bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of Inositol hexakisphosphate Kinase 1 (IP6K1) Does Not Increase Akt or mTOR Activity in vitro.
Authors: Barclay Richie , Mackenzie Richard
Published: June 2021

Background

Inositol hexakisphosphate Kinase 1 (IP6K1) has been identified as a key signalling enzyme that inhibits Akt activity in hepatic cell lines and diabetic rodent models (Chakraborty et al., 2010; 2012; Goshal et al., 2016). Given both glucose transport and protein synthesis require Akt activation, we investigate if IP6K1 altered the IGF ‐ Akt – mTOR signalling cascade.

Methods

C2C12 mouse skeletal myoblast cells were cultured in 12mL growth media (500mL DMEM, 10% FBS, 1% anti ‐ anti, 1% penicillin ‐ streptomycin) under standard conditions, changing media every 24 – 48 hours. Once 80% confluence was reached myoblasts were incubated in 12mL differentiation media (500mL DMEM, 2% donor equine serum, 1% anti ‐anti, 1% penicillin ‐ streptomycin) for 96 hours, changing media every 24 – 48 hours, to form mature myotubes. Myotubes were treated with IGF −1 (10ng/ml) +/− N2‐(m‐(trifluoromethy)lbenzyl) N6‐(p‐nitrobenzyl)purine (TNP; 10μm) for 24 hours before being lysed (1X cell lysis buffer supplemented with protease and phosphatase inhibitor cocktail) and total protein content determined using Lowry method. Western blot analysis was used to quantify t‐Akt, p‐Akt308, p‐Akt473, t‐mTOR, p‐mTOR2448 and IP6K1.

Results

A significant condition x IP6K1 effect was noted (P < 0.05) between IGF – 1 treatment and control in C2C12 myotubes, whilst IGF – 1 + TNP decreased IP6K1 content compared to IGF – 1 alone (P < 0.05). Significant differences were observed between IGF – 1 + TNP and control in p‐Akt308 and p‐Akt473 (P < 0.05), however no significant differences were observed between IGF – 1 + TNP and control in p/Akt308, p/Akt473 and p/mTOR2448 (P > 0.05).

Conclusions

This research is the first to characterize IP6K1 signalling in the IGF – 1, Akt – mTOR signalling cascade in C2C12 muscle cells. A reduction in IP6K1 did not increase p‐Akt or p‐mTOR activity.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modulating Cytoskeletal Structure And Cellular Signaling To Target Neuron Cell Membrane Repair
Authors: Paleo Brian J. , Bhattacharya Sayak , Weisleder Noah
Published: June 2021

Most cells in the human body have the capacity to reseal their cellular membranes following disruption of the lipid bilayer. The cell signaling response regulating membrane repair is dependent on changes in the mechanical tension on the plasma membrane. Increased mechanical tension on the membrane can lead to membrane disruptions remaining open and thus preventing resealing of the membrane. Disruption of these membrane repair processes can result in a number of diseases including muscular dystrophy, Alzheimer's and neurodegeneration. Conversely, if membrane repair can be elevated in striated muscle or neuronal tissues, this response could potentially minimize cell death in neuromuscular diseases and alleviate the progression of these disease states. We have previously shown that tripartite motif (TRIM) 72, also known as mitsugumin 53 (MG53), can increase plasma membrane repair in skeletal and cardiac muscle and in non‐muscle cell types where it is not usually expressed. Our recent results indicate that there are distinct TRIM72/MG53 regulated cellular signaling events that control the membrane repair response in multiple cell types. This observation led us to screen for novel TRIM family proteins that may be able to mediate membrane repair in neuronal cells. We found that TRIM2 transfected cells show an increased capacity for membrane resealing following multi‐photon laser injury and that knockdown of TRIM2 decreases membrane repair. Because TRIM2 is highly expressed in the nervous system and was previously shown to regulate neurofilaments (NFL), the objective of this study is to prove that TRIM2 protects against membrane damage in neurons through regulation of neurofilament ubiquitination and that disruption of TRIM2 or NFL leads to compromised membrane repair and neuron cell death. Through confocal microscopy, we have shown that TRIM2 co‐localizes with NFL in neurons. Following injury to the cell this association is altered, leading to changes in the subcellular localization of TRIM2 and an apparent reorganization of the neurofilament cytoskeleton. Given the extensive expression of TRIM2 in the nervous system, it may represent a good candidate for targeting membrane repair in the treatment of a variety of neurodegenerative diseases.

Support or Funding Information

American Heart Association, National Institutes of Health, National Institute of Neurological Disorders and Stroke (NINDS)‐sponsored T32, and Jain Foundation

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Damage Response in Doxorubicin‐Exposed Cardiac Fibroblasts
Authors: Mancilla Trevi Ann , Aune Greg
Published: June 2021

