bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Spirulina Extracts on Vasodilator Function of Arteries from Hypertensive Rats
Authors: Villalpando Diva M. , Verdasco Carlos , Plaza Ignacio , Rivas Juan Gómez , de Bethencourt Fermin R. , Otero Cristina , Ferrer Mercedes
Published: June 2021

Hypertension is a multifactorial disorder that is considered one of the major causes of premature death worldwide. This pathology is associated with functional and structural vascular alterations in which oxidative stress and decreased nitric oxide (NO) bioavailability are involved. On the other hand, the use of natural compounds, including extracts of microalgae, for the treatment of cardiovascular diseases is gaining attention. Based on the anti‐oxidant and anti‐inflammatory properties of Spirulina, the aim of this study was to investigate the effect of aqueous Spirulina extract on the release and function of NO, as well as on the expression of the antioxidant protein hemoxygenase‐1 (HO‐1), phosphorylated Akt (p‐Akt) and phosphorylated eNOS (p‐eNOS) that regulate the production of NO. For this purpose, aortic segments from male Spontaneously Hypertensive rats (SHR) and its normotensive counterparts, Wistar‐Kyoto (WKY) rats, were isolated. Aortic segments exposed to the aqueous Spirulina extract (0.1 % w/v, for 3 h) and those not exposed were used to analyze the following parameters: (i) the release of NO; (ii) the NO‐dependent response to acetylcholine (ACh) and the vasodilator response to the NO donor, sodium nitroprusside (SNP); and (iii) the expression of HO‐1, p‐eNOS and p‐Akt. The results showed that Spirulina extracts increased the release of NO more in arteries from SHR than that of WKY rats. Spirulina extracts also increased the ACh‐ and SNP‐induced response in arteries from both WKY and SHR. Additionally, Spirulina increased the expression of HO‐1, p‐Akt and p‐eNOS in arteries from SHR. These results suggest that Spirulina extracts exert benefits for the treatment of hypertension by increasing the release and function of NO in which the increased levels of HO‐1, p‐eNOS and p‐Akt can be involved.

Support or Funding Information

Supported by Comunidad de Madrid (S2013/ABI‐2783, “INSPIRA1‐CM”) and Fondo Europeo de Desarrollo Regional.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Distinct Biaxial Mechanical Properties between Right and Left Ventricles in Healthy Adult Sheep
Authors: Nguyen‐Truong Michael , Labus Kevin M. , Liu Wenqiang , McGilvray Kirk , Puttlitz Christian M. , Wang Zhijie
Published: June 2021

Right ventricle failure (RVF) is a serious disease with a 40% mortality rate where the right heart is unable to sufficiently pump blood to the body. RVF is associated with biomechanical changes such as extracellular matrix deposition and right ventricle (RV) wall stiffening. It has been well established that the RV is distinct from the left ventricle (LV) in embryologic origin, anatomy and function, and the two chambers' pathologies are significantly different. However, unlike left ventricle failure (LVF), the mechanisms of RVF are not well understood and it is partly due to the lack of knowledge in the difference in biomechanical properties and mechanobiology between the two chambers.

To investigate the mechanical properties of the LV and RV, ex vivo biaxial tensile mechanical tests were performed in adult, healthy sheep hearts. Using homogenous, square slices of fresh ovine ventricles, 1:1 biaxial tests were performed and the stress‐strain responses were recorded. The elastic modulus at high and low strain regions were then derived as E1 and E2, respectively. The LV and RV showed distinct anisotropic mechanical properties: the longitudinal elastic moduli are larger than those of circumferential axis in the RV, while the opposite trend was observed in the LV. Moreover, we found that the RV was stiffer than the LV in the longitudinal direction (which is parallel to the outflow tract direction), whereas in the circumferential direction the RV was more compliant than the LV (Figure and Table). To our knowledge this is the first characterization and comparison of biaxial mechanical properties of LV and RV in a large animal model. Future work will test more heart samples and examine the histology to correlate structure to mechanical function. Overall, our finding that the healthy RV was stiffer than the healthy LV in the outflow tract direction may shed light on the biomechanical changes under pathological conditions, such as pressure overload induced RVF.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Albumin and palmitate overload increases osteopontin production in renal tubule epithelial cells
Authors: Cobbs Alyssa , Ballou Kristopher , Chen Xiaoming , George Jasmine , Zhao Xueying
Published: June 2021

