bluuebunny/crossref_metadata_2025 huggingface dataset:
Science Education: A Basic Scientist's View of Translational Medicine.
Authors: Schwartz‐Bloom Rochelle D.
Published: June 2021

The big buzz‐word these days is "translational medicine". Basic scientists must demonstrate how their research findings can lead to the improvement of human health. However, the general population knows relatively little about what scientists really do, and given problems in our K‐12 science education, we are facing a danger of becoming a scientifically‐illerate society. As basic scientists, have an obligation to help society understand what we do and what it means. What good is it if we make wonderful discoveries and publish our findings in scientific journals, but the public doesn't understand what we discover? Shouldn't our society have the basic tools to understand science so they can make informed decisions about their lives? A lack of science literacy at the K‐12 level creates future adults who are science illiterate. And when these folks go to the polls, they may make ill‐informed decisions as they cast their ballots. So, to me, translational medicine also means articulating what we do as scientists to the K‐12 students, the public, and legislators, to help them better understand our science, and how it affects their lives and their health. My approach to "translating science" is embodied in science education activities that I have developed for the high school student & teacher population. I will present some examples, and show how such approaches can have real impact on learning and interest in science.
RNF4 acted as an ubiquitin E3 ligase involved in ubiquitin‐dependent degradation of Sumoylated‐Sp1.
Authors: Wang Yi‐ Ting , Chang Wen‐Chang , Hung Jan‐Jong
Published: June 2021

Sp1 is a ubiquitously expressed transcription factor that plays important role in the regulation of many GC‐rich genes encoding for housekeeping proteins. The transcriptional activity of Sp1 is mainly determined by three elements: transcriptional activity, DNA binding ability and its protein stability. Our previously study revealed that modification of Sp1 by SUMO‐1 altered its localization and ubiquitination and then result in Sp1 degradation. Recently, mammalian RNF4, a Ring finger‐containing ubiquitin E3 ligase, targets sumoylated protein for ubiquitin‐dependent degradation. Our preliminary data showed that sumoylated‐Sp1 is prone to recruit RNF4 than wild‐type Sp1. Ectopic expression of RNF4 markedly decreased the protein level of Sp1 in a RNF4 dose dependent manner. By overexpressed the RNF4/CS1, a Ring domain mutant which has been reported that devoid of ubiquitin E3 ligase activity, the protein level of Sp1 did not change. In vitro pulldown assay also revealed that sumoylation increased its interaction with RNF4. Moreover, In vitro ubiquitination assay was used to determine the ubiquitination level of Sp1 in the presence of RNF4, we found that Sp1 is access to ubiquitination with RNF4. Therefore, we infer that RNF4 acted as an ubiquitin E3 ligase targets sumoylated‐Sp1 for ubiquitin‐dependent degradation.
Trichostatin A Inhibits LPS‐induced Cyclooxygenase 2 Expression Through Inhibition of C/EBPδ Gene Expression in RAW264.7 Cells
Authors: Hung Jan‐Jong
Published: June 2021

Cyclooxygenase 2 (COX‐2) is an important inflammatory factor. Previous studies have indicated that COX‐2 could be induced under LPS treatment, and could be inhibited by trichostatin A (TSA). Our previous research reveals that C/EBPδ induced by LPS is through the activation of the pathway of ERK/JNK1/2/p38 and NF‐κB, followed by the recruitment of c‐Rel and c‐Jun to the C/EBPδ promoter. Herein we found trichostatin A (TSA) could inhibit LPS‐induced Cox‐2 gene expression is due to the inhibition of C/EBPδ. Furthermore, the inhibition of C/EBPδ was through preventing c‐Jun recruitment by Sp1 to the C/EBPδ promoter, but not due to the repression of ERK or NF‐κB pathway. In addition, we also found that the interaction of Sp1 and c‐Jun was inhibited under TSA treatment. At last, it is interesting that c‐terminus of c‐Jun was phosphorylated under TSA treatment. To detect the level of PP2B under TSA treatment found that the level of PP2B was reduced. Taken together, inhibition of LPS‐induced COX‐2 by TSA was due to repression of C/EBPδ, and C/EBPδ down‐regulation was through the inhibition of PP2B led the highly phosphorylation at c‐terminus of to repress the interaction of c‐Jun and Sp1.
Comparison of two homogeneous cell‐based kinase assays for JAK2 V617F: SureFire™ pSTAT5 and GeneBLAzer® FRET assays
Authors: Qian Jie , Mason Jennifer L. , Holskin Beverly P. , Murray Kristen A. , Meyer Sheryl L. , Ator Mark A. , Angeles Thelma S.
Published: June 2021