Pediatric cancers survivors exposed to doxorubicin during treatment are at an increased risk for cardiovascular disease. This study investigates two known responses to doxorubicin injury, the p53‐DNA‐damage pathway and the Parkin‐mitophagy pathway in the cardiac fibroblast. Doxorubicin is known to upregulate p53 expression. However, mitophagy of the mitochondria damaged by doxorubicin may be inhibited by this upregulation. Previous studies in cardiac myocytes have shown that cytosolic p53 can bind to Parkin and prevent it from localizing to damaged mitochondria, an early step of mitophagy. This study investigated if this also occurs in cardiac fibroblasts. Expression levels of p53 and Parkin were measured in primary cardiac fibroblasts exposed to doxorubicin. Immunoblotting and fluorescent microscopy both demonstrated increases in p53 and Parkin after doxorubicin exposure (p<0.05). Furthermore, the pattern of Parkin staining remains diffuse throughout the cytoplasm of the doxorubicin‐treated cells. Fibroblasts treated with a mitochondrial uncoupler had a linear staining pattern that may be due to the localization of Parkin with the mitochondria. As p53 is not upregulated by the uncoupler, Parkin localization would not inhibited. Future studies will determine if doxorubicin‐induced upregulation of p53 inhibits mitophagy and the effect this has on the cardiac fibroblast phenotype. Since cardiac fibroblasts mediate adverse cardiac remodeling, stress‐induced phenotypic changes to the cell will impact this process. Therefore, effects on the cardiac fibroblast are essential to understanding the development of the dilated cardiomyopathy seen in these patients.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Improved cardiac remodeling after in vivo knockdown of type VI collagen following myocardial infarction
Authors: Baker Nicholas J. , Minasyan Ashot , Adapala Ravi , McShannic Christian M. , Gan Connie , Hom Brian L. , Thodeti Charles K. , Meszaros J. Gary
Published: June 2021

Rationale

Our previous studies have shown that hearts of Col6−/− mice are protected from myocardial infarction (MI) injury, as these hearts exhibit reduced myocyte apoptosis, fibrosis and wall thinning leading to preserved long‐term cardiac function. The current project was designed to test whether knockdown of Col6 is feasible in the rat MI model and carries a similar cardioprotective effect during post‐MI remodeling.

Objective

To utilize a novel, non‐viral delivery method to deliver and determine whether siRNA‐mediated knockdown of Col6 is effective in the infarcted rat heart and affords cardioprotection in vivo in the weeks following MI injury.

Methods and Results

MI was induced via ligation of the left anterior descending artery (LAD) in adult Sprague‐Dawley rats and cardiac function was measured by transthoracic echocardiography. Calculated changes in endocardial volume, percent ejection fraction (%EF), and percent fractional shortening (%FAC) were performed at 1–6 weeks post‐MI. We witnessed a decrease in mean endocardial %EF of 44.7 ± 1.4 % in vehicle treated (control) rats, a parameter that was higher in siRNA treated hearts at 47.4 ± 1.8 %. A similar improvement was also seen in endocardial %FAC, which was 30.1 ± 1.4 % in vehicle treated rats versus 34.0 ± 1.3 % in siRNA treated rats. Knockdown of Col6 in vivo resulted in lower endocardial diastolic volume in siRNA treated rats (654.1 ± 81.3 μl in the vehicle group compared to 549.8 ± 8.4 μl for the siRNA treated group) and lower systolic volume in siRNA treated rats (362.7 ± 49.9 μl in the vehicle group compared to 291.6 ± 9.6 μl in the siRNA treated group).

Conclusions

siRNA mediated Col6 knockdown using JET‐PEI as a novel delivery method was effective in vivo, reducing Col6 expression levels over 75% in the anterior wall of the LV. Echocardiography revealed that the siRNA injections preserved cardiac function and prevented left ventricular chamber dilation following MI, indicating that this approach may have a positive therapeutic benefit in the recovery from MI injury.

Support or Funding Information

NIH R15HL132312

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
AMP‐Activated Protein Kinase (AMPK) Modulates Myoendothelial Junctions (MEJ) in Murine Microvessels
Authors: Qiu Jiehua , Tahir Sibgha , Schubert Kai‐Michael , Schneider Holger , Pohl Ulrich
Published: June 2021

Myoendothelial junctions (MEJ) in microvessels are not only the site of direct cell communication via gap junctions but also represent a signaling microdomain critical for regulating vascular processes such as endothelium dependent hyperpolarization (EDH). The 5′‐adenosine monophosphate‐activated protein kinase (AMPK) has been shown to influence vascular tone. It has not yet been studied, however, whether AMPK could also affect MEJ. Thus, the aim of this project was to elucidate the impact of AMPK on MEJ dynamics and its potential mechanism of action.

We studied isolated small mesenteric and skeletal muscle resistance arteries of mice using a pressure myograph system developed in our laboratory. Using confocal and two photon excitation fluorescence microscopy we identified the internal elastic lamina, which in these vessels exhibits small holes that allow direct contact between endothelial and smooth muscle cells. On average, we found that in about 35% of the holes, structures with a bright actin core as assessed by using F‐actin fluorescence in LifeAct mice or by staining with phalloidin which we could define as sites of MEJ. These structures showed expression of Cx37 and Cx43 suggesting that gap junctions (MEGJs) were also localized in these structures. Over observation times of up to 2 hours, we found that the number and localization of these MEJ varied, while they always positioned in the area of holes of IEL only. Thus, for the first time we could investigate MEJ dynamics. AMPK negatively regulates MEJ formation, since in AMPK α1, but not α2 knockout (KO) mice, the number of MEJ was significantly increased. In accordance, in wild type mice, incubation with the AMPK inhibitory compound C (CC) significantly augmented the number of MEJ. Furthermore, KO of AMPK α1, in intact mice as well as in cultured human and porcine smooth muscle cells, went along with an increase in PAI‐1 expression. Accordingly, incubation of arteries with exogenous PAI‐1 also increased the number of MEJ. The negative effect of AMPK on PAI‐1 could be explained by enhanced expression of the silencer of the PAI‐1 gene, small heterodimer partner (SHP). Since MEJ are also involved in EDH, we studied whether an enhanced formation of MEJ correlated with the magnitude of EDH induced vasodilation. Indeed, a higher amount of MEJs in α1 KO mice went along with a left shift of acetylcholine (ACh) ‐induced dilation dose effect curve.

