bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of the Reproductive Toxicity of L‐Histidine Methyl Ester Hydrochloride of Amphotericin‐B In Vivo
Authors: Jaimes Paola , Reyes‐Esparza Jorge , Blake Ivan Ortega , Rodriguez‐Fragoso Lourdes
Published: June 2021

Systemic mycosis has increased in recent years. The drug that is used for the treatment of these mycoses is amphotericin‐B, which is a highly toxic drug generating hepatotoxicity and nephrotoxicity. These adverse effects are very serious, which causes an abandonment of the treatment, there are some alternatives such as new lipid formulations of amphotericin‐B. Unfortunately these types of medications are not available for all people because of their high cost. Recently this group has found that derivative AmB (A21) has similar pharmacological effects of AmB but not induce nephotoxicity and hepatotoxicity. However we unknown if it induces other side effects than AmB. Therefore, the aim of this work was to evaluate the effect of a derivative of AmB (A21) on the structure and function of the male and female reproductive system of Wistar rats

Derivative A21 was given intraperitoneally at a dose of 2 mg/Kg daily to female and male rats for a period of 45 days, 120 days and a third group received during 120 days and 65 days without treatment with the analog A21 in order to study the reversion of the injury.. Samples were collected after sacrificed and tissues were fixed with formalin 4% then were cut to histology which were stained with hematoxylin‐eosin. Blood was obtained by cardiac puncture and seric hormone levels were measured. Testosterone levels for male rats and progesterone for female rats, levels were measured by an Enzyme‐linked Immunosorbent Assay (ELISA).

The analog A21 did not cause structural and functional damage in male and female reproductive system from rats when were treated for a period of 45 days, since the morphology was integrated and hormone levels were normal compared with their respective control. The analog A21 caused structural and functional damage when was given during 120 days. Testosterone levels was increased (3‐fold) and were observed several alterations in the structure of testis. While in female rats in this period of treatment did not generate structural and functional damage. When rats were treated during 120 days, suspended the treatment and sacrificed 65 days without treatment, the structures of the male reproductive system was reestablished and the levels of testosterone reduced. Female Wistar rats dis not show any functional and structural damage.

Derivative A21 is able to induce structural and functional damage in male reproductive system after 120 days of treatment, however, this effect is reverted after 65 days after stopping the treatment. Derivative did not show any alteration in female reproductive system. Therefore, derivative could be used as treatment for mycosis with the advantage that the side effects produced are reversible.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Stromal Vascular Fraction Vasculogenesis, Vessel Incorporation, and Integration with Intact Angiogenic Microvascular Networks in an Ex Vivo Cultured Tissue Model
Authors: Hodges Nicholas A , Shang Hulan , Guevara Rocio R , Porvasnik Stacy L , Schmidt Christine , Katz Adam J. , Murfee Walter L
Published: June 2021

Angiogenesis, the growth of new vessels from pre‐existing vessels, is vital to wound healing in the cases of myocardial infarction and peripheral arterial disease. Adipose‐derived stromal vascular fraction (SVF), consisting of endothelial cells, endothelial progenitor cells, pericytes, smooth muscle cells, fibroblasts and immune cells, can be transplanted to assist in wound healing as seen in various clinical trials. However, the spatiotemporal incorporation of the SVF within living tissues remains largely unknown. As such, understanding where and how SVF cells contribute to microvascular growth and remodeling will help guide their therapeutic use. The objective of this study was to evaluate the impact of SVF cells and their fate during microvascular growth utilizing a novel tissue culture model that enables time‐lapse observation of cell dynamics across intact networks. SVF was isolated from adipose of an adult male Wistar rat, labeled with DiI, and seeded onto mesentery tissues harvested from the same rat before culturing in MEM + 10% fetal bovine serum and 1% PenStrep for 3 days. Tissues were then labeled for lectin to identify vessels. Alternatively, SVF was isolated from adipose harvested from adult male or female SD‐EGFP rats, seeded onto lectin labeled mesentery from adult male Wistar rats, placed under the same culture conditions, and imaged every 24 hours. Vascularized area was increased in SVF treated versus control tissues (SVF = 84.5±1.3%; Control = 56.5%; p < 0.001; n = 4 tissues per group). The SVF related microvasculature was characterized by asterisk‐shaped clusters. GFP tracking of SVF revealed vessel segment formation via vasculogenesis containing endothelial cells and pericytes in previously avascular areas. In pre‐vascularized regions, SVF cells contributed to new vessel growth and integration with and along native vessels. SVF cells were observed both in apparent endothelial cell and wrapping pericyte locations. Interestingly, examples of new vessel segments derived from SVF cells were observed along LYVE‐1 positive lymphatic vessels. These results suggest that SVF transplantation increases vascular area through vasculogenesis and contributes to angiogenesis of pre‐existing vasculature. The results also support the use of the rat mesentery culture model as a novel tool for elucidating SVF cell transplant dynamics in an intact tissue environment.