Osteopontin (OPN) is one of the proinflammatory cytokines upregulated in the kidneys of diabetic animals and patients with diabetic nephropathy (DN). An increase in albuminuria and albumin‐bound fatty acids presents a proinflammatory environment to the proximal tubules in proteinuric kidney diseases including DN. This study was designed to examine if fatty acid overload or albumin itself could stimulate OPN production by renal tubular epithelial cells. Renal tubular OPN expression and secretion were examined in Zucker diabetic (ZD) rats and NRK‐52E cell line exposure to bovine serum albumin (BSA) or BSA overloaded with saturated fatty acid palmitate. Gene microarray and real‐time PCR confirmed an albuminuria‐associated upregulation of OPN transcripts in kidney cortex of ZD rats at the age of 7–20 weeks. Immunofluorescence staining of kidney sections revealed an increase in OPN protein in damaged proximal tubules of diabetic rats. Western blot analysis detected a progressive increase in urinary OPN levels in the ZD rats, which was prevented by antiproteinuric treatment with losartan, an angiotensin II receptor blocker. When exposed to fatty acid‐free bovine serum albumin (BSA, 10 mg/ml), NRK‐52E cells exhibited an increase in OPN protein expression. Palmitate (250 μM) further enhanced albumin‐induced OPN in NRK‐52E cells. These results demonstrate a stimulatory effect of albumin and its conjugated fatty acids on OPN expression in renal tubule epithelial cells. Thus, besides lowering albuminuria/proteinuria, mitigating circulating fatty acid levels may be an important factor in preventing renal inflammation in proteinuric diseases.

Support or Funding Information

NIH SC1DK112151, NIH/NCRR/RCMI 8G12MD007602 & 8U54MD007588

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Organelle Segregation and Quality Control during Meiotic Differentiation
Authors: Ünal Elçin , Sawyer Eric , King Grant , Goodman Jay
Published: June 2021

Meiotic differentiation is a complex developmental program that involves extensive cellular remodeling. In budding yeast, under nutrient deprived conditions, a diploid mother cell undergoes two successive meiotic divisions to generate four nuclei. After meiosis, the four nuclei are packaged with a subset of the organelles and cytoplasm originating from the mother to form four haploid gametes. Our lab has found that during the latter process, certain components of the mother cell are not inherited by the gametes and are subsequently produced de novo. For example, the nuclear pore complexes and components of the nucleoli do not segregate with any of the four nuclei, whereas only a subset of the mitochondria is inherited by the gametes. A programmed pathway that entails vacuole permeabilization subsequently degrades the excluded cellular materials. We posit that the restricted inheritance of organelles and other cellular sub‐compartments is required for cellular rejuvenation that occurs as part of gametogenesis.

Support or Funding Information

Pew Charitable Trusts (00027344), Damon Runyon Cancer Research Foundation (35‐15), March of Dimes (5‐FY15‐99), National Institute of Health (DP2 AG055946‐01) and Glenn Foundation for Medical Research

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cold‐Inducible RNA Binding Protein (CIRP) In Inflammation and Breast Cancer
Authors: Lujan Daniel A. , Perrone‐Bizzozero Nora , Beswick Ellen J. , Kusewitt Donna F. , Hartley Rebecca S.
Published: June 2021

RNA binding proteins (RBPs) post‐transcriptionally regulate gene expression by associating with the regulatory sequences in the untranslated regions of mRNAs. RBPs are often disrupted in cancer, but current knowledge of their role in regulating cancer‐associated genes is largely based on in vitro studies. This study seeks to determine the in vivo role of cold‐inducible RBP (CIRP), which is overexpressed in some human breast cancers. CIRP promotes cell proliferation and survival and inhibits apoptosis in vitro and promotes inflammation in vivo, all hallmarks of cancer. We tested the hypothesis that CIRP promotes breast tumorigenesis in vivo by crossing a transgenic mouse overexpressing CIRP in the mammary epithelium with the PyMT mouse model of breast cancer. Effects on both early (7‐week‐old mice) and late (14‐week‐old mice) tumorigenesis were assessed. Unexpectedly, mammary glands from 7‐week‐old CIRP/PyMT mice had 50% fewer mammary intraepithelial neoplasias than PyMT mice. Consistent with this, histopathological analysis showed decreased mitosis and increased apoptosis in CIRP/PyMT vs. PyMT mammary glands. Late stage tumorigenesis was also inhibited. CIRP/PyMT mice had slower tumor growth kinetics, smaller tumors, decreased tumor burden and decreased pulmonary metastasis compared to PyMT mice. To assess the mechanism of inhibition, mammary glands from 7 and 14‐week‐old mice were incubated in medium overnight and Luminex cytokine assays were performed on the medium. Results showed a decrease in cytokines that promote inflammation, angiogenesis and metastasis. Altogether, our results suggest that CIRP inhibits proliferation and promotes apoptosis in vivo to suppress early tumorigenesis, potentially via decreasing a protumorigenic inflammatory response.

Support or Funding Information

This work was funded by the University of New Mexico Research Allocation Committee, the American Association of Anatomists Fellows Grant Award Program and by the National Institutes of Health under the Ruth L. Kirschstein National Research Service Award (1 F31 CA213933‐01A1)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Reduced Renal Primary Cilia Expression in Humanized Sickle Cell Mice
Authors: Taylor Crystal Monique , Kasztan Malgorzata , Yoder Bradley , Pollock Jennifer S. , Pollock David
Published: June 2021