This work describes for the first time a role of the α1 subunit of AMPK as a potent modulator of MEJ dynamics which is probably mediated by controlling PAI‐1 expression. Our data also for the first time demonstrate a dynamic regulation of MEJ expression in intact blood vessels.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Envigo's Female Salt‐Sensitive Rapp Rats are now Spontaneously Hypertensive and have Higher Frequencies of CD4+, CD4+ CD25+ and CD4+ CD25+ FoxP3+ T cells compared to Normotensive Salt‐resistant Rapp Rats
Authors: Pai Amrita V. , West Crystal A. , Souza Aline , Kadam Parnika S. , Pollner Emma J. , West David A. , Ji Hong , Wu Xie S. , Baylis Chris , Sandberg Kathryn
Published: June 2021

Introduction

We and others have found female Rapp (Jr) salt‐sensitive (SS) rats recently purchased from Envigo, formerly Harlan (Hsd) and maintained on a low sodium diet are spontaneously hypertensive. To further investigate the role of T cells in the new phenotype of spontaneous hypertension (SH) observed in the Envigo's Rapp strain (SHSS/JrHsd), we compared the T cell profile in SHSS/JrHsd with that of the Rapp normotensive salt‐resistant rats (NSR/JrHsd).

Methods

Mean arterial pressure (MAP) was measured by radio‐telemetry in juvenile (j) 1 month old (mo) SHSS/JrHsd, and in young (y) adult 4 mo SHSS/JrHsd and NSR/JrHsd rats. Helper (CD4+) and cytotoxic (CD8+) T cells were assessed in the kidney by multicolor flow cytometry in 1 and 4 mo SHSS/JrHsd and NSR/JrHsd rats.

Results

Radio telemetry confirmed that 1 mo jSHSS/JrHsd rats had normal MAP [(mmHg): 100±1; n=13]. Blood pressure rapidly increased with age and was significantly higher in 4 mo ySHSS/JrHsd rats compared to age matched normotensive yNSR/JrHsd [(mmHg): yNSR/JrHsd, 103±2 vs. ySHSS/JrHsd, 160±11; p<0.007; n=3/group]. The frequencies of CD4+ (1.3 fold; p<0.0001), CD4+ CD25+ (1.5 fold; p=0.002) T cells and the CD4+/CD8+ ratio [(CD4+/CD8+): jNSR/JrHsd, 1.1±0.07 vs. jSHSS/JrHsd, 2.1±0.1; p<0.0001; n=8/group] were significantly higher in jSHSS/JrHsd compared to jNSR/JrHsd. The frequencies of CD4+ (1.4 fold; p<0.0001), CD4+CD25+ (2.1 fold; p<0.0001) T cells and the CD4+/CD8+ ratio [(CD4+/CD8+): yNSR/JrHsd, 1.6±0.07 vs. ySHSS/JrHsd, 5.6±0.6; p<0.0001; n=7–8/group] were even higher in ySHSS/JrHsd compared to yNSR/JrHsd. Moreover, the ySHSS/JrHsd rats had a higher frequency of CD4+CD25+ FoxP3+ T cells compared to the yNSR/JrHsd at 4 mo (2 fold; p=0.0009).

Conclusions

Female Rapp SS rats from Envigo (formerly Harlan; Hsd) are normotensive as juveniles but get spontaneous hypertension by the time they are 4 mo; they should more appropriately be referred to as SHSS/JrHsd Rapp rats. Our T cell findings support previous studies indicating CD4+ T cells and subtypes contribute to hypertension.

Support or Funding Information

TL1‐TR001431 (AVP), UL1‐TR001409 (KS) and R01‐HL119380 (KS, HJ)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Computational Modeling of the Impact of Inflammation on Renal Hemodynamic Function
Authors: Dent Elena , Clemmer John , Ryan Mike , Pruett William
Published: June 2021

Systemic lupus erythematosus (SLE) is a chronic multi‐system autoimmune disease characterized by inflammation and alternating periods of flares and remission. Hypertension is a common comorbidity observed in SLE affecting up to 74% of this patient population. Lupus nephritis (LN) is a common manifestation of SLE but whether flare and remission impacts renal hemodynamic function or contributes to hypertension remains unclear. Mathematical modeling is an experimental approach that can provide valuable insight for predicting pathophysiological mechanisms. We hypothesize that developing a mathematical model of SLE flares will provide predictive mechanistic insight into renal hemodynamic changes that drive subsequent hypertension via inflammatory pathways. This study utilizes physiological simulation to test whether episodes of inflammation impair renal hemodynamic function and increase long‐term blood pressure. A previously described model of LN and renal inflammation by Budu‐Grajdeanu et al. (PMID: 20478032) was modified and incorporated into HumMod, a large integrative model of human physiology. To model SLE flares and remission, moderate or high inflammatory insults were given for 28 days followed by remission for 28 days for a total of 12 months. Glomerular filtration rate decreased during inflammatory episodes, and returned towards normal as inflammation subsided; however, it trended downward throughout the year (Figure 1). The number of sclerotic and non‐functional nephrons increased over the course of the simulation. Blood pressure was unchanged in both cases (Figure 2). These results suggest repeated inflammatory insults can impact renal hemodynamics and renal injury and may have the potential to eventually increase blood pressure. Longer simulations and further validation are needed to accurately model the chronic damage and loss of nephrons during SLE flares and remission and the impact of inflammatory mediators on renal hemodynamics, tubular function, and long‐term blood pressure control.

