bluuebunny/crossref_metadata_2025 huggingface dataset:
Gintonin Suppresses Fat Accumulation in 3T3‐L1 cell
Authors: Oh Jae-Hoon , Lee Kippeum , Jin Heegu , Chei Sungwoo , Oh Hyun-Ji , Lee Boo-Yong
Published: April 2020

Obesity is caused by an imbalance between energy intake and energy expenditure. Furthermore, obesity is closely relational to many metabolic diseases such as type 2 diabetes and hypertension. Excess fat is stored in adipose tissue, which contains of fat cells, or adipocytes, which have an capacity for saving surplus energy in the form of lipid. In our sturdy, Anti‐obesity effect of gintonin was investigated in 3T3‐L1. 3T3‐L1 is adipocytes which play critical roles in health and disease as a master regulator of energy homeostasis and metabolic health. Gintonin is practically a glycolipoprotein component of ginseng. Gintonin consists of large amounts of amino acids, the principal being phenylalanine. In the components of carbohydrates of gintonin, glucose is the major component. Ginseng in a broader classification consists of saponin fraction also called as ginsenosides and nonsaponin fractions. Gintonin was found to be different from ginsenosides classified with non‐saponin fractions. According to our that gintonin decreased the expression of adipogenesis proteins such as C/EBP‐α, and fatty acid binding protein‐4 (aP2). Gintonin substantially increased the expression of lipolytic enzymes such as ATGL and p‐HSL. Gintonin decreases adipogenesis and promote lipolysis in 3T3‐L1. In conclusion, we suggest that gintonin might have the protential to beneficially suppress body fat accumulation in the obesity.
Transcranial Direct Current Stimulation Transiently Increases Solute Transport in Brain Tissue
Authors: Fu Bingmei , Xia Yifan , Khalid Wasem , Huang Guangyao , Yin Zhaokai , Bikson Marom
Published: April 2020

Introduction

Transcranial direct current stimulation (tDCS) is a non‐invasive electrical stimulation investigated to treat a broad range of brain disorders and to enhance memory and cognition in healthy individuals. Our prior study has shown that tDCS with proper dosage can achieve non‐invasive and temporary increase in the blood‐brain barrier (BBB) permeability (P). To further investigate whether tDCS also increases the material transport in brain tissue, we employed high resolution multiphoton microscopy to collect the spreading images of fluorescently‐labeled solutes in the rat brain tissue after tDCS treatment. The effective solute diffusion coefficient in the brain tissue (Deff ) was determined by using our previously developed method. Deff is a quantitative indicator for solute transport in a medium.

Methods

After 20 min 1 mA tDCS treatment, sodium fluorescein (MW 376), or FITC‐dextran 70k in 1% BSA mammalian Ringer was injected into the rat (SD, 250–300g) cerebral circulation via the ipsilateral carotid artery by a syringe pump at a constant rate of ~3 ml/min. Simultaneously, the 3‐D images of a post‐capillary vessel and its surrounding area in the rat brain tissue 100–200 mm below the pia mater were collected by laser scanning multiphoton microscopy with 820 nm excitation wavelength. The cerebral microvessel permeability (P) and the effective solute diffusion coefficient in the brain tissue Deff were determined from the rate of dye spreading images. Specifically, Deff was estimated by curve fitting the spatio‐temporal solute concentration (fluorescence intensity) distribution by using an unsteady diffusion transport model.

Results and Discussion

It was found that in 5–10 min post 20 min–1mA tDCS, Deff/Dfree increases from 0.45 to 0.50 for sodium fluorescein (n=5) and from 0.11 to 0.25 for dextran 70k (n=6), all the increased Deff returned to the control level in 25–30 min post tDCS. tDCS indeed transiently makes the brain tissue less restrictive to solute transport, especially for larger solutes.

Conclusions

tDCS not only transiently increases the BBB permeability but also enhances the solute transport in brain tissue.

Support or Funding Information

Supported by NIH RO1 NS101362‐01
The Role of the Gut‐Brain Axis in the Capacity for Learning in Zebrafish
Authors: Deutsch Colin J , Uno Jennifer K
Published: April 2020

Starved (low calorie) and overfed (high calorie) diets are common in food insecure areas and can shift gut microbiota diversity. While there is increasing evidence linking the gut microbiota composition to psychiatric disorders including anxiety and depression, little is known regarding the impact gut microbes have on learning behavior and capacity. Current literature supports that alterations in intestinal microbiota diversity and richness can influence serotonergic and dopaminergic neurotransmission. The aim of this study is to examine the influence of gut‐brain axis on learning capacity. We hypothesize that a decrease in microbial diversity and abundance will negatively impact learning in zebrafish. Fish were grouped and fed a control diet, a starved low caloric diet and an overfed high caloric diet. Afterwards microbial DNA and RNA from zebrafish brain tissue was isolated. Bacterial abundance was quantified with primers designed against 16s ribosomal RNA. Dopamine and serotonin receptor expression was examined using TaqMan primer probes. Overfed fish exhibited a significant increase weight relative to starved fish, in addition starved fish displayed a 7.0 mg decrease in weight (p<.001, n=5). Both overfed and starved fish exhibited a decrease in intestinal microbial abundance. Associative learning was assessed using a plus‐maze protocol. Interestingly we observed modest, though not significant increases in learning in the starved fish relative to overfed and control fish. Coinciding with this data, overfed fish showed no significant changes in serotonin or dopamine receptor gene expression compared to control diets, however starved diets illustrated an increasing trend in serotonin receptor expression and a significant increase in dopamine D1 receptor expression (p<.04, n=11). These results indicate that starved and overfed diets decrease gut microbial abundance and may influence learning capacity through the gut‐brain axis in starved fish.