Support or Funding Information

Funded by NIH R01AG049821

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anti‐angiogenesis Triggers VEGF‐enriched Exosomes Release from Endothelial Cells
Authors: Zeng Ye , Fu Bingmei M
Published: June 2021

Although anti‐angiogenic therapies (AATs) have some effects against multiple malignancies, they are limited by subsequent tumor vasculogenesis and progression. To investigate the mechanisms by which tumor vasculogenesis and progression following AATs, we transfected microRNA (miR)‐9 into human umbilical vein endothelial cells (HUVECs) to mimic the tumor‐associated endothelial cells in hepatocellular carcinoma and simulated the AATs in vitro and in vivo. We found that administration of the angiogenesis inhibitor vandetanib completely abolished miR‐9‐induced angiogenesis and promoted autophagy in HUVECs, but induced the release of vascular endothelial growth factor (VEGF)‐enriched exosomes. These VEGF‐enriched exosomes significantly promoted the formation of endothelial vessels and vasculogenic mimicry in hepatocellular carcinoma and its progression in mice. Anti‐autophagic therapy is proposed to improve the efficacy of AATs. However, similar effects by AATs were observed with the application of anti‐autophagy by 3‐methyladenine. Our results revealed that tumor vasculogenesis and progression after AATs and anti‐autophagic therapies were due to the cross‐talk between endothelial cells and tumor cells via VEGF‐enriched exosomes. These findings provide a critical insight into a new interpretation of why AATs have not achieved expected outcomes. They also suggest that control of exosome release or alteration of exosome cargo composition to inhibit tumor vasculogenesis may augment the anti‐angiogenic and anti‐autophagic therapies for tumors.

Support or Funding Information

Supported by National Natural Science Foundation of China (Grant no.11402153).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Amyloid‐β Peptide‐mediated Disruption of Ca2+ dynamics in Contractile Pericytes
Authors: Gonzales Albert L. , Nelson Mark
Published: June 2021

Alzheimer's diseases and Cerebral amyloid angiopathy are diseases centered on the accumulation of toxic amyloid‐β‐containing plaques leading to the slow and progressive deterioration of the brain. Recent work has linked cardiovascular pathologies with these forms neurodegeneration, whereby vascular dysfunction potentiates the inability to efficiently clear toxic amyloid‐β from ageing brains. Our recent work describes how brain capillaries, the point of nutrient delivery and waste removal between circulating blood and surrounding neurons, and contractile pericytes act in concert to detect and respond to neural activity, increasing blood flow to meet local demand (functional hyperemia). Capillary pericytes closest to the terminal arteriole possess multiple vessel‐wrapping projections and are capable of exerting subcellular Ca2+‐dependent contraction. We tested the hypothesis that amyloid‐β peptides form Ca2+‐permeable pores within the plasma membrane of pericytes, leading to membrane permeabilization and pericyte Ca2+ dysregulation. Using transgenic mice expressing genetically encoded Ca2+ biosensors in contractile pericytes and smooth muscle cells, we found that free amyloid‐β oligomers (1–40) selectively increased the frequency of Ca2+ events in pericytes but not on neighboring arteriole smooth muscle cells. In addition, administration of amyloid‐β oligomers prevented the membrane depolarization‐induced contraction of pericytes by 60 mM KCl. These data suggest amyloid‐β peptide‐mediated increases in Ca2+ event frequency “short‐circuits” pericyte Ca2+ signaling, leading to Ca2+ overload and disruption of pericyte‐mediated control of junctional blood flow.

Support or Funding Information

Totman Medical Research Trust, Foundation Leducq, Horizon 2020, NIH R01HL131181, R01HL121706, R37DK053832, and K01HL138215.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Macrophage Insulin‐Like Growth Factor 1 Downregulated Matrix Metalloproteinases and Promoted Features of a Stable Atherosclerotic Plaque
Authors: Snarski Patricia , Danchuk Svitlana , Sukhanov Sergiy , Yoshida Tadashi , Higashi Yusuke , Shai Shaw Yung , Chandrasekar Bysani , Delafontaine Patrice
Published: June 2021

We have shown that insulin‐like growth factor I (IGF‐1) reduced atherosclerosis in Apoe‐null mice. We also reported that deficiency for macrophage (MF)‐specific IGF‐1 receptor (IGF1R) upregulated matrix metalloproteinases (MMPs), decreased MMP inhibitor RECK (reversion inducing cysteine rich protein with kazal motifs) and promoted atherogenesis. To study whether increase in MF IGF‐1 will suppress MMPs and decrease atherosclerosis, we generated Apoe‐null mice with IGF‐1 overexpression driven by MF‐specific promoter (scavenger receptor A) called SRA mice and we also performed experiments with cultured THP‐1 macrophages. THP‐1 MFs were incubated with IGF‐1 (0, 20, 100ng/mL) for 48 hrs. IGF‐1 (20 ng/ml) decreased expression of MMP8 (34+/−13% decrease, P<0.05) and MMP14 protein (35+/−17% decrease, P<0.05) as quantified by ELISA. To demonstrate that IGF‐1 effect on MMPs was mediated via IGF1R‐dependent mechanism, we used Picropodophyllin (PPP), a specific IGF1R inhibitor. MF pre‐treatment with PPP (1 ug/ml, 2hrs) completely abolished IGF‐1‐induced MMP8 downregulation in MF. We generated two mouse lines overexpressing IGF‐1 in MF: SRA12 and SRA17. IGF‐1 level was increased in peritoneal MF isolated from SRA12 (21±4% increase) and from SRA17 mice (52±9% increase) compared to control Apoe‐null mice (both are P<0.05). SRA12 and SRA17 mice were fed with a high fat diet for 12 weeks. MF‐specific IGF‐1 overexpression significantly decreased atherosclerotic burden (SRA12, 23±3% decrease, n=20, and SRA17, 28±6% decrease, n=16, vs. control, n=29, en face analysis) and increased plaque collagen levels (11.1±6%, SRA12 and 66.5±11%, n=11, SRA17, Trichrome staining). Atherosclerotic plaques in both SRA12 and SRA17 mice had reduced necrotic cores suggesting elevated plaque stability. Of note, the increased levels of plaque collagen in SRA17 mice correlated with a significant decrease of MMP1, MMP2, MMP8, MMP12, MMP13, and MMP14 protein levels in peritoneal MF (immunoblotting, 30–72% decrease vs. control). Protein level of RECK was increased in both SRA12 and SRA17 peritoneal MF. In summary, we show that IGF‐1 reduced MMPs in vitro and MF IGF‐1 decreased MMPs, upregulated collagen and suppressed atherosclerotic burden in vivo. Thus, we identified the novel MMP‐ (and collagen) dependent mechanism potentially mediating MF IGF‐1 atheroprotective effect.

