bluuebunny/crossref_metadata_2025 huggingface dataset:
An Asymmetry Fractal Tree Model of the Spatial Heterogeneity of Myocardial Flow
Authors: Li Jiahang , Huo Yunlong
Published: June 2021

Objective

The blood flow in the myocardium exists obvious spatial heterogeneity, and the spatial heterogeneity has significant fractal features. The objective of this work is to develop an asymmetry fractal tree model based on the branch features calculated from anatomical data to explain the fracture of myocardium flow heterogeneity.

Method

In this work, we use a developed asymmetry fractal tree model in a previous work to analyze the fractal features of the myocardial flow heterogeneity based on the anatomical data of mice of different ages. In addition, the model predictions are compared with experimental results in mice hearts grouped by ages using nonradioactive fluorescent microsphere measurements.

Results

The results from the experimental measurements show good agreement with the prediction from our model. The fractal dimensions (D) of the myocardium heterogeneity in our model and the experimental measurement are 1.22 and 1.25, respectively.

Conclusions

This validated asymmetry fractal tree model will implement a persuasive elucidation of the fractal nature of myocardium heterogeneity, and give a theoretical view for the understanding of the interaction between branching features and coronary hemodynamics.

Support or Funding Information

This research is supported in part by the National Natural Science Foundation of China Grant 11372010, 11672006 (Y Huo); Shenzhen Science and Technology Innovation Institution (China) Grant JCYJ20160427170536358 (Y Huo).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Novel Substrates for DNA Cytidine Deaminase APOBEC3B with Possible Implications for Alkylating Agent Chemotherapy
Authors: Chapman Joseph , Custance Michael , Shen Birong , Furano Anthony V
Published: June 2021

Mutations mediated by APOBEC3B (A3B), a single‐strand specific cytidine deaminase, are common in various human cancers, often occurring in long, strand‐coordinated clusters, but also as isolated lesions. The clusters are thought to result from transient single‐stranded DNA generated during replication, recombination, transcription, or, as we recently adduced, DNA repair. A3B has a strong preference for C in a 5′‐TpC‐3′ (TpC) context and catalyzes hydrolysis of the primary amine to generate uracil. Subsequent repair results in mutation of C's to approximately equal numbers of transitions (T) and transversions (A or G), by which A3B‐mediated mutations are identified. We report here that A3B, in some instances, can deaminate cytosine residues in a double‐stranded DNA context in vitro. For example, a C paired to O6‐methylguanine (O6meG), if in a TpC context, is deaminated by A3B, likely due to weak hydrogen bonding between the cytosine primary amine and the guanine carbonyl group. Additionally, an abasic (AP) site opposite a C, or adjacent to the G of a C/G pair, renders the C susceptible to deamination. These results could account for the isolated, non‐clustered A3B mutations observed in many cancers. However, AP sites are common DNA lesions, and O6meG levels increase under alkylating conditions caused by environmental nitrosamines and certain chemotherapeutic treatments. Therefore, these foci could contribute to A3B‐mediated mutations in all cells.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The effects of triptolide on inflammatory responses in endothelial cells
Authors: Wang Youping , Song Chundong , Liu Suxiao , Cui Lin , Gao Yuan , Shen Si , Zhu Minjun , Xu Hui
Published: June 2021

It is widely held that endothelial dysfunction plays a crucial role in various pathological inflammatory conditions. Although triptolide, an active ingredient of Traditional Chinese Medicine Tripterygium wilfordii Hook.F., has been shown to possess anti‐inflammatory effects in vivo, little information is available that describes the effects of triptolide on inflammatory responses in endothelial cells (ECs). In this study, we determined whether triptolide suppresses inflammatory responses of ECs via inhibition of NF‐κB signaling. In human umbilical vein endothelial cells (HUVECs), treatment with lipopolysaccharide (LPS) enhanced cytokine and chemokine production, adhesion molecule expression, and monocyte adhesion. These effects were associated with increased activation of NF‐κB pathway. Furthermore, we found that pretreatment with triptolide dose‐dependently attenuated LPS‐induced actions. Our results indicate that triptolide suppresses the inflammatory responses of ECs via the inhibition of NF‐κB signaling. [This work was supported by the National Natural Science Foundation of China (No. 81673734) and Henan Provincial Science and Technique Foundation for International Cooperation Program (No. 162102410048)].

Support or Funding Information

This work was supported by the National Natural Science Foundation of China (No. 81673734) and Henan Provincial Science and Technique Foundation for International Cooperation Program (No. 162102410048)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Implications for Growth Differentiation Factor – 11 in Cardiovascular Disease and Metabolic Syndrome
Authors: Jamaiyar Anurag , Xu Yanyong , Wan Weiguo , Enrick Molly , Kolz Christopher L , Zhang Yanqiao , Chilian William M , Yin Liya
Published: June 2021

