bluuebunny/crossref_metadata_2025 huggingface dataset:
Late‐In‐Life Treadmill‐Training Ameliorates the Decline in Cardiac Autophagy Associated with Aging in Mice
Authors: Cho Jae Min , Ramous Caroline , Mattera Maria Sara , Hansen Michele , Pires Karla Maria , Ferhat Maroua , Whitehead Kevin , Carter Kandis , Buffolo Marcio , Park Seul‐Ki , Boudina Sihem , Symons J. David
Published: June 2021

Protein aggregates accumulate and organelles become damaged and/or dysfunctional during the process of aging. A progressive loss of the cellular quality control mechanism autophagy contributes to this age‐associated decline in cellular function in many organs. Evidence for an age‐associated repression in cardiac autophagy is not consistent. We hypothesized that 24‐month old (old) male C57Bl6/J mice exhibit repressed autophagosome formation in the heart, myocardial dysfunction, and reduced exercise capacity vs. 6‐month old (adult) mice. First, cardiac lysates from old mice displayed reduced (p<0.05) accumulation of LC3II/GAPDH and degradation of p62 vs. adult animals (n=12 per group). Second, the lysosomal acidification inhibitor chloroquine (CQ) induced accrual (p<0.05) of LC3II/GAPDH and p62 in hearts from adult but not old mice (n=7 per group). Third, left ventricular mass was greater (p<0.05), and indices of systolic, diastolic, and global left ventricular function (transthoracic echocardiography) were impaired, in old vs. adult animals (n=12 per group). Finally, maximal workload performed during a treadmill‐test was less (p<0.05) in aged (n=11) vs. adult (n=12) mice. To determine whether late‐in‐life exercise training induces cardiac autophagy, separate cohorts of male mice completed a progressive‐resistance treadmill‐running program (old‐ETR) or remained sedentary (old‐SED) from 21–24 months. Body composition, exercise performance during a maximal workload test, soleus muscle citrate synthase (CS) activity, indices of cardiac antioxidant enzyme activity, markers of cardiac autophagy, and indices of myocardial function, all improved (p<0.05) in old‐ETR (n=11) vs. old‐SED (n=12) mice. These data are the first to demonstrate that markers of cardiac autophagy are elevated, and indices of myocardial function are improved, in old mice that complete a treadmill‐training regimen that is sufficient to increase skeletal muscle CS activity and maximal exercise capacity. These data provide strong proof of concept to evaluate cause and effect relationships among exercise‐training, myocardial.

Support or Funding Information

UU Research Fellowship (JMC); APS UGRF (CR); AHA17POST33670663 (SKP); NIHRO1DK098646‐01A1, NIHRO1DK099110, AHA16GRANT30990018 (SB); AHA16GRNT31050004, NIH RO3AGO52848, NIH RO1HL141540 (JDS).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Obesogenic diet in aging mice disrupts gut microbe composition and alters neutrophil:lymphocyte ratio leading to inflamed milieu in acute heart failure
Authors: Kain Vasundhara , Van Der Pol William , Mariappan Nithya , Ahmad Aftab , Eipers Peter , Morrow Casey , Gibson Deanna L , Gladine Cecile , Vigor Claire , Durand Thierry , Halade Ganesh V
Published: June 2021

Calorie dense obesogenic diet (OBD) is a prime risk factor for cardiovascular disease in aging. However, increasing age coupled with changes in the diet can affect the interaction of intestinal microbiota impacting immune system that can lead to chronic inflammation. How the age and calorie enriched OBD interact with microbial flora and impacts leukocyte profiling is currently under‐investigated. Here, we tested whether OBD in young and aging mice alters the gut microbe composition and the splenic leukocyte profile in acute heart failure. Young (2 month‐old) and aging (18 month‐old) mice were supplemented with STD‐(standard chow diet ~4% safflower oil diet) and OBD‐(obesogenic diet‐10% safflower oil) diet for 2 months and then subjected to coronary artery ligation to induce myocardial infarction (MI). Fecal samples were collected pre‐ and post‐diet and the microbial flora analyzed using 16S V4 rRNA gene DNA sequencing and QIIME informatics. The STD and OBD in aging mice resulted in an expansion of the genus Allobaculum in the fecal microbiota. However, we found a pathological change of neutrophil: lymphocyte ratio in aging mice in comparison to their young counterparts. Thus, calorie‐enriched OBD dysregulated splenic leukocytes by decreasing immune responsive F4/80+/CD169+ macrophages in aging. OBD programmed neutrophil swarming with an increase in isoprostanes levels with dysregulation of lipoxygenases and cytokines. In summary, calorie dense OBD in aging mice disrupted the composition of the gut microbiota that correlates with the development of integrative and system‐wide non‐resolving inflammation in acute heart failure.