Support or Funding Information

VA Merit Award (BX002604‐01A2)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Enteric Neuroinflammation Driven by Exposure to Pyridostigmine Bromide is a Possible Contributing Factor to Gulf War Illness
Authors: Hernandez‐Rivera Siomara , Grubišiæ Vladimir , Fried David , Gulbransen Brian D.
Published: June 2021

Gulf War Illness (GWI) is a chronic disorder characterized by a spectrum of six symptoms that include gastrointestinal disorders. Exposure to the anti‐nerve gas drug pyridostigmine bromide (PB) is linked with the development of GWI, but the exact mechanisms remain unclear. We hypothesize that PB disrupts gut functions by creating persistent neuroinflammation within the enteric nervous system (ENS). We tested the acute effects of PB on the ENS in situ with calcium imaging recordings and in vivo by exposing mice to 9 μg/mL or 90 μg/mL PB (low‐ or high‐PB) for 7 days and subsequently assessing gut function using in vivo and ex vivo tests of colonic motility and barrier function. Neurochemistry of the ENS was assessed by immunohistochemistry. Our results show that glial calcium responses were significantly increased in the presence of PB (181.0%, p<0.001) when compared to ADP response. Mice exposed to high‐PB exhibited increased fecal pellet output (80.7%, p<0.05), higher fecal fluid content (19.1%, p<0.05), slower colonic bead expulsion (218.3%, p<0.001), altered neuromuscular control, and neurodegeneration in the ENS (18.3% neuronal loss, p<0.05) in comparison with control. Mice exposed to low‐PB showed an increase in the proportion of inhibitory enteric neurons (35%, p<0.05), while PB treatment caused reduced colon length (10.4%, p=0.0704 low‐PB; 9.8%, p=0.0908 high‐PB) and increased colonic fluid transport (31.0% low‐PB, 32.7% high=PB, p<0.05 both) regardless of the dose. Our results show that the acute exposure to PB significantly alters the anatomy and functions of the ENS. We propose that these changes contribute to the pathophysiology of GWI.

Support or Funding Information

Department of Defense (DOD) W81XWH1610631

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ca2+‐ activated Cl− secretion in murine distal colon is inhibited in acute dextran sulfate sodium (DSS)‐induced inflammation via down‐regulation of TMEM16A
Authors: Nickerson Andrew , Minor Emily , Rottgen Trey , Rajendran Vazhaikkurichi
Published: June 2021

Background

Mouse colonic epithelium exhibits Ca2+‐activated Cl− conductance (CaCC), which is mediated in large part by a Cl− channel known as TMEM16A (a.k.a. ANO1). Dextran sulfate sodium (DSS)‐ treated mice have been previously shown in the literature to have a dramatically reduced CaCC when treated with carbamoylcholine (CCH; a muscarinic receptor agonist). However, this finding predates the identification of TMEM16A as the primary mediator of CaCC, and was thus attributed to dysfunctional Na‐K ATPase and/or basolateral K+ channel activity. To date, the effects of DSS treatment on TMEM16A itself have not been determined. Furthermore, a cytokine involved in DSS pathology, transforming growth factor β (TGF‐β), has been shown to suppress TMEM16A expression in epithelial cell lines (T84 and HAEC). It is therefore plausible that DSS treatment causes a decrease in TMEM16A protein expression in mouse colon, and this phenomenon is potentially mediated by the effects of TGF‐β.

Hypotheses

1.) DSS‐induced inflammation causes a down‐regulation of TMEM16A expression, contributing significantly to reduced CaCC. 2.) Increased TGF‐β signaling in DSS‐induced inflammation results in suppression of TMEM16A.

Methods

BALB/c mice were given water with or without 5% DSS for 7 days, followed by 1 day of regular water. Disease Activity Index (DAI) scores, colonic length measurements and inflammatory cytokine profiles were recorded. Distal colonic epithelium was harvested either for short‐circuit current (ISC) recordings in an Ussing chamber or for qRT‐PCR and Western blot analyses. T84 cells were grown into monolayers and treated with one of a series of DSS‐associated cytokines for 48 hours. ISC recordings from T84 monolayers were taken and cells were subsequently used for qRT‐PCR and Western blot analyses.

Results

We found that DSS‐treated mice exhibit a substantially reduced secretory response to CCH compared to control (−1.4 vs 13.4 μA/cm2), which was corroborated by a decrease in TMEM16A protein (70%) and mRNA (81%) expression. T84 cells treated with TGF‐β, but not other DSS‐associated cytokines, also showed a reduction in CCH‐induced Cl− secretion (109.71 vs 170.1 μA/cm2), as well as TMEM16A protein expression (32%) compared to control.

Conclusions

DSS‐induced inflammation suppresses the expression of TMEM16A in mice at the transcript and protein level. We speculate that this effect is potentially mediated by TGF‐β signaling in the colonic epithelium.

Support or Funding Information

NIH 10019896.1.1006856R

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Curcuma longa L. Supplementation Ameliorates Hepatic Steatosis and Insulin Resistance in Rats Submitted to Fructose Consumption
Authors: Hasimoto Fabiana Kurokawa , Francisqueti Fabiane Valentini , Garcia Jessica Leite , Alves Pedro Henrique Rizzi , Ferron Artur Junio Togneri , Corrêa Camila Renata
Published: June 2021

Aim

To evaluate the effect of Curcuma longa L. supplementation on hepatic steatosis and insulin resistance in animals submitted to fructose consumption.