While cardiovascular disease is the largest cause of mortality in the United States, its severity is especially pronounced in metabolic syndrome patients. In previous studies, we have shown that coronary collateral growth (CCG) was impaired in Zucker Obese Fatty (ZOF) rats, a rat model of metabolic syndrome while coronary collateral grew extensively in Zucker Lean (ZLN) rats after a 10‐day repetitive ischemia (RI) protocol. To understand the underlying mechanism, we performed microarray and RNA‐seq to profile the gene expression of ischemic heart during coronary collateral growth in ZLN and ZOF rats which underwent the RI protocol. One of the most downregulated genes is Growth Differentiation Factor 11 (Gdf11), which was found to be significantly reduced in ZOF rat hearts compared to ZLN rat hearts and its expression was validated by quantitative RT‐PCR (n = 6). Moreover, Gdf11 mRNA expression is significantly decreased in both diabetic (db/db) and obese (ob/ob) mouse hearts compared to age‐matched wild type mice. To study the role of Gdf11 in vascular dysfunction in metabolic disorder, vascular endothelial cells and induced vascular progenitor cells (iVPCs) were treated with high glucose and high lipid (linoleic, oleic and palmitic acid). Interestingly, Gdf11 expression was decreased significantly under these treatments. Furthermore, we study the role of Gdf11 in the cardiovascular development and diseases using Gdf11 knockout mice with a LacZ gene knock in, which could track the gene expression of Gdf11 by LacZ staining. Whole‐mount of X‐gal staining showed that that Gdf11 is mostly expressed in the vasculature of the heart. Histological analyses confirmed that Gdf11 is expressed in blood vessels throughout the myocardium. These preliminary data suggested that 1) Gdf11 is downregulated in the heart of rodent models of metabolic syndrome and it might contribute to impaired coronary collateral growth in ZOF rats; 2) high glucose or high fatty acid affects the Gdf11 expression; 3) genetic studies show that Gdf11 is highly expressed in myocardial blood vessels, which suggests the role of Gdf11 in vascular growth. Gdf11 is controversial in its function in aging. However, our study is more focused on the function of Gdf11 in cardiovascular development and diseases. Using a Gdf11 knockout model will enable us to interrogate this controversy.

Support or Funding Information

The research is funded by National Institutes of Health grant 2R01HL103227‐05(YZ, LY), 1R01HL135110‐01 (WMC, LY), 1 R01 HL137008‐01A1 (LY), 1R15HL115540‐01 (LY) and 14BGIA18770028 from American Heart Association.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interactions of Angiotensin Converting Enzyme 2 with Fascin‐1 as a Novel Mechanism in Regulation of its Enzymatic Activity
Authors: Ogunlade Blessing , Guidry Jesse J , Lazartigues Eric , Filipeanu Catalin
Published: June 2021

Angiotensin converting enzyme 2 (ACE2) catalyzes the conversion of Angiotensin (Ang)‐II to Ang‐(1–7), attenuating the deleterious effects of angiotensin type 1 receptor (AT1R) activation on the cardiovascular functions. Our group originally demonstrated that increased Ang‐II levels stimulate interactions between ACE2 and AT1R followed by enzyme degradation into lysosomes, overall leading to a diminished compensatory activity against overactive renin‐angiotensin system. Although we showed that lysosomal inhibitors largely prevent Ang‐II mediated hypertension in vivo, the cellular mechanisms regulating ACE2 degradation by elevated Ang‐II remain largely unknown.

To start elucidating these aspects, we performed a proteomic experiment in Neuro2A cells to find binding partners of ACE2 which interactions are modulated by Ang‐II. Surprisingly, only fascin‐1, an actin‐bundling protein displayed differential interactions with ACE2 in these conditions, suggesting that it may be involved in ACE2 internalization and degradation. The interactions between ACE2 and fascin‐1 were confirmed by confocal microscopy. These experiments also demonstrated that in control conditions ACE2/fascin‐1 complexes are localized at the plasma membrane. Treatment with Ang‐II (100 nM, 4 h) induced internalization of these complexes within cytoplasm. Preliminary experiments determining the total cellular levels of ACE2 in transfected HEK293T by western‐blot demonstrated that Ang‐II treatment for 18 h significantly decrease the cellular enzyme levels. In contrast, in cells overexpressing fascin‐1, ACE2 cellular levels were preserved despite treatment with Ang‐II. Similarly, in HEK293T cells co‐transfected with ACE2 and AT1R, treatment with Ang‐II (100 nM, 4 h) diminished the basal enzymatic activity (18382 ±2430) by 36 ±12%. Overexpression of fascin‐1 did not significantly change ACE2 basal activity (86 ±24%), but in these cells Ang‐II was unable to decrease enzymatic activity (85 ± 28% ).

In conclusion, our results reveal for the first time a potential role of fascin‐1 in cardiovascular diseases. As an ACE2 binding partner, fascin‐1 may prevent enzyme degradation in presence of elevated Ang‐II and it appears that its cellular levels are critical in prevention of the deleterious effects of overactive renin‐angiotensin system. Also, these data highlight fascin‐1 as an additional possible therapeutic target in the treatment of cardiovascular diseases.

Support or Funding Information

5 P30 GM106392‐03 (JJG). NHLBI HL093178 (EL) and a Bridge Grant from Howard University (CMF).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protective Mechanisms of Physical Activity in Vascular Dysfunction in High‐Fat Diet‐Induced Obesity Via Attenuation of NLRP3 Inflammasome Activation in Mouse Aorta
Authors: Lee Jonghae , Hong Junyoung , Umetani Michihisa , LaVoy Emily , Park Yoonjung
Published: June 2021

PURPOSE

Inflammasomes play a pivotal role in inflammatory milieu in vascular pathology via caspase‐1 mediated pyroptosis and IL‐1β processing, but physical activity ameliorates the obesity‐associated inflammation and vascular dysfunction. Thus, we investigated the protective effect of physical activity on the NLRP3 inflammasome‐associated vascular dysfunction and its potential underlying mechanisms.

METHODS

: C57BL/6J male mice were randomly divided into four groups: (1) control low‐fat diet (LF‐SED), (2) LF diet with free access to a voluntary running wheel (LF‐RUN), (3) high‐fat diet (HF‐SED; 45% of calories from fat), and (4) HF‐RUN. To test effects of HF diet and physical activity in NLRP3 inflammasome‐related signaling pathways, nitric oxide (NO), and oxidative stress in mouse aorta, RT‐qPCR and immunofluorescence staining were performed.