Sickle cell disease (SCD) is an autosomal recessive hematologic disorder that afflicts millions of people across the globe. As longevity improves due to therapeutic advancements, end‐organ damage, namely sickle cell nephropathy (SCN), has emerged as a major contributor to mortality and morbidity in these patients. Renal primary cilia are mechanosensory organelles that extend from the apical membrane of epithelial cells throughout the nephron and contribute to structural homeostasis. Studies have shown that renal primary cilia display alterations in response to renal ischemic injury and are essential in the epithelial repair process. Defects in renal primary cilia expression and signaling have been shown to contribute to the development of renal cystogenesis. In addition, recent studies have demonstrated an increased prevalence of renal cyst formation in sickle cell disease patients. Therefore, we hypothesized that renal primary cilia expression is reduced in a mouse model of sickle cell disease. We further predicted that acute hypoxia challenge will reduce renal primary ciliogenesis in SCD animals compared to controls. Male 14‐week old humanized sickle cell mice (HbSS) and their genetic controls (HbAA) were subjected to 3 hr exposure to hypoxia (10% O2) prior to the assessment of renal primary cilia formation. Cilia formation was assessed using Western blot analysis on kidney section lysates. Prior to hypoxia exposure, acetylated □‐tubulin expression, a protein involved in cilia architecture, was reduced in the cortex of HbSS mice compared to the genetic controls (0.45 ± 0.02 vs. 1 ± 0.02 a.u. [arbitrary units], respectively, p= 0.03, n=4). In addition, IFT88 expression, a protein involved in cilia assembly, was also reduced in the cortex of HbSS mice compared to HbAA mice (0.59 ± 0.00 vs. 1 ± 0.001, respectively, p= 0.02, n=4). After a 3 hr hypoxia challenge, HbSS mice maintained a significant reduction in cortical acetylated □‐tubulin expression compared to hypoxia‐ challenged HbAA mice (0.38 ± 0.02 vs. 1 ± 0.03 a.u., respectively, p= 0.008, n=5). However, there was no significant difference in cortical IFT88 expression in HbSS mice compared to controls following hypoxia (1.1± 0.002 vs. 1 ± 0.002 a.u., respectively, p= 0.69, n=5). These data suggest that renal primary cilia expression is reduced in SCD and acute hypoxia exposure may promote renal ciliogenesis. These findings provide a potential mechanism by which renal injury is potentiated in SCN.

Support or Funding Information

William Townsend Porter Pre‐ doctoral Fellowship from the American Physiological Society to C. Taylor

U01 HL117684 to DMP

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lithium Induces Cilia Expression in Renal Intercalated Cells
Authors: Nair Anil V. , Franken Gijs , Paunescu Teodor , Capen Diane , Brown Dennis
Published: June 2021

Lithium remains a first line drug for treatment of bipolar disorder (BD). It has demonstrated efficiency for the prevention of recurrence of mood episodes, and also as an enhancement strategy in major depressive disorder. Current research and a growing number of publications predict a wider role for Li in the coming years for various neurological disorders. One major limitation of Li treatment is the short‐ and long‐term adverse effects it has on the kidney in a subpopulation of its recipients. Up to 40% of Li receiving patients report various degrees of acute increase in urine production ‐ Nephrogenic Diabetes Insipidus (NDI). Long‐term Li treatment has a more severe side effect known as Chronic Kidney Disease (CKD), characterized by reduced glomerular filtration rate, interstitial nephritis, renal fibrosis, microcyst formation, ultimately leading to end‐stage renal disease. Renal cyst formation has also been implicated as a ciliopathy in polycystic kidney disease. Here, we studied the effect of Li on cilia in a mouse model treated for 3 months. Using immuno‐fluorescence, we imaged cilia in 16 μm thick sections of paraformaldehyde‐fixed mouse kidney. Cilia were labeled using an antibody against acetylated alpha‐tubulin. The 3D confocal images were deconvolved using Volocity software, and ciliary length from control and Li‐treated animals was measured in different tubule segments. In the different tubule segments, the mean length of cilia was different, but was not affected by Li except in the cortical collecting duct, where Li treatment significantly increased ciliary length from 6.6±0.4 μm to 7.7±0.3 μm. Collecting ducts are composed of two major cell types, intercalated (ICs) and principal cells (PCs), which were identified by the expression of cell type‐specific markers, the vacuolar proton‐pumping ATPase and aquaporin‐2, respectively. Among epithelial cells in the kidney, ICs are the only cell type that was never shown to express primary cilia. Remarkably, we found that Li treatment induces cilia expression in ICs. Li treatment is known to increase the ratio of ICs to PCs. Whether bona fide ICs are growing cilia de novo in response to Li treatment, or we are detecting PCs that have converted into ICs but still have a residual cilium that is not completely reabsorbed, remains to be elucidated.

Support or Funding Information

National Institutes of Health Grant DK096586 (D. Brown)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Influence Of “Priming” Exercise On Pulmonary Oxygen Uptake Kinetics During Heavy‐Intensity Cycle Exercise From An Elevated Baseline In Type 2 Diabetes.
Authors: Gildea Norita , Rocha Joel , Green Simon , O'Shea Donal , Egaña Mikel
Published: June 2021

In healthy individuals phase II oxygen uptake (V̇ O2) kinetics (τ V̇ O2p) is slower when heavy‐intensity exercise is initiated from a moderate‐intensity exercise baseline (work‐to‐work) than unloaded cycling; but τ V̇ O2p is not enhanced when work‐to‐work exercise is preceded by a prior bout of heavy‐intensity priming exercise (PE). This negates an insufficient muscle O2 availability as the primary culprit in healthy individuals.