Methods

36 male Wistar rats (30 days of age) were used. Initially, the animals were assigned to receive standard diet + water (C group, n = 12) or standard diet + water with 30% fructose (F group, n = 24) for 16 weeks. After this period, rats were assigned into 3 groups to receive water or Curcuma longa L. (80 mg) via gavage for 8 weeks: Control (C group, n=12), Fructose (F group, n=12) and Frutose + Curcuma longa (F+CL group, n=12). Food (FC) and water (WC) consumption were measured daily and body weight (BW) weekly. At week 24, all animals were euthanized. Blood samples were collected for biochemical assays: plasma triglycerides (TG). Insulin resistance was calculated according to homeostasis model assessment of insulin resistance (HOMA‐IR). Steatosis was assessed in hematoxylin‐eosin stained liver sections. Statistical analysis were performed with ANOVA one way, p<0,05.

Results

There was no significant difference among the final weight (FW) and TG of the groups (p>0,05). About the comorbidities, fructose elevated the levels of HOMA‐IR in F group and Curcuma longa L. supplementantion improved this parameter (C = 9,80 ± 5,5; F = 25,72 ± 10,41; F+CL = 12,77 ± 5). Histological analysis revealed steatosis in F group characterized by the presence of larger drops. At the same time, in F+CL group supplementation was able to improve steatosis since it was possible to verify smaller drops. C group didn't show liver alterations.

Conclusion

The results showed that Curcuma longa L. is effective in ameliorating hepatic steatosis and insulin resistance leaded by fructose consumption.

Support or Funding Information

Fapesp 2016/18461‐2
Potential prebiotic effects of rice wine on Lactobacillus and Streptococcus
Authors: Xu Xiao , Yuan Biao , Xia Xiudong , Zhou Siduo , Han Yanhui , Xiao Hang , Dong Mingsheng
Published: June 2021

Rice wine has been associated with multiple health benefits due to various functional compounds it contains such as short‐chain fatty acids, organic acids, oligosaccharides and peptides. Lactobacillus and Streptococcus are considered as beneficial probiotics with protective effects on gastro‐intestinal health. In this study, rice wine was subjected to the simulated gastrointestinal digestion consisting of mouth, stomach and small intestinal phases. The final digesta was used to determine its potential prebiotic effects on probiotics, including Lactobacillus helveticus MB2‐1, Lactobacillus plantarum 70810, Lactobacillus plantarum D1501, Streptococcus thermophilus M5, Lactobacillus bulgaricus 1‐4, Lactobacillus plantarum 17‐1, Lactobacillus casei subsp. rahamnosus LS‐8, Lactobacillus plantarum B1‐6. Chemical analysis revealed that rice wine contained a considerate amount of citric acid, succinic acid, malic acid, lactic acid, acetic acid, propanoic acid and formic acid. The rice wine digesta dose‐dependently promoted the growth of probiotics. For example, the lag phase of Lactobacillus helveticus MB2‐1 was shortened from 10 h to 7 h, and μmax was increased by 16% due to the treatment of rice wine digesta. Similarly, the growth of Streptococcus thermophilus M5 and Lactobacillus bulgaricus 1‐4 was increased by addition of rice wine digesta, which was evidenced by shortened lag phase and increased maximum growth rate and maximum OD600. This is the first report on the prebiotic effects of rice wine digesta, which provided a basis to use rice wine and related products as prebiotics.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of mood after cognitive fatigue on gait in older adults
Authors: Avolio Abby , Goodwin Aurora , Fulk George , Martin Rebecca , Boolani Ali
Published: June 2021

Objective

The objective of this study was to examine the relationship between self reported mood (fatigue, vigor, motivation, trait physical energy (TPE), trait physical fatigue (TPF), trait mental energy (TME), trait mental fatigue (TMF)) and gait.

Methods

Subjects (N=12, age= 75.39±6.67 yrs, height=176.01±51.94 cm, weight=61.97±12.61 kg)) were involved in 2 days of a 2‐hour protocol of cognitively fatiguing tasks, after which they were administered the Profile of Mood Survey (POMS) and Trait Mental and Physical Fatigue and Energy surveys (TS), tasked to complete a 6MWT at their normal speed, and then the POMS and TS were re‐administered. Gait speed (GS), step length (SL) and stride length (StL) were measured using the OptoGait™ during the 6MWT. Gait was measured the first and last 30 seconds of the 6MWT. A bi‐variate Pearson correlation was used to determine the relationship between mood scores before the 6MWT to GS, SL, and StL in the first 30 seconds of the 6MWT and mood scores after the 6MWT to GS, SL, and StL in the last 30 seconds of the 6MWT.

Results

There were no relationships between all self reported mood variables and gait variables during the first 30 seconds of the 6MWT. However, there was a significant relationship between self reported fatigue and GS (p=.044, R=−.371); vigor and GS (p=.016, R=.437), SL (p=.020, R=.422), StL (p=.023, R=.415); TME and GS (p=.029, R=.400), SL (p=.023, R=.415), StL (p=.024, R=.410) in the last 30 seconds of the 6MWT. Which ones were there no relationship between last 30 seconds of the 6MWT.