Support or Funding Information

Authors acknowledge the support from National Institutes of Health (NIH)‐NCCIH (formerly known as NCCAM) AT006704, HL132989‐01 to GVH and American Heart Association postdoctoral fellowship [POST31000008] to V. K. The authors also acknowledge support of Microbiome Resource at the University of Alabama at Birmingham: Comprehensive Cancer Center (P30AR050948), Center for Clinical Translational Science (UL1TR001417), University‐Wide Institutional Core, Heflin Center for Genomic Sciences and Microbiome Center to CM

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gamma‐mangostin of Garcinia mangostana peels ameliorates hyperglycemia in synergism of insulin
Authors: Chen Shi‐Pei , Lue Sheng‐I , Fu Yaw‐Syan , Cheng Hui‐Wen , Wang Chun‐erh , Weng Ching‐Feng
Published: June 2021

Type II diabetes mellitus is diagnosed because of obese having high sugar, high salt and high fat in diet, and is a heterogeneous group of disorders characterized by hyperglycemia due to an absolute or relative deficit in insulin synthesis or action. Mangostins, isolated from Garcinia mangostana peels, contain various isoforms α, β and γ, however, the medicinal values of γ‐mangostin particular in anti‐diabetic as an alternative medicine or folk medicine remain to be explored. This study attempted to investigate hypoglycemic efficacy of γ‐mangostin through execution in cellular and mouse levels. In vitro, the vascular smooth cells isolated from the external carotid artery of SD rat and FL83B mouse hepatocytes were applied to determine the cytotoxicity and glucose uptake. The cell viability data showed that 0.5 μM, 2.5 μM and 5 μM γ‐mangostin had non‐toxicticity in both cell types. The hypoglycemic effectiveness of 5 μM γ‐mangostin treatment was greater than those of 0.5 and 2.5 μM γ‐mangostin during two hours test of two cell types. Compared with 10−7 M insulin, the potency of γ‐mangostin plus insulin in synergistic glucose uptake reached plateau within 90 min. Furthermore, the hypoglycemic efficacy of oral administration 0.1, 0.2 and 0.4 mg γ‐mangostin was similar to 0.2 U insulin intraperitoneal injection and oral 50 mg/kg metformin for 120 min of oral glucose tolerance test (OGTT) in diet‐induced diabetic ICR mice. The hypoglycemic ability of oral 0.4 mg γ‐mangostin was more effective than that of 12 mg/kg Acarbose for 180 min in oral starch tolerance test. Diabetic ICR mice were daily fed 0.2 mg γ‐mangostin, 0.4 mg γ‐mangostin and vehicle for 4 weeks, the data revealed that γ‐mangostin caused the lowing blood glucose at fasting level and OGTT. Taken altogether, γ‐mangostin plays an insulin sensitizer to dampen the hyperglycemia with non‐ cytotoxicity in cell line tested.

Support or Funding Information

Ministry of Science and Technology (107‐2813‐C‐259‐061‐B), Taiwan.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Melatonin Supplementation Decreases Inflammatory Adipokines Expression from Adipose Tissue of Obese Animals
Authors: da Silva Mendes Farias Talita , Paixão Regislane Ino , Cruz Maysa Mariana , Dourado Roberta
Published: June 2021

The incidence of obese people worldwide is increasing every year and accumulating evidence indicates that dysregulated expression of adipokines, caused by excess adiposity and adipocyte dysfunction, has been linked to the pathogenesis of obesity‐linked complications. Melatonin, a hormone responsible for the synchronization of mammalian circadian rhythms, is related to beneficial effects on the control of obesity and its complications. In this work we evaluated the effects of melatonin supplementation on the production of adipokines in epididydimal (EPI) and inguinal (ING) fat depots from obese animals induced by a high‐fat‐diet (HFD). Male C57Bl/6j mice were divided into 3 groups: CO, animals fed with control diet; Obese, animals fed with HFD and Obese + melatonin, animals fed with HFD and supplemented with melatonin (1mg/kg). Melatonin presented a preventive effect on body weight gain and slowed down the effects on food and energy efficiency triggered by HFD. Adiponectin, IL‐6, leptin, and TNF‐α gene expression in ING and EPI fat depots were analysed. It was observed an increase (by 55 %, p <0.05) in adiponectin mRNA in ING depot when the animals received melatonin. There was an increase (3,7‐fold, p <0.05) in the IL‐6 mRNA expression in the EPI depot (but not in ING) in HFD group, that was prevented by melatonin supplementation. The same increase was observed on leptin expression in EPI adipose depot (6‐fold, p <0.05) from animals fed with HFD. Again, melatonin supplementation completely abolished this effect. Similarly, there was an increase (by 23‐fold, p <0.05) in the expression of this gene in the ING depot from HFD animals, and herein, we observed only a prevention trend (p = 0.0514) by melatonin on leptin mRNA in this fat depot. In both, EPI and ING depots, TNF‐α mRNA was significantly increased (by 5‐fold and 7‐fold respectively, p <0.05) in the HFD as compared to the CO group, but the melatonin supplementation not decreased this parameter. The protein analysis in these tissues (ING and EPI) performed by Elisa, showed that IL‐6 protein was increased (68%, p <0.05) in EPI fat depot from HFD group when compared to CO. This elevation was completely prevented when the animals were supplemented with melatonin. By the other side, no changes were observed between the groups concerning IL‐6 secretion in ING depot. The results obtained herein suggest that melatonin supplementation improves the adipokines expression in animais with obesity induced by a HFD, that is, it increases adiponectin while reducing the expression of inflammatory cytokines in adipose tissues. Together, these results show that melatonin supplementation had an protetive action on obesity complications

Support or Funding Information

Fapesp (2015/03554‐2)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cardiovascular Effects of Pyridoxamine in Type 1 Diabetic Female Rats
Authors: Delbin Maria A , Passos Matheus P , Ruberti Olivia M , Jarrete Aline P , Sousa Andressa S
Published: June 2021