Purpose

Given that type 2 diabetes (T2D) induces a lengthened τ V̇ O2p during unprimed submaximal exercise compared with healthy controls, which is attributed, at least in part, to limitations in O2 supply, the purpose of the study was to test the hypothesis that heavy‐intensity PE would subsequently accelerate τ V̇ O2p in T2D.

Methods

Seven middle‐aged individuals with T2D (46 ± 8 yr) and 7 controls (ND) (41 ± 10 yr) performed four bouts of constant‐load, heavy‐intensity exercise each commencing from a baseline of moderate‐intensity exercise. Two of these work‐to‐work bouts were completed without PE and two bouts were undertaken preceded by PE. V̇O2 kinetics was calculated from continuously measured breath‐by‐breath V̇O2 data, whilst the rate of muscle deoxygenation (i.e., deoxygenated haemoglobin and myoglobin, HHb+Mb) was continuously measured by near‐infrared spectroscopy at the vastus lateralis muscle. Diabetes and priming comparisons were made using two‐way ANOVA with repeated measures.

Results

The amplitude of the V̇O2 slow component was reduced (P < 0.05) in both groups following PE (T2D: from 0.13 ± 0.15 to 0.08 ± 0.10 l/min; ND: from 0.20 ± 0.18 to 0.12 ± 0.11 l/min). In addition, PE tended to reduce τ V̇ O2p during work‐to‐work cycling transitions in T2D (from 54 ± 14 to 42 ± 17 s) but not in ND (42 ± 11 vs. 41 ± 11 s), bringing the τ V̇ O2p in T2D on a par with their ND counterparts, in the absence of significant changes in the dynamic response of (HHb+Mb). It is likely inter‐individual variability in response precluded the attainment of statistical significance.

Conclusion

In middle‐aged individuals with T2D, the enhanced V̇ O2 kinetics response was likely consequent to a priming‐induced increase in muscle blood flow and/or improved muscle perfusion to metabolic rate.

Support or Funding Information

This publication has emanated from research conducted with the financial support of the Health Research Board of Ireland (Grant No HRA_POR/2073/274)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inorganic nitrate supplementation improves exercise tolerance and calf vascular function in patients with peripheral artery disease
Authors: Bock Joshua , Treichler David , Ueda Kenichi , Hughes William , Casey Darren
Published: June 2021

Peripheral artery disease (PAD) is characterized by widespread atherosclerotic development and endothelial dysfunction in the vasculature. This pathological process leads to hypoperfusion of local tissues, increases dependence on anaerobic metabolism, and induces the hallmark symptom of PAD; ischemic claudication. Additionally, PAD is associated with reductions in the bioavailability of nitric oxide (NO), a known regulator of vasodilator function. Together, ischemic claudication and reduced NO bioavailability attenuate exercise tolerance in PAD patients. Inorganic nitrate (NO3−) supplementation boosts NO bioavailability independent of NO‐synthase enzymes, subsequently improving vasodilator function and enhancing skeletal muscle oxygenation. Thus, inorganic NO3− supplementation may improve exercise tolerance in PAD patients. We therefore investigated if eight weeks of sodium NO3− (NaNO3) supplementation would improve exercise tolerance (six‐minute walk test, 6MWT) as well as calf blood flow or vasodilation in response to reactive hyperemia (strain gauge plethysmography) in patients with PAD. Twenty‐one patients diagnosed with moderate‐to‐severe PAD (ankle‐brachial index 0.80±0.07) were randomly assigned to NaNO3 (1g/day; n=13) or placebo (microcrystalline cellulose; n=8) supplementation groups. Eight weeks of NaNO3 supplementation elevated plasma NO3− (32.3±5.6 to 379.8±56.7μmol/L) and nitrite (192.2±15.0 to 353.1±38.7 nmol/L), improved 6MWT distance (387±26 to 425±24 m), peak calf blood flow (BFPeak, 11.6±1.5 to 14.1±1.5 mL/100 mL tissue/min), and peak calf vascular conductance (VCPeak, 11.1±1.3 to 14.2±1.5 mL/dL tissue/min/mmHg, P<0.05 for all). Moreover, improvements in 6MWT distance correlated with calf BFPeak (r=0.70) and VCPeak (r=0.61, P<0.05 for both). Placebo supplementation did not change any of the outcome measures (P=0.22 to 0.92). Neither NaNO3 nor placebo supplementation changed total calf blood flow (P=0.11) or conductance (P=0.18). These data demonstrate that eight weeks of inorganic NO3− supplementation elicits a clinically‐significant improvement in exercise tolerance which correlated to improvements in blood flow and vasodilator function in the lower limbs of patients with PAD.