RESULTS

Voluntary running decreased key markers of NLRP3 inflammasome activation, caspase‐1 and IL‐1β, in whole aortic vessel of obese mice. HF‐RUN restored diet‐induced reduction of NO production compared to HF‐SED. Intracellular superoxide production was elevated in HF‐SED mice compared to LF‐SED and LF‐RUN while it was reduced in HF‐RUN mouse aorta.

CONCLUSION

Our findings suggest that physical activity protects diet‐induced aortic vascular dysfunction by suppressing NLRP3 inflammasome activation and improved NO bioavailability via reduced oxidative stress.

Support or Funding Information

This study was supported by grants from the Cardiovascular Research Foundation Asia (#16215 to Y.P.) and the University of Houston (Small Grant Program to Y.P.).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Changes in Peak Oxygen Uptake (VO2peak) Following Renal Transplant: Results after 3‐year Follow‐up
Authors: Bushroe Cody M , Chakkera Harini A , Angadi Siddhartha S
Published: June 2021

Background

Data regarding changes in markers of cardiovascular and all‐cause mortality, namely, VO2peak and minute ventilation/carbon dioxide production (VE/VCO2) slope following renal transplantation past 1‐year post‐transplant are lacking. We examined changes in VO2peak and VE/VCO2 before and after ~3‐years after renal transplantation.

Methods

Eighteen patients (age 56.2 ± 2.5 years; 82.9 ± 4.9 kg; 9 male) completed an exercise test to exhaustion using a ramp treadmill protocol. Subjects underwent pre‐transplant exercise testing (PreTx) following which they were placed on the transplant wait‐list. Average time on the waitlist was 15.6 ± 2 months. Following transplant, subjects underwent follow‐up maximal exercise testing using the same protocol (PostTx). Mean duration to post‐transplant exercise testing was 35 ± 2 months. Gas exchange data, heart rate and respiratory exchange ratios were recorded throughout exercise testing. All variables are reported as mean ± standard error. α was set at 0.05 and a paired t‐test carried out to examine changes in VO2peak and maximal VE/VCO2 slope before and after transplant.

Results

Significant reductions in relative VO2peak were observed from PreTx = 15.2 ± 0.8 mL/kg/min to PostTX = 12.2 ± 0.4 mL/kg/min (p = 0.002 ). Also, significant increases in Ve/VCO2 were observed from PreTx = 30 ± 1.4 to PostTx = 43 ± 1.6 (p < .001).

Conclusions

Long term follow‐up (~3‐years) in this small cohort of patients following renal transplantation demonstrated significant reductions in VO2peak (20%) and worsening of the VE/VCO2 slope (30%) which are modifiable markers of mortality risk. This underscores the need for larger cohort studies to be undertaken in this population given that these markers are independently linked to cardiovascular and all‐cause mortality risk. These data may have implications for cardiovascular stratification and cardiovascular risk reduction in this population.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Acetylation on Liquid‐Liquid Phase Separation and Amyloid Fibril Formation of Tau
Authors: Zhao Ziting , Gao Meng , Huang Yongqi
Published: June 2021

The function of proteins depends on their phase behavior. When the concentration of a protein exceeds a critical value, it may undergo liquid‐liquid phase separation, leading to demixing of proteins from solution. The resulted liquid droplets are dense and protein‐rich. The microtubule associated protein tau can enrich tubulin by forming liquid droplets, which promotes the formation of microtubule bundles. On the other hand, aberrant aggregation of Tau into neurofibrillary tangles correlates with the progressive destruction of nerve cells. Therefore, studying the aggregating mechanism of tau and drug design targeting tau fibrils are critical for the therapy of Alzheimer's disease. Recent studies have shown that lysine acetylation plays an important role in modulating tau aggregation and microtubule assembly. Considering the potential correlations between liquid‐liquid phase separation and amyloid formation, here we investigated the effect of lysine acetylation on the phase behavior and function of tau in vitro. We used site‐directed mutagenesis, light scattering, thioflavin T florescence assay, and microrheology to study the liquid‐liquid phase separation and aggregation of tau. Consistent with previous observations, acetylation on K274/K321 (mimicked by K274Q/K321Q) significantly impaired tau aggregation whereas acetylation on K280/K281/K369 (mimicked by K280Q/K281Q/K369Q) significantly promoted tau filament formation as compared with the WT protein. Although acetylation on K274/K321 and K280/K281/K369 differentially affected tau aggregation, both acetylation patterns promoted liquid‐liquid phase separation of tau. Moreover, microrheology characterization and protein concentration determination showed that the droplets formed by acetylated tau were denser than the WT protein, suggesting that more intermolecular interactions were form upon lysine acetylation. These results show that lysine residues within the MT‐binding repeats regulate the phase behavior of tau via complex interactions.

Support or Funding Information

This work was supported by National Natural Science Foundation of China (Grant number: 21603121) and Hubei University of Technology.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Resuscitation from Hemorrhagic Shock with High Molecular Weight Polymerized Hemoglobin
Authors: Williams Alexander Thomas , Palmer Andre , Cabrales Pedro
Published: June 2021

Background

Hemoglobin (Hb) based oxygen carriers (HBOCs) have been proposed as an alternative to red blood cells (RBCs) for transfusion medicine. Lack of mechanistic analysis lead to clinical evaluation of low molecular weight (MW) HBOCs, which failed in phase III clinical trials due to a myriad of side effects not seen in pre‐clinical trials. Increasing the MW of HBOCs can decrease these side effects by confining HBOCs to the vascular space and increasing nitric oxide generation. HBOCs are advantageous to blood due to their non‐immunogenic properties and potential to use non‐human hb sources, among others. In this study we compare the efficacy of fresh blood, freshly synthesized polymerized bovine hb (PolybHb), and PolybHb stored for 2 years in restoring cardiac function after hemorrhagic shock.