Support or Funding Information

NIH/NHLNI 2R01HL070241‐13A1

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Adipose Specific Loss of PCPE2 Promotes Microvascular Endothelial Dysfunction and Decreased Angiogenic Potential
Authors: Brand Lukas , Hader Shelby N , Sorci‐Thomas Mary , Beyer Andreas M
Published: June 2021

Procollagen C‐endopeptidase enhancer 2 (PCPE2) is an extracellular matrix protein which regulates the oligomerization of the high and low‐density lipoprotein (HDL/LDL) receptors that enable cellular uptake of HDL/LDL from the extracellular matrix under healthy conditions. To evaluate Effect of adipose PCPE2 floxed mice were crossed with mice expressing adipose specific cre recombinase. We hypothesized elevated lipid levels induced by PCPE2 KO mice vascular function and angiogenic potential.

In Adipose specific PCPE2 KO we found that due to the inability of lipoprotein lipids in HDL to be taken up by the adipocytes, the concentration of HDL in the plasma compartment significantly increased. Using this model vascular function was evaluated using pressure mayo graph setup. Endothelial function was determined by dose response curves to ACH or increasing levels of shear stress. Smooth muscle function was tested by response to Papaverine. Angiogenic potential was evaluated by embedding sections of brown adipose tissue in Matrigel and vascular growth measured by sprout formation. Statistical significance was established using a student t‐test P<0.05 vs. CTRL.

Adipose specific PCPE2 KO resulted in elevated plasma lipid levels as elevated by insert numbers. Homozygous loss of Ad‐PCPE2 resulted in decreased endothelial function compared to control mice. Interestingly heterozygote deletion of Ad‐PCPE2 deletion trended toward increase in endothelial mediated dilation (Fig. 1A and B). Smooth muscle function was not affected by either genotype (Fig. 1C). Angiogenic Potential was significantly decreased in both heterozygote Ad‐PCPE2 KO mice compared to CTRLs (% growth Mean, SEM, N: CTRL 3.05±1.31, 6; Het 1.07±0.59*, 4)

In conclusion, our data suggest that ad‐PCPE2 loss of function elevated plasma lipid levels that promote endothelial dysfunction as assessed by decreased vasodilator capacity and angiogenic potential.

Support or Funding Information

Funded by MST HL127649 and HL138907; AMB R01‐HL133029‐02

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Reduced expression of inflammation regulatory proteins, ETS‐1 and TNIP‐1, may cause lupus in humans
Authors: Econome Jason , Malkiel Susan , Diamond Betty
Published: June 2021

Systemic Lupus Erythematosus is a painfully crippling autoimmune disorder whereby its victims suffer from chronic inflammation, and an increased susceptibility to kidney failure if left untreated. The illness can in part be triggered by a number of known environmental factors including ultraviolet light but its genetic roots remain unclear. Identifying the causal risk alleles would greatly aide in earlier detection and the administration of effective immunotherapies as well as a longer life for the lupus sufferer. Suspected risk alleles for genes, ETS‐1 and TNIP‐1, normally associated with regulating inflammation, were tested for allelic expression imbalance. Leukocytes were isolated from middle age females, of Asian descent who were heterozygous for either suspected gene. After cells were isolated from differential centrifugation, total RNA was collected, cDNA synthesized, and a Taqman quantitative PCR assay performed in order to detect reduced levels of expression for either gene. Neither B cells nor T cells showed Ct values (Ct ≥ 1 or 2x difference in mRNA) outside the standard deviation range in comparison to negative controls. Adjustments to the current procedure to enrich for a larger, more stable population of total RNA, leading to more reliable qPCR results, are needed to corroborate a role for these two risk alleles in the cause of this devastating disease.

Support or Funding Information

This research was funded by the American Physiology Society and hosted by Dr. S. Malkiel and Dr. B. Diamond at The Feinstein Institute, Manhasset, NY.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
REAVER: An Improved Image Analysis Pipeline for Quantifying Microvascular Networks
Authors: Corliss Bruce A. , Doty Richard , Matthews Corbin , Peirce Shayn M.
Published: June 2021

Introduction

Alterations in microvessel networks, including angiogenesis and capillary regression, play key roles in disease, injury, and development. Imaging of microvascular networks can capture their spatial structures, but effective study of network architecture requires methods to accurately quantify them. We present REAVER (Rapid Editable Analysis of Vessel Elements Routine), a tool to analyze blood vessel networks, and demonstrate that REAVER has superior performance compared to the state‐of‐the‐art1–3.

Material and Methods

Retinas from C57Bl/6J mice were labeled with IB4 Lectin Alexa Flour 647 and imaged with a Nikon TE‐2000E point scanning confocal microscope. A total of 36 z‐stacks from a variety of tissues were flattened to 2D images with a maximum intensity projection and used as a benchmark dataset. To establish ground truth, all images were manually analyzed in ImageJ. REAVER is written in MATLAB 2018a (Figure 1A).