Support or Funding Information

Elon University
Collaborative Work Mediated by the Use of Google Drive as a Teaching Strategy Learning in Subjects of Research in Students of the Health Area
Authors: Arroyo Ana Gabriela Campos , Matías Flora María Cabrera , Escobar Omar David Orlanzzini
Published: April 2020

Scientific research in nutrition is essential to improve the quality of life of populations, identify health problems, propose health strategies and prevention plans for the most prevalent diseases. So, it is necessary to encourage students to interest in subjects related to research using innovative teaching and learning strategies that favor individual and collaborative learning.

Objective

Identify how the use of Google Drive contributes as a teaching‐learning strategy in the subject of research and development of degree projects in students in the area of health.

Materials and methods

The approach is mixed, concurrent and exploratory. 117 students from the Faculty of Nutrition Sciences of the University of Morelia were considered. At the beginning of the course, all students were asked to create a Gmail account in order to have access to all Google Drive tools. Once this was done, they were explained in face‐to‐face classes how to access and what elements the Google Drive platform has, individual folders with last name and first name were created and during the whole semester the students worked personally in their folders, developing activities in a way individual or in teams according to the programmed topic. One of the main activities carried out was the development of its own research protocol. Once the semester was completed, a questionnaire was applied through Google Forms, which consisted of two sections. The first section with identification questions and the second section with 15 closed and open questions to identify how the use of Google Drive. Contributes as a teaching‐learning strategy in research subjects.

Results

97.9% of the students considered that Google Drive is a good teaching‐learning strategy, 95.7% indicated that it helped them learn and facilitated the realization of their research projects, and 97.9% indicated that it helped them to work on collaborative form.

Conclusion

The use of Google Drive can be a teaching‐learning strategy that contributes to working in a collaborative, real‐time, comfortable, dynamic way and that provides a new way in which the teacher and the student can keep a personalized follow‐up in each of the research projects that are carried out.

Support or Funding Information

Research supported and endorsed by the Santander University and University of Morelia.
G Protein Gamma Subunits: Phosphorylation‐Mediated Governors of G protein Signaling in Humans?
Authors: Toosi Zahra Nassiri , Choudhury Ajanta , Su Xinya , Torres Matthew P.
Published: April 2020

Intracellular heterotrimeric G proteins (consisting of Gα, Gβ, and Gγ subunits), interface with 7‐transmembrane G protein coupled receptors (GPCRs) and function as the primary transducers of extracellular signals such as hormones, neurotransmitters, as well as pharmaceutical compounds. Upon binding their cognate ligands, GPCRs promote the dissociation of Gα and Gβγ subunits, which are then free to interact with effector proteins that promote intracellular signaling responses to the stimulus. Within this context, Gγ subunits are thought of primarily as membrane anchors for Gβ subunits, which must reside at the plasma membrane to facilitate functional protein interactions. However, emerging evidence suggests that Gγ subunits serve as phosphorylation‐dependent governors of G protein signaling. We have recently demonstrated this in the budding yeast model system, in which negative feedback phosphorylation of the intrinsically disordered N‐terminal tail of Gγ/Ste18 disrupts Gβγ/effector interactions and inhibits the normal GPCR‐dependent activation of MAPKs (Choudhury S. et al, Cell Rep. 2018). More recently, we have discovered that the yeast Gγ subunit undergoes combinatorial phosphorylation in response to several different forms of extracellular stimuli, including osmotic stress, pH stress, glucose deprivation, and cell cycle progression, in addition to GPCR activation. In light of these discoveries, we have hypothesized that Gγ phosphorylation plays a similar role in mammalian GPCR pathways. Indeed, we show that, like Gγ/Ste18, all mammalian Gγ subunits have an intrinsically disordered N‐terminus and most also undergo multi‐site phosphorylation. Here we provide new evidence to evaluate the hypothesis that Gγ phosphorylation is a governor of G protein signaling in mammals by demonstrating that phosphorylation is readily detectable on many different human Gγ subunits (Gγ3, 4, 5, 7, 10, 12) and by systematically testing the requirement for phosphorylation in proper Gβγ‐specific signaling pathways including the activation of PKD1 kinase by phospholipase C β2/3.

Support or Funding Information

NIH
Anti‐photoaging Activity of Ginseng Seed in UVB‐irradiated Human Skin Fibroblasts
Authors: Heo Huijin , Sung Jeehye , Kim Younghwa , Lee Junsoo
Published: April 2020

Ultraviolet (UV) radiation from the sun is the most common environmental stressor to damage the skin. Excessive UV radiation usually causes photoaging, inflammation, and even skin carcinogenesis. The seed of Korean ginseng (Panax ginseng C.A. Meyer) contains various phytochemicals including phytosterols, fatty acids, and ginsenosides. Ginseng seed is known to exhibit antioxidant, hepatoprotective, and melanin inhibitory activities. However, not much is known about the anti‐photoaging activities of ginseng seed. The aim of this study was to investigate the protective effect of ginseng seed embryo (GSE) and ginseng seed coat (GSC) on UVB‐induced photoaging in human fibroblasts cell line (Hs68 cells). The contents of total polyphenols and total flavonoids were higher in GSC compared to GSE. Linoleic acid was the major fatty acid in GSE (57.69%) and GSC (39.95%). The content of ginsenoside Rh4 of GSC was about 13 times higher than that of GSE. GSE and GSC significantly attenuated UVB‐induced expression of matrix metalloproteinases (MMPs) and increased the production of collagen in Hs68 cells. Furthermore, UVB‐induced ROS generation was markedly attenuated by pretreatment with GSE and GSC in a dose‐dependent manner, demonstrating that ginseng seed is to be a potent ROS scavenger. These results indicate that ginseng seed protects human skin fibroblasts from UVB‐induced photoaging and therefore has the potential for use in cosmeceutical preparations.
Alteration of KCa Channel Function by Homocysteine in Human Internal Mammary Artery
Authors: Sun Wen-Tao , Xue Hong-Mei , Hou Hai-Tao , He Guo-Wei , Yang Qin
Published: April 2020

Objectives

Hyperhomocysteinemia is an independent risk factor for cardiovascular disease. We previously in porcine coronary arteries demonstrated that disruption of KCa channel activity is involved in homocysteine‐induced impairment of dilatory function. Whether homocysteine affects KCa channels in human arteries remains uninvestigated. In the present study, we aimed to investigate the effect of homocysteine on the expression and functionality of the members of KCa channel family, including large‐ (BKCa), intermediate‐ (IKCa) and small‐ (SKCa) conductance KCa channels in human internal mammary artery (IMA).