Methods

PolybHb was synthesized in the low oxygen affinity (T) state with a 35:1 ratio of glutaraldehyde to bovine Hb, and then subjected to 8–9 cycles of diafiltration. This resulted in a PolybHb solution containing only polymerized Hb molecules < 0.2 uM and >500kDa. PolybHb was then frozen at −80°C and stored until used. Fresh red blood cells were prepared from the blood removed during hemorrhage by centrifugation at 1000g for 7 minutes, removal of the buffy coat, hemoconcentration to a hematocrit of 50%. Rats (200–250g) were hemorrhaged for 50% of their blood volume, held in hypovolemia for 30 minutes, and resuscitated to recover blood pressure to 90% pre‐hemorrhage with fresh autologous blood (fresh blood), PolybHb generated within 3 months of the study (PolybHb 2017), or PolybHb generated two years prior to the study (PolybHb 2015).

Results

Shock resulted in impaired oxygen delivery and cardiac function. Resuscitation with fresh blood, PolybHb 2017, and PolybHb 2015 restored cardiac function and systemic vascular resistance. Slightly more PolybHb was infused before restoration of 90% pre‐hemorrhage MAP was achieved, but the volume was not statistically significant. Likely as a result of the higher resuscitation volume, stroke volume and stroke work increased more in animals resuscitated with PolybHb than fresh autologous blood. PolybHb did not impair cardiac contractility nor cause excessive vasoconstriction.

Conclusions

Studies indicate that PolybHb is as efficacious as fresh autologous blood in restoring cardiac function in rats after hemorrhagic shock. Additionally, PolybHb can be safely stored at −80°C for two years and retain efficacy, unlike red blood cells.

Support or Funding Information

NIH Grants from the National Heart Lung and Blood Institute, T32‐HL105373, P01‐HL110900, R01‐HL126945

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Letrozole Treatment of Pubertal Female Mice Results in Impaired Insulin Action in Skeletal Muscle
Authors: Hernandez‐Carretero Angelina Marie , Martins Vitor , Riopel Matthew , Ho Bryan , Schenk Simon , Thackray Varykina
Published: June 2021

Approximately 6–15% of women worldwide are affected by polycystic ovary syndrome (PCOS). Although PCOS was originally described as a reproductive disorder with associated obesity, over 30 years of clinical research have confirmed that it is also a metabolic disorder. Hyperandrogenism is the strongest predictor of metabolic dysfunction associated with PCOS. In addition to infertility, women with PCOS diagnosed with hyperandrogenism and ovulatory dysfunction have an increased risk of developing gestational diabetes, type 2 diabetes, cardiovascular disease, and non‐alcoholic fatty liver disease. We previously developed a mouse model that uses the aromatase inhibitor, letrozole to elevate endogenous testosterone levels in female mice. This mouse model has hallmarks of PCOS including hyperandrogenism, anovulation and polycystic ovaries. In addition, we showed that letrozole treatment results in increased weight, abdominal adiposity, dysglycemia, hyperinsulinemia, and insulin resistance. In the current study, we further characterized the metabolic dysfunction that occurs after letrozole treatment to determine whether systemic impairment in insulin sensitivity is associated with lower insulin‐stimulated glucose disposal in peripheral insulin target tissues, such as liver or skeletal muscle. We performed hyperinsulinemic‐euglycemic clamp studies and observed that letrozole‐treated mice were indeed insulin resistant independent of body weight. Additionally, letrozole‐treated mice had an overall lower glucose infusion rate and glucose turnover, when normalized to body weight. Moreover, letrozole‐treated mice exhibited an impaired insulin‐stimulated glucose disposal rate, which is indicative of insulin resistance in skeletal muscle. In contrast, there was no difference observed in insulin‐stimulated suppression of hepatic glucose production. Lastly, ex vivo 2‐deoxyglucose uptake into skeletal muscle upon insulin stimulation was significantly reduced in letrozole‐treated mice compared to placebo. These results are in agreement with our previous findings that indicated skeletal muscle‐specific insulin signaling (as measured by AKT phosphorylation) was blunted in letrozole‐treated mice. In summary, we demonstrated that the letrozole‐induced PCOS mouse model exhibits impaired insulin action in skeletal muscle. Future studies will focus on mechanisms involved in skeletal insulin resistance in this mouse model.

Support or Funding Information

Supported by NIH Grant P50 HD012303, AHA 16POST29990015 to M.R., and NIH NIGMS K12GM068524 to A.H‐C.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Hepatic MDM2 in the Regulation of Energy and Lipid Metabolism in Obesity
Authors: Wang L. , Liu Z. , Hui X. , Xu A. , Cheng K.K.Y.
Published: June 2021

Introduction

Very‐low‐density‐lipoprotein (VLDL) is a major vehicle for transporting lipids from liver to peripherals in plasma. VLDL‐triglyceride secretion is an important pathway for disposition of hepatic fat, and its inhibition results in hepatic steatosis. VLDL assembly and secretion are controlled by lipid availability and insulin resistance. MDM2, an E3 ligase mediating the final step of ubiquitination. We previously found that MDM2 regulates glucosestimulated insulin secretion via regulation of pyruvate carboxylase (PC) in the pancreatic islet. As PC is a key metabolic enzyme that mediates gluconeogenesis in the liver and MDM2 has been shown to control hepatic fatty acid oxidation. Therefore, we here investigated the role of MDM2 in hepatic energy and lipid metabolism using a hepatocyte‐specific MDM2 knockout mouse model.