Results and Discussion

REAVER demonstrated markedly higher performance in segmentation and quantification of vessel networks compared to currently available software packages. For vessel length, REAVER had lower mean relative error to ground truth (−1.4% ± 0.091) compared to Angioquant (62.8% ± 0.10), Angiotool (27.0% ± 0.08), and RAVE (43.1% ± 0.17) (Figure 1B). REAVER also had lower error to ground truth with branchpoint count (10.1% ± 0.3) compared to Angioquant (−649% ± 5.06), Angiotool (−341.2% ± 2.90), and RAVE (−1884% ± 12.36) (Figure 1C). Higher accuracy in quantification of vessel networks will allow for improved ability to discern alterations in vessel architecture for microvascular research.

Support or Funding Information

Funding: This work was supported by NIH R21 EY028868‐01 and The Hartwell Foundation (to S.M.P.).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ROS Play a Role in Long‐term Gamma Radiation‐induced Heart Damage
Authors: de Oliveira Amanda Almeida , Somasekhar Likitha , Mitra Kunal , Bowles Dawn Elizabeth , Nunes Kenia Pedrosa
Published: June 2021

Cardiovascular problems are common side effects of cancer therapy, and the heart is one of the major organs that are of concern for potential damage following radiation exposure. Radiation directly affects biological molecules. However, the most impact of radiation‐mediated cell death derives from the generated free radicals, which increase oxidative stress. Reactive oxygen species (ROS) are a hallmark in cardiovascular diseases (CVD), mainly because of its interplay with nitric oxide (NO). NO offers cardioprotection against ROS‐induced damage and ROS, in turn, limits the beneficial effects of NO. Furthermore, radiation induces adaptive alterations in the vasculature leading to vascular dysfunction. Although pathways triggering cardiovascular damage following radiation exposure involve vascular damage in humans, the long‐term mechanisms by which radiation‐induced oxidative stress affects the heart have not yet been clearly defined. We hypothesized that increased ROS contribute to long‐term heart damage following radiation exposure. Flash frozen heart (apex) samples from male C5BL/6J mice (5 months) sacrificed nine months' post radiation was provided by the Department of Surgery at Duke University. Briefly, live animals were subjected to whole body radiation (dose: 200 cGy) at the Brookhaven National Lab. Control animals received Sham treatment. Transverse sections (10 μm) of heart from irradiated and non‐irradiated mice were obtained in a cryostat. Representative images of cardiac fibers were revealed with H&E staining. Additionally, ROS and NO production were evaluated by confocal microscopy. Briefly, the fluorescent probes, Dihydroethidium (DHE) and DAF‐FM Diacetate, were diluted in PBS (5 μM) and the samples were incubated in a humidified chamber at 37° C for 30 minutes. Slides were imaged using a 20x objective and analyzed using ImageJ software (NIH, Bethesda, MD, USA). Protein expression for NOS isoforms was also assessed using a standard Western blotting technique. Our histological analysis indicates loss of cardiac tissue (heart ‐ apex) in irradiated animals compared to Sham animals, suggestive of anatomical remodeling. Confocal microscopy showed an increase in intracellular ROS in irradiated tissue (n=5; p≤0.05), but no significant difference was observed in NO production (n=5; p>0.05). Different NOS isoforms evaluation showed elevated iNOS protein expression levels in irradiated animals. In conclusion, gamma radiation stimulates sustained ROS production in heart tissue and activation of iNOS might be a mechanism for counterbalancing the disruption in the heart antioxidant response system.

Support or Funding Information

NASA Florida space Grant Consortium 2018

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Acute Effect of Electronic Hookah Smoking on Endothelial Function in Young Healthy Male Adults
Authors: Ikharo Ebahi , Rezk‐Hanna Mary
Published: June 2021

Smoking is a major cause of cardiovascular morbidity and mortality worldwide. Over recent years, alternative forms of traditional tobacco smoking have emerged, including electronic (e‐) cigarettes and e‐hookah bowls, with claims that they are safer tobacco alternatives. Unlike e‐cigarettes, e‐hookah bowls are used through traditional water‐pipes, allowing the vapor—containing aerosolized nicotine, propylene glycol, glycerin, and flavorings—to pass through a water‐filled base, potentially altering the vapor, before it is inhaled to the user's mouth. While e‐cigarettes have been shown to cause acute endothelial dysfunction, the effect of e‐hookah is unknown. In eight overtly healthy, young, male adult hookah smokers who do not smoke cigarettes (age 25±3 years; body mass index, BMI 23.9±2.7 kg·m2 mean±SD), we measured heart rate, blood pressure and endothelial function (as measured by brachial artery flow‐mediated dilation (FMD)) before and after a 30‐minute e‐hookah smoking session. Hookah smoking increased heart rate (Δ heart rate: +10±13 beats.min−1) and blood pressure (ΔSBP: +12±7 mm Hg; ΔDBP: +8±5 mm Hg). Importantly, after e‐hookah smoking, FMD decreased from 6.59 ± 2.61 to 5.33 ± 1.97 %Δ, indicating impaired endothelial function. As functional impairment of the endothelium is one of the first recognizable signs of development of atherosclerosis, this new preliminary data provide an initial line of scientific evidence to counter claims that smoking e‐hookah is a safe tobacco alternative. Future studies are indicated to elucidate the long‐term effects of e‐hookah smoking on vascular health, which overtime could accelerate the development of cardiovascular diseases.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Reversal of Endothelial Exosomes‐induced Smooth Muscle Phenotype Transition by 7‐ ketocholesterol Stimulation: Role of NLRP3 Inflammasome Activation
Authors: Yuan Xinxu , Bhat Owais M. , Lohner Hannah , Huang Dandan , Li Pin‐Lan
Published: June 2021