The Janus kinase (JAK)/Signal Transducers and Activators of Transcription (STAT) signaling pathway plays an important role in cellular responses to cytokines and growth factors. Aberrant, constitutive activation of JAK2 signaling has been implicated in myeloproliferative disorders (MPDs) with a single, activating somatic V617F mutation in the JH2 pseudokinase domain of JAK2 as the prevalent molecular lesion. In order to discover small molecule inhibitors for this target, cell‐based kinase assays are important for identifying potential inhibitors. Here, we present two homogeneous 384‐well plate cell‐based assays utilizing Invitrogen's CellSensor® JAK2 V617F irf1‐bla HEL cell line: (1) SureFire(tm) pSTAT5 AlphaScreen® assay from PerkinElmer; and (2) GeneBLAzer® fluorescence resonance energy transfer (FRET) assay from Invitrogen. The SureFire(tm) assay measures levels of phosphorylated STAT5 while the GeneBLAzer® assay monitors ?‐lactamase activity in cells. The S/B ratios for the SureFire(tm) and GeneBLAzer® assays were 10 and 3, respectively. The experimental procedures will be compared, as well as the potency of select compounds. In conclusion, both assays are suitable for compound profiling.
The role of nitric oxide synthase in regulation of ultraviolet light‐induced phosphorylation of the alpha‐subunit of eukaryotic initiation factor 2
Authors: LU WEI , LASZLO CSABA FERENC , WU SHIYONG
Published: June 2021

Ultraviolet light (UV) induces phosphorylation of the alpha subunit of the eukaryotic initiation factor 2 (eIF2α) and inhibits global protein synthesis. Both eIF2 kinases PKR (RNA‐dependent protein kinase)‐like ER (endoplasmic reticulum) kinase (PERK) and general control non‐derepressible‐2 (GCN2) have been shown to phosphorylate eIF2α in response to UV‐irradiation. However, the roles of PERK and GCN2 in UV‐induced translation inhibition are controversial. The upstream signaling pathway(s) that leads to the activation of PERK or GCN2 remains unknown. This report contains data that show PERK and GCN2 contribute equally for UV‐induced eIF2α phosphorylation in human keratinocyte (HaCat) cells. Reducing expression of PERK or GCN2 by small interfering RNA (siRNA) decreases phosphorylation of eIF2α after UV‐irradiation. The data also show that nitric oxide synthase (NOS) mediated oxidative stress plays a role in regulation of eIF2α phosphorylation in HaCat cells upon UV‐irradiation. Treating the cells with a broad NOS inhibitor NG‐methyl‐L‐arginine, a free radical scavenger N‐acetyl‐L‐cysteine or an NOS substrate L‐arginine partially inhibits the UV‐induced eIF2α phosphorylation. The same treatments also partially rescued the translation inhibition induced by UV‐irradiation. The results presented above led to a hypothesis that NOS mediates UV‐induced eIF2α phosphorylation by activation of both PERK and GCN2 via oxidative‐stress and L‐arginine starvation signaling pathways.
Effects of curcumin on Lung cancer; Metabolomic Approach
Authors: Goja Arvind , Zaheeruddin Shaistha , Mateescu Andreea , Sadaat Nadia , Gupta Smiti V.
Published: June 2021

Prognosis of lung cancer, the leading cause of cancer mortality in the US improves if detected at an early stage. Curcumin, a dietary phytochemical has proven anti‐inflammatory, antioxidant, and anti‐carcinogenic activity in several cancer types. This study aims to evaluate the effect of dietary curcumin in lung cancer induced hamster model, using a metabolomics approach. 32 male Golden Syrian hamsters were randomized into treatment (n=20; cancer induced by injecting N‐nitrosobis‐2‐(oxopropyl) amine (BOP);12 weeks), and control (n=12) groups. Each group was subdivided into no (T0, C0) or 2% curcumin (T2, C2) diet groups. Proton nuclear magnetic resonance spectroscopy was conducted on 24h urine samples, in Deuterium oxide using a 700 MHz instrument at 25ºC. The NMR spectra were processed and analyzed using Principal Component Analysis (PCA; SIMCA P+, Umetrics), an unsupervised multivariate analysis technique. The PCA score plots showed clear separation between spectra from(i) the treatment (T0) and control (C0) groups (ii) groups with (T2) and without (T0) dietary curcumin (p < 0.05). The metabolites responsible for the inter‐group differences were quantified using the CHENOMX metabolite database. These metabolites, in addition to providing a mechanistic insight into cancer progression and/or inhibition, may serve as biomarkers for early diagnosis, progression and treatment of lung cancer.
Prostate cancer cells home to bone in a new in vivo model of bone metastasis
Authors: Reeves Kimberley Jayne , van der Pluijm Gabri , Cecchini Marco G , Eaton Colby L , Hamdy Freddie C , Brown Nicola J
Published: June 2021

This study aimed to establish the dorsal skinfold chamber (DSC) model of prostate cancer metastasis. Metatarsals from newborn mice are engrafted into a DSC implanted a SCID mouse (5‐6 weeks old). We have demonstrated that metatarsals are rapidly vascularised by inