bluuebunny/crossref_metadata_2025 huggingface dataset:
Novel Anti‐inflammatories: Effect of Amidation Inhibitors on Inflammatory Pathways in Macrophages
Authors: Lucrezi Jacob , Hiremath Thejas , May Sheldon W.
Published: June 2021

Chronic inflammation is responsible for numerous life‐threatening diseases. Substance P (SP), a pro‐inflammatory peptide, is synthesized by the post‐translational amidation of its glycine‐extended precursor (SP‐Gly). SP has been implicated in chronic inflammation along with inducible nitric oxide synthase (iNOS) and the cytokine TNF‐α. In primary mouse macrophages exogenous SP stimulates production of iNOS and cytokines, including TNF‐α, and enhances other stimulators such as the endotoxin LPS. Drugs designed to interfere with TNF‐α signaling have been shown to be an effective treatment for chronic inflammatory diseases. Therefore, elucidation of signaling pathways involving TNF‐α are of great importance to understanding and treating these diseases. We have previously shown that a prototypical amidation inhibitor, 4‐ phenyl‐3‐butenoic acid (PBA), and the ester of the highly potent amidation inhibitor, N‐acetyl‐(L)‐phenylalanylacrylic acid (NAPAA), are effective against chronic and acute rat models of inflammation, respectively. We report the effects of these amidation inhibitors on the activities of SP, SP‐Gly and LPS in inflammatory pathways in a commonly used macrophage‐like mouse cell line, RAW 264.7. We report that SP but not SP‐Gly stimulates TNF‐α production from RAW 264.7 macrophages, that PBA inhibits this SP stimulation and that NAPAA inhibits LPS stimulation of iNOS activity.
Unraveling the Molecular Mechanism behind Thrombin Induced Myofibrobast Transition
Authors: Meyer Jennifer Ashley , Strande Jennifer
Published: June 2021

TGF‐β1 is a mediator in fibrosis by activating its downstream Smad signaling pathway, where they are translocated to the nucleus. TGF‐ β has also been shown to inhibit thrombin signaling. Thrombin has been shown to act via the PAR1 receptor, a G‐protein coupled receptor. Activation of PAR1 by thrombin results in many profibrotic effects of thrombin via the induction and activation of secondary mediators including upregulation of TGF‐β and procollagen. Therefore, we hypothesize that thrombin promotes myofibroblast formation through a TGF‐ β independent/dependent pathway.

Cells treated with thrombin showed an increase in α‐SMA compared to control. Inhibition of all G‐protein coupled receptors resulted in a complete abolition of fluorescent intensity, indicating that the mechanism by which thrombin induces fibrosis is via G‐proteins, yet inhibition of individual G‐proteins did not have this effect, indicating a potential additive effect. Thrombin treatment also results in increased translocation of Smad2 and 3 to the nucleus. Our results suggest a possible transactivation of the TGF‐ β receptor by the PAR1 receptor. Ongoing experiments will further elucidate the mechanism behind thrombin induced fibrosis.
Epidermal growth factor receptor transactivation and fibronectin matrix assembly by the G‐protein coupled receptor, GPER, requires a transmembrane signaling complex consisting of PTPN12, integrin α5β1, and MMP‐3
Authors: Magruder Hilary Thompson , Filardo Edward J
Published: June 2021

G‐protein estrogen receptor (GPER) positively associates with breast tumor progression, including tumor size and extramammary metastases. Via activation of integrin α5β1, GPER coordinates two cellular activities that facilitate breast tumor cell survival, fibronectin matrix assembly and the release of membrane‐tethered proHB‐EGF. RNAi knockdown of either MMP‐3 or PTPN12 inhibits both of these cellular processes regulated by effectors on opposing sides of the plasma membrane. Moreover, PTPN12 knockdown also negatively influences the ability of GPER to promote the centripetal movement of integrin α5β1 from focal to fibrillar adhesions, the site of fibronectin fibril formation, and results in the accumulation of the Shc adaptor protein on the cytoplasmic tail of integrin α5β1. In support of the concept that PTPN12 is necessary for activated integrin α5β1, SKBR3 cells stably expressing PTPN12 shRNA showed reduced formation of anchorage independent colonies. Efforts to further understand the influence of PTPN12 knockdown on “inside‐out” integrin signaling events required for EGFR transactivation are underway with particular interest in understanding its impact on MMP‐3 activation and HB‐EGF release. A better understand of the molecular mechanism of GPER induced EGFR transactivation may lead to the development of future therapeutic strategies that target this receptor and prevent metastatic disease.
Vasoactive Intestinal Peptide Receptor 1 and 2 Expressing Cells Result in Cellular Migration through Alternative Signaling Pathways
Authors: Van der Steen Travis , Meyer Megan , Dorsam Sheri , Dorsam Glenn
Published: June 2021

Vasoactive intestinal peptide (VIP) influences cell proliferation, apoptosis, and cellular migration. Two G‐protein coupled receptors called, vasoactive intestinal peptide receptors 1 (VPAC1) and VPAC2, bind VIP with a similar affinity and elicits G protein signaling, including Gαs. It has been reported that T cell lines exclusively expressing VPAC1 or VPAC2 show opposite effects on chemotaxis. In contrast, primary human T cells show similar chemotactic effects by VPAC1 or 2 specific agonist. Moreover, VIP receptor splice variants have been recently discovered to have altered signaling properties making it difficult to determine the biology of these receptors. We hypothesized that full‐length VPAC1 and 2 receptors couple through Gαs, which mediate different magnitudes of intracellular cAMP resulting in chemotaxis. Utilizing two T cell lines that exclusively express VPAC1 or VPAC2 receptors that were validated by qPCR and immunoblots, we investigated the extent to which VIP effected chemotaxis through Gαs signaling. These experiments have identified that VIP induces equivalent chemotaxis through a cAMP‐dependent (VPAC2) and independent (VPAC1) mechanism. Collectively our data suggest that VIP receptors differ not in the cellular effect (chemotaxis) but rather their signaling. Funding: (COBRE) 2P20RR015566
IL‐1beta Loss Contributes to Reduced Toxicity of the TRIF‐biased TLR4 agonist Monophosphate Lipid A
Authors: Embry Chelsea Anne , Ganhapudhi Siva , Desta Dereje , Mitchell Thomas
Published: June 2021

Monophosphate Lipid A (MPLA) is a TRIF‐biased TLR4 agonist that retains the immuno‐stimulatory properties of its parent molecule lipopolysaccharide (LPS) while displaying 0.1–1% as much toxicity. We previously determined that TRIF‐biased signaling downstream of TLR4 activation by MPLA is responsible for loss of IL‐1beta maturation activity (C.A. Embry et al. Science Signaling 2011). While this mechanism was fully elucidate in vitro, the role of IL‐1beta in mediating differential inflammatory processes in vivo between the two TLR4 agonists, MPLA and LPS, has yet to be determined. In this study, IL‐1RI knock‐out mice (that are unable to induce inflammation in response to IL‐1beta) were compared to wild type in their ability to induce a pro‐inflammatory response after challenge with either MPLA or LPS. Serum and tissue samples from treated mice were analyzed for levels of IL‐RI/MyD88‐dependent factors at 1hr and 6hrs. Results suggest that strong IL‐1beta production induced by LPS can account for most of the differential activation of MyD88‐dependent inflammatory factors compared to MPLA at later time points in vivo. These results indicate that MPLA's TRIF‐biased TLR4 activation results in loss of factors that potentiate inflammation and toxicity.
Quantitative proteomic analysis of proteins and post‐translational modifications during the metaphase to anaphase transition after altered O‐GlcNAcylation
Authors: Whelan Stephen Alan , Tan Ee Phie , Costello Catherine E. , McComb Mark , Slawson Chad
Published: June 2021

Post‐translational modifications (PTMs) are essential in regulating the function, stability, and localization of proteins. One such modification is the O‐GlcNAc. O‐GlcNAc is the attachment of a single N‐acetylglucosamine moiety to serine and threonine residues on nuclear and cytoplasmic proteins. The modification is dynamically processed on proteins by the enzymes O‐GlcNAc transferase (O‐GlcNAc addition) or O‐GlcNAcase (O‐GlcNAc removal). Our objective is to explore how alterations in O‐GlcNAcylation alter several different types of PTM's during mitosis. We employed SILAC (Stable Isotope Labeling of Amino Acids in Cell culture) labeling to quantitate differences in cells with either an O‐GlcNAc transferase or O‐GlcNAcase gain of function. These samples were combined, digested, and fractionated to enrich for O‐GlcNAc, phosphate, acetyl, or ubiquitinated peptides. By mass spectrometry analysis, we have identified numerous proteins and PTMs altered by aberrant O‐GlcNAcylation. Multiple bioinformatic and systems biology tools has allowed a comprehensive analysis of the interplay between multiple post‐translational modifications providing valuable insight into how these modifications control cellular functions such as mitotic progression. NIH‐NHLBI contract N01 HV‐00239 (current CPC), S10 RR020946 (Thermo LTQ‐Orbitrap), and P41 RR010888 (Mass Spectrometry Resource) to C.E.C.
Comparative analysis of M. pneumoniae models using the CytoSEED plugin for metabolic model visualization
Authors: Hazekamp Nicholas Lee , Kammeraad Joshua , Bockstege Benjamin , DeJongh Matthew
Published: June 2021

We have developed a Cytoscape plugin for exploring genome‐scale metabolic models in the Model SEED database (www.theseed.org/models). We have used this plugin to compare our metabolic model for Mycoplasma pneumoniae M129 (Mp) with the one published by Yus et al. in Science in 2009. Mp is missing an annotated gene for the transaldolase (TAL) activity, which converts erythrose‐4‐phosphate into sedoheptulose‐7‐phosphate. However, it is known that Mp produces phospho‐ribose‐diphosphate, indicating that the pentose phosphate pathway is functional. The Yus model asserts that the TAL activity is present; however, our software shows that two reactions present in our model associated with fructose‐bisphosphate aldolase and phosphofructokinase bridge this gap using sedoheptulose‐1,7‐bisphosphate as an intermediary compound. Our software also shows differences in asserted transport reactions, glycerophospholipid metabolism, and maintenance of oxidation‐reduction balance. Our software enables comparing the results of flux variability analysis for the two models on different growth media, and the determination of which reactions are essential on the minimal media discovered by Yus et al.

NSF 0745100, 0850546
Identifying drug response signatures using genome‐scale metabolic network analysis of human cell line expression data
Authors: Zielinski Daniel , Mo Monica , Palsson Bernhard
Published: June 2021

There is an increasing awareness in the drug development community of the need to extend traditional single target‐centric approaches to network level perturbation assessment. We present a study of systems‐level drug response phenotypes using a genome‐scale reconstruction of the human metabolic network as a context for interpreting high‐throughput data sets. The analysis presented shows that: (1) metabolic response phenotypes can be obtained from “omics” data types, (2) a metabolic network approach can be applied to expression profiling data from drug‐treated cells in culture, (3) predicted results can be experimentally verified and recapitulated in cell culture. Example case studies presented include diabetic (metformin) and psychiatric (haloperidol) therapies. This study represents notable progress toward the prospective use of systems biology in the drug discovery and development process.
FTY720‐PHOSPHATE (GILENYA) SELECTIVITY OF S1P1 GPCR REQUIRES MOTIFS IN INTRACELLULAR LOOP 1 AND TRANSMEMBRANE DOMAIN 2
Authors: Tigyi Gabor , Valentine Wiliam J , Godwin Virginia I , Osborne Daniel A , Liu Dianna , Fujiwara Yuko , Van Brocklyn James , Bittman Robert , Parrill Abby
Published: June 2021