Methods

Residual IMA segments collected from patients undergoing coronary artery bypass grafting (CABG) were used for myography study for KCa subtype‐mediated relaxation and contraction. Western blot and immunohistochemistry were employed to detect the expression and distribution of KCa subtypes in IMA.

Results

BKCa subtype in the KCa channel family significantly regulated the vasoreactivity of IMA. Blockade of BKCa channels inhibited acetylcholine‐induced relaxation and enhanced U46619‐induced contraction. In comparison, both IKCa and SKCa subtypes played a minor role in the control of relaxation and contraction of IMA. Exposure to homocysteine compromised the vasodilating activity of the BKCa subtype in IMA, evidenced by the suppressed relaxant response to the BKCa channel opener and the decreased impact of the BKCa channel blocker on acetylcholine‐induced relaxation and U46619‐evoked contraction. The functional loss of BKCa was associated with a downregulation of the protein expression of BKCa β1 subunits in the smooth muscle of IMA. Inhibition of endoplasmic reticulum (ER) stress showed protective effect against homocysteine on the function and expression of BKCa channels. In contrast, the role of IKCa and SKCa subtypes in mediating endothelium‐dependent relaxation was augmented by homocysteine though no alterations in the regulatory capacity on vasoconstriction were observed. Homocystine did not alter the protein expression level of IKCa and SKCa subtypes.

Conclusions

Homocysteine compromises the vasodilating activity of BKCa channels in IMA, which may be attributed to the loss of BKCa β1 subunits mediated by ER stress. Unlike BKCa, IKCa and SKCa subtypes are unessential for vasoregulation in IMA, the loss of BKCa functionality in hyperhomocysteinemia enhances the role of IKCa and SKCa in mediating endothelial dilator function. Targeting BKCa may form a strategy to improve the postoperative graft performance in CABG patients with hyperhomocysteinemia who receives IMA grafting.

Support or Funding Information

Supported by NSFC 81870227 & 81870288, Tianjin Science and Technology Commission (18PTZWHZ00060) & Binhai New Area Health Bureau (2018BWKZ005), and Non‐profit Central Research Institute Fund of Chinese Academy of Medical Sciences 2018TX31002 & 2019XK310001.
Protective Effect of Persimmon Calyx Extract against UVB‐induced Skin Photoaging in Human Skin Fibroblasts
Authors: Jeong Taekyoung , Park Jeong Hwan , Hong Seonghwa , Lee Junsoo
Published: April 2020

The persimmon is the edible fruit, which has been widely cultivated in the countries of East Asia. The persimmon calyx is a by‐product generated during persimmon processing. It has been reported that persimmon calyx possesses anti‐oxidative, anti‐inflammatory, and anti‐coagulant properties. However, few reports have been published on the protective activities of persimmon calyx on photoaging in skin fibroblast cells. Therefore, the purpose of this study was to evaluate the protective activities of persimmon calyx methanol extract (PCME) on UVB‐induced photoaging in human skin fibroblasts (Hs68 cells). The cells were treated with PCME at concentrations ranging from 5 to 20 □g/mL for 24 h and then the cells were exposed to UVB (30 mJ/cm2) irradiation. The results revealed that PCME showed cytoprotective effects in a dose‐dependent manner. PCME also significantly inhibited the release of matrix metalloproteinase 1 and 3. Furthermore, PCME reduced UVB‐induced production of reactive oxygen species and collagen degradation in Hs68 cells. Taken together, our data suggest that PCME has strong protective activity against UVB‐induced skin photoaging and thus can be a useful agent for anti‐photoaging cosmetic ingredient.
Role of Nrf2 in Regulation of Expression of CYP 2C11 and CYP 4X1 Epoxygenases and Production of Epoxyeicosatrienoic Acids (EETs) in the Rat Brain
Authors: Gebremedhin Debebe , Guerts Aron , Lombard Julian H.
Published: April 2020

Nuclear factor (erythroid‐derived 2)‐like‐2 (Nrf2) is the master antioxidant and cell‐protective transcription factor native in the rat brain. The CYP epoxygenases 2C11 and 4X1 are expressed in the brain and metabolize arachidonic acid to epoxyeicosatrienoic acids (EETs) that are known to induce cerebral vasodilation and increase nutritive blood flow to areas of heightened neuronal activity and also provide antioxidant and anti‐inflammatory actions. The influence of Nrf2 gene deletion on CYP 2C11 and 4X1 epoxygenase expression and activity in the rat brain is not known. In this study we used isoform specific antibodies and Western blotting to determine the effect of Nrf2 gene knockout (KO) on the expression of proteins for CYP 2C11 and CYP 4X1 epoxygenases in whole brain and isolated cerebral arteries. We also measured the production of epoxyeicosatrienoic acids (EETs) in homogenates of whole brain and isolated cerebral arteries of Nrf2 wild type (WT) and Nrf2 KO rats. CYP epoxygenase expression and EETs production were reduced in whole brain and arteries of the NRF2 KO rats compared to WT controls. These findings indicate that knockout of the Nrf2 gene leads to a marked reduction in the expression of proteins for CYP 2C11 and CYP 4X1 epoxygenases and in the production of 8, 9‐, 11,12‐ and 14,15‐EETs compared to those measured in the Nrf2 WT rat brain issues and cerebral arteries. The results of the current study suggest that CYP 2C11 and 4X1 epoxygenases may be components of the Nrf2‐induced phase 2 genes that are known to limit oxidative stress and provide cellular defense in the brain. We postulate that Nrf2 may regulate functions of the cerebral circulation and provide protection against oxidative stress, in part, via modulation of the expression level of CYP 2C11 and 4X1 epoxygenase proteins and the production of EETs.