The formation of advanced glycation end products (AGEs) is an important pathway involved in cardiovascular complications of diabetes mellitus (DM). The pharmacologic approaches in clinical use have remained unsatisfactory in preventing the diabetic cardiovascular complications and more innovative therapies are needed. Therefore, the aim of the study was to investigate the coronary vascular responsiveness and oxidative stress in the left ventricle in type 1 diabetic female rats and the effects of the combined treatment with insulin and pyridoxamine (AGEs inhibitor). This study was approved by the Ethical Committee for Animal Research Use (CEUA UNICAMP 4532/2017). Female Wistar rats (180–200g) were divided into four groups: control (CTR), type 1 DM (DM1, alloxan 170 mg/kg IP), DM1 treated with insulin (DM1+INS, 2U morning + 4U evening SC/day) and DM1 treated with insulin and pyridoxamine (DM1+INS+PDX, 180 mg/kg/day oral gavage). One week after DM1 induction the DM1+INS and DM1+INS+PDX groups initiated the respective treatment that lasted for three weeks. Concentration‐response curves to acetylcholine (ACh), sodium nitroprusside (SNP) and thromboxane A2 analog (U46619) were obtained in septal coronary artery. The generation of reactive oxygen species (ROS) were evaluated in left ventricle in basal condition and in the presence of superoxide dismutase mimetic (MnTMPyP). Biochemical parameters were measured. One‐way ANOVA followed by a Tukey's test was performed. As expected, the body weight was reduced and the blood glucose was increased in DM1 as compared with CTR group. In DM1+INS and DM1+INS+PDX groups the blood glucose was reduced and the body weight values were normalized. The maximal response (EMAX) to ACh was decreased in DM1 (72±6%) and DM1+INS (78±8%) groups compared with CTR (97±8%), the combined treatment in DM1+INS+PDX (99±5%) restored this alteration with no changes in potency (pEC50). Neither the EMAX nor pEC50 were modified for SNP and U46619 in all experimental groups. At basal condition, ROS measurement in the left ventricle was not modified in all groups. However, the reduction of ROS generation in the presence of MnTMPyP was impaired in DM1 group (approximately 50%) as compared with CTR group. Only the combined treatment with insulin and pyridoxamine was effective in preventing this alteration in DM1+INS+PDX group. In conclusion, our results show that insulin associated with pyridoxamine treatment prevented the coronary dysfunction and cardiac oxidative stress in type 1 diabetic female rats.

Support or Funding Information

FAPESP (2016/16351‐5; 2017/22670‐9) and CAPES.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of Acute Dietary Nitrate Supplementation on Exercise Blood Flow in Hypertension: Does Medication Status Matter?
Authors: La Salle D. Taylor , Broxterman Ryan M , Ratchford Stephen M. , Richardson Russell S. , Trinity Joel D.
Published: June 2021

Dietary nitrates are effective at lowering resting blood pressure (BP) in humans and improving skeletal muscle blood flow (BF) during exercise in animals. However, evidence of improved BP status and BF during exercise in hypertension is lacking. The purpose of this investigation was to determine if acute (3‐day) nitrate supplementation with beetroot juice (BR) lowers BP and augments BF in hypertensive individuals on and off antihypertensive medication during progressive handgrip (HG) and single‐leg knee extension (KE) exercise. In a double blind crossover study, placebo (nitrate depleted BR) and BR were administered in 14 subjects with controlled hypertension (ON‐MEDS; 53±11 yrs., 11 men, 3 women) and 14 subjects not taking antihypertensive medications (OFF‐MEDS; 49±13 yrs., 11 men, 3 women). Following either 3 days of BR or placebo supplementation, participants performed HG (15, 30, 45% of maximal voluntary contraction [MVC]) and single‐leg KE exercise (40, 60, 80% of work rate maximum [WRmax]). When OFF‐MEDs, BR significantly reduced resting diastolic BP (DBP) (BR: 84 ± 3 vs. PL: 88 ± 3 mmHg; p < 0.01) and mean arterial pressure (MAP) (BR: 101 ± 3 vs. PL: 106 ± 3 mmHg; p < 0.01), with systolic BP (SBP) trending lower (BR: 136 ± 4 vs. PL: 141 ± 5 mmHg; p = 0.067). BR did not change resting BP for the ON‐MEDS group. During HG exercise, BR supplementation only lowered SBP at 15% MVC (p < 0.05) and 30% MVC (p = 0.05) when OFF‐MEDS, with no effect on BF or vascular conductance (VC). During KE exercise, in the OFF‐MEDS group, BR significantly lowered SBP and MAP at 80% WRmax, but not at the lower intensities. When OFF‐MEDS, BR significantly increased leg BF at 40% WRmax (BR: 2667 ± 173 vs. PL: 2461 ± 193 ml/min; p = 0.04) and leg VC at 40% (BR: 24 ± 2 vs. PL: 21 ± 1 ml/min/mmHg; p < 0.01) and 60% (BR: 27 ± 2 vs. PL: 25 ± 2 ml/min/mmHg; p < 0.03) WRmax. When ON‐MEDS, BR did not alter BP, BF or VC during HG or KE exercise. These findings reveal that BR can lower BP at rest and during exercise, as well as augment the blood flow response during leg exercise in unmedicated hypertensive individuals, suggesting a potential benefit of BR to this population.