FTY720 phosphate (FTY720P) is a high potency agonist for all the EDG‐family S1P receptors except S1P2. To map the distinguishing features of S1P2 ligand recognition we applied a computational modeling‐guided mutagenesis strategy that was based on the high degree of sequence homology between S1P1 and S1P2. S1P2 point mutants of the ligand‐binding pocket were characterized. The residues R3.28, E3.29, and K7.34 were essential for activation. Mutation of residues A3.32, L3.36, V5.41, F6.44, W6.48, S7.42, and S7.46, predicted to interact with the S1P hydrophobic tail, impaired activation by S1P. Replacing individual or multiple residues in the ligand‐binding pocket of S1P2 with S1P1 sequence did not impart activation by FTY720P. Chimeric S1P1/S1P2 receptors were generated and characterized. The S1P2 chimera with S1P1 sequence from the N‐terminus to TM2 was activated by FTY720P and the S1P2(IC1‐TM2)S1P1 domain‐insertion chimera showed S1P1‐like activation. Twelve residues in this domain differ between S1P1 and S1P2, distributed in four motifs a–d. Insertion of 78RPMYY in motif b alone or simultaneous swapping of five other residues in motifs c and d from S1P1 into S1P2 introduced FTY720P responsiveness. Molecular dynamics calculations indicate that FTY720P binding selectivity is a function of the entropic contribution to the binding free energy rather than enthalpic contributions and that preferred agonists retain substantial flexibility when bound.
Importance of Very Long Chain Acyl‐CoA Synthetase 3 (ACSVL3) in cholesterol homeostasis and lipid raft signaling in U87 glioma cells
Authors: Kolar Elizabeth Anne , Richard Jean-Philippe , Pei Zhengtong , Laterra John , Watkins Paul A.
Published: June 2021

We previously showed that ACSVL3 is overexpressed in human malignant gliomas and glioma cell lines. ACSVL3 knockdown (KD) U87 glioma cells had significantly lower rates of cholesterol synthesis than control cells. Real‐time PCR results suggest that expression of genes important in fatty acid and cholesterol synthesis is lower in ACSVL3 KD U87 cells. Depleting U87 cells of ACSVL3 decreased their malignant growth properties in vitro and altered growth factor‐stimulated Akt signaling. Plasma membrane signaling complexes are localized in cholesterol‐ and sphingolipid‐rich detergent‐insoluble microdomains, or lipid rafts. To assess whether changes in Akt signaling correlated with raft cholesterol, total cell membranes from control and ACSVL3 KD U87 cells were separated into Triton X‐100‐soluble and ‐insoluble (raft) fractions. Raft fractions contained raft markers e.g. caveolin‐2 by immunoblot. The cholesterol content of raft fractions from ACSVL3 KD cells was significantly lower than that of control cells. Treatment of control cells with methyl‐beta‐cyclodextrin, which decreased raft cholesterol, produced changes in Akt signaling similar to those seen in KD cells. These results suggest that the downstream effects of ACSVL3 influence lipid raft biochemistry possibly by altering cholesterol homeostasis. Supported by NS062043.
IGFBP‐3 Forms Multimers in Physiologic Conditions
Authors: Mireuta Matei , Hancock Mark , Baxter Robert , Pollak Michael
Published: June 2021

Insulin‐like growth factors 1 and 2 (IGF‐1 and IGF‐2) are peptide hormones similar in molecular structure to insulin, and they regulate a variety of cellular activities, including metabolism, proliferation, and growth. IGF actions are tightly modulated by a family of proteins called insulin‐like growth factor‐binding proteins (IGFBPs), of which IGFBP‐1 through 6 have been characterized. Most IGFBPs in the blood originate from the liver, but they are also expressed in many other tissues. IGFBP‐3 and ‐5 are the most abundant IGFBPs in the circulation. They form a ternary complex with IGFs and a third protein termed acid‐labile subunit.

It has been suggested that IGFBP‐3 and ‐5 can form homomultimers, possibly through disulfide bond exchange, but this effect has been regarded mainly as an artifact. In this work, we show that IGFBP‐3 multimers can occur in physiologic conditions. We demonstrate by surface plasmon resonance that the affinity of the IGFBP‐3 self‐association reaction is in the low nanomolar range. Additionally, we demonstrate by native gel electrophoresis, that IGFBP‐3 multimers of various sizes occur at physiologic concentrations and in several physiologic settings. Moreover, we show that most of these species are SDS‐stable. Our work provides evidence of a previously unrecognized complexity in the IGF‐ 1/IGFBP‐3 system.
Human Steroidogenic Cytochrome P45017A1: Roles of Key Active Site Residues
Authors: Jackson Michelle R. , Petrunak Elyse M. , Scott Emily E.
Published: June 2021

Cytochrome P450 17A1 (CYP17A1) is a monooxygenase required for the biosynthesis of androgens and estrogens, which promote prostate and breast cancer growth, respectively. A new approach is to inhibit CYP17A1. However, CYP17A1 performs not only the 17,20‐lyase reaction needed only for androgen/estrogen production, but also a 17α‐hydroxylation reaction important for the synthesis of androgens, estrogens, and glucocorticoids. CYP17A1 inhibitor drugs currently available block both reactions and have side effects due to disruption of glucocorticoid synthesis. Our goal is to investigate CYP17A1 enzyme function to facilitate the design of inhibitors that only block the lyase reaction. Based on new CYP17A1 structures, active site amino acids Y201, N202, D298, R239, E305, I371, and V482 are proposed to play key roles in catalysis. Site‐directed mutagenesis was used to examine the roles of each amino acid in binding substrate or the prostate cancer drug abiraterone. Results show that abiraterone still binds tightly without hydrogen bond interaction to N202. Evaluating the roles these amino acids play in ligand binding will help understand enzyme function and direct drug development. Future studies will focus on evaluating the roles these amino acids play in the enzymatic catalysis of progesterone and pregnenolone via hydroxylase and lyase reactions. Supported by NIH NIGMS (GM078441, GM103420) and NCRR (RR017708).
AMP‐activated protein kinase (AMPK) activity in Leigh Syndrome French Canadian Type (LSFC) patient fibroblasts: differences between primary and immortalized cell lines
Authors: Mukaneza Yvette , Rivard Marie-Ève , Rosiers Christine Des , Coderre Lise , Consortium LSFC
Published: June 2021