Support or Funding Information

NIH R01 #HL‐128242 and NIH #R21‐OD018309
Estrogen Replacement Improves Mesenteric Arterial Relaxation in UC Davis Pre‐diabetic Rats: Possible Involvement of Estrogen Receptor Beta
Authors: Razan Md Rahatullah , Akther Farjana , Karimi Habibullah A. , Amissi Said , Graham James L. , Stanhope Kimber L. , Havel Peter J. , Rahimian Roshanak
Published: April 2020

The risk of cardiovascular diseases (CVD) is lower in premenopausal women compared to men, and this lower risk is related to the presumptive cardioprotective effects of female sex hormones, estrogen in particular. However, premenopausal women may lose this protection with the onset of menopause and/or when they become diabetic. Little is known about the vascular function in presence or absence of estrogen at the early stage of diabetes.

The objective of this study was to investigate the effects of 17β‐estradiol (E2) replacement on mesenteric arterial (MA) function in ovariectomized (ovex) UC Davis type 2 diabetes mellitus (UCD‐T2DM) rats at the pre‐diabetic stage. MA were obtained from four groups of ovex rats implanted with a subcutaneous pellet for 45 days: (1) Control (Ctrl), ovex + placebo, (2) Ctrl, ovex + E2 (1.5mg/pellet), (3) pre‐diabetic, ovex + placebo, (4) pre‐diabetic, ovex + E2. Endothelium‐dependent vasodilation (EDV) to acetylcholine (ACh, 10−8 to 10−5 M) in MA were measured before and after indomethacin (Indo, 10μM), a cyclooxygenase (COX) inhibitor; followed by addition of L‐NAME (200μM), a nitric oxide synthase (NOS) inhibitor. The MA expression of estrogen receptors (ERs) and plasma E2 concentration were determined by Western blotting and ELISA assay, respectively.

E2 treatment significantly increased plasma estradiol concentration in both Ctrl and Prediabetic groups. Analysis of ERs showed a higher expression of ER‐beta only in E2‐treated prediabetic group. ACh responses were similar in Ctrl groups, regardless of E2 replacement. ACh induced relaxation was, however, markedly reduced in MA of placebo treated pre‐diabetic group compared with other groups. Treatment of pre‐diabetic with E2 enhanced relaxation to the same level as those from Ctrl rats. Blocking of both COX and NOS completely abolished the relaxation responses to ACh in pre‐diabetic groups, regardless of E2 replacement, indicating no contribution of EDHF‐type relaxation in these groups. However, the added effect of L‐NAME, in presence of Indo, in blocking ACh‐mediated vasodilation was much more prominent in E2‐treated prediabetic group when compared with placebo treated group, indicating an enhanced role of NO. On the other hand, both NO and EDHF remained important for EDV in Ctrl groups. Addition of L‐NAME, however, caused a much smaller reduction of EDV in E2 treated than in placebo treated arteries, suggesting EDHF is the predominant mediator of mesenteric vasorelaxation in this groups.

Here, we show that EDV is impaired at pre‐diabetic stage, may be due to loss of EDHF. E2 treatment restores EDV in MA of pre‐diabetic group, possibly via elevated contribution of NO‐mediated relaxation by increased E2 signaling (via ER beta) in this group. The role of ER alpha remains to be investigated.

Support or Funding Information

Supported by NHLBI.
Exercise training to increase stress?
Authors: Valencia Miles , Bowens Jayda , Azhand Bilal , Monroy Jenna , Horner Angela
Published: April 2020

Tendons are series elastic structures that connect bone to muscle and transmit energy by absorbing, transferring, and dissipating energy. Remodeling can be induced from overuse, disuse, or aging, and as a result may change the tendon’s stiffness over an individual’s lifetime. The rate and degree of intrinsic changes in tendons are not well understood. In this study, we investigate the effects of different exercise types on tendon remodeling. We measured tendon gross morphology and mechanics in mice (Mus musculus) using exercise regimens with varying mechanical loading: voluntary wheel running, treadmill running, and repeated jumping. After 4 weeks of training, tendons were harvested for mechanical testing. When individuals were pooled across exercise treatments, there was a significant decrease in stiffness with increasing cross‐sectional area (CSA). Surprisingly, although jump mice had the smallest CSA of the three training regimens, they exhibited the greatest failure stress and stiffness. Treadmill‐training mice had the largest CSA but also the lowest failure stress and stiffness. These data suggest that exercise contributes to physical remodeling of tendon in mice, but the mechanical properties are not correlated strictly with CSA. Our future work will investigate the role of remodeling on intrinsic tendon structure and material properties.
Efficacy Assessment and Mechanism Exploration of Kruppel‐Like Factor 2‐Targeting Intervention Strategy for Non‐Small Cell Lung Cancer Therapy
Authors: Wu Chieh-Hsi , Pan Chun-Hsu , Wang Jieyu
Published: April 2020