Recent studies have indicated that endothelial Nod‐like receptor pyrin domain 3 (NLRP3) inflammasome activation during hypercholesterolemia (HC) triggers inflammatory inflammation and consequent atherosclerosis. However, the cell‐cell communication mechanisms between endothelium and smooth muscle cells (SMCs) during inflammation remain poorly understood. The present study hypothesized that activation of coronary endothelial NLRP3 inflammasomes during HC enhances the phenotypic transition of SMCs via exosomes to accelerate atherogenesis. In the primary cultures of carotid endothelial cells (CECs), we found that 7‐ketocholesterol (7‐Ket) treatment activated NLRP3 inflammasome as we showed previously and at the same time it led to a dose‐dependent increase in exosome release as measured by Nanoparticle Tracking Analysis (NTA). The increased exosomes from CECs were found to contain inflammasome products such as IL‐1β, which was associated with the decreased interaction of lysosome with multivesicular bodies (MVBs), as shown by the colocalization of MVB markers VPS16 with IL‐1β or Lamp‐1 under the confocal microscope. Mechanistically, we demonstrated that 7‐keto‐induced exosome secretion with IL‐1β and reduced interaction of lysosomes with MVBs could be enhanced by acid ceramidase (AC) inhibitor‐carmofur, lysosome inhibitor‐bafilomycin A1, or AC gene deletion. When arterial SMCs were incubated with the exosomes collected from CECs with inflammasome activation by 7‐Ket, their phenotypic transition to more synthetic was observed, as shown by increased vimentin or vimentin/SM22 ratio in the Western blot analysis and by enhanced cell migration as measured using wound healing assay method. These effects of purified exosomes from CECs upon NLRP3 inflammasome activation by 7‐Ket were opposite to that induced by exosomes from vehicle‐treated CECs, where decreased vimentin level and cell migration occurred. These results suggest that activation of the NLRP3 inflammasome in CECs by 7‐Ket releases its products via exosome secretion. These exosomes act on SMCs to promote their phenotypic transition to synthetic and also to enhance SMC migration. However, exosomes from CECs without treatment of 7‐Ket control SMC differentiation and reduce their migration, which may maintain normal SMC phenotype and function.

Support or Funding Information

Supported by HL057244, HL075316, and DK120491

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hyperglycemia impairs mitochondrial respiration in human brain microvascular endothelial cells
Authors: Chen Allen L. , Sakamuri Siva S. V. P. , Sure Venkata N.. , Evans Wesley R. , Zheng Sufen , Katakam Prasad V.G.
Published: June 2021

Objective

Diabetes is an important risk factor for the stroke. Hyperglycemia‐induced changes in the cerebral vasculature play critical role in adverse effects of diabetes on stroke. Previously, we reported the regulation of vascular function by mitochondria. In this study, we characterized for the first time the effect of hyperglycemia on mitochondrial respiratory function in human brain microvascular endothelial cells (HBMECs) in vitro using the Seahorse XFe24 instrument.

Methods

HBVECs were plated on seahorse cell pates and exposed to endothelial cell media containing normal glucose (5 mmol/L), high glucose (25 mmol/L) and normal glucose (5 mmol/L) with mannitol (20 mmol/L) for 24 hours. Basal respiration, ATP production and maximal respiration were measured along with the spare respiratory capacity, proton leak and non‐mitochondrial respiration. Parameters were reported as oxygen consumption rate normalized to the protein concentration of cell lysates in each well (OCR, picomoles/L of O2 per microgram protein). Experiments were repeated three times with 7 to 8 technical replicates for each treatment in each experiment.

Results

Basal respiration was not significantly altered by hyperglycemia, whereas maximal respiration was decreased by 27.5 % when compared to HBMECs treated with normal glucose (7.31±0.32 vs 10.1±0.29, p=0.0002). Spare respiratory capacity was decreased by 47.5% (2.20±0.30 vs 4.25±0.24, p=0.0003) by hyperglycemia, whereas no significant changes were observed in ATP production and non‐mitochondrial respiration. Proton leak showed a strong trend towards decrease (1.09±0.07 vs 1.45±0.03, p=0.06) in high glucose‐treated HBMECs when compared to normal glucose‐treated cells. The observed respiratory changes were due to hyperglycemia but not due to elevated osmotic pressure, as HBMECs treated with mannitol did not exhibit similar respiratory changes.

Conclusions

Hyperglycemia diminishes the ability of endothelial cells to meet the increased energy demands placed by cellular stress such as neurovascular coupling (physiological) stroke (pathological). Thus, mitochondrial respiratory dysfunction in HBMECs is a novel link between hyperglycemia and cerebrovascular dysfunction.