Conclusion

Results indicate that self‐reported feelings of fatigue and energy play a role in gait performance during prolonged walking activity. These results suggest that mood in older adults who are cognitively fatigued may be related to gait. It seems that feelings of mental energy impact gait however, feelings of physical energy don't. We need to further evaluate why feelings of physical energy and fatigue are not related to gait. We also need to further evaluate whether changes in mood also change gait speed, stride length and step length.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Relationship between higher and lower level serial subtraction task performance and gait during a 6‐minute walk test
Authors: Rogers Emily , Avolio Abby , Goodwin Aurora , Vaccaro Rachael , Fulk George , Martin Rebecca , Boolani Ali
Published: June 2021

Purpose

The purpose of this study was to examine the relationship between cognitive fatigue as measured by performance on the serial subtract 3 (S3) and 7 (S7) at the end of a two‐hour protocol and walking performance as measured by step length (SL), gait speed (GS), and stride length (StL) in the first 30‐seconds of a 6‐minute walk test (6MWT) immediately following a cognitively fatiguing protocol.

Methods

Healthy older adults (N=12, age= 75.39±6.67 yrs, height=176.01±51.94 cm, weight=61.97±12.61 kg) were involved in 2 days of a 2‐hour protocol of cognitively fatiguing tasks (S3 and S7, continuous performance task (CPT), rapid visual input processing (RVIP),) after which they were tasked to complete a 6MWT at their normal speed. Gait speed, step length and stride length were measured using the OptoGait™. A bi‐variate Pearson correlation was used to determine the relationship between scores on the serial subtract 3 and 7 (after 2‐hours of cognitively fatiguing tasks) and GS, SL and StL for the first 30 seconds of the 6MWT.

Results

There was a positive significant relationship between S3 number correct and SL (R=.512, p=.018), GS (R=.457, p=.037), and StL (R=.481, p=.027) and S3 number of attempts and SL (R=.474, p=.030), GS (R=.434, p=.050), StL (R=.450, p=.041). There was no significant relationship between S3 number incorrect and S7 number correct, number incorrect and total attempts and gait parameters (p>.05).

Conclusion

Results suggest that after cognitive fatigue there is a relationship between lower level cognitive tasks and gait. Results suggest that showing lower level unique and characteristic errors in arithmetical reasoning may reflect underlying impairment in certain cognitive functions involved in gait. However, more challenging cognitive tasks (S7) was not related to gait. This may suggest that lower level executive function is involved in walking performance in older adults. Further research needs to be conducted to explore the exact relationship between higher and lower level cognitive tasks and single task gait and the neural circuitry involved.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sex Based Variability in Spatial Memory and Synaptic Transmission Occur Following Chronic Intermittent Hypoxia (CIH)
Authors: Barnard Jasmine Elizabeth , Nwakudu Chinwendu , Khuu Maggie , Garcia Alfredo
Published: June 2021

Sex is a key biological variable that has been shown to influence both physiology and disease. A growing body of research suggests that sex differences occur in respiration and brain physiology. This study examines how sex impacts changes in spatial memory and hippocampal synaptic plasticity following CIH, a model of sleep disordered breathing. Adult mice (>P30) of both sexes were either exposed to 0 days (control) or 10 days of CIH. On the final four days of CIH, mice were trained and tested in a Barnes maze apparatus. Glutamatergic synaptic transmission in area CA1 was recorded in hippocampal brain slices harvested from control and CIH mice. CIH exposure increased the latency for locating the exit in both sexes during the probe trial; however, only females showed increased variability in performance. Following CIH, long term potentiation (LTP) was depressed in both male and female mice. While no sex differences were observed in LTP, our ongoing experiments suggest that paired pulse facilitation (PPF) after CIH increases in females.

Support or Funding Information

The University of Chicago, Faculty Diversity Career Advancement Grant

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dark Chocolate (70% Cacao) Modulates Gamma Wave Frequencies in Vigorously Active Individuals
Authors: Silver Sasha , AlTikriti Ranea , Jin Nicole , Hodgkin Georgia , Bains Gurinder , Dhuri Sayali , Patel Krisha , Bradburn Jessica , Miller Josh , Bruhjell Kristin , Berk Lee
Published: June 2021

Introduction

Visualization and the demand to increase cognitive function to improve performance is an area of growing interest in athletes and physically active individuals. Modulatory control of acute action from cacao flavonoids on brain state remains unknown. The cacao flavonoid, epicatechin, can cross the blood brain barrier and accumulate in the hippocampal region of the brain, having a direct association with cognition, learning, and memory. Gamma (25–48 Hz) waves, the fastest of the brainwave frequencies, are optimal for cognitive function.

Purpose

Determine if savoring and ingestion of dark chocolate (70% cacao) can modulate gamma wave frequencies between visualization at a state of rest and a state of exercise performance via electroencephalography (EEG) in vigorously active individuals.

Methods

Ten vigorously active, healthy adults were enrolled. Participants consisted of 5 females and 5 males with a mean age of 23.7 ± 2.2 years. Vigorous activity was defined by the Centers for Disease Control and American College of Sports Medicine. Participants' visualization of rest and exercise were assessed by EEG Power Spectral Density (PSD) during three 60‐seconds trials: without cacao (1.4g of 70% cacao bar, Parliament Chocolates, Redlands, CA) consumption, savoring cacao, and after fully ingesting cacao. EEG wave band activity was recorded from nine cerebral cortical scalp locations using the EEG B‐Alert X10 System, Carlsbad CA. A Z‐score, using a reference baseline at visualization of rest without cacao, was utilized.