Background & Objective

LSFC is an autosomal recessive disease caused by mutations in the leucine‐rich pentatricopeptide repeat containing protein (LRPPRC) gene resulting in decreased LRPPRC expression. This leads to a tissue‐specific cytochrome c oxidase deficiency and a reduction in energy production. AMPK is a key enzyme in the regulation of energy homeostasis. To test the hypothesis that AMPK activity may differ in LSFC and control cells, we evaluated the phosphorylated and active form of AMPK in primary (P) fibroblasts and, because of the limited availability of P cells, we also tested their immortalized (I) counterparts.

Results

Under basal conditions, P fibroblasts from LSFC patients and controls showed marginal differences in the levels of phosphorylated AMPK (pAMPK) as well as acetyl‐CoA carboxylase (ACC), one of many AMPK substrates. However, compared to P cells, I cells from both controls and LSFC patients showed a 60% and 80% decrease in pAMPK, respectively. Consistent with pAMPK levels, phosphorylated ACC levels were also decreased in I cells by 58 % in controls and 69% in LSFC patients. The total levels of these proteins were similar in all cases.

Conclusion

These results highlight an important decrease in AMPK activity level in I vs. P cells, which may compromise the capacity of these cells to palliate an energy deficit.

Supported by: Association de l'acidose lactique and CIHR Emerging Team Grant
Modulation Of Sympathetic Nervous, Renin Angiotensin And Kallikrein Kinin Systems On Youth Obesity
Authors: Fernandes Fernanda Barrinha , Fernandes Adriana Barrinha , Febba Andreia Cristina Silva , Leite Cleber Aparecido , de Souza Vitalle Maria Sylvia , Arita Daniele Yuri , de Sousa Cunha Tatiana , Jung Flavia Fernandes , Casarini Dulce Elena
Published: June 2021

The aim of this study was to evaluate the relationship between obesity and Sympathetic Nervous (SNS), Renin Angiotensin (RAS) and Kallikrein Kinin (KKS) Systems. Subjects were divided in groups: 43 normal weight (NW), 14 overweight (OV), 37 obese (OB) and 10 MO (MO). The Ang I, Ang II, Ang‐(1–7), Bradykinin (BK), Vasopressin (VP), norepinephrine (NE), epinephrine (EP), serotonin (SE) plasmatic levels had been measured by HPLC. Ang (1–7) levels were lower in MO and OB compared with NW (0.06±0.04; 0.24±0.17 and 0.52±0.21nmol/mL). Ang I levels were higher in MO compared with NW (0.06±0.03; 0.03±0.02). OB had higher values of VP than NW (0.107±0.0217; 0.087±0.016ng/mL). OB had higher values of EP and NE than NW (48.3±20.3 vs 34.7±17.2 and 85.6±27.9 vs 45.9±28.3 pg/mL respectively). The NW had higher concentration of SE than the OV (281.2±113.0 vs 200.1±117.1 ng/mL). MO had lower BK levels than NW (0.367 vs 0.885). Our results showed a reduction of Ang (1–7) and increased of Ang I associated with weight gain in adolescents suggesting a modulation of ACE2 and NEP. According to our results, obesity in youth seems to be related with high levels of EP, NE, blood pressure and low Ang (1–7). These alterations in the SNS, RAS and KKS, associated with low levels of Ang 1–7 and BK may contribute to the development of hypertension in obese adolescents, since these can resulting in cardiovascular target organ damage such as the heart, the vascular wall and the kidney.
Characterizations of the Middle Meatus and Hiatus Semilunaris for the purposes of Endoscopic Procedures
Authors: Dahlstrom Kelly Anne , Olinger Anthony Boniface
Published: June 2021

Anatomical variation of the lateral nasal wall, including the pathway from the frontal, ethmoidal and maxillary sinuses may affect the communication between the paranasal sinuses and the nasal cavity. The middle meatus and hiatus semilunaris are areas where variations can occur which predispose patients to recurring sinusitis. Endoscopy allows for visualization of the entire nasal cavity, for both diagnostic and therapeutic uses. The purpose of this study was to provide an accurate characterization of the middle meatus and hiatus semilunaris for the purposes of endoscopic procedures. 116 cadaveric lateral nasal walls were observed. The middle meatus was measured at three distinct points. Cranial length was also recorded. A 3D digitizer was used to record spatial relationships between the openings of the paranasal sinuses and the hiatus semilunaris. The average widths for the middle meatus were 1.69mm anteriorly, 2.83mm at the transition angle and 4.74mm posteriorly. The posterior width was significantly larger than either of the two other measurements. The hiatus semilunaris was categorized according to the varying morphologies. Given that the middle meatus was significantly wider posteriorly, our findings support the recommendation that entering the middle meatus posteriorly may allow an endoscopic surgeon easier access to the structures of the lateral nasal wall.
14‐3‐3σ synergizes Wnt and Akt to promote embryonic stem cell proliferation via regulating GSK‐3β/β‐catenin
Authors: Liou Jun-Yang
Published: June 2021