Methods

MDM2floxed/floxed mice were crossed with transgenic mice with Cre expression under the control of albumin promoter to generate hepatocyte‐specific MDM2 knockout (HMDM2‐ KO) mice. Mice were fed with a high‐fat‐high‐cholesterol (HFHC) diet for 16 weeks to induce obesity, insulin resistance and hepatic steatosis. Basic metabolic parameters and VLDL secretion rate were monitored.

Results

Hepatocyte‐specific deletion of MDM2 has no obvious effect on body weight, insulin sensitivity, glucose tolerance and energy expenditure under HFHC diet feeding. HMDM2‐KO mice are characterized by postprandial hypertriglyceridemia, which results from increased hepatic VLDL secretion. Moreover, HMDM2‐KO mice displayed attenuated hepatic steatosis and downregulated mRNA and protein level of inflammatory markers in the liver when compared to their wild‐type littermates.

Conclusion

Our study proposes a novel role of hepatic MDM2 in regulation VLDL‐triglyceride secretion independent of insulin resistance.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Transcutaneous Application of Carbon Dioxide on diabetic muscle atrophy
Authors: Kondo Hiroyo , Tani Tomomi , Naito Kana , Niimi Yurina , Nakane Moemi , Sugiyama Yui , Okayasu Kanako , Takemoto Hatsumi , Matsumoto Tomohiro , Takuwa Miho , Hirabayashi Takumi , Ikeji Takuya , Ono Kohei , Fujino Hidemi
Published: June 2021

Introduction

Diabetes results in muscle atrophy, also called diabetic myopathy. Both the increase in muscle proteolysis and the decrease in muscle protein synthesis contribute to diabetic muscle atrophy. Transcutaneous application of carbon dioxide (CO2) has been used for cardiac disease and skin problems for a long time. In addition, it has been reported the CO2 treatment leads to an increase of muscle mass in normal rats. Therefore, the transcutaneous application of CO2 would have a therapeutic potential for diabetic muscle atrophy. The aim of the present study was to investigate the effects of transcutaneous application of CO2 on diabetic muscle atrophy.

Methods

Male Goto‐kakizaki rats were divided into control and CO2 treatment groups. The hair on the lower limbs was shaved and the hydrogel was applied. The CO2 adaptor was attached to the limbs and sealed, and CO2 was administered into the adaptor for 30 min. This treatment was performed everyday for 8 weeks.

Results

The body weight, epididymal and retroperitoneal fat were lower in the CO2 treatment group than in the control. In addition, the level of fasting blood glucose in the CO2 treatment group was significantly decreased compared with the control. Furthermore, the muscle masses of lower limbs in the CO2 treatment group were higher than those in the control.

Conclution

These results indicate that the transcutaneous CO2 treatment may have a therapeutic potential for diabetic muscle atrophy.

Support or Funding Information

Supported by Grants‐in‐Aid for Science Research from the Japanese MEXT.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Appendicular Muscle Mass is Lower in Late‐ Compared to Early‐perimenopausal Women
Authors: Park Young‐Min , Jankowski Catherine M , Ozemek Cemal , Hildreth Kerry L , Kohrt Wendy M , Moreau Kerrie L
Published: June 2021

OBJECTIVE

The age‐related decline in skeletal muscle mass (i.e. sarcopenia) and bone mineral density (BMD; i.e. low bone mass, osteoporosis) contribute to physical disability in older women. Although menopause‐related loss of BMD is well documented, whether menopause influences muscle mass and sarcopenia is unclear. We determined the extent to which appendicular lean mass (ALM) and BMD differ across the stages of menopause in healthy women.

METHODS

145 women (aged 30–70 years) classified as premenopausal (Pre; regular menstrual cycles; n=30, 38±6yrs; mean ± SD), early perimenopausal (Eperi; > 2 cycles with cycle length changes of ≥7 days; n=31, 50±3yrs), late perimenopausal (Lperi; ≥2 but <12 months of amenorrhea; n=30, 50±4yrs), early postmenopausal (Epost; ≤5 yrs since menopause; n=26, 55±3yrs), or late postmenopausal (Lpost; >5 yrs since menopause; n=28, 62±4yrs) were enrolled. Dual‐energy x‐ray absorptiometry (DEXA) was used to assess ALM (kg/m2), BMD, total body lean mass, and fat mass.

RESULTS

ALM was lower (p<0.05) in Lperi and Lpost compared to Eperi with no significant differences between other groups (Pre, 6.6±0.6; Eperi, 6.8±0.8; Lperi, 6.1±0.8; Epost, 6.5±1.1; and Lpost, 6.2±0.9 kg/m2). The prevalence of sarcopenia (ALM ≤5.67 kg/m2) was 7, 3, 30, 27, and 32% in Pre, Eperi, Lperi, Epost, and Lpost, respectively. BMD tends to be lower (p=0.05, both) in Lperi and Epost compared to Eperi, and was lower (p<0.05) in Lpost compared to Eperi (Pre, 1.108±0.100; Eperi, 1.164±0.104; Lperi, 1.092±0.100; Epost, 1.088±0.107; and Lpost, 1.045±0.113 gm/cm2). There was a consistent increase in total fat mass and decrease in total lean mass from Pre to each subsequent stage (both, p<0.05 for Pre vs. Lpost).