Support or Funding Information

This study was supported by the American Heart Association 13GRNT16490002 (D.P.C.) and National Institutes of Health Clinical and Translational Science Award U54TR001356.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exercise Induces Depot‐Specific Adaptations to White and Brown Adipose Tissue
Authors: Baer Lisa A. , Lehnig Adam C. , Dewal Revati S. , Kitching Kathryn M. , Sindeldecker Devin A. , Goodyear Laurie J. , Stanford Kristin I.
Published: June 2021

Exercise has profound effects on whole body metabolism through adaptations to various tissues including adipose tissue. Several recent studies have investigated exercise‐induced adaptations to two white adipose tissue (WAT) depots, inguinal WAT (ingWAT) and perigonadal WAT (pgWAT), and one brown adipose tissue (BAT) depot, interscapular BAT (iBAT), and shown that each adipose tissue (AT) depot has a distinct response to exercise. Here, we investigated the effects of exercise on each of the 14 AT depots in the body. Ten week old male C57BL/6 mice housed in static cages (sedentary) or were given open access to a running wheel (exercise‐trained) for 3 weeks, and each adipose depot was removed. Exercise increased mitochondrial activity in ingWAT, and pgWAT, and decreased mitochondrial activity in iBAT. Genes and proteins involved in glucose metabolism were upregulated with exercise in ingWAT, and decreased in iBAT, anterior subcutaneous WAT (asWAT), and retroperitoneal WAT (rpWAT). Exercise increased genes and proteins involved in fatty acid oxidation in pgWAT, iBAT, asWAT, cervical BAT (cBAT) and perirenal WAT (prWAT). These data indicate that each adipose tissue depot has a unique response to exercise. Identifying the depot specific adaptations to adipose tissue in response to exercise is essential to determine how adipose tissue contributes to the overall beneficial effect of exercise.

Support or Funding Information

National Institutes of Health Grants R01‐HL138738 and K01‐DK105109

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Moderate Intensity Endurance Exercise Training Increases Muscle Glycogen Content but Does Not Alter Substrate Oxidation in C57BL6 Mice
Authors: Fuller Scott , Simon Jacob , Huang Tai‐Yu , Batdorf Heidi , Scott Matthew , Waskom Callie , Essajee Nabil , Burke Susan , Collier Jason , Noland Robert
Published: June 2021

INTRODUCTION

Aerobic exercise training induces a range of desirable cardiovascular and metabolic adaptations but questions remain regarding the minimum thresholds of intensity, duration, and frequency of training required to elicit measurable changes in health outcomes. We examined the effects of a moderate intensity treadmill endurance training protocol in mice in order to determine whether this intensity level would induce changes in body composition and serum parameters. Furthermore, we investigated the extent to which any such changes would be accompanied by similar changes in muscle and liver substrate oxidation in an effort to better characterize the suite of metabolic adaptations that occur in response to moderate intensity exercise.

METHODS

20 male C57BL6 mice, approximately 12 weeks of age, were randomly assigned to either an exercise (EX) or sedentary (CON) group. All animals were maintained on low‐fat (13%) rodent chow (LabDiet 5001) and group housed. The EX group underwent a 6‐week program daily treadmill exercise at speeds no greater than 20 meters/min at an incline of 5°. Moderate‐intensity training sessions elicited a 50% increase in circulating lactate, but remained below a lactate threshold level. Body composition was measured weekly by NMR. Mice were euthanized following the cessation of the exercise training protocol. Muscle and liver tissues were collected and substrate oxidation was assessed using 14C labeled substrates. Glycogen content was assayed in liver and quadriceps muscle. Serum parameters were also assessed.

RESULTS

The EX group exhibited reduction in body weight and fat mass. Glycogen storage in muscle was significantly increased (p<0.05) by exercise training, whereas intramuscular triglycerides were decreased (p<0.05). There were no statistically significant changes observed in pyruvate, leucine, palmitate, or lignocerate oxidation between the groups, which was supported by a relative lack of changes in gene/protein expression of relevant pathways.

CONCLUSION

A moderate intensity 6‐week treadmill endurance exercise program was sufficient to induce a set of “traditional” training adaptations (reduced body weight and fat mass, increased glycogen); however, the stimulus was not sufficient to alter substrate oxidative capacity in skeletal muscle. Collectively these data suggest that in order for exercise training to induce remodeling of substrate metabolism pathways at a molecular level, protocols using higher intensities may be required.

Support or Funding Information

This work was supported by NIH R01 DK103860‐01 (R.C.N.).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Muscular Strength in Well Healed Burn Survivors is Similar to Controls
Authors: Jaffery Manall , Romero Steven , Moralez Gilbert , Kennedy Naomi , Crandall Craig
Published: June 2021

Recovery from severe burn injuries involves prolonged hospitalization and accompanying bed rest. Additionally, burn survivors are often physically inactive for years after recovery. Inactivity leads to reduced muscle cross sectional area and a reduction in muscular strength. Previous studies have shown that lower body muscular strength is decreased 6 months after discharge from the hospital in burn survivors. However, it is unknown whether well‐healed burn survivors regain the loss of muscular strength by two years post burn injury.

Purpose

Using peak torque per body weight (tq/bw) as an indicator of muscular strength, the purpose of this study was to test the hypothesis that muscular strength is reduced in well healed burn survivors compared with non‐burned control subjects.