Support or Funding Information

This study was supported by Veteran Affairs Rehabilitation Research and Development Grant IK2RX001215 CDA2 (to J. D. Trinity) and American Heart Association Grant 14SDG1850039 (to J. D. Trinity)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of Mitochondrial Fission Improves Mitochondrial Respiratory Capacity in Primary Myotubes Derived from Obese Humans
Authors: Kugler Benjamin A , Hale Margaret , Eugene Nancy , Garcia Jessica , Houmard Joseph A , Zou Kai
Published: June 2021

Obesity induces mitochondrial fragmentation in skeletal muscle causing reduced mitochondrial respiratory capacity, which has been linked with the development of insulin resistance and type 2 diabetes. Recent evidence suggests that excessive mitochondrial fission is the culprit behind the mitochondrial fragmentation in skeletal muscle from obese humans. The purpose of this study was to determine if inhibiting mitochondrial fission in human skeletal muscle cells derived from severely obese individuals reduces mitochondrial fragmentation and improves mitochondrial respiratory capacity. Human skeletal muscle cells were isolated from muscle biopsies obtained from severely obese, insulin‐resistant subjects (n=6, BMI = 47.3 ± 2.8 kg/m2, HOMA‐IR = 3.4 ± 0.4), pooled and differentiated into myotubes. On day 6 of differentiation, myotubes were treated with either mitochondrial fission inhibitor Mdivi‐1 (20μM) or DMSO for 12 hrs. Mitochondrial respiratory capacity was measured using Seahorse XFp Analyzer. Live cells were stained with MitoTracker, and mitochondrial morphology was examined by confocal microscopy. Mitochondrial fission, fusion, and OXPHOS protein expressions were assessed by immunoblot. All experiments were repeated on three different occasions. Preliminary data indicate that mitochondrial fission protein dynamin related protein (Drp1) was reduced (20.9%, P = 0.093), with no changes in mitochondrial content or mitochondrial fusion proteins. Inhibiting mitochondrial fission resulted in a reduction of mitochondrial networks (9.7%) while increasing mitochondrial branch length (14.7%). Reserved mitochondrial respiratory capacity, as measured by the mitochondrial spare capacity, was significantly increased (31.4%, P < 0.05) with Mdivi‐1 treatment. These results demonstrate that inhibition of mitochondrial fission improves mitochondrial morphology and respiratory capacity in skeletal muscle from obese humans, suggesting mitochondrial fission may be a novel therapeutic target for treating metabolic disorders that are typically linked to mitochondrial dysfunction.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Altered skeletal muscle redox status, structural integrity, and ryanodine receptor are associated with contractile dysfunction following systemic iron loading in mice
Authors: Liang Chen , Mickey Marisa C , Receno Candace N , Atalay Mustafa , DeRuisseau Keith C
Published: June 2021

We previously reported that a moderate degree of systemic iron loading caused muscle functional impairment in mice. Such deterioration was not attributed to oxidative stress since it did not incur protein or lipid oxidative damage. On the other hand, muscle contractile dysfunction was associated with increased thioredoxin protein levels, suggestive of altered redox signaling. Thus, the purpose of this study was to further examine skeletal muscle redox status and potential meditators of contractile dysfunction following a physiologically‐relevant degree of iron loading. Twelve‐week‐old male CD‐1 mice received subcutaneous injections of iron (4 mg iron dextran/200 μL) or vehicle 5 days/week for 2 weeks (n=10/group). One day following the last injection the gastrocnemius muscles were harvested for measurement of biochemical parameters. Systemic iron loading elevated the muscle non‐heme iron (NHI) concentration by approximately 4‐fold (P < 0.001). Although reduced glutathione GSH (P = 0.39) and glutathione peroxidase (GPX) activity (P = 0.82) measures were similar between the two groups, iron‐loaded mice displayed greater glutathione disulfide (GSSG; P = 0.03) and GSSG/GSH ratio (P = 0.03), which are sensitive markers of oxidative stress and redox status. In addition, redox regulator glutaredoxin‐2 (GRX2) protein expression was lower in the muscle of iron‐loaded mice compared with controls (P = 0.04). Muscle structural integrity marker αII‐spectrin at 240 kDa was lower in the iron‐loaded muscles compared with controls (P = 0.02). No differences were observed in its cleavage products at 150 kDa (P = 0.41) or 120 kDa (P = 0.11). Similar to αII‐spectrin, protein levels of focal adhesion α‐actinin were also lower in the iron‐loaded gastrocnemius (P = 0.04). Levels of calstabin‐1 associated with ryanodine receptor (RyR1) were lower in iron‐loaded muscles compared with controls (P = 0.04), while total RyR1 (P = 0.99) was similar between groups. In summary, contractile dysfunction that results from moderate iron loading may be mediated by a disturbance in the muscle redox balance and from changes arising from an increased proteolytic response and aberrant sarcoplasmic reticulum Ca2+ release.

Support or Funding Information

Supported by the Syracuse University SOE

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
F‐box and WD‐40 Domain Protein 5 (Fbxw5) and F‐box Protein 44 (Fbxo44) are Expressed in Skeletal Muscle
Authors: White Adrianna , Parke Jane , Waddell David
Published: June 2021

Skeletal muscle atrophy is a physiological condition that is caused by a myriad of conditions, including immobilization, denervation, spinal cord injury and corticosteroid use and results in decreased muscle size and strength. The molecular genetic events of neurogenic atrophy were analyzed in a previous study using gastrocnemius muscle isolated from mice following 3 days and 14 days of denervation. The gene expression profile in the denervated muscle tissue was analyzed by microarray and compared to control muscle tissue to identify novel genes that are differentially expressed in response to neurogenic atrophy. The microarray data revealed for the first time that F‐box and WD‐40 Domain Protein 5 (Fbxw5) and F‐box Protein 44 are expressed in skeletal muscle and are differentially regulated in response to denervation. Fbxw5 and Fbxo44 are classified as E3 ubiquitin ligases and both have been found to interact with cullin 4 and DNA damage binding protein 1 (DDB1) complexes. Furthermore, Fbxo44 has been found to ubiquitinate and destabilize both BRCA1 and regulator of G protein signaling 2 (RGS2), while Fbxw5 has been shown to destabilize DLC1, a RhoA GTPase‐activating protein, and tuberous sclerosis complex 2 (TSC2), a negative regulator of mTOR. To confirm that Fbxw5 and Fbxo44 are expressed in muscle, quantitative PCR (qPCR) was used to assess the expression levels of these two E3 ligases in both proliferating and differentiated muscle cells and the results demonstrate that expression levels appear to remain relatively constant in proliferating myoblasts and differentiated myotubes. Furthermore, Western blot analysis supported the qPCR data and showed that Fbxw5 and Fbxo44 protein levels remain relatively constant as cultured myoblasts differentiate to myotubes. The discovery that Fbxw5 and Fbxo44 are expressed in skeletal muscle and are differentially regulated in response to neurogenic atrophy helps further our understanding of the molecular genetic events of muscle wasting and may eventually lead to the identification of new therapeutic targets for the treatment and prevention of atrophy.