Support or Funding Information

Support: National Institute of Health: National Institute of General Medical Sciences and National Institute of Neurological Disorders and Stroke (Katakam: R01NS094834).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exposure to WTC Dust Alters Rat Cardiac Gap Junctional Proteins
Authors: Tanwar Vineeta , Bhattacharya Sayak , Rustagi Sachit , Bulgart Hannah , Cohen Mitchell , Wold Loren STATE
Published: June 2021

Objective

First responders (FR) present at Ground Zero during the first 72 h after the World Trade Center (WTC) towers collapsed have exhibited increased incidences of cardiovascular disease (CVD) risk. Despite increasing evidences of CVDs, the mechanistic underpinnings behind adverse effects of WTC dust exposure are not well studied. Using a system that generated/delivered supercoarse (10–53 μm) WTC dusts (in a manner that mimicked FR exposures), this study sought to examine potential toxicities in the heart.

Methods

In this study, rats were exposed on two consecutive days for 2 h/day by intratracheal inhalation (ITIH) to supercoarse (10–53 μm) WTC dusts (while under isoflurane [ISO] anesthesia) or an air/ISO (naïve) mixture. This dose conservatively modeled likely exposures to nearly ≈250 mg dust/m3 FR exposure. Hearts were harvested 30 & 60 days post‐exposure and processed for qualitative and quantitative protein expression analyses.

Results

Data obtained from right ventricular samples demonstrate decreased desmin expression in the WTC exposed group as compared to the control groups (naive and ISO). Interestingly, although no change in the expression of connexin‐43 was observed between the exposed and control groups, localization studies using confocal microscopy revealed disrupted organization of connexin‐43 at the cardiomyocyte junction. Additionally, confocal imaging suggested reduction of F‐actin expression in the WTC exposed group compared to control groups. Further, expression profiles of the PI3K/Akt signaling axis using western blot analysis revealed a marked decrease in activated phospho‐Akt in the WTC dust exposed group in comparison to control groups. Although blunted activation of Akt was observed, phospho‐PI3K levels were unaltered.

Conclusion

Exposure to WTC dusts caused significant remodeling of gap junctions and altered expression of selected mechanical junction proteins at the intercalated disks. These results illustrate that a defect in a structural/gap junction protein can cause alterations in structural integrity leading to electrical and mechanical perturbations that can contribute to cardiac 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.
Evolution of Human Pulmonary Hemodynamics during Severe Sustained Hypoxia
Authors: Hoffmann Fabian , Limper Ulrich , Zaha Vlad , Reuter Hannes , Zange Leonora , Schulz‐Menger Jeannette , Hein Marc , Sadek Hesham , Baldus Stephan , Levine Benjamin , Jordan Jens , Tank Jens
Published: June 2021

Background

Susceptible individuals experience hypoxia‐induced pulmonary hypertension when ascending to high altitude. Descent is considered the primary intervention to treat this condition. A human feasibility study testing influences of severe sustained normobaric hypoxia corresponding to >7,000 m altitude on the heart allowed us to follow the evolution of pulmonary hypertension using state‐of‐the‐art cardiovascular imaging.

Material & Methods

Two healthy, professional mountaineers (subject A, male 57 years, subject B, female 50 years) first pre‐acclimatized for one week in the field at 4559 m altitude. Then, they stayed at the :envihab facility of the German Aerospace Center in Cologne where normobaric hypoxia was achieved by nitrogen dilution starting at 13.5% O2 to a minimum of 8% O2 over three weeks. Afterwards, 8.5% O2 during the daytime and 8.8% O2 at night was kept for another 14 days. Velocity encoded single plane phase contrast MRI of the pulmonary artery (PA) and echocardiography were performed 1 month before, during and 1 and 3 months after hypoxia for assessment of right ventricular (RV) function, PA pressure, area, flow and distensibility.

Results

Systolic PA pressure (PAPsys) was closely related to the decrease in oxygen during acclimatization. In subject B PAPsys peaked at 67 mmHg, 48 mmHg higher than baseline. PAPsys decreased steadily to a minimum of 44 mmHg after two weeks while still at 8.55% oxygen. Peak values occurred with clinical symptoms of acute mountain sickness and a drop in exercise capacity but without signs of cerebral or pulmonary edema. In subject A peak PAPsys was 44 mmHg and remained stable over the entire hypoxia exposure.

Conclusion

In healthy humans, severe normobaric hypoxia induces pulmonary hypertension with paradoxically preserved right ventricular function. Pulmonary hypertension may improve over time in some individuals, even in the face of sustained hypoxia. The findings attest to the remarkable physiological reserve of healthy human beings allowing for adaptation to massively increased pulmonary pressure.

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 transcriptional regulation of periostin by scleraxis in cardiac myofibroblasts
Authors: Czubryt Michael P , Nagalingam Raghu S , Schwartz Leah Y , Al‐Hattab Danah S , Aroutiounova Nina
Published: June 2021

Extracellular matrix (ECM) remodeling resulting in cardiac fibrosis is a crucial factor in determining the severity of myocardial dysfunction during heart failure. In response to stress or damage, cardiac fibroblasts undergo phenotype conversion to myofibroblasts, the primary cell type responsible for ECM synthesis and remodeling in the heart. Periostin, a marker of the myofibroblast phenotype, is a matricellular secreted protein that contributes to cardiac remodeling. Periostin plays important roles in mediating cell–matrix signal transduction and in promoting pro‐fibrotic TGF‐β signaling via coupling with matrix associated lysyl oxidase; it is highly expressed in myofibroblasts, but is not expressed in fibroblasts. We have shown that the transcription factor scleraxis induces the phenotype conversion of cardiac fibroblasts to myofibroblasts by directly transactivating the expression of proteins highly up‐regulated in myofibroblasts, including collagen 1α2, fibronectin and α‐smooth muscle actin. Here we report that the treatment of primary adult rat cardiac myofibroblasts with angiotensin II significantly increases periostin gene expression in parallel with increased scleraxis expression. Over‐expression of scleraxis in myofibroblasts increased periostin expression nearly 19‐fold at the mRNA level. TGF‐b treatment induced periostin expression in cardiac myofibroblasts isolated from wild type mice, but failed to show a similar effect in cells isolated from scleraxis null mice. Using in silico analysis, we identified putative E‐box sites within the periostin gene promoter to which scleraxis may bind to potentially regulate promoter transactivation. Our data suggests that scleraxis is necessary and sufficient for TGF‐β mediated periostin expression. Our findings are consistent with a potential role for scleraxis in cardiac fibrosis, where periostin and fibrillar collagen gene expression are elevated in parallel with scleraxis.