Methods

Peak tq/bw was assessed during leg extension at 90°/sec and 120°/sec using Biodex isokinetic dynamometer in nine non‐burned adults (4 females; age 34 ± 9 years; height 172 ± 19 cm; mass 81 ± 24 kg; mean ± SD) and 19 burn survivors (6 females; age 39 ± 13 years; height 172 ± 10 cm; mass 80 ± 22 kg; mean ± SD). Average body surface area burned was 49 ± 23% (range: 16 – 87%). Subjects were at least two years post burn injury.

Results

Neither tq/bw at 90°/sec (controls: 57.4 ± 16.4 %; burn survivors: 60.4 ± 17.9 % P = 0.6) nor tq/bw at 180°/sec (controls: 49.5 ± 18.8 %; burn survivors: 47.9 ± 12.8 % P = 0.8) differed between groups.

Conclusions

These data suggest that by at least two years post burn injury, burn survivors achieve similar levels of leg muscular strength as non‐burned individuals.

Support or Funding Information

Funding provided by the National Institute for Health

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification and Characterization of TSSK6 Activating Co‐chaperone (TSACC) in Skeletal Muscle
Authors: Deluna Jonnaly , Waddell David
Published: June 2021

Skeletal muscle atrophy is a physiological condition that is caused by a variety of conditions, including immobilization, denervation, spinal cord injury and aging and results in decreased muscle size and strength. In order to better characterize the molecular genetic events of neurogenic atrophy, the gastrocnemius muscle was isolated from mice following 3 days and 14 days of sciatic nerve denervation. The gene expression profile in the denervated muscle tissue was then analyzed by microarray and compared to control muscle tissue in order to identify novel neurogenic atrophy‐induced genes. The microarray data revealed for the first time that TSSK6 Activating Co‐chaperone (TSACC) is expressed in skeletal muscle and is significantly induced in response to denervation. To confirm that TSACC is expressed in muscle cells, we cloned the predicted TSACC cDNA from muscle cells, as well as a novel TSACC splice variant that contains an additional exon. Furthermore, quantitative PCR (qPCR) was used to assess TSACC expression levels in both proliferating and differentiated muscle cells and the results demonstrate that TSACC expression levels are relatively low in proliferating myoblasts, but show significantly elevated expression in differentiated myotubes. In order to characterize the transcriptional regulation of TSACC, fragments of the proximal promoter located immediately upstream of the start of transcription were cloned and fused to a reporter gene. The reporter plasmids were then transfected into C2C12 mouse muscle cells in combination with myogenic regulatory factor (MRF) expression plasmids, which resulted in significant activation of reporter gene activity. Interestingly, there are several predicted MyoD binding elements in the proximal promoter region of the TSACC gene further suggesting that TSACC may be regulated by muscle specific transcription factors. The discovery that TSACC is induced in response to neurogenic atrophy helps further our understanding of the molecular genetic events of muscle atrophy and may eventually lead to the identification of new therapeutic targets for the treatment and prevention of muscle wasting.

Support or Funding Information

The work was support by University of North Florida Transformational Learning Opportunity grants to D.W.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mitochondrial Respiration In Skeletal Muscle Mitochondria From Desert Pupfish Suggests Limitation Of Metabolic Substrates As A Mechanism For Paradoxical Anaerobism
Authors: Hillyard Stanley , Jastroch Martin , van Breukelen Frank
Published: June 2021

Skeletal muscle mitochondria were isolated from desert pupfish (Cyprinodon spp) that were acclimated to ecologically relevant temperatures of 28 °C (M28) and 33 °C (M33). The respiratory control ratio (RCR) of M33 pupfish was ~2.6X lower than that of M28 pupfish, initially suggesting mitochondria were damaged by the warmer temperature. However, state 4 respiration was below expected values in both M28 and M33 pupfish suggesting little proton leak. Further, membrane potential was lower in the M33 pupfish than M28 pupfish and did not change with assay temperature. These data suggested a reduced proton motive force in M33 pupfish. Analyses of respiration as a function of membrane potential suggest a block in substrate utilization in M33 pupfish. M33 pupfish also experience reduced production of reactive oxygen species (ROS) from the Q site of complex I. Taken all together, the data suggest pupfish may be limiting mitochondrial use to avoid ROS production by restricting access of substrates to the mitochondrion. These results are consistent with our measurements of oxygen consumption in intact fish. Specifically, fish acclimated to 33 °C demonstrate extended periods of paradoxical anaerobism in which oxygen consumption is absent despite the presence of oxygen.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determining the Effect of a Novel Disease‐Inducing Rodent Diet on Outcomes of a Murine Model of Polymicrobial Sepsis
Authors: Joshua Jessica , Pogarcic Damir , Convissar Jacob , Johnson Alex Walker , Gigliotti Joseph Christopher
Published: June 2021