Support or Funding Information

The work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of sex and exercise training on bone volume and density in Nrf2‐deficient rats
Authors: Wahl John Andrew , Park Hyerim , Barsoum Mina‐Michael , Restrepo Alexis , Dewell Isabella , Wahl Jake , Maraj Joshua , Peterson William , Reid‐Foley Emily , Ballehr Kelsey , Bedwell Kristin , Medarev Steven , Schartz Kaley , Sadowsky Dylan , Delp Michael , Geurts Aron , Muller‐Delp Judy
Published: June 2021

Nuclear factor (erythroid‐derived 2)‐like 2 (Nrf2) is a transcription factor that is crucial in maintaining cellular redox homeostasis. In female mice, Nrf2 deficiency has been shown to increase bone resorption in the femur. In contrast, treadmill exercise training has been shown to increase bone density in the femur of aged rats. We tested the hypothesis that the absence of Nrf2 would reduce or negate increases in bone density induced by exercise training. We exercise trained male and female wild type Sprague‐Dawley (WT) rats, and their littermates lacking the Nrf2 gene (Nrf2KO). Rats were either exercise trained (EX) on a motor‐driven treadmill 5 days/wk or remained sedentary (SED) in their cages for 10–12 weeks. A mixed moderate aerobic and high‐intensity interval training protocol was utilized. At the end of the training period, femurs were cleaned, then scanned and analyzed using a Scanco Medical uCT35 micro‐CT with a resolution of 12 micron‐voxels. Total volume, bone volume, bone volume/total volume (BV/TV), connectivity and number of trabeculae were evaluated in trabecular bone. In both WT and Nrf2KO male rats, exercise training increased trabecular number (EX: 2.8 ± 0.1 vs. SED: 2.4 ± 0.1; P<0.05) and BV/TV (EX: 0.17± 0.01 vs. SED: 0.13 ± 0.01; P=0.06), and tended to increase connectivity (EX: 31.9 ± 2.5 vs. SED: 24.7 ± 2.5; P=0.09). Surprisingly, total volume, bone volume, BV/TV, connectivity and number of trabeculae were all more than 100% higher in femurs from females as compared to males (female vs. male; P<0.0000001 for all parameters). There were no significant effects of training or Nrf2KO genotype on bone parameters in females. These data suggest that female hormones are powerful regulators of bone growth and remodeling, masking the effects of either deletion of the Nrf2 gene or treadmill exercise.

Support or Funding Information

NIH Grant Number R15AG055029

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 Exercise Training and Exercise Training plus Kinesio Taping on Cartilage and Synovial Markers and Pain in Patients with Knee Osteoarthritis
Authors: Okudan Nilsel , Belviranli Muaz , Oguz Ramazan
Published: June 2021

Establishing prevention and therapeutic strategies for osteoarthritis (OA) is necessary to minimize functional disability and the impact of the disease on society. Therefore, the aim of this study was to investigate the effect of exercise training and exercise training plus kinesio taping technique on cartilage and synovial markers and pain at rest and 20 minutes after the walking exercise in patients with knee OA. The study protocol was approved by Local Ethical Committee of Faculty of Medicine, Selçuk University. Twenty‐two female patients aged between 38–60 years who were diagnosed with OA according to the criteria of the American Rheumatology Association participated in this study. The participants were divided into exercise training and exercise training plus kinesio taping groups randomly. The functional status of the patients was assessed using the Western Ontario and McMaster Universities OA index (WOMAC). Pain assessment was performed on a visual analogue scale (VAS). Participants were invited to the laboratory twice before and after treatment. The blood samples were taken at rest and immediately after the 20‐min walking exercise before treatment and 6‐week after the application of exercise training or exercise training plus kinesio taping technique. Cartilage oligomeric matrix protein (COMP), matrix metalloproteinase (MMP)‐1 and ‐3 concentrations were measured in these samples. In patients with knee OA, VAS and WOMAC scores significantly decreased after the exercise training or exercise training plus kinesio taping (P < 0.05). Plasma MMP‐1 and MMP‐3, and serum COMP concentrations significantly increased immediately after 20‐min of walking exercise both before and after treatment (P < 0.05). Resting serum COMP concentrations did not significantly differ after 6 weeks of exercise training when compared to the pre‐treatment (P > 0.05). However, resting serum COMP concentration decreased with exercise training plus kinesio taping technique (P < 0.05). Plasma MMP‐3 concentration decreased with both exercise training or exercise training plus kinesio taping technique (P < 0.05). However, plasma MMP‐1 concentrations did not differ between groups after treatment (P > 0.05). As a result, exercise training and exercise training plus kinesio taping reduced pain and improved functional capacity in patients with knee OA. Additionally, exercise training plus kinesio taping showed a positive effect on cartilage and synovial destruction, but only exercise training showed a positive effect on synovial destruction.