Non‐small cell lung cancer (NSCLC), the most common type of lung cancer, has the problems such as less sensitivity to chemotherapy and resistance to tyrosine kinase inhibitors. Kruppel‐like factor 2 (KLF2) has been reported to have the potential activities in tumor suppression and immune regulation. The expression level of KLF2 is lower in NSCLC and negatively correlated with the stages of tumor progression and lymph node metastasis. To date, not only no KLF2‐promoting compounds have been developed, but no related studies on its anti‐tumor efficacy in vivo have been verified. Therefore, the purposes of this study were focused on the potential and related mechanisms of KLF2 applied as a targeted molecule for NSCLC therapy. Our data indicated that the intervention of lovastatin markedly up‐regulated protein expression of KLF2 in A549 human non‐small cell lung cancer cells. Moreover, KLF2 siRNA intervention could significant inhibit serum‐induced cell growth in A549 cells. The analysis result of the Ingenuity pathway analysis (IPA) showed that several intracellular molecules, such as AKT, mTOR and FOXO1, maybe the potential upstream regulators to involve in KLF2 expression. Besides, the experimental data also evidenced that lovastatin can suppress activations of AKT and mTOR proteins as well as to reduce protein expression of FOXO1. In summary, these results suggested that intensifying KLF2 expression could apply as a feasible therapeutic strategy to improve NSCLC therapy, and statinoid compounds have potential to be developed as KLF2 activator. In addition, activations or expressions of AKT‐mTOR pathway and FOXO1 may be attributed to suppression of KLF2 in NSCLC cells, such as A549 cells.
Prenatal chronic intermittent nicotine aerosol exposure programming a sex dependent hypertensive phenotype via vascular eNOS uncoupling
Authors: Liu Bailin , Liang Xia , Jie Jian , Li Yong , Dasgupta Chiranjib , Huang Xiaohui , Zhang Lubo , Shao Xuesi M. , Xiao DaLiao
Published: April 2020

Objective

Maternal cigarette smoking/nicotine use during pregnancy is associated with an increased risk of cardiovascular diseases and development of hypertensive phenotype in later life. However, the involved molecular pathway is remain unclear. The present study tested the hypothesis that cigarette smoking‐relevant nicotine inhalation during pregnancy causes a gender‐dependent change the balance of resistance vasculatures eNOS and prooxidant functions in adult offspring, leading to development of hypertensive phenotype in adult male offspring.

Methods

The time‐dated pregnant rats were exposed to chronic intermittent nicotine aerosol (CINA) via a novel lung alveolar region‐targeted aerosol method during pregnancy (E4 to E20). Experiments were performed in adult (~6 month‐old) offspring.

Results

CINA exposure enhanced vascular H2O2 production which was inhibited by eNOS inhibitor, L‐NAME in male offspring. CINA exposure selectively enhanced vascular NOX2 expression. CINA exposure enhanced angiotensin II (Ang II)‐induced BP response in male offspring, which was not affected by NOX2 inhibitor (gp91 ds‐tat) but significantly reversed by L‐NAME or BH4 treatment in vivo. In pressure myography experiments. CINA exposure had no effect on pressure‐induced myogenic tone but decreased the Acetylcholine (Ach)‐induced relaxation in adult male but not female offspring. In the presence of L‐NAME or H2O2 scavenger (PEG‐catalase), there were no differences of Ach‐induced relaxation between CINA and saline control offspring. Furthermore, pretreatment of eNOS cofactor BH4 enhanced Ach‐induced relaxation in CINA‐exposed male offspring, while no effect in saline control group. In addition, exogenous NO donor (SNP)‐induced mesenteric artery relaxations were similar between CINA exposed and saline control male offspring.

Conclusions

These findings provide novel evidence that maternal inhaled nicotine exposure could impair fetal vascular development and induce a gender‐dependent development of endothelial dysfunction phenotype in adult male offspring via enhancing vascular eNOS uncoupling. The uncoupled eNOS‐mediated ROS plays a predominant role in CINA‐induced hypertensive response in male offspring

Support or Funding Information

Supported in part by the NIH grants: R01HL135623, R03DA041492, and R01HD088039; TRDRP grants: 29IR0437 and T30FT0936
Combination of dapagliflozin and statins attenuate renal lipotoxicity in high‐fat high‐fructose diet‐induced insulin resistant rats
Authors: Thongnak Laongdao , Pongchaidecha Anchalee , Chatsudthipong Varanuj , Lungkaphin Anusorn
Published: April 2020

High‐fat high‐fructose diet (HFF) can induce dyslipidemia and lipid accumulation in the kidney which are related to insulin resistance and metabolic syndrome. Dyslipidemia and lipid accumulation in obesity can lead to lipotoxicity and cellular damage particularly kidney injury by activating oxidative stress, fibrotic and apoptotic pathways. This study investigated whether dapagliflozin alone or combined with atorvastatin could improve metabolic parameters, lipid metabolism and lipid accumulation induced kidney injury in HFF‐induced insulin resistant rats. Male Wistar rats were fed with high‐fat diet with 10% fructose in drinking water for 16 weeks. After that, these rats were received drug treatments for 4 weeks; vehicle (HFF), SGLT2 inhibitor dapagliflozin (HFFD), atorvastatin (HFFA) and SGLT2 inhibitor combined with atorvastatin (HFFDA) groups. HFF rats demonstrated insulin resistance, dyslipidemia, and renal dysfunction associated with impaired renal lipid metabolism and lipid accumulation. Lipid accumulation in the kidney could promote renal dysfunction in HFF rats via the activation of oxidative stress, fibrosis and apoptosis. Dapagliflozin treatment could improve insulin resistance, lipid synthesis and lipotoxicity‐related renal oxidative stress, fibrosis and apoptosis leading to reversal of kidney dysfunction in HFF model. The combined treatment (HFFDA group) could also improve metabolic parameters which was not different from single drug treatment (HFFD or HFFA group). Notably, the reduced lipid accumulation in kidney which were linked to the improvement in lipid oxidation was better in the combined treatment group when compared to a single drug treatment. In summary, dapagliflozin combined with atorvastatin treatment exert the beneficial effects on metabolic parameters, lipid metabolism, and lipid accumulation induced kidney injury via the attenuation of oxidative stress, fibrosis and apoptosis in high‐fat high‐fructose diet‐induced insulin resistant model.