Results

During visualization of rest, there was a 541.5% increase in gamma wave frequency when comparing no cacao to savoring cacao and a 169.3% increase when comparing no cacao to after ingestion of cacao. During exercise visualization, there was a 207.5% increase in gamma wave frequency when comparing no cacao to savoring cacao and a 45.7% increase when comparing no cacao to after ingestion of cacao. When savoring cacao, there was a 13.8% increase in gamma wave frequency when comparing rest visualization to exercise visualization. After fully ingesting cacao, there was a 28.5% increase in gamma wave frequency when comparing rest visualization to exercise visualization.

Conclusion

We suggest that EEG gamma waves are heightened during savoring and ingestion of 1.4 g of cacao, when visualization of rest and exercise take place. Our study demonstrates the potential increase in cognitive function for vigorously active individuals to achieve a level of peak performance. Further research is needed to expand these positive findings.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular mechanisms of spinal cord regeneration
Authors: Echeverri Karen
Published: June 2021

Humans have very limited regenerative capacity especially in the central nervous system. In contrast, salamanders like the axolotl can functionally regenerate nervous tissue after injury throughout their life. After spinal cord injury (SCI) in the axolotl a population of cells, called glial cells adjacent to the injury site proliferates and migrates to reconnect the injured spinal cord and direct subsequent axon regeneration. This is in stark contrast to the mammalian response to SCI whereby injured astrocytes become reactive and contribute to glial scar formation and inhibition of axon regeneration. A major gap in knowledge exists regarding the molecular mechanism promoting a pro‐regenerative glial cell response to injury. Previously we identified the transcription factor c‐Fos as an essential regulator of the pro‐regenerative glial cell response to injury. However, c‐Fos functions as an obligate heterodimer with JUN family members to regulate gene expression. In mammalian astrocytes, up‐regulation of c‐Fos and c‐Jun after injury promotes reactive astrogliosis. Here I will present molecular data that identifies the molecular circuitry that must be carefully regulated to direct axolotl glial cells towards a regenerative response.

Support or Funding Information

Work in the Echeverri lab is funded by grants from NIH NICHD and Regenerative Medicine Minnesota.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Astrocyte Specific Insulin Receptor Deletion Contributes to Reproductive and Metabolic Dysregulation in Mice
Authors: Manaserh Iyad H. , Ravi Samyuktha , Hill Jennifer W.
Published: June 2021

Insulin resistance and obesity are associated with infertility and reduced GnRH release. It has been previously shown that male and female mice lacking insulin signaling in all cell types of the brain during development exhibit subfertility and develop diet‐sensitive obesity, mild insulin resistance and elevated insulin and leptin plasma levels. To determine whether astrocyte insulin sensing plays an important role in the regulation of fertility, we generated mice lacking insulin receptors in astrocytes (GLIRKO mice). GLIRKO males and females showed a delay in balanopreputial separation or vaginal opening and first estrous, respectively. GLIRKO female mice also exhibited longer estrus cycle lengths and irregular estrous cyclicity. In addition, these mice exhibited decrease in pregnancy rate and litter size, and thus decreased fertility. Histological examination of testes and ovaries showed impaired spermatogenesis and ovarian follicle maturation. GLIRKO mice also show hypothalamic hypogonadism, but normal sexual behavior, which confirms that the observed subfertility phenotype is due to HPG axis dysregulation. GLIRKO male mice displayed higher body weight and growth, and significant differences in body composition at 7 months of age. These mice also showed insulin resistance at 2 months and both insulin and glucose intolerance at 7 months of age. Histological analysis showed altered liver morphology with higher fat droplets. GLIRKO mice displayed significant decrease in energy expenditure, and a striking decrease in body temperature during fed state and overnight fasting conditions. However, when fasted mice were exposed to 4C, they were able to mount a robust thermogenic response and rapidly increase body temperature. These observations support the idea that astrocytic population that integrate information about energy stores to regulate body temperature is different from those required to respond to a cold challenge. Overall, our findings suggest that impaired insulin sensing in astrocytes delays the initiation of puberty and affects adult reproductive function. They also suggest that that astrocytic insulin signaling regulate body weight, systematic glucose metabolism and regulate thermogenic responses to nutritional and cold challenges

Support or Funding Information

This project is supported by NIH R01 HD081792 grant to Dr. Jennifer W. Hill

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Non‐invasive, Repeated Measurement of Estrogen Levels within Subjects in Prairie Voles
Authors: Francis Amie , Curtis J. Thomas
Published: June 2021

Prairie voles are a good model for the studying the formation of social bonds; in particular, the formation of monogamous pair bonds. Previous studies in our lab have shown that successful pair bond formation hinges on pregnancy onset within a short time after mating. Female prairie voles only enter estrous when exposed to a potential mate. A surge in estrogen levels induces behavioral estrous and progressive increases in estrogen are seen throughout the course of pregnancy. Measurements of estrogen are generally taken from blood plasma, but collection methods are highly invasive or require the termination of subjects. To better measure estrogen levels in female prairie voles over a time course, a non‐invasive means of collecting samples is needed.