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 3‐bormopyruvate on the growth of castration‐resistant prostate cancer
Authors: Wu Yen‐Hsuan , Wu Hsin‐En , Li Chia‐Yang
Published: June 2021

Prostate cancer (PCa) is the second most frequently diagnosed cancer and the sixth leading cause of cancer death among men worldwide. Androgen deprivation therapy has been the mainstay of treatment for patients with advanced PCa; however, the majority of patients will eventually develop a poor prognosis stage of castration resistant prostate cancer (CRPC) and lead to mortality. For the treatment of CRPC, the therapeutic efficacy of currently treated drugs for CRPC is limited. Therefore, development of new agents for treating CRPC is urgently needed.

3‐bormopyruvate (3‐BP) 3‐bromopyruvic acid (3‐BP) is a pyruvate analogue with anti‐tumor activity in breast cancer, pancreatic cancer, colon cancer, liver cancer. However, the effect of 3‐BP on the growth of CRPC remains unclear. In the present study we tested the effects of 3‐BP in CRPC including cell growth, cell cycle progression, cell apoptosis, migration, and the therapeutic efficiency of combination therapy with chemotherapy. Our results showed that 3‐BP inhibited the growth rate of cells, induced G2/M phase arrest, increased cell apoptosis, attenuated cell migration, _and promoted the therapeutic efficiency of combination therapy with chemotherapy in prostate cancer cells. These results suggest that 3‐BP has potential to be a therapeutic agent for treating CRPC.

Support or Funding Information

This study was supported by the Ministry of Science and Technology, Taiwan, R.O.C. (Grant No. MOST 107‐2320‐B‐037‐019).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of differential expression of cytokines secreted by rhesus monkeys bone marrow mesenchymal stromal cells on age‐related bone aging and autophagy
Authors: Liu Fei , Li Yuanmin , Zhang Jie , Lu Yanrong , Chen Younan , Cheng Jingqiu
Published: June 2021

Background

Aging is a gradual process characterized by a deterioration in the function of the organism. Studies have shown that there may be a close relationship between organ/tissue aging and stem cells. In addition, progressive DNA damage and mitochondrial dysfunction are considered to be major causes of natural aging. Moreover, autophagy plays an important role in maintaining skeletal homeostasis, and changes in this pathway are associated with osteoporosis. Therefore, the search for age‐related bone aging mechanisms can explain the progressive decline of organs and the development of diseases associated with advanced age, and can clarify the ways in which the relevant therapeutic methods are developed.

Methods

We isolated bone marrow mesenchymal stem cells (rBMSCs) from aged, middle‐aged, and young rhesus monkeys and cultured them to passage 3, followed by transcriptome sequencing of three different age‐derived cells to explore aging‐related GO term and KEGG Pathway. Further analysis of hub genes were performed by gene co‐expression network analysis. In addition, we also measured telomerase activity, cell death and β‐galactosidase staining in rBMSCs of different ages.

Results

We successfully isolated and cultivated old, middle and young rBMSCs. As shown in Figure 1D, aged rBMSCs stained the most prominent positive galactosidase, while Figure 1C showed that the telomerase activity of rBMSCS gradually decreased from young to old, and the same results also be obtained by cell death analysis and cell culture supernatants (Figure 1A, B). In addition, we found the enrichment of aging‐related genes through both GO term and KEGG pathway analysis (Figure 2). Eleven hub genes were further obtained by the protein interaction map, including LRRC17, VILL, PENK, IFIT1B, MX1, PTGFR, GJB2, IFI44L, CHI3L, LOC718478, GBP2 (Figure 3).

Among them, LRRC17 has been reported to participate in and maintain bone homeostasis of osteoblasts and osteoclasts.

Conclusions

We initially explored the effects of age on rBMSCs and possible hub genes in the aging process. The aging characterization of rBMSCs in different age groups all showed a significant decreasing trend. Eleven key factors, including LRRC17, may be involved in the process of bone aging. Taken together, these results suggest several new key factor in bone aging and related aging pathways.

Support or Funding Information

This work was supported by the Chengdu Giant Panda Breeding Research Foundation (CPF2015‐08).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
IP3‐evoked Ca2+ release in Endothelial Cells is Modulated by the Immunosuppressant FK506‐binding Protein (FKBP12)
Authors: Buckley Charlotte , Wilson Calum , McCarron John
Published: June 2021

The endoplasmic reticulum (ER) is integral to the regulation of cytoplasmic Ca2+ concentration in endothelial cells. Ca2+ release from the ER is controlled by two intracellular receptor/channel complexes, the inositol 1,4,5‐triphosphate receptor (IP3R) and the ryanodine receptor (RyR). These receptors have been shown to be regulated by the accessory FK506‐binding protein (FKBP). FK506 is a potent immunosuppressant, which is routinely used after organ transplant to lower the risk of rejection, and has also been used in drug‐eluting stents. FK506 has been shown to act on smooth muscle cells either directly by binding to the channel, or indirectly via FKBP modulation of two targets, calcineurin or the mammalian target of rapamycin (mTOR). However very little is known on the role FKBPs play in endothelial cell Ca2+ signaling.