Support or Funding Information

Thailand Research Fund (RSA6080015 AL); the Royal Golden Jubilee PhD program (PhD/0063/2560 LT and AL); the Faculty of Medicine Research Fund, Chiang Mai University (AL and AP); the Graduate Research Scholarship Chiang Mai University (LT); and the Functional Food Research Center for Well‐being, Chiang Mai University (AL).
Structural and Molecular Characteristics of Axons in the Long Head of the Biceps Tendon
Authors: Streicher Johannes , Boesmueller Sandra , Gesslbauer Bernhard , Hirtler Lena , Bormann Daniel , Pastor Angel M. , Mittermayr Rainer , Blumer Roland
Published: April 2020

Objectives

Although the long head of the biceps tendon (LHBT) is a major source of shoulder pain, the innervation is not sufficiently documented. Thus, the present study aimed to characterize structural and molecular features of nervous elements in the LHBT.

Methods

The proximal part of eleven LHBTs was harvested intraoperatively. There were 8 female and 3 male specimens. Age ranged from 66 to 86 years. For structural analyses, nervous elements were viewed in the transmission electron microscope. For molecular characterization, we used a series of neuronal markers: This included general neuronal markers [antibodies against neurofilament and protein gene product 9.5 (PGP9.5)] as well as specific neuronal markers[antibodies against myelin basic protein (MBP), calcitonin gene related product (CGRP), substance P (SP), tyrosine hydroxylase (TH), and growth associated protein 43 (GAP43)]. Anti‐neurofilament and anti PGP9.5 visualized the overall innervation. Anti‐MBP visualized myelination, anti‐CGRP and anti‐SP nociceptive fibers, anti‐TH sympathetic nerve fibers, and anti‐GAP43 nerve fibers during development and regeneration. Immunolabelled sections were analyzed in the confocal laser scanning microscope.

All procedures performed in this study were approved by the local medical ethics committee (Austrian workers compensation boards’ research ethics committee and institutional review board of the Medical University of Vienna, No. 06/2017) and in accordance with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all persons included in this study.

Results

We show that the LHBT contains unmyelinated as well as myelinated nerve fibers which group in nerve fascicles and follow blood vessels. Many myelinated and unmyelinated axons exhibit molecular features of nociceptive nerve fibers. Another subpopulation of unmyelinated axons exhibits molecular characteristics of sympathetic nerve fibers. Unmyelinated sympathetic fibers and unmyelinated nociceptive fibers express proteins that are found during development and regeneration.

Conclusion

Present findings show that the LHBT of aged persons is richly innervated. The occurrence of growth protein GAP43 in nociceptive nerve fibers supports the hypothesis that in‐growth of nociceptive fibers is the source of chronic tendon pain.
The Cholesterol Synthesis Enzyme Lanosterol 14α‐Demethylase is Post‐Translationally Regulated by the E3 Ubiquitin Ligase MARCH6
Authors: Scott Nicola A. , Sharpe Laura J. , Capell-Hattam Isabelle M. , Gullo Samuel J. , Luu Winnie , Brown Andrew J.
Published: April 2020

Cholesterol synthesis is a complex process with over twenty enzymes involved. Each enzyme within the pathway undergoes distinct regulation to maintain cholesterol homeostasis. Typically, enzymes within the pathway are degraded in response to a high sterol status, but some are highly stable. In this study, we demonstrate that an enzyme at the branch point in the pathway, lanosterol 14α‐demethylase (LDM), undergoes unique regulation. We show that the preceding enzyme, lanosterol synthase (LSS) is stable, whilst LDM is rapidly degraded. Surprisingly, this degradation is not triggered by sterols but by nitric oxide. Here we identify that the important E3 ubiquitin ligase, MARCH6, is responsible for facilitating the ubiquitination and proteasomal degradation of LDM and also the terminal cholesterol synthesis enzyme, 24‐dehydrocholesterol reductase (DHCR24). Our work highlights MARCH6 as the first example of an E3 ubiquitin ligase which targets multiple steps in a single biochemical pathway and indicates new facets in the control of cholesterol synthesis.

Support or Funding Information

This work was supported by the Australian Research Council Grant DP170101178. The PhD students were funded by the Research Training Program from the Australian Government Department of Education.
Polyhexamethyleneguanidine phosphate‐induced cytotoxicity is mediated by endoplasmic reticulum stress
Authors: Jung Young-Suk , Kim Sou Hyun , Lee Seunghyun , Son Seung Won
Published: April 2020

Polyhexamethyleneguanidine phosphate (PHMG‐P) is a widely used polymeric antimicrobial agent to induce significant pulmonary toxicity. Several studies have reported that the liver also can be a target organ of PHMG toxicity, but the exact effect of this compound on liver cells is not well understood. To identify the mechanism of PHMG hepatotoxicity, HepG2 cells were exposed to PHMG‐P for 72 h. Cell viability was significantly decreased by PHMG‐P in a time‐and concentration‐dependent manner. Mitochondrial membrane potential was markedly reduced by PHMG‐P and the apoptotic signaling cascade was activated. The increases observed in CHOP, p‐IRE, and p‐JNK levels in PHMG‐P‐treated cells indicated the induction of ER stress. To verify the role of ER stress in PHMG‐P‐induced cytotoxicity, HepG2 cells were pretreated with the chemical chaperone, TUDCA and then co‐treated with TUDCA and PHMG‐P for 24 h. Interestingly, TUDCA inhibited PHMG‐P‐induced ER stress and cytotoxicity in a dose‐dependent manner. Apoptotic cell death and mitochondrial depolarization were also prevented by TUDCA. Proteins involved in the apoptotic pathway were all normalized to their control levels in TUDCA‐treated cells. In conclusion, the results suggest that PHMG‐P induced significant cytotoxicity in liver cells and ER stress‐mediated apoptosis may be an important mechanism mediating this hepatotoxicity.