Pluripotent embryonic stem (ES) cells are considered to be an unlimited cell source for tissue regeneration and cell‐based therapy. Investigating the molecular mechanism underlying the regulation of ES cell expansion is thus important. 14‐3‐3 proteins are implicated in controlling cell division, signaling transduction and survival by interacting with various regulatory proteins. However, the function of 14‐3‐3 in ES cell proliferation is unknown. In this study, we show that retinoid acid (RA) suppressed selectively the expression of 14‐3‐3σ isoform. Knockdown of 14‐3‐3σ with siRNA reduced mESC proliferation, while 14‐3‐3σ transfection increased mESC growth and rescued RA‐induced growth arrest. The growth‐enhancing action of 14‐3‐ 3σ was abrogated by β‐catenin knockdown. 14‐3‐3σ bound GSK‐ 3β and increased GSK‐3β phosphorylation in a PI‐3K/Akt‐dependent manner. Furthermore, 14‐3‐3σ enhanced Wnt3ainduced β‐catenin level and GSK‐3β phosphorylation. DKK abolished Wnt3a‐induced effect but did not interfere GSK‐3β/14‐ 3‐3σ binding. These findings show for the first time that 14‐3‐3σ plays an important role in regulating mESC proliferation by binding and sequestering phosphorylated GSK‐3β and enhancing Wnt‐signaled GSK‐3β inactivation. 14‐3‐3σ is a novel target for ES cell expansion.
Role of cinobufotalin in the pathogenesis of preeclampsia: in vivo and in vitro studies
Authors: Horvat Darijana , Kelso Kelsey , Allen Steven , Fothergill Russell , Zawieja David , Kuehl Thomas , Uddin Mohammad Nasir
Published: June 2021

Objective

Preeclampsia (preE) is a hypertensive disorder of pregnancy. Cardiotonic steroids (CS) are endogenous inhibitors of Na+/K+ ATPase. We have shown that one CS, marinobufagenin (MBG), is elevated in preE prior to the development of symptoms. We have also demonstrated that MBG impairs cytotrophoblast (CTB) function, which is critical for placental development. We evaluated the effect of cinobufotalin (CINO), another CS, on pregnant rats and CTB cells to assess its role in preE.

Methods

Four groups of rats were studied: normal pregnant (NP, n=10) rats, NP injected with MBG (NPM, n=10) rats, NP injected with CINO (NPC, n=10) rats, and a rat model of preE (PDS, n=10). CTB cell function and signaling were assayed after CINO treatment.

Results

NPC rats showed hypertension and proteinuria similar to PDS and NPM rats. CINO at ≥ 1 nM inhibited CTB cell proliferation, migration, and invasion but had no effect on cell viability. A higher percentage of cells were in the G0/G1 phase in groups treated with ≥ 1 nM CINO. CINO caused an increase in stress signaling p38 MAPK and a positive annexin‐V stain in CTB cells, indicating the activation of apoptotic signaling. However the CINO induced apoptotic signaling was prevented by p38 inhibition.

Conclusions

We have demonstrated that: 1) CINO induces preE in NP rats and 2) CINO impairs CTB cell function. Therefore, CINO may play a role in the pathogenesis of preE.
ANATOMICAL MODEL IN WAX OF THE SYMPATHETIC NERVOUS SYSTEM: OBSERVATION AND NUMERICAL PHOTOGRAPHY IN ROTATION
Authors: Le Floch-Prigent Patrice Pierre , DARNAUD Isabelle
Published: June 2021

An anatomical model in wax of the right sympathetic nervous model was mounted on a real skeleton with the osseous elements in connection. It was photographed in rotation every 5 degree on 360 degree in its whole and on its three main, vertical, stages in order to get a 3‐dimensional numerical reconstruction and to deliver it freely on the net for every interested public. The attentive observation of the sample allowed to appreciate its anatomical veracity which was excellent. A numerical D‐40 Nikon camera was used and a rotational device made of two thick, circular, identical marble disks assembled by a ball‐bearing. The upper disk was graduated on its periphery. The sample belonged to the Spitzner collection from the Orfila's museum (Paris), closed and evacuated in december 2009. Its number was N° 1232 (Surgical and Radiological Anatomy catalog, 1995, supp. 1).
Protective effect of vitamin C against carbofuran‐induce testicular
Authors: AL-AMOUDI WAEL MOHAMMED
Published: June 2021

The effect of the carbamate insecticide “carbofuran” on testes of male albino mice and the possible protective role of vitamin C against the insecticide toxicity was assessed. Treating mice with carbofuran for 8 weeks induced a significant decrease in the diameters and germinal epithelial heights of the seminiferous tubules. The histological evidence showed inhibition of spermatogenesis. There was also a decline in sperm count. Histochemical results revealed that animals given carofuran had decreased contents of carbohydrates and total proteins in the testicular tissue. Treating mice with carbofuran and vitamin C showed an improvement in the testicular damage. The seminiferous tubules appeared normal and the different stages of spermatogenic cells showed an advanced degree of activation. The present work is supported by Umm Al‐Qura University
12,13‐dihydroxyoctadecenoic acid regulates hematopoietic stem cell and progenitor cell function in zebrafish and mouse
Authors: Froemel Timo , Jungblut Benno , Barbosa-Sicard Eduardo , Trouvain Caroline , Hu Jiong , Popp Rüdiger , Dimmeler Stefanie , Hammock Bruce D. , Fleming Ingrid
Published: June 2021

The soluble epoxide hydrolase (sEH) is a fatty acid epoxide metabolizing enzyme and its deletion and/or inhibition have been linked with altered cardiovascular homeostasis. Epoxyeicosatrienoic acids, main substrates of the sEH are known to promote angiogenesis. The morpholino (MO) mediated downregulation/inhibition of the sEH in zebrafish results in a defective caudal vein plexus (CVP). Beside the vascular defects a reduced number of hematopoietic stem and progenitor cells (HSPC) could be observed. A screen of fatty acids revealed that the sEH metabolite 12,13‐dihydroxyoctadecenoic acid (12,13‐ DiHOME) was able to rescue these HSPC defects. In a CFU‐S12 assay a reduced number of colonies could be detected in spleens of those mice that received bone marrow cells from sEH−/− mice in comparison to cells derived from wild type mice, whereas 12,13‐ DiHOME was able to rescue these phenotype. Furthermore sEH−/− mice showed less HSPC mobilization after G‐CSF treatment. Summarized, the sEH activity regulates vascular sprouting of the caudal vein plexus in the zebrafish and its product 12,13‐DiHOME influences hematopoietic stem and progenitor cell function in zebrafish and mouse.
The acute effects of Montbretin A (MbA), an á‐glucosidase inhibitor, on plasma glucose levels in the Zucker Diabetic Fatty rat
Authors: McNeill John Hugh , Yuen Violet G , Brayer Gary , Withers Stephen G , Andersen Raymond J , Coleman John
Published: June 2021