We measured estrogen levels in fecal samples collected from female prairie voles over the course of pairing, mating, and pregnancy. Estrogen was extracted from fecal pellets using an organic solvent and ELISAs were run to measure changes in 17b‐estradiol concentrations. Animals that became pregnant showed progressively increasing levels of fecal estrogen over the course of mating and pregnancy, while animals that did not achieve pregnancy showed distinctly less fecal estrogen over the same time course. Our results suggest that measurement of fecal estrogen provides an alternative to more invasive means of measuring changes in sex hormones across time within subjects.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Calorie Restriction In The Preeclamptic‐like BPH/5 Mouse Reduces Adipose Tissue Gene Expression of Preeclampsia‐Related Inflammatory Genes
Authors: Reijnders Dorien , Olsson Kelsey , Redman Leanne , Sones Jennifer
Published: June 2021

Background

Preeclampsia (PE) is a hypertensive disease of pregnancy that is a leading cause of fetal and maternal morbidity/mortality. Obesity is a risk factor for developing PE, however, the underlying mechanism is unknown. Using the PE‐like BPH/5 mouse model, we previously demonstrated that cyclooxygenase (Cox)‐2 and interleukin (IL)‐15 dysregulation at the maternal‐fetal interface plays a causal role in poor maternal and fetal outcomes. Because BPH/5 mice are obese, we hypothesized that the maternal reproductive white adipose tissue (rWAT) is a source of these pro‐inflammatory molecules, and that reducing rWAT mass by calorie restriction (CR) would decrease the inflammatory uterine milieu in this model.

Methods

Using qRT‐PCR, we measured mRNA expression of Ptgs2 (the gene encoding Cox2), IL‐15 and 18S (housekeeper gene) within the rWAT and implantation sites of non pregnant and pregnant (embryonic day (e) 7.5) ad libitum fed BPH/5 and C57Bl/6J control mice. In addition, we determined Ptgs2 and IL‐15 mRNA expression in BPH/5 rWAT and implantation sites after 7 days CR (pair‐feeding with pregnant C57Bl/6J mice) beginning the day of copulation (e0.5).

Results

Ptgs2 expression in non‐pregnant rWAT was comparable between BPH/5 and C57Bl/6J. However, there was a 2‐fold increase in Ptgs2 expression at e7.5 in BPH/5 rWAT versus C57Bl/6J. After CR, Ptgs2 expression in BPH/5 rWAT was significantly lower compared to ad libitum fed BPH/5 mice (p=0.038, n=6). Also in implantation sites, the expression of Ptgs2 was significantly lower in CR as compared to ad libitum fed BPH/5 mice (p=0.046, n=6). IL‐15 expression was 30‐fold higher in BPH/5 rWAT and 8‐fold higher in implantation sites at e7.5 compared to C57Bl/6J control mice (p=0.0015, n=6), but this expression was not significantly affected by CR in either tissue.

Conclusion

These data show that 7 days CR in BPH/5 obese mice during early pregnancy reduces the expression of Cox‐2 coding genes in rWAT and at the maternal‐fetal interface, suggesting that WAT‐derived inflammatory mediators might be involved in the development of PE and poor pregnancy outcomes in obese BPH/5 mice. Future studies should investigate the molecular phenotype of WAT in women that are at risk to develop PE, to provide further insight in the mechanisms that make obesity a risk factor and possible interventions to prevent PE.

Support or Funding Information

This study has been funded by the Lousiana Biomedical Collaborative Research Program.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Early life stress reduces in vivo lipolysis efficiency in female mice fed a high fat diet
Authors: Herald Joseph Blake , Leachman Jackeline , Loria Analia S.
Published: June 2021

Previously, we have shown that female mice exposed to maternal separation and early weaning (MSEW), a model of postnatal neglect, increases fat mass and worsens the cardiometabolic function in response to high fat diet (HF) feeding. We also found that β3 adrenergic receptor mRNA expression, involved in local lipolysis stimulation, was reduced in gonadal white adipose tissue (gWAT). Thus, the aim of this study was to test the hypothesis that female MSEW mice display reduced lipolysis in‐vivo and in‐vitro. MSEW was performed by separating the pups from the mother for periods of 4–8 hr during postnatal days 2–16. Mice were weaned at P17. Control mice remained undisturbed in the home cage all times and weaned at P21. At weaning, mice were placed on a low fat (10% kcal from fat) or high fat (60% kcal from fat) diet for 16 weeks. At week 14, an in‐vivo lipolysis test was performed by injecting with 1mg/kg BW of isoproterenol, IP. Free fatty acids (FFA) were measured in plasma obtained after 15 minutes of the injection. At week 16, an in‐vitro lipolysis test was performed in gWAT. As such, gWAT was removed from female control and MSEW mice, weighed, and washed with ice‐cold PBS. In‐vitro lipolysis was performed in the presence or absence of isoproterenol (10uM, Sigma Aldrich). Free glycerol kit (Sigma Aldrich) was used to measure lipolytic products and results were normalized to tissue weight. In‐vivo, FFAs were reduced in female MSEW mice fed a HF (102±13 uM/ml) when compared with controls (144±24 uM/ml, p=0.066). In‐vitro, we found similar free glycerol levels in control and MSEW mice fed a LF (9.2±0.8 vs. 10.5±1.4 mg/dl/g tissue, respectively). Although HF reduced isoproterenol‐induced free glycerol levels, control and MSEW mice showed not statistically significant differences (4.4±0.6 vs. 3.5±0.6 mg/dl/g tissue, respectively). Taken together, these data indicate that HF reduced lipolysis capacity either in control and MSEW mice. Also, MSEW reduced lipolysis when determined in‐vivo conditions, suggesting that neuroendocrine and autonomic factors may be implicated in the mechanism by which MSEW mice are prone to increase fat mass.

Support or Funding Information

This study was supported by funds from the NIH National Heart, Lung, and Blood Institute to A.S.L (R00 HL111354), start‐up funds from the University of Kentucky to A.S.L, and the pilot project from the University of Kentucky Center of Research in Obesity and Cardiovascular Disease COBRE P20 GM103527‐06 to A.S.L.

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