Support or Funding Information

This study was funded by Nano‐Material Technology Development Program through the National Research Foundation of Korea (NRF) funded by Ministry of Science and ICT (NRF‐2018M3A7B4071233).
Effects of FPMINT derivative on Human Equilibrative Nucleoside Transporters
Authors: Mak Wing Shum Winston , Leung George P.H.
Published: April 2020

Equilibrative nucleoside transporters (ENTs) are the proteins responsible for transporting nucleosides across cell membrane. The role of ENT 1 in cardioprotection has been reported but the biological functions of ENT 2 remains unclear. To study the functions of ENT 2, pharmacological approach using selective ENT 2 inhibitors is an option but such inhibitors are not available. Our laboratory has recently developed a compound called FPMINT, which is slightly more selective to ENT2 than ENT1 but the difference between its IC50 on ENT1 and ENT2 is not very distinctive. Therefore, modification of the structure of FPMINT to increase its selectivity to ENT 2 is required. In this study, the effects of a FPMINT derivative was examined. PK15 nucleoside transporter‐deficient cells transfected with ENT1 or ENT2 were used as the models. Cell viability and protein expression level were studied by MTT assay and Western blotting, respectively. The nucleoside transport was studied by measuring the [3H]uridine and [3H]adenosine uptake in the cells. The results showed that the viability of PK15NTD/ENT1 and PK15/NTD/ENT 2 cells were not affected after incubation with FPMINT derivative (0.5 μM to 50 μM) for 24 to 48 hr. The protein expression level of both ENT 1 and ENT 2 was also unchanged. The ENT1‐mediated [3H]uridine and [3H]adenosine transport were inhibited by FPMINT derivative in a concentration‐dependent manner with IC50 values of 2.458 μM and 7.113 μM, respectively, compared to the IC50 values of 0.5697 μM and 2.571 μM respectively in the case of ENT 2. In addition, FPMINT derivative reduced the Vmax of ENT1 and ENT2‐mediated [3H]uridine transport without significant change on the apparent Km. Furthermore, the inhibitory effects of FPMINT derivative on both ENT 1 and ENT 2 could not be washed away. In conclusion, our study has provided evidence that FPMINT derivative is an irreversible and non‐competitive inhibitor which is more selective to ENT2 than ENT1, when compared to its parent compound FPMINT.
Vitamin D Receptor Deficiency Impairs Pial Collateral Circulation in Mice
Authors: Pal Eva , Hricisak Laszlo , Lekai Agnes , Nagy Dorina , Fulop Agnes , Benyo Zoltan
Published: April 2020

Vitamin D deficiency is a global health problem, which can increase the risk of cerebrovascular diseases including ischaemic stroke. However, the pathophysiological mechanism underlying this association is not well understood yet. The goal of the present work was to examine the impacts of vitamin D deficiency on the mouse pial collateral circulation which might provide a compensatory pathway after unilateral common carotid artery occlusion (CAO). The morphology of the leptomeningeal collaterals and the distance of the anastomotic line (half distance between the nearest branching points of the anterior cerebral artery (ACA) and the middle cerebral artery (MCA)) from the midline were evaluated after staining of the cerebral vasculature in adult male mice carrying a functionally inactive vitamin D receptor (KO, n=6) and in their wild‐type littermates (WT, n=6). The acute cerebrocortical blood flow (CoBF) changes after CAO was measured in anesthetized WT (n=8) and KO (n=8) mice using laser speckle imaging to evaluate regional differences in the territories supplied by the ACA and MCA, i.e. the frontoparietal and temporal cortices, respectively. Vitamin D deficiency decreased the number of leptomeningeal collaterals between the ACA and MCA, whereas the tortuosity of the collaterals was enhanced. The anastomotic line measured at 4 mm caudally from the frontal pole was located cc. 400 μm closer to the midline in KO than in WT mice, therefore the territory supplied by the ACA decreased. The maximal CoBF reductions were similar between the two groups in the temporal cortex, whereas in the frontoparietal cortex more severe hypoperfusion was developed in the KO animals, as the blood flow in the ipsilateral frontoparietal region showed a decrease of 13% compared to the contralateral region, whereas this difference was only 6% in WT mice. In conclusion, vitamin D deficiency leads to impaired development of leptomeningeal collaterals, as well as enhanced CoBF reduction in the frontoparietal cortex after CAO. These morphological and functional alterations can potentially compromise the cerebral circulation, therefore it may increase the risk of cerebrovascular diseases and worsen the outcome of stroke.