To investigate this, Ca2+ signals were measured in the endothelium of second order mesenteric arteries using Cal‐520 (5 μM). IP3Rs were activated by localised photo‐uncaging of IP3 (5 μM), and RyR by the application of Caffeine (10 mM). FK506 (10 μM), which displaces FKBP from each receptor to inhibit calceinurin, increased the IP3‐evoked Ca2+ rise. The phosphatase inhibitors cypermethrin and okadaic acid also increased the IP3‐mediated Ca2+ release. Cypermethrin and okadaic acid did not prevent FK506‐induced increase in Ca2+ release. These results suggest that while calceinurin may modulate IP3‐evoked Ca2+ release, the phosphatase is not necessary for the FK506‐mediated potentiation of IP3‐evoked Ca2+ increase.

Rapamycin (10 μM), which disrupts the binding of FKBP12 to the receptor and inhibits mTOR, did not significantly alter IP3‐induced Ca2+ release in endothelial cells, indicating that mTOR does not exert a significant role in endothelial IP3‐induced Ca2+ release.

Interestingly Caffeine, the activator of RyR‐mediated Ca2+ release, did not activate Ca2+ signals in the endothelium either in intact tissue or in freshly isolated endothelial patches, suggesting that Caffeine‐activated RyRs do not play a role in endothelial Ca2+ release.

These results indicate that FKBPs play an integral role in regulating endothelial IP3‐mediated Ca2+ release, and may underlie the severe endothelial dysfunction associated with FK506 immunosuppressants.

Support or Funding Information

Wellcome Trust

British Heart Foundation

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mitochondrial dysfunction increases bronchopulmonary C‐fiber excitability via PKC alpha signaling
Authors: Bahia Parmvir K , Hadley Stephen H , Taylor‐Clark Thomas E
Published: June 2021

Airway sensory nerve excitability is a key determinant of respiratory disease‐associated reflexes and sensations such as cough and dyspnea. One notable feature of peripheral terminals of sensory nerves is that they are packed with mitochondria. Mitochondrial dysfunction and subsequent oxidative stress has been reported for a variety of cell types in inflammatory diseases. We have previously shown that reactive oxygen species (ROS) produced by Antimycin A (a mitochondrial complex III Qi site inhibitor) increase excitability in bronchopulmonary nociceptive C‐fibers. This increased excitability decreases the mechanical threshold for activation and increases action potential firing in response to a P2X2/3 agonist. This Antimycin A–induced hyperexcitability is dependent on mitochondrial ROS and is blocked by intracellular antioxidants.

Given that ROS are known to activate protein kinase C (PKC), we hypothesized that this increased excitability was due to PKC activation. We used dissociated vagal ganglion neurons to study activation of the PKC isoforms: alpha, beta1, beta2, delta and epsilon in response to Antimycin‐A. Cells were stained with antibodies against the specific isoforms to determine their cellular location. Of these isoforms, only PKC alpha translocated to the region of the cell membrane in response to Antimycin A treatment.

We also examined the effects of Antimycin A induced ROS production on sensory nerve terminals. We recorded action potential firing from the terminals of individual bronchopulmonary C‐fibers using a mouse ex vivo lung‐vagal ganglia preparation. C‐fibers were characterized as nociceptors or non‐nociceptors based upon conduction velocity and response to transient receptor potential (TRP) channel agonists. Antimycin A–induced hyperexcitability was inhibited by the PKC inhibitor bisindolylmaleimide (BIM) I, but not by its inactive analog BIM V. Further, this BIM I inhibition occurred at a concentration that specifically blocks activity of PKC alpha over other isoforms. In conclusion, ROS evoked by mitochondrial dysfunction caused nociceptor hyperexcitability via the translocation and activation of PKC alpha.

Support or Funding Information

This work was supported by the National Heart Lung and Blood Institute in Bethesda, USA (R01‐HL119802)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mouse models of human craniofacials spliceosomopathies: Are they neurocristopathies?
Authors: Jerome‐Majewska Loydie , Beauchamp Marie‐Claude , Djedid Anissa , Daupin Kevin , Clokie Kayla , Kumar Shruti , Alam Sabrina , Majewski Jacek
Published: June 2021

Splicing, the removal of introns from pre‐messenger RNA is an essential step for expression of most genes in multicellular organisms and for expanding the number of proteins coded by genomes. Recently, exome sequencing revealed that mutations in splicing factors and small nuclear ribonucleoprotein particles (snRNPs) which form core components of the spliceosome, are responsible for craniofacial malformations. Our group used in situ hybridization to examine expression of three core components of the spliceosome: Eftud2 – mutated in patients with mandibulofacial dysostosis, Guion‐Almeida type (MFDGA); SnrpB – responsible for Cerebrocostomandibular Syndrome (CMS); and Sf3b4 which is responsible for Nager syndrome. Herein, we report expression of Eftud2, Sf3b4 and SnrpB during craniofacial development. In addition, we used CRISPR/Cas9 to generate mice with deletion (Eftud2 del) and conditional mutation of exon 2 of Eftud2 (Eftud2 flox). At embryonic days (E) 7.5 and 8.5, Eftud2 was highly expressed in ectodermal and mesodermal components of the future craniofacial region, by E9.5 expression was also found in the body wall and developing heart. Eftud2 del heterozygous mice were viable and fertile, though these embryos showed reduced levels of Eftud2 mRNA and protein. In contrast, Eftud2 del homozygous mutant embryos arrest at E3.5 and failed to grow and hatch ex vivo. To examine the requirement for Eftud2 in neural crest cells, the Wnt1‐Cre2 transgenic line was used to delete exon 2 of Eftud2 specifically in that lineage. Eftud2 flox homozygous mutant embryos carrying the Wnt1‐Cre2 transgene displayed hypoplasia of the midbrain and pharyngeal arches starting at E9.5. By E11.5, most embryos also had an open neural tube and all embryos showed exencephaly at E14.5. Cartilage preparations revealed an absence of cartilage in the head, reduction/or absence of Meckel's cartilage, and abnormal inner ear development. Since deletion of exon 2 is predicted to generate a truncated protein with partial function, our data suggest that normal levels of Eftud2 is crucial in neural crest cells for normal craniofacial development. Future studies are focused on elucidating the molecular and transcriptional basis of MFDM using this mouse model.