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 salt‐sensitivity in C57Bl/6J mice is linked to increased renal protein expressions of SLC4A4 and SLC4A5
Authors: Zhu Gangyi , Yu Yanting , Ren Zhiyun , Asico Laureano , Jose Pedro , Wang Xiaoyan
Published: June 2021

Background

Renal regulation of sodium reabsorption largely depends on the function and protein expression of exchanger /transporters /channels for sodium and other ions along the nephron. Decreased protein expression of NCC mediated aldosterone‐escape, a reflection of pressure natriuresis phenomenon. However, the role of renal ion transporters in salt‐sensitive mice is not well understood. Therefore, we studied the effect of varying amounts of sodium intake on the blood pressure (BP) and renal expression of exchanger /transporters/channels for sodium and other ions in salt‐sensitive C57Bl/6J mice and salt‐resistant BALB/c mice.

Methods

Male C57BL6 and Balb/c mice were 3 months old, fed normal salt (0.6% NaCl, NS), low salt (<0.02% NaCl, LS) and high salt (4% NaCl, HS) diet, respectively, for 1 week. The mice were placed in metabolic cages for 24 hr urine collection. BP was measured under anesthesia before sacrificing the mice. The kidneys were removed for immunoblotting and immunostaining. Data were analyzed with one‐way ANOVA, Holm‐Sidak test (P<0.05 is significant).

Results

4% NaCl diet increased the BP (ΔBP=+20 mm Hg) of C57Bl/6J (n=5) but not of sex‐and agematched BALB/c mice (n=5). Additional studies show that C57Bl/6J mice on high NaCl intake (1.6%, 4%, 6%) had higher BP (ΔBP=+20 mm Hg) relative to when they were on low or normal NaCl intake (<0.09%, 0.6%, 0.8%). The dietary salt‐induced increase in BP was observed in anesthetized and conscious, un‐anesthetized mice, measured by telemetry. There were no differences in water/food intake, urinary excretion, and serum concentrations of Na+, K+, and Cl− between the two mouse strains. High salt intake increased the renal protein expressions of SLC4A5 (NBCe2) (162±8% vs. 100±5%) compared to normal salt diet in C57Bl/6J mice, which was not observed in BALB/c mice. High salt intake increased the renal protein expressions of SLC4A4 (NBC1) (163±12%) compared to normal salt diet in C57Bl/6J mice. By contrast, the 4% NaCl diet increased renal SLC26A6 (Pendrin L1) in both C57Bl/6J (190±1%) and BALB/c (161±8%) mice. Increased NaCl intake did not affect the renal protein expression of NHE3 in either mouse strain. Renal protein expressions of NCC and αENaC, but not of NKCC2 and γENaC, were decreased by high salt diet in BALB/c mice (normal response to HS intake), but not in C57Bl/6J mice. SLC4A5 was located in proximal tubules and distal convoluted tubules, colocalized with NHE3 and NCC respectively in the mouse renal cortex. We conclude that the salt‐sensitivity of BP of C57Bl/6J mice is associated with increased renal protein expressions of SLC4A4 and SLC4A5 and impaired salt‐mediated down‐regulation of NCC and αENaC.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Metabolic and cardiovascular responses to chronic intermittent hypoxia and hypercapnia
Authors: do Carmo Jussara M. , da Silva Alexandre A. , Moak Sydney P. , Wang Zhen , Hall John E.
Published: June 2021

Obstructive sleep apnea exposes individuals to chronic intermittent hypoxia‐hypercapnia (CIHH) episodes and it is associated with increases in sympathetic nervous system activity and blood pressure (BP). Previous experimental studies utilized mainly exposure to chronic intermittent hypoxia (CIH) alone without hypercapnia. In this study we examined if CIHH potentiates the pressor and metabolic responses compared to CIH alone. Sprague‐Dawley rats were instrumented with telemetry probes for measuring mean arterial pressure (MAP) and heart rate (HR) 24‐hrs/day. Rats (n=10) were exposed to CIH (7% O2, 8 hrs/day from 8:00–16:00 hours for 5 consecutive days) or CIHH (7% O2 plus 8% CO2, 8 hrs/day from 8:00–16:00 hours for 5 consecutive days). CIHH and CIH reduced food intake similarly (21±1 to 17±1 vs. 16±1 to 14±1 g/day) leading to a net cumulative decrease in food intake of −22 and −21g, respectively. No changes were observed in plasma glucose concentrations. However, MAP response to CIHH was attenuated by 30% compared to CIH alone (+7±1 vs. +10±1 mmHg). CIHH also induced bradycardia when compared to CIH alone (−24±4 vs. +3±4 bpm, respectively). These results show that combining hypoxia with hypercapnia attenuates the rise in MAP and reduces HR without altering blood glucose and food intake when compared to hypoxia alone. Thus, chronic intermittent hypoxia, but not intermittent hypercapnia, appears to contribute to elevated MAP in rats (NHLBI‐PO1HL51971, NIGMS P20GM104357, NIGMS U54GM115428)