CONCLUSION

In addition to the menopause‐related decline in bone mass, the menopause transition appears to be a vulnerable period for the loss of skeletal muscle mass that may begin during the late perimenopausal transition. Potential mechanisms underlying menopause‐related loss of muscle mass such as estrogen receptor signaling and protein metabolism should be explored in future studies.

Support or Funding Information

NIH R01AG049762, R01AG027678, R56HL114073, P30DK048520, P50HD073063, NCATS Colorado CTSA UL1TR001082, and Eastern Colorado VA GRECC

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
FTY720P Increases, at 2 hrs, the Activity of the Na+/K+ ATPase in Caco‐2 cells via PKC and PI3K
Authors: Noureddine Maysoon , Kreydiyyeh Sawsan
Published: June 2021

Fingolimod phosphate (FTY720P), an analogue of sphingosine‐1‐phosphate (S1P), was shown previously to modulate, in a time‐dependent manner, the activity of hepatic Na+/K+ ATPase with an inhibitory effect appearing at 15 min and a stimulatory one at 2hrs. Because in the colon, the ATPase plays a pivotal role in the absorption of sodium and water, a time response study was conducted in human epithelial colorectal adenocarcinoma (Caco‐2) cells to examine if FTY720P regulates the activity of the ATPase in a similar fashion. An inhibitory effect was already demonstrated at 15 min. This work investigates the effect of FTY720P at 2 hrs and attempts to determine the major signaling molecules involved. The activity of the Na+/K+ ATPase was assayed by measuring the amount of inorganic phosphate released in presence and absence of ouabain, a specific inhibitor of the enzyme. FTY720P exerted a significant and prominent stimulatory effect on the ATPase at 2 hours. This effect disappeared completely in presence of JTE‐013, a blocker of S1P receptor 2 (S1PR2), as well as in presence of each of calphostin and wortmannin, respective inhibitors of protein kinase C (PKC) and phosphoinositide 3‐kinase (PI3K). Moreover, Phorbol‐12‐Myristate‐13‐Acetate (PMA), an activator of PKC, simulated the stimulatory effect of FTY720P on Na+/K+ ATPase further confirming the involvement of PKC. BAPT/AM, a chelator of calcium ions, cancelled the effect of FTY720P on the ATPase, indicating thus an involvement of calcium ions in the signaling pathway. Since only conventional PKC isomers require the presence of calcium ions for their activation, FTY720P probably acts via a conventional PKC isomer. The stimulatory effect of PMA disappeared in presence of wortmannin, suggesting that PKC is upstream of PI3K. It was concluded that at 2 hours, FTY720‐P binds to S1PR2 leading to the activation of conventional PKC which, in turn, activates directly or indirectly PI3K, resulting in Na+/K+ ATPase stimulation.

Support or Funding Information

University Research Board

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
PGE2 and NO Mediate the Inhibitory Effect Exerted by FTY720P at 15 Minutes on Colonic Na+/K+ ATPase
Authors: Rida Reem , Kreydiyyeh Sawsan
Published: June 2021

Inflammatory Bowel Disease was shown to be accompaniedwith higher levels of sphingosine‐1‐ phosphate (S1P) and lower Na+/K+ ATPase activity. This work was undertaken to see if a cause‐effect relationship exists between S1P and the ATPase, and in case it does to determine the main mediators involved, using Caco‐2 cells as a model and fingolimod phosphate (FTY720P), a S1P analogue. The activity of the Na+/K+ ATPase was assayed by measuring the amount of inorganic phosphate liberated in presence and absence of ouabain, a specific inhibitor of the Na+/K+ATPase. Western blot analysis showed that sphingosine‐1‐phosphate receptor 2 (S1PR2) is the mostly expressed receptor in Caco‐2 cells. FTY720P, applied for 15 min, reduced significantly the activity of the Na+/K+ ATPase, with a maximal inhibition observed at a concentration of 7.5nM. The effect of FTY720P disappeared completely in presence of JTE‐013 (a specific blocker of S1PR2) and indomethacin (an inhibitor of cyclooxygenase enzymes), and was mimicked by CYM5520 (a S1PR2 agonist), prostaglandin E2 (PGE2), and sulprostone (an agonist of E prostanoid receptor 3 (EP3)). Carboxy‐PTIO a nitric oxide scavenger, abrogated the effect of FTY720P as well as that of PGE2. Treating the cells with Glyco‐SNAP (nitric oxide donor) reduced significantly the ATPase activity. It was concluded that FTY720‐P inhibits the ATPase by binding to S1PR2 leading to PGE2 production. PGE2 binds to EP3 receptors and increases nitric oxide levels resulting in an inhibitory effect on the Na+/K+ ATPase.