Sepsis affects more than a million Americans each year and is responsible for more deaths in the United States than prostate cancer, breast cancer, and AIDS combined. Obesity has been positively associated with the severity of sepsis amongst patients in the ICU, suggesting diet significantly influences the pathophysiology of disease. Animal studies support the interaction of diet with sepsis pathogenesis and severity; however the diets used have a drastically different nutrient profile as compared to the reported nutritional intake of Americans. Therefore, the goal of the current application was to determine how a novel rodent diet that adequately models the complex nutrient composition of Americans (Americanized diet ‐ AD) influences the severity of sepsis in mice. All animal experiments were performed following animal protocols approved by the Liberty University IACUC and conform to the FASEB Statement of Principles for the use of animals in research and education. Weanling male C57Bl/6 mice were fed either chow (n=5) or the AD (n=5) ad libitum for 8 weeks with body weights and 24‐hour dietary intake collected weekly. After 8‐weeks, mice underwent cecal‐ligation and puncture to induce polymicrobial peritonitis. 24 hours later blood was collected, animals were euthanized, and kidney and spleen tissues were collected. Circulating interleukin‐6 (IL6) and blood urea nitrogen (BUN) concentrations were determined using commercially available assays. Kidney and splenic IL6, TNFα, and C3 gene expression were measured using RT‐PCR and expression normalized to GAPDH using the delta‐Ct method. All data were analyzed using general linear model analyses in SPSS. Mice consuming the AD tended (P=0.05) to have significantly greater body weights starting after 4 weeks of feeding as compared to mice consuming chow. Following CLP, systemic IL6 values were significantly greater (P=0.04) in mice fed the AD (80±22 μg/mL plasma) as compared to chow fed mice (18±10). There was also a numerical increase in BUN in the AD fed mice following CLP (120±22 mg/dL) as compared to mice fed chow (70±35); however this did not reach statistical significance (P=0.16). No difference was observed in TNFα or C3 mRNA expression in the kidney or spleen; however, there was a 4‐fold increase in splenic (but not renal) IL6 mRNA expression in mice fed the AD as compared to mice fed chow (n=3). Taken together, these data highlight the significance of diet to the immunopathology of polymicrobial sepsis in mice and suggest that diet has a direct impact on IL‐6 production in the spleen.

Support or Funding Information

This work was supported by Liberty University, College of Osteopathic Medicine intramural grant #2016‐07.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Transcriptome Response of Lipid Droplet and Mitochondrial Dynamics in Zebrafish (Danio rerio) During Lipid Absorption
Authors: Francis Elizabeth Anne , Johnson Jeffery , Walters James William
Published: June 2021

Gastro‐intestinal diseases such as Crohn's disease and colon cancer have increasingly been shown to involve disruption of fusion and fission pathways linking cell growth and metabolism. Increased cell stress, Reactive Oxidative Species (ROS), and dysregulated lipid homeostasis may result from the disruption of mitochondrial dynamics. The objective of this research is to identify transcription level changes in mitochondrial fusion and fission genes within enterocytes in response to the increased metabolic load during dietary lipid absorption. Preliminary data from our lab show ultrastructural changes within enterocytes as lipid is absorbed from the lumen of the intestine. In these experiments, mitochondria increase in size presumably through fission events. Additionally, we observed an increase in apposition events between mitochondria and lipid droplets following an egg yolk diet. To investigate these phenomena, we compared dissected intestines of six days post‐fertilization zebrafish (Danio rerio) larvae after a high‐fat meal of 1 mM oleic acid and 1 mM oleic acid with cholesterol. After three hours of feeding with either diet, dissected intestines were processed for extraction of total RNA. Isolated RNA was then amplified with Qiagen RT2 Profiler Array ™ specific for zebrafish mitochondrial genes. Our results show 3.3 and 1.28 fold upregulation of the mitochondrial fusion genes Opa1 and Mfn2, respectively, in the presence of oleic acid and a 1.42 fold upregulation in FIS1 in the presence of cholesterol. These data support the hypothesis that mitochondrial fusion genes play a role in the processing of dietary fat within enterocytes.

Support or Funding Information

Research reported in this presentation was supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number P20GM103434 to the West Virginia IDeA Network of Biomedical Research Excellence (WV‐INBRE).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Using a Physiological Model to Understand Water and Electrolyte Disturbances Following Transsphenoidal Pituitary Surgery
Authors: Blair Evan T. , Clemmer John S. , Harkey H. Louis , Hester Robert L. , Pruett W. Andrew
Published: June 2021

Disturbances in water and electrolyte homeostasis are common following transsphenoidal surgery. These disturbances are variable and unpredictable, increasing patient risk and complicating postsurgical treatment decisions. Clinically, it is generally accepted that damage to the pituitary stalk is the source of these disturbances, but the mechanisms behind the response variability and underlying pathophysiology remains unknown. The objective was to test the hypothesis that stepwise damage to the posterior pituitary produces a spectrum of water and electrolyte disturbance which can account for the full variety of disease presentation. We used HumMod, a large mathematical model of human physiology, to simulate pituitary stalk damage at differing fractions: 20, 40, 60, and 80%. The damaged neurons were modeled to undergo a 5‐day countdown to degeneration and release their stored ADH as they die, as is described in the literature. Pituitary damage changed the balance of water and electrolytes to various degrees. Lower damage (20%) resulted in transient polyuria, while intermediate damage (40%) was associated with a delayed polyuria and diabetes insipidus. Higher levels of damage (60 and 80%) resembled triphasic diabetes insipidus. We postulate that the results from this model can serve as a mechanistic basis for many varieties of postsurgical water and electrolyte disturbances, where increasing cellular damage to the pituitary potentiates the likelihood of a full triphasic response. However, our simulation suggests that water and electrolyte disturbances following pituitary surgery cannot be grouped into a spectrum of disease that depends only on the level of pituitary damage. Other mechanisms which are still unclear and not a part of this model are likely at play.