Postprandial hyperglycemia is a risk factor for cardiovascular disease in diabetic patients. Acarbose and Miglitol, current therapeutic α‐glucosidase inhibitors, reduce postprandial hyperglycemia but, due to side effects, are not well tolerated. Research is ongoing to develop products that will minimize side effects and improve patient compliance. MbA, a purified glycosylated acyl flavonol, was isolated from extracts of the plant, Crocosmia crocomiflora. In vitro studies have shown MbA to be an inhibitor of both exquisite specificity and affinity for human pancreatic α‐amylase. The objective of this study was to determine if MbA was an orally effective glucose lowering agent in an animal model of type 2 diabetes. Animals were challenged with a starch load (2 g/kg) immediately followed by MbA (dose range: 0.5 to 10 mg/kg) administered by oral gavage. Plasma glucose levels were measured immediately prior to and at 30, 60, 90 and 120 minutes post‐administration. Acarbose 10 mg/kg was used as a control for glucose response. MbA prevented the starch‐induced increase in glucose levels at doses greater than 5 mg/kg. MbA (10 mg/kg) reduced plasma glucose levels by 38±3% as compared to 26±2% for acarbose in fatty treated animals. These data show that MbA was effective in lowering plasma glucose levels in vivo. Future studies will examine the chronic effects of oral MbA treatment on diabetes mellitus. Supported by CDRD and CIHR.
Effects of Herb Water Extracts on Enzyme activities Related to Glucose Metabolism in Hep G2 Cell
Authors: Kim Kyoungkon , Lee Jaesung , Heo Jeongwon , Baek Jongmi , Kim Taehyuk , Kim Daejung , Kim Taewoo , Choe Myeon
Published: June 2021

As a major threat to human well‐being, diabetes mellitus is one of the most complex disorder characterized by hyperglycemia. Drugs have been developed to cure diabetes mellitus with several principle, such as pancreatic beta cell stimulator and hypoglycemic action, but the side effects were expected from most of drugs in long‐term administration. Therefore, there has been a growing interest in alternative therapeutic use of natural products for diabetes, especially those derived from plants. This study was carried out to investigate beneficial effects of herbal plants water extract on enzymes activities of glucokinase(GK), pyruvate dehydrogenase‐α and acetyl‐CoA carboxylase(ACC).
Stromovascular Cell Differentiation in Collagen Matrix Induced by High Fat Medium
Authors: Ettinger Susan , White Joshua , Brane Luke , Kunza Brittany , Agaronov Alan
Published: June 2021

Stromovascular Cells (SVCs) derived from adipose tissue have potential to differentiate into cells of mesodermal origin such as adipocytes, cartilage, bone, and skeletal muscle. To study the influences of extracellular matrix and substrate on SVC differentiation, we cultured SVCs in a three‐dimensional (3D) collagen‐I matrix or on a two dimensional tissue culture surface (2D) with lipid supplemented and low lipid medium. Samples taken from three fat depots (inguinal, epididymal, and retroperitoneal) of adult male rats were treated with collagenase and the SVCs plated on either 3D or 2D tissue culture plates. Cells were incubated in 10% marine‐derived lipid supplemented media (HL) or low‐lipid control media (LL) for 4 weeks. SVCs in LL media proliferated more rapidly in 3D than 2D matrix; no evidence of differentiation was observed. Differentiation was only seen in SVCs cultured in HL medium on 3D matrix. (see figure). SVC differentiation to mature adipocytes has previously been shown to require dexamethasone, insulin and other growth stimulators. In conclusion, in our 3D model, SVCs grown in 10% lipid supplemented medium underwent robust adipocyte differentiation without growth stimulators. We propose that the 3D model, by replicating matrix binding and stresses in the in vivo environment, may allow a more physiologically relevant examination of factors that regulate SVC proliferation and differentiation.
Whey protein sweetened beverages attenuate plasma glucose, hunger and calorie intake in young women
Authors: Zafar Tasleem Ara , Ghazaii Carol Waslien
Published: June 2021

Whey protein has potential as a functional food component to regulate blood glucose and body weight. Beverages may prove an effective vehicle for delivering whey protein for consumption. The current study investigated the effect of whey‐glucose mixed drinks relative to glucose or whey alone on a) glycemic response, b) appetite and c) calorie intake. Test beverages were given once a week on fasting as 50g or 75g glucose, 25g whey or 25g whey mixed with 50g glucose. Blood glucose and appetite were measured at 0, 15, 30, 45, 60, 90, 120, 150 and 180 min. Pizza was served at 180min and calories consumed were calculated. Both blood glucose and calories intake were reduced after whey protein drinks with or without glucose. The change in blood glucose was associated with reduced appetite and calorie intake. This study was supported by Kuwait University research grant WF02/09.
A study of consumers’ perceptions and preferences of regional wine served with traditional dishes
Authors: Kim Song-Suk
Published: June 2021