Support or Funding Information

University Research Board

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Thyroid Transcription Factor‐1 modulation of secretome alters sensitivity to cisplatin in lung cancer
Authors: Phelps Cody Alan , Mu David
Published: June 2021

Thyroid Transcription Factor‐1 (TTF‐1, alternatively known as NKX2‐1) is essential for lung development. Mice homozygous null for TTF‐1 fail to develop lungs and will die in utero. Like many developmental genes, TTF‐1 has also been shown to play a role in cancer development. Its role was originally elucidated when our lab and others showed that TTF‐1 resides in the 14q13.3 region that undergoes focal amplification in roughly 15% of lung adenocarcinomas, and knockdown of TTF‐1 retarded cell growth, suggesting a role as an oncogene. Since this finding, many tumor suppressive roles have been discovered, and TTF‐1 positivity in lung cancer is correlated with a better patient outcome. Our lab is interested in how TTF‐1 can alter lung cancer sensitivity to cisplatin, a common chemotherapy drug used in lung cancer. Our findings reveal that TTF‐1 can sensitize A549 cells to cisplatin. Furthermore, this sensitivity can be found in the secretome of TTF‐1 expressing cells, as conditioned media collected from TTF‐1 transfected cells can sensitize recipient cells to cisplatin relative to controls. We dissected the secretome between vesicle and soluble fractions. Exosomes failed to alter sensitivity to cisplatin. By probing the soluble secreted factors altered by TTF‐1 we found that GM‐CSF was the most differentially regulated. By adding a GM‐CSF neutralizing antibody to A549 cells expressing TTF‐1, we found a conferral of resistance to cisplatin. Conversely, by adding a recombinant GM‐CSF to A549 control cells not expressing TTF‐1, we found that it had the ability to confer sensitivity to cisplatin. Patient data from The Cancer Genome Atlas and the Director's Challenge Lung Study have found a strong positive correlation with TTF‐1 and GM‐CSF in patients. Intriguingly other chemotherapy drugs including carboplatin and gemcitabine did not show a differential effect with TTF‐1, suggesting that TTF‐1 may confer a special sensitivity to cisplatin and could serve as a guide for deciding patient treatment strategies.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The oculocerebrorenal syndrome of Lowe protein (OCRL) inhibits the Na/bicarbonate cotransporter NBCe1 expressed in Xenopus laevis oocytes.
Authors: Michenkova Marie , Thornell Ian M. , Peng Ji‐Ben , Bevensee Mark O.
Published: June 2021

The electrogenic Na/HCO3 cotransporter NBCe1 regulates intracellular pH in many tissues and contributes to solute reabsorption/secretion in epithelia. NBCe1 is modulated by hormones, classic second messengers, and other signaling molecules/ions. We previously reported that both the phospholipid PIP2 itself and its hydrolysis to IP3 (Ca2+) stimulates the activity of NBCe1 expressed in oocytes. In this preparation, activating a co‐expressed voltage‐sensitive phosphatase (VSP) that dephosphorylates PIP2 to PIP without stimulating IP3/Ca2+ signaling decreases NBCe1 activity. In the current study, we examined the effect of the PIP2‐lowering phosphatase, inositol polyphosphate 5‐phosphatase encoded by the oculocerebrorenal syndrome of Lowe gene (OCRL) on the activity of NBCe1 variants expressed in oocytes. Mutations and variants of OCRL lead to Dent disease type 2, as well as Lowe Syndrome, which is characterized by cataracts, intellectual disability, basal ganglia calcification, and proximal renal tubular acidosis (pRTA). Using the 2‐electrode voltage‐clamp technique, we found that oocytes co‐expressing human OCRL with NBCe1‐A (both HA tagged) displayed a characteristic NBC‐mediated, HCO3–‐induced outward current (INBC) that was 59% smaller than oocytes expressing NBCe1‐A. We also observed OCRL inhibition of HA‐tagged NBCe1‐B and ‐C. We obtained similar results after replacing the HA tag on NBCe1‐A with a FLAG tag. The OCRL‐induced decrease in INBC could be explained by a decrease in NBCe1 expression at the plasma membrane. Co‐expressing OCRL caused a 44% decrease in INBC and a parallel 33% decrease in FLAG‐tagged NBCe1‐A oocyte surface expression as determined by single‐oocyte chemiluminescence. Thus, a physiological phosphatase that lowers PIP2 can regulate the activity and/or expression of NBCe1. Altered activity/expression of NBCe1 caused by mutant OCRL variants seen in Dent disease and Lowe Syndrome may contribute to observed patient symptoms, especially pRTA.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CHRONIC NEURO ‐ SPINAL DEGENERATIVE DISEASE AND EFFECTIVE INTEGRATIVE DYNAMIC SIGMA QUANTUM REGENERATION NEURAL CELLS SIGNALING STIMULATION THERAPIES ‐ Case Study
Authors: TULP ORIEN LEE , Einstein George P , Konyk Carla M
Published: June 2021

Background

Complex spinal conditions may be the result of multiple disorders and pathologies, that interact together to generate back pain, discomfort and depression. At Einstein Medical Institute USA, ‐ University of Science Art and Technology developing research in Integrative therapies by applications Mathematical Functions of Sigma Quanta Coherence E.M. Energy and Einstein Matrix Dose Protocol in diverse case study treatments.

Materials and Methods

Progressive degenerative diseases affect nerve cells in the brain and the spinal cord. Motor neurons reach from the brain to the spinal cord and from the spinal cord to the muscles throughout the body. Patient had failed back syndrome of 16 years duration with chronic pain, eight spine surgeries, 100 spinal injections, nerve blocks, an implanting pump, a spinal simulator and countless prescribed medications. The multiple diagnostic matrix approach was performed. For major analysis of the electrical signals generated by the brain represent not only the brain function but also the status of the whole body was applied. The treatment protocol included detox, acupuncture, different frequency low laser therapies ‐ LLT, Sigma Quanta Coherence E.M.E and G.P. Einstein Matrix Dose Protocol for stimulation neural signaling in Brain and Spinal cord.

