indica Against Fusarium oxysporum
Authors: DeJesus Kaitlin
Published: June 2021

As of 2018, there are over 7.8 billion people in the world. With an increase in population, it does not guarantee an increase of resources (de Oliveira et al. 2014; Chan, Tong, and Ng 2016). There are currently less than 3% of direct‐farming in the United States alone. With an advance in medicine, technology, and intake of new information, we can help stabilize the economy with our all in one products such as genetically modified organisms like BT corn where it contains its own pesticides to void away any creatures but still look ripe and delicious for the customer. With increase population and new advancements comes with new discoveries such as viruses, bacteria, and overall pathogens. (Agtmaal et al. 2017; Tümen et al. 2018; de Oliveira et al. 2014)

Fusarium oxysporum, a soil‐borne fungus that is best known for causing Fusarium wilt disease to many agricultural crops such as bananas, onions, peas, potatoes and tomatoes. Essential oils have beneficial ingredients that contain antibacterial, antifungal, and protective coating properties that aid in the process of shielding plants from these plant pathogens. Fusarium oxysporum f.sp. has been shown to have less mycelial growth when the topical use of Azadirachta indica essential oil is used (Aznar et al. 2015; Díaz‐Cruz and Cassone 2018). Azadirachta indica or the common name, Neem Oil, is a native of south‐eastern Asia, mainly cultivated in Indian subcontinent and used for the development of industrial products(Mohanty, Raghavendra, and Prasad Dash 2008; El‐Khatib, Ali, and El‐Mohamedy 2017).

This could positively impact society by providing more information regarding in understanding plant pathogens and to use essential oils like A. indica as a topical treatment against the growth of fungal infections. The need to test various concentrations of A. indica oil when exposed to Fusarium oxysporum, will help population to understand and adapt which concentrations are best against plant pathogens.

The experiment was ran through different combinations of neem oil, surfactant, and water to prevent the growth of 1 cm discs of Fusarium oxysporum. Results showed a negative correlation of fungal growth and neem oil where the higher concentration of neem oil, the less mycelial growth from Fusarium oxysporum. The effectiveness of neem oil against the fungi that causes Fusarium wilt effects the life span of agricultural crops.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison of T Cell Immune Response to Murine Cytomegalovirus (MCMV) in “Dirty” and Inbred Mice
Authors: Yocky Alyssa
Published: June 2021

Human cytomegalovirus (HCMV) is a herpes virus that is estimated to infect roughly half of the population of the United States and has been shown to cause congenital birth defects as well as disease in AIDs patients and immunocompromised patients. Due to this, extensive research has been done in mice using its ortholog, murine cytomegalovirus (MCMV). Although many breakthroughs have been made, the transfer of data over from the laboratory into human studies has proven difficult. Here, we examine the use of “dirty” mice as an alternative for laboratory mice as models of the human immune system. We injected “dirty” mice from the pet store and inbred laboratory mice with MCMV and measured the T cell immune response. Using flow cytometry, we quantified levels of activated cytotoxic T cells (CD8+CD25+) and activated helper T cells (CD4+CD25+) in the blood. We observed that there was no significant difference in the circulating levels of CD8+CD25+ or CD4+CD25+ T cells between dirty mice and clean mice 3 days after viral infection. In contrast, 9 days after viral infection, we observed significantly (p<0.05) higher percentages of CD8+CD25+ and CD4+CD25+ in dirty mice than in clean. These results show that dirty mice have significantly greater immune response that more closely mirrors the human immune system than inbred mice do. This suggests that dirty mice could be a more viable option for modeling the human immune system response to viral infection than our current mouse models.

Support or Funding Information

Fort Lewis College

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effect of Quorum Sensing by Staphylococcus epidermis on Blood Meal Choice in the Mosquito Aedes aegypti
Authors: Smith Ashley A. , Hechtel Laura J.
Published: June 2021

Female mosquitoes utilize volatile organic compounds (VOCs) elicited from the host in order to determine host preference. These VOCs include metabolites produced directly by the host or altered by the skin microflora and from products of bacterial quorum sensing. This research examined what bacterial density is required for quorum sensing to elicit enough VOCs that can be utilized for host selection in mosquitoes. The treehole mosquito, Aedes aegypti was used in a series of blood meal assays where females were given the choice of feeding on an artificial membrane with varying concentrations of Staphylococcus epidermis vs a control where only the medium was present on the artificial membrane. The number of females on the experimental or control artificial membranes were counted and recorded every minute. Blood meal preference was determined by finding the difference between the # of females on the experimental membrane vs. control membrane and then dividing by the total number of females observed. Results indicate that there is a positive correlation between bacterial concentration and mosquito blood meal preference. Female mosquitoes showed a high preference for blood meals where bacterial concentrations were greater than 106 CFU's. Since quorum sensing is utilized by bacteria typically at higher densities only, this research seems to indicate that VOCs produced through quorum sensing are utilized by mosquitoes in host selection. This research also supports differences among human host preferences in mosquitoes and may help to find alternatives to the use of insect‐repellents such as DEET.

Support or Funding Information

American Association of Anatomists (AAA)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Circulating exosomes of long non‐coding RNA DANCR correlated to prognosis of Hepatitis C virus‐related hepatocellular carcinoma
Authors: Wang Shu‐Chi , Tu Wen‐Yu , Hsieh Meng‐Hsuan , Huang Jee‐Fu , Chuang Wan‐Long , Dai Chia‐Yen , Yu Ming‐Lung
Published: June 2021

Hepatitis C virus (HCV) is one of major etiologies of hepatocellular carcinoma (HCC) worldwide. HCC is a highly prevalent and deadly cancer because of high risks of recurrence and/or metastasis. Identification of potential prognostic markers and/or target molecules could help in the preventing and managing HCV‐related HCC after the success of DAA treatment. Extracellular exosomes contain numerous signaling molecules including lncRNAs and play an important role in intercellular communication. To develop of liquid biopsy for prognostic markers of Hepatitis C Virus‐related hepatocellular carcinoma after surgical resection, we determined DANCR expression level in circulating exosomes. Total exosomes were extracted from 200 μL of serum by exosome precipitation solution and quantification DANCR by qPCR analysis. Results indicates exosomes DANCR levels was significantly higher among HCV‐related HCC patients with post‐resection recurrence than those without. Higher versus low exosomes DANCR with cut‐off at −7.5dCt [lncDANCR‐U6]) could predict HCV‐HCC recurrence (area under curve: 0.83). The 5‐y cumulative incidence of recurrence and mortality was 90.0% and 37.7%, respectively, among patients with higher exosomes DANCR, compared to 39.5% and 20.6%, respectively, among those with lower exosomes DANCR with harzard ratio (CI) of 3.8 (2.52–5.84) and 2.2 (1.29–3.89), respectively, by Cox regression model. The diagnostic power for HCC recurrence among HCV‐HCC patients was better with exosomes DANCR than with hepatic tumor DANCR or Tumor/Non‐tumor DANCR ratio. By knockdown or overexpression of DANCR in JFHlo (long‐term HCV‐infectious) cell models, we observed that increased DANCR enhanced cell migration and invasion ability by regulating mesenchymal‐associated β‐catenin expression. In this study, we demonstrated that circulating exosomes lnc‐DANCR could serve as a liquid biopsy for prognostic biomarker of HCC recurrence after surgical resection for HCV‐HCC.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.