The study was carried out to investigate the demand of local native wine when consumers visited traditional restaurants in Kyungbuk area. The total number of subjects were 377 adults who are residents in Gyeongbuk area. The questionnaire survey was performed by the trained interviewer. About 83.8% of the participants recognized the importance of development of native local wine and preferred to enjoy it with traditional dishes. Approximately 87.0% of men and 82.5% of women among subjects had strong positive opinions that traditional restaurants could serve with local wine, when alcoholic drinks were served. Four age groups of subjects were formed in order to observe the demand of the native local wine in the different ages of participants. The age groups were as follows; 20~29(20s), 30~39(30s), 40~49(40s), 50~59(50s), 60 and over(over 60s). During traditional meal time, the degree of preferences of native local wine in the age groups was that 84.8% for 20s, 77.2% for 30s, 88.6% for 40s, 87.2% for 50s, and 100% for over 60s. Exceptionally, subjects in the young ages(20s) showed as much as higher demand of native wine compared to the other age groups. The level of preference in age of 30s was the lowest among the age groups. It was interesting to note that the young age such as 20s presented a strong demand of the native local wine when having traditional dishes. The findings of results from the present study indicate that the favorite local wine to go with the traditional dishes should be developed, and the local resources as ingredients of wine production could be used.
Comparison of the efficacy of popular weight loss programs in sedentary overweight women III: fasting blood lipids and glucose
Authors: Koozehchian M , Baetge C , Lockard B , Mardock M , Simbo S , Jung Y , Galvan E , Levers K , Byrd M , Khanna D , Sanchez B , Jagim A , Oliver J , Kresta J , Horrell K , Leopold T , Cho M , Rasmussen C , Kreider R
Published: June 2021

126 sedentary women (44±12 yr, 44.8±5% body fat, 35.4±6 kg/m2) were randomized to participate in a control group (C) or the Curves Complete® 90‐day Challenge (CC), Weight Watchers® Points Plus (WW), Jenny Craig® (JC), or Nutrisystem® Advance Select™ (NS) weight loss programs for 12‐wks. Fasting blood samples were obtained at 0, 4, 8, and 12 wks and analyzed by MANOVA. Data are presented as percent changes from baseline after 12‐wks. Participants in the CC group had greater changes in TG (C −1.9±23; CC −8.3±28; WW −2.7±32; JC 15.1±43; NS 11.1±29 %, p=0.03) with TG levels increased in the JC and NS groups. Although some trends were observed, no significant interactions were observed among groups in total CHL (C −1.5±11; CC −5.9±9; WW 1.0±12; JC −2.3±11; NS −3.8±15 %, p=0.22), HDL‐c (C −3.7±9; CC −0.6±14; WW 1.1±13; JC −5.0±16; NS 2.9±6.3 %, p=0.008) and hip 4.0±13 %, p=0.45), LDL‐c (C −3.3±17; CC −7.0±9; WW 11.0±52; JC −2.7±12; NS −6.1±21 %, p=0.12), the ratio of CHL/HDL‐c (C 2.9±6.3 %, p=0.008) and hip 2.4±24; CC −4.0±11; WW −0.3±13; JC 4.8±16; NS 0.1±12 %, p=0.48), or blood glucose (C −1.3±9; CC −2.4±16; WW −4.8±19; JC −1.6±10; NS −2.9±16 %, p=0.93). Results indicate that participation in the CC program promotes more favorable changes in TG levels than other popular weight loss programs and that adherence to popular diets may affect blood lipids differentially. Supported by Curves International (Waco, TX)
Fruit‐vegetable ferment attenuates high fat‐fructose‐induced inflammation and oxidative stress in rats
Authors: Tsai Jyun-Han , Korivi Mallikarjuna , Chao Huan-Chun , Yang Jo-Hsuan , Li I-Huan , Huang Hui-Yu
Published: June 2021

This study investigated the anti‐inflammatory and antioxidant activities of fruit‐vegetable ferment (FVF) in chronic high fat‐fructose diet‐induced insulin resistance (IR) rats. Thirty rats were equally assigned into three groups, including control (CON), high fat‐fructose diet (HFD), and HFD plus FVF (HFD+FVF). Third group received FVF (178.5 mg/kg b.w., orally) for 8‐week. HFD‐induced elevated serum inflammatory cytokines, including interleukin‐1£] (IL‐£]), IL‐6 and tumor necrosis factor‐£\ (TNF‐£\) were significantly (P<0.01) controlled in FVF treated group. Furthermore, decreased antioxidant enzyme activities, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were significantly (P<0.01) restored by FVF supplementation. Our data concludes that fermented fruit‐vegetable substance is capable to prevent the high fat‐fructose‐induced inflammation and oxidative stress by decreasing pro‐inflammatory cytokines and increasing antioxidant enzymes.
Analysis of LIPC ‐514C>T and APOA5 ‐1131T>C polymorphisms in Mexican Mestizos and Mexican Amerindians
Authors: Vargas-Martinez Carolina , Contreras Alejandra V , Fernandez-Lopez Juan Carlos , Silva-Zolezzi Irma , Tejero Maria Elizabeth
Published: June 2021

Description of frequencies and haplotypes in Mexican Mestizo (MM) and Mexican Amerindian (MA) populations, of two SNPs implicated in lipid metabolism, which have been associated to lipid levels in white populations. For this purpose we analyzed DNA extracted from blood samples of 372 MA and 300 MM that participated in the Mexican Genome Diversity Project by allelic discrimination, we also constructed haplotypes and identified tag SNPs.

Results

The frequency for LIPC rs1800588 was 0.430 in MM and 0.262 in MA, whereas for APOA5 rs662799 the frequency was 0.168 in MM and 0.262 in MA. Three haplotypes (>5%) were identified on the LIPC region on MM, one of them including the rs1800588 (8.3%). Six haplotypes (>5%) were identified on the APOA5 region. The rs662799 SNP was included in haplotypes showing similar frequencies in MM and MA (14% and 14.8% respectively). The APOA5 rs662799 is in high LD with tag SNPs that are present in BUD13 and ZNF259. The LIPC rs1077834, that has been associated to higher high density lipoprotein concentrations in white populations, showed high linkage desequilibrium with our SNP of interest.