Support or Funding Information

EFOP‐3.6.3‐VEKOP‐16‐2017‐00009, OTKA K‐112964, OTKA K‐125174, NVKP‐16‐1‐2016‐0042
Traumatic Brain Injury Alters PAQR7‐induced Vasorelaxation in Pressurized Cerebral Arteries
Authors: Goldsmith Jacob A. , Evanson Kirk W. , Levenson Cathy W.
Published: April 2020

Traumatic brain injury (TBI) induces acute changes in cerebrovascular function, including alterations to cerebral artery contractility, which ultimately increase the incidence of both ischemic and hemorrhagic events. Cerebrovascular tone is controlled by hormonal, neurogenic, endothelial, and myogenic responses. Progesterone, a steroid hormone, has been evaluated as a potential treatment to improve TBI outcome; however, progesterone has failed to translate to patient benefits in clinical trials, potentially due to its pleiotropic effects on multiple receptors. Progestin & adipoQ receptor 7 (PAQR7) is a plasma membrane progesterone receptor involved in the vasorelaxation of human blood vessels. However, little is known about the smooth muscle‐specific role of PAQR7 in resistance size cerebral arteries following TBI. Here, we examined PAQR7 function in pressurized endothelium‐denuded cerebral resistance size arteries 24 hours following moderately severe TBI (controlled cortical impact). Baseline myogenic tone of middle cerebral arteries (MCA) was attenuated following TBI. PAQR7‐induced vasorelaxation in pressurized MCA from TBI rats was attenuated in a concentration‐dependent manner as compared to sham controls (10−9M‐10−7M). Moreover, pre‐incubation with inhibitors of adenylyl cyclase (AC), PKA, or PKG attenuated PAQR7‐induced vasorelaxation in both groups. In conclusion, smooth muscle‐specific PAQR7 vasodilation involves an AC‐PKA/PKG pathway in cerebral resistance size arteries, and moderately severe TBI attenuates cerebral artery myogenic tone, which reduces overall arterial vasodilatory capacity.

Support or Funding Information

Supported by The Florida State University Graduate School Dissertation Award Grant.
GC‐MS based comparative metabolomic analysis of MCF7 and MDA‐MB‐231 cancer cells treated with Tamoxifen and/or Taxol
Authors: Semreen Mohammad , Alniss Hasan , Al-Awady Raafat , Mousa Muath , Almehdi Ahmed , Soares Nelson
Published: April 2020

Metabolomics of cancer cells is a powerful tool to understand the development, progression and treatment outcome of different types of cancer. Therefore, a highly sensitive, selective and reproducible platform that provides deep information about modifications of metabolism of cancer cells undergoing chemotherapy is required. In the present study, Tamoxifen (5μM) and/or Taxol (1μM) were added to MCF7 and MDA‐MB‐231 cancer cells to investigate the effect of these drugs on the metabolism of the treated cells. In the breast cancer cells (MCF7), of the 68 metabolites identified, 20 were differentially abundant in treated cells with treatment (p< 0.01). Subsequent enriched metabolic pathway analysis (MetaboAnalyst) revealed that tamoxifen had a significant impact in the steroid biosynthesis. Specifically, there was a 32‐fold increase of Lathosterol with tamoxifen, while Zymosterol had a 2.7‐fold decrease with tamoxifen and Desmosterol has decreased by 27‐fold under treatment with both drugs. Additionally, treatment with both drugs had an impact on amino acid pathways (Glycine, Phenylalanine, Threonine, Valine). On the other hand, 59 metabolites have been identified in MDA‐MB‐231 cells. Fourteen of them were differentially abundant in treated cells (p< 0.01). Unlike the MCF7, drug treatment had no effect in the aa pathway in MDA‐MB‐231 cells and seemed to mainly impact sugar metabolism. For example, treatment with tamoxifen lowered the levels of sugars like L‐Rhamnose, Glycerol, while Xylose decreased only upon treatment with combination of the two drugs. Our data showed that a set of metabolites was affected by the presence of either of the drugs, however there was a number of metabolites that responded to a specific drug treatment. This result suggests that a combination of tamoxifen and taxol may be considered as an alternative therapy against cancer cells.
Resistance Training with or without Blood Flow Restriction Improves Short‐Term Memory in Rats
Authors: Rebelo Macario Arosti , Garcia Nádia Fagundes , Puggina Enrico Fuini , de Moraes Camila
Published: April 2020

Low load resistance training with blood flow restriction is usually used for improvements in muscle strength in special care conditions. However, there is limited information regarding the effects of this exercise approach on memory function. The aim of this study was to investigate the effect of resistance training with or without blood flow restriction, carried out three times a week, on short‐term memory. Male Wistar Hannover rats (70 days old) were divided into four groups: Sedentary Sham (S/S, n=10), Trained Sham (T/S, n=10), Sedentary with blood flow restriction (S/BFR, n=11), Trained with blood flow restriction (T/BFR, n=9). Animals from the S/BFR and T/BFR had their right femoral artery flow restricted (blood arrival and venous return) about 36–38% and animals from sham groups were submitted to the same surgical procedures except blood restriction. Experimental protocol was approved by local ethic committee (protocol: 2016.5.80.90.4). Exercise training began two weeks after surgery procedures. The exercise sessions were carried out three times a week and consisted of climb a vertical ladder (110 cm high, 18 cm wide, 2 cm grid, 80° inclination), carrying a load (50% of 1 Maximum Repetition) for 6 sets of 10 repetitions with 1‐minute rest between sets. After four weeks of exercise training, memory performance was assessed using the Object Recognition Test and the discrimination index (dI) was calculated (dI= [(time spent exploring the novel object minus time spent exploring the familiar object)/ total exploration time]). The interval between testing sessions 1 and 2 was 30 minutes. Femoral blood flow restriction did not cause impairment on memory performance (S/SdI=0.33±0.08 vs S/BFRdI=0.37±0.03). On the other hand, exercise training improved memory performance in both groups (T/SdI=0.76±0.07 vs T/BFRdI=0.59±0.04). The resistance training, carried out 3 times a week, was effective to improve short‐term memory either with or without peripheral blood flow restriction.

Support or Funding Information

Supported by FAPESP Grant # 2017/13348‐6; Grant # 2018/16712‐3