Support or Funding Information

This work is supported by the Hearin Foundation through the Medical Student Research Program as well as T32 HL105324.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Treatment of Infectious Endometritis with a Novel Protein, VPI‐O22, in Cows
Authors: Talukder Jamil , Srivastava Ajay , Ray Abhijit , Lall Rajiv
Published: June 2021

Background

Endometritis, usually caused by an infection in 7–20% of dairy cows within 10 days of postpartum, deteriorates health, decreases milk production, and causes a huge economic loss to farms. In addition, treatment of infectious endometritis with antibiotics raises concern about the development of bacterial resistance. Therefore, an alternative remedy of antibiotics is desirable.

Aims

A natural, novel protein, VPI‐O22 could inhibit bacterial infection and restore uterine tone and health.

Methods

Cows with clinical endometritis (n=4) were identified based on the clinical signs, rectal temperature, score of mucus characteristics, total polymorph numbers, pH of uterine discharge, uterine swab analysis, and bacterial culture, along with healthy cows (control, n=4). Two hundred ml of VPI‐O22 (5%) were infused intrauterine with a special pipette attached to a pump in the treatment group. Seven days later, all the above parameters were reexamined. Total number of polymorph in uterine discharge was counted using a hemocytometer, stained with Giemsa and Gram stain. Bacterial culture was performed on blood agar, nutrient broth, and nutrient agar media and were identified using Gram stain. Zone of inhibition studies were performed with different concentrations of VPI‐O22 and 0.1% of chlorhexamide as a positive control.

Results

Rectal temperature was higher in cows with endometritis and was found to be normal after one week of VPI‐O22 treatment. Mean pH of uterine discharge from metritis cows was 8.8 (control 6.8) and changed to 7.0 after one week of VPI‐O22 treatment. Mucus score was 6 times higher, and total number of polymorph was more than 70 times higher in cows with metritis and significantly (p<0.01) reduced (2 and 4 times higher, respectively, compared with control) after VPI‐O22 treatment. Bacterial culture of uterine swab demonstrated the presence of different types of bacteria including gram positive, gram negative, α‐ and β‐hemolytic, rods, and cocci. Zone of inhibition studies with different concentrations of VPI‐O22 demonstrated that 5% would be more than enough to kill all types of bacteria found in these studies.

Conclusion

VPI‐O22 is a potent antimicrobial protein and has the ability to kill different types of bacteria in in‐vitro and clinical applications.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Whole‐Body Carbon Monoxide Production In Normal Adult Human Volunteers
Authors: Goktepe Nazli , Johnson Fruzsina Klara , Johnson Robert Alan
Published: June 2021

BACKGROUND

Humans and other mammals break down heme to generate carbon monoxide (CO) in a reaction that is catalyzed by heme oxygenase (HO). It is well documented that CO can bind to nitric oxide synthase to inhibit nitric oxide formation. Numerous studies have also shown that increased HO expression and subsequent CO formation inhibit nitric oxide production resulting in endothelial dysfunction. These findings have been demonstrated in numerous pathological animal models and in intubated trauma patients. Past clinical studies, measuring whole body CO formation have required patients to be placed in sealed chambers or intubated. Such restrictions have made it difficult to readily assess CO formation in patients. In an attempt to overcome these limitations, we have developed a technique to measure whole‐body CO excretion without the complexity of sequestration or intubation.

APPLICATION

The earliest studies noting CO generation in humans reported that carboxyhemoglobin levels are elevated in females, specifically during estrus. It has also been reported that that estrogen will induce the expression of HO. Accordingly we hypothesized that females will generate higher rates of whole‐body CO generation in comparison to males; furthermore rates of CO generation will be additionally exaggerated during estrus.

METHODS

Male and female volunteers were fitted with masks to measure minute volume ventilation using a non‐recirculating circuit. Samples of exhaled breath were collected and assayed for CO content via solid‐phase gas chromatography. For each subject, the whole‐body CO excretion was calculated as the product of the CO concentration and minute volume ventilation; excretion was normalized to body weight.

RESULTS

Our measurements of whole‐body CO excretion using our newly developed technique were in accordance with those previously reported in intubated male trauma patients. However, in this series we learned that CO excretion in females is not higher than in males (p > 0.05). Furthermore, rates of CO excretion during estrus are not elevated when compared to the rest of the menstrual cycle.

CONCLUSIONS

Whole body CO formation is not elevated during estrus so reports of elevated carboxyhemoglobin levels may be the result of changes in hemoglobin activities, but not the result of enhanced CO formation per se.

Support or Funding Information

William Carey University College of Osteopathic Medicine

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