Result

EEG has been found to be a very powerful tool in the field of neurology and clinical neurophysiology. The electrical signals generated by the brain represent not only the brain function but also the status of the whole body. The patient after several diagnostic EEG with Integrative therapies by applications Mathematical Functions of Sigma Quanta Coherence E.M. Energy and Einstein Matrix Doses Protocol in neuro therapeutic sessions was able to walk again with low pain. The general equation for stimulation neural signaling is Sigma Quantum E= h · ∑ fN. where h represents the Planck's constant, and ∑fN is sum all frequencies in stimulation the cell signaling in brain and in neural system of total body.

Conclusion

The G.P. Einstein Mathematical Functions of Sigma Quanta Coherence E.M. Energy with Einstein Matrix Doses is safe, can be applied and designed for personal therapies by MD with extensive training in Quantum Physics, Electromagnetics Energy Systems and electro‐electronic bio‐currents in human neuro‐physiological system.

Support or Funding Information

Support from Institutional Resources of USAT Montserrat and the Einstein Medical Institute.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Plasticity of sympathetic post‐ganglionic neuron after spinal cord injury
Authors: Li Yaqing , McKinnon Michael L. , Hochman Shawn
Published: June 2021

Autonomic dysreflexia (AD) is a hypertensive crisis seen in high‐thoracic spinal cord injured (ht‐SCI) patients normally triggered by noxious stimuli below the injury level. Previous animal research on mechanisms underlying AD have mostly focused on neural plasticity within the injured spinal cord. No studies have assessed whether there is also plasticity in the sympathetic post‐ganglionic neurons (SPNs) ‐ the final neural element responsible for vasoconstriction.

Thoracic sympathetic post‐ganglionic neurons (tSPNs) are located in paravertebral ganglia and provide the dominant sympathetic control of blood pressure. Their cellular properties are understudied. The few previous ex vivo studies used sharp electrodes that likely leads to significant impalement injury‐induced changes [1, 2]. We undertook whole‐cell patch clamp recordings in an adult mouse tSPN ex vivo preparation. Compared to earlier reports using sharp electrodes, we observed an order of magnitude greater passive membrane properties (cell resistance and membrane time constant) and repetitive rather than phasic firing properties. Together these observations support much greater excitability and integrative actions of tSPNs than previously assumed.

Using the same methodology we then studied whether these neurons undergo plastic changes after high‐thoracic (T2) spinal cord transection (ht‐SCI), an animal model of AD. Recordings were obtained from T3‐T9 tSPNs at 3 weeks and 6 weeks post injury. Preliminary data suggest that early after injury (3 weeks), tSPNs are hypo‐active compared to normal animal, including more hyperpolarized resting membrane potential and higher rheobase and threshold voltage. At 6 weeks post‐SCI, tSPN passive membrane properties are similar to control mice. However, several neurons show spontaneously unstable membrane potential (55% of recorded neurons), which is rarely seen in control. Evidences that tSPNs may become more excitable include lower rheobase and threshold voltage values on average and an increased incidence of nifedipine sensitive persistent inward currents (PIC; 69% of recorded neurons after SCI has PIC comparing to 22% in control). Also, tSPNs appear to receive more frequent spontaneous cholinergic synaptic input after SCI.

Support or Funding Information

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Development and Progression of Mechanical Allodynia in UC, Davis Type 2 Diabetic Rats
Authors: Crawford Charles Kikutaro , Ybarbo Kai , Grotle Ann‐Katrin , Huo Yu , Graham James , Stanhope Kimber , Havel Peter J , Stone Audrey J
Published: June 2021

Type 2 diabetes (T2DM) is a metabolic disorder that can lead to many neuropathic complications such as mechanical allodynia, which is perceived pain caused by a normally non‐painful stimulus. T2DM is also associated with exaggerated pressor and cardioaccelerator responses to skeletal muscle contraction, namely the exercise pressor reflex. The mechanisms by which diabetes elicits these effects are not fully understood; however, both mechanical allodynia and the exercise pressor reflex are evoked by thin‐fiber afferent activity. The purpose of this study was to utilize noninvasive techniques to monitor the development and progression of diabetes‐induced neuropathy in thin‐fiber afferents. We used male UC Davis T2DM (n=9) rats from the age of 4 to 7 months to model the progression of T2DM. We took monthly measurements of blood glucose, HbA1c, and body weight. We also measured changes in mechanical sensitivity using von Frey filaments. Filaments were applied to the L5 dermatome on the plantar side of the left and right hind paws, between the footpads; the lowest force eliciting an abrupt paw withdrawal was measured and averaged between paws. We found that blood glucose (4mo: 255±28 mg/dl, 5mo: 366±50 mg/dl, 6mo: 417±58 mg/dl, 7mo: 453±49 mg/dl) and HbA1c (4mo: 5.7±0.2%, 5mo: 6.6±0.4%, 6mo: 9.3±1.2%, 7mo: 10.3±1.2%) were above the diabetic threshold (300 mg/dl, 6.5%) at 5, 6, and 7 months of age; they were significantly higher at 6 and 7 months, compared to 4 months, p<0.05. Body weight was significantly higher at 5, 6, and 7 months compared to 4 months (4mo: 525±4g, 5mo: 609±3g, 6mo: 620±21g, 7mo: 610±31g; p<0.05). Sensitivity to von Frey filaments was significantly greater, indicating mechanical allodynia, at 6 and 7 months compared to 4 months (4mo: 29±3.2 g, 5mo: 26.3±4.5 g, 6mo: 14.1±3.3 g, 7mo: 9.7±1.7 g; p<0.05). We conclude that mechanical allodynia progresses in severity as the rats develop and progress with T2DM. Moreover, these observations may provide insight to diabetes‐induced changes in thin‐fiber dependent reflexes, such as the exercise pressor reflex.

Support or Funding Information

N/A

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