Support or Funding Information

NHLBI‐PO1HL51971, NIGMS P20GM104357, NIGMS U54GM115428

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Disproportional and paradoxical changes in blood pressure, heart rate and baroreflex under isoflurane anesthesia
Authors: Tsai Ching‐Yi , Chan Samuel H.H.
Published: June 2021

For the biomedical researchers, the mandatory employment of anesthetics in experiments involving animals is often regarded as a major limitation because the results may be different from those obtained under a conscious state. For the anesthesiologists, whether patients may have been subjected to unnecessarily deep anesthesia that might jeopardize functions of the cardiovascular system and create inadvertent damage to the nervous system is a recurrent concern. Using isoflurane as the representative anesthetic agent, the goal of this study was to address the issue of whether general anesthetics induce their depressive actions equally on all aspects of cardiovascular functions and their regulations and whether the degree of depression is directly related to the doses. To mimic the clinical setting during surgery, changes of blood pressure (BP), heart rate (HR) and baroreflex in mice instrumented for radiotelemetry were recorded under a conscious state, to be followed by the delivery of incremental concentrations of isoflurane. We found that there was a significant but disproportional reduction in the cardiovascular parameters when C57BL/6 mice were subjected to 1.5% isoflurane anesthesia. Compared with the conscious state, the averaged reduction in BP or HR amounted to 23.5% or 20.5%. On the other hand, the experimental indices for baroreflex‐mediated sympathetic vasomotor tone or cardiac vagal baroreflex manifested a precipitous decline of 90.0% or 63.4%. On incremental concentrations of isoflurane up to a maximum of 4%, mice exhibited sustained BP and HR, alongside returning of both arms of the baroreflex towards the conscious level. We further found that bilateral adrenalectomy exerted minimal effects on the changes in BP, HR and baroreflex in mice subjected to incremental concentrations of isoflurane. It is therefore unlikely that norepinephrine and epinephrine released from the adrenal medulla may account for the disproportional and paradoxical changes in BP, HR and baroreflex under isoflurane anesthesia. These surprise observations offer at least two challenges to the general assumptions on general anesthesia. The effects of isoflurane on cardiovascular functions and their regulations are not exerted proportionally, and both baroreflex‐mediated sympathetic vasomotor tone and cardiac vagal baroreflex exhibit large functional capacity and plasticity in terms of regulating BP and HR.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hypothalamic‐Pituitary‐Adrenal Axis Responses to an Acute Bout of Moderate Intensity Aerobic Exercise in Breast Cancer Survivors
Authors: Evans Elizabeth S , Little Jonathan , McNeill Kara T , Poole Cara , Bailey Stephen P
Published: June 2021

The purpose of this investigation was to examine the effect of one 30‐minute bout of moderate intensity aerobic exercise on hypothalamic‐pituitary‐adrenal (HPA) axis hormone responses in women with and without a history of breast cancer. Study participants included 21 women between the ages of 40–75 years, 11 of whom had completed treatments for Stage I–III invasive breast cancer (breast cancer survivors) and 10 of whom did not have a history of cancer diagnosis or treatment (controls). Participants completed a 30‐minute session of aerobic exercise on the cycle ergometer at a workload corresponding to 60% of VO2peak. Blood samples were taken pre‐exercise, immediately post‐exercise, and 30 minutes post‐exercise. Plasma concentrations of corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and cortisol were measured at each time point using enzyme‐linked immunosorbent assay (ELISA) techniques (Biotang, Inc., Lexington, MA). Data were compared between groups using Mann‐Whitney U tests and across time using Wilcoxon Signed‐Rank tests. Physical characteristics (e.g. age, height, body mass, BMI, percent body fat and VO2peak) were similar between breast cancer survivors and controls (56 ± 8 years vs. 56 ± 7 years; 163.1 ± 5.3 cm vs. 165.7 ± 5.7 cm; 66.8 ± 9.9 kg vs. 77.0 ± 19.0 kg; 25.1 ± 4.1 kg/m2 vs. 28.1 ± 7.3 kg/m2; 31.1 ± 5.3% vs. 33.4 ± 6.6%; and 21.7 ± 6.8 mL/kg/min vs. 24.3 ± 9.8 mL/kg/min, respectively; p = 0.193–0.973). Cortisol was somewhat elevated immediately post‐exercise in breast cancer survivors relative to controls (Table 1, p = 0.085). CRH decreased significantly at 30 minutes post‐exercise relative to immediately post‐exercise values in breast cancer survivors (Table 1, p= 0.003). ACTH decreased somewhat at 30 minutes post‐exercise relative to immediately post‐exercise values in both groups (p = 0.091 and 0.059 for breast cancer survivors and controls, respectively). All other CRH, ACTH, and cortisol responses were similar between groups and across time (p = 0.114–0.929). These results suggest that one 30‐minute session of moderate intensity aerobic exercise has largely similar effects on hormonal responses of the HPA axis in women with and without a history of breast cancer. These preliminary findings continue to demonstrate that moderate aerobic exercise is well tolerated in breast cancer survivors, thus supporting current exercise recommendations for this population.

Support or Funding Information

This work was supported by Elon University Faculty Research and Development funds.

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