Support or Funding Information

Canadian Institute of Health Research (MOP#142452)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Alterations in adipocyte morphology and leucocyte infiltration in adipose tissue in mice supplemented with non‐nutritive sweeteners
Authors: Contreras Irazú , Zepeda‐Muñoz Jesus , Estrada José Antonio
Published: June 2021

Consumption of non‐nutritive sweeteners has been related to changes in feeding behavior and energy metabolism that may lead to weight gain and insulin resistance under specific circumstances. The molecular mechanisms related to these effects have not been thoroughly determined, but chronic low‐grade inflammation in adipose tissue is known to affect leukocyte recruitment and adipocyte functions in these pathologies.

The main objective of this study was to determine the morphological characteristics of adipocytes and leucocyte recruitment to gonadal and mesenteric adipose tissue in mice supplemented with commercial non‐nutritive sweetener formulations. Adult male and female BALB/c mice were randomly assigned into experimental (sucralose or steviol glycosides) and control (no supplementation) groups. Non‐nutritive sweeteners were supplemented for 6 weeks in daily drinking water. At the end of treatment, the animals were sacrificed and gonadal and mesenteric tissues were obtained to assess adipocyte morphology and leucocyte infiltration by histochemical staining and bright‐field microscopy.

The results show that gonadal and mesenteric tissue from both male and female mice treated with either sucralose or esteviol glycosides had altered adipocyte morphology, with increased adipocyte size compared to controls. Additionally, a decreased number of adipocytes was observed in male mice from both sweetener groups. Male mice supplemented with steviol glycosides also had decreased leucocyte infiltration in gonadal and mesenteric adipose tissue, compared to controls.

Altogether, our results suggest that frequent intake of commercial non‐nutritive sweeteners induces changes to adipocyte morphology and leucocyte infiltration in adipose tissue that may be relevant for metabolic changes associated to weight gain/loss in individuals that use these compounds in their regular diet.

Support or Funding Information

Consejo Mexiquense de Ciencia y Tecnología

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Minocycline Normalizes the Hypertension in Gastrin−/− Mice
Authors: Asico Laureano D , Hunt Jessica , Wang Xiaoyan , Wank Stephen A , Yagnesh Tailor H. , Wang Timothy C , Raj Dominic , Jose Pedro A
Published: June 2021

The advent of cutting edge technology has paved the way for the identification of new players for sodium homeostasis and blood pressure (BP) control. The role of gut microbiome in metabolic disorders has been the subject of a growing number of metagenomic and metabolomic studies. Dysbiosis, or the alteration in the gut microbiota composition and function, affects the development of hypertension. Gut microbiota, comprised of mainly of Firmicutes (F) and Bacteroidetes (B), constantly adapt to lifestyle modifications. An increased F/B is observed in rodent and human hypertension. Minocycline selectively inhibits M1 polarization of macrophages, which results in anti‐inflammatory and anti‐hypertensive effects and normalizes the microbiome ratio and thus, the BP. We reported that the hormone gastrin responds to increased dietary sodium load and promotes the excretion of excess sodium through the interaction of renal dopamine receptors (D1R and D5R), and cholecystokinin B receptor (CCKBR), the so‐called “gastrorenal axis”. Gast−/− mice on a salt‐resistant background develop hypertension while wild‐type littermates have normal BP. Gast−/− mice have negligible gastric acid production, which results in increased bacterial survival and dysbiosis of the gut microbiota. Therefore, we tested the hypothesis that minocycline alleviates the high BP of Gast−/− mice. Gast−/− mice and their wild‐type littermates fed a high salt diet (4% NaCl) were given minocycline (50 mg/kg/day, p.o., 28 days). Results showed that minocycline decreased the high BP of conscious Gast−/− mice both at nighttime (130.7±0.4 vs. 123.2±0.3, P<0.05, Dunn's test, 4/group) and daytime (126.1±0.5 vs. 117.5±0.4 P<0.05), and the nighttime (122.4±0.4 vs. 118.4±0.5 P<0.001, Dunn's test, 4/group), but not the daytime BP of the wild‐type littermates. These results imply that inhibition of M1 macrophages decreases BP. In another set of Gast−/− mice, we tested the effect of doxycycline (43 mg/kg/day, p.o., 28 days) which selectively inhibits M2 polarization of macrophages leading to pro‐inflammatory and pro‐hypertensive effects. Doxycycline decreased the BP of conscious Gast−/− mice at nighttime (134.7±0.7 vs. 123.8±8.0 mm Hg, P<0.05, Dunn's test) and at daytime (120.7±0.6 vs. 113.1±2.8 mm Hg), conceivably by normalizing the dysbiosis expected in the knockout mice. By contrast, doxycycline increased BP in wild‐type littermates at nighttime (97.6±1.2 vs. 110.5±5.1 mm Hg) but not at daytime, which can be explained by the inhibition of the M2 macrophage polarization. Thus, the resultant hypertensive phenotype is influenced by the dynamic interplay among genetics, state of inflammation, and gut microbiota.

Support or Funding Information

R01DK039308, R01HL092196, and P01HL074940

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