Conclusions

The genetic information obtained from this study will be useful in future studies where associations between lipid metabolism phenotypes, environmental factors, and disease onset need to be determine in Mexican population. Source of research support CONACYT.
The rs9842733 of the adiponectin Q gene confers risk for multiple predisposing traits for atherosclerosis
Authors: Dzimiri Nduna , Muiya Paul , Al-Najai Mohammed , Vigilla MaryGrace , Andres Edith , Elhawari Samar , Gueco Daisy , Mazhet Nejat , Alshahid Maie , Meyer Brian F
Published: June 2021

Adiponectin (ADIPOQ) plays an important role in various processes that may lead to atherosclerosis. Following a linkage study which pointed to the ADIPOQ gene as a potential locus for early onset of coronary artery disease (CAD) in familial hyperlipidaemia, we evaluated by rtPCR the role of ADIPOQ rs9842733A>T in CAD and its risk traits, including hypertension (HTN), type 2 diabetes (T2DM) and dyslipidaemia disorders in 4650 Saudi individuals (2343 CAD versus 2307 angiographed controls). While the recessive TT genotype was modestly association with CAD [Odds ratio(95% Confidence Interval = 1.23(1.05–1.45); p=0.012), this relationship correlated well with the severity of vessel disease (p=0.022) as determined by the number of affected coronary vessels. Interestingly, the T allele was also implicated in both HTN (3544 vs 1106) [1.44(1.03–2.00); p = 0.034 and T2DM (2562 vs 2088) [1.30(1.00–1.74); p=0.05]. Particularly noteworthy was that the association of the T allele with the degree of CAD became more significant in individuals with low HDL levels (p=0.004) and T2DM (p=0.009), and appeared to exclusively implicate the male gender (p=0.004). These results demonstrate that rs9842733A>T confers risk for CAD and its predisposing traits. Furthermore, the fact that this SNP resides in the 3′‐UTR of the gene suggests that gene regulatory, rather than protein functional mechanisms, might be involved in these actions.
High‐Resolution Electrocardiography in Assessment of Atrial Electrical Instability
Authors: Urgenishbayevich Eleuov Almasbek , Derbisalievich Dalenov Erbolat , Andrei Dubitsky Andreievich
Published: June 2021

The study involved 685 patients aged 32 to 75 years. 457 people ‐ patients with PAF and 90 people ‐ the control group. Equipment used: electrocardiograph MAC‐1, company «Marquet» USA, tredmil‐test firm «Marquet» USA, monitoring of ECG by Holter, Toshiba 3805 ultrasonic device. Parameters were calculated: the interval PQ, filtered wave P‐(FiP), and the difference between FiP and UnFiP, the duration of high‐frequency signal at the end of P wave at the last 5 mV and the rms amplitude of the last 20 ms of P wave (RMS 20). Comparison the duration FiP rate revealed that it was significantly higher in patients with PAF than in the control group: 142 ±7.1 and 124±6.2 ms. Index Under 5 uV was significantly higher in patients with PAF, than in the control group of 29 ±4.2 ms and 18 ±2.4ms, and the index «delta» P in the group with PAF was significantly higher than in the control group of 46 ±1.4 ms and 25±1.3ms. The data obtained revealed the presence of PPP in patients with PAF, 66%, while in the control group at 10%.
STAT3 is a positive regulator of endothelial function in the brain
Authors: Davis Catherine Monica , Young Jennifer M , Alkayed Nabil J
Published: June 2021

Signal Transducer and Activator of Transcription 3 (STAT3) is protective to the brain following ischemic injury. The function of STAT3 in cerebral endothelial cells (ECs) in response to ischemic injury has not been investigated. Since ablation of STAT3 specifically in neurons does not alter infarct size, endothelial STAT3 may be responsible for the STAT3‐mediated protection observed in the brain. We therefore hypothesized that STAT3 protects ECs from ischemic injury. We used primary mouse cerebrovascular ECs and subjected them to oxygen and glucose deprivation (OGD), an in vitro model of ischemia. Using Western blot analysis, we found that STAT3 protein levels and phosphorylation are regulated by OGD; they are decreased immediately following OGD, increasing by 24 hours of reoxygenation to exceed pre‐OGD levels. We show that attenuation of STAT3 signaling, by pharmacological inhibition, induces EC death and reduces both nitric oxide release and trans‐endothelial electrical resistance (TEER). STAT3 is therefore a positive regulator of correct endothelial function in the brain, playing an important role in determining susceptibility to ischemic damage. Since endothelial dysfunction is a contributing factor, as well as an outcome of pathological states, regulation of endothelial STAT3 may have important consequences in cerebrovascular disease. Supported by the American Heart Association.
The tryptophan hydroxylase 2 (Tph2) polymorphism C2432T mediates wound repair and behavioral responses to SSRIs in a sex‐specific manner
Authors: Brookshire Bethany , Hill-Smith Tiffany E. , Balu Darrick T , Turner Jill , Blendy Julie A , Lucki Irwin
Published: June 2021

Current therapies for Major Depressive Disorder involve manipulation of brain monoamine systems, but mechanisms behind successful treatment responses remain unclear. We examined whether a novel polymorphism C2432T in the Tph2 gene mediates differences in wound repair and response to citalopram between two mouse strains, MRL/MpJ (MRL; antidepressant‐responding) compared to C57Bl/6J (C57; antidepressant‐nonresponding) (Balu et al, 2009, Clark et al, 1998). We cross‐bred the MRL and C57 strains for two generations to isolate C2432T and determine functionality using wound closure and the behavioral response to SSRIs in the tail suspension test (TST). MRL mice showed an enhanced wound healing response (p<0.001, n=10) and increased response to 20 mg/kg i.p. citalopram in the TST (p<0.001, n=10) compared to C57 mice. In the F2 generation, we observed a sex‐specific effect of C2432T on wound healing and the response to citalopram. “MRL‐like” females showed enhanced wound healing (p<0.05, n=5) and citalopram response (p<0.001, n=10), while males showed no genotype‐specific effects. The C2432T polymorphism may mediate serotonin involvement in wound repair and antidepressant response in a sex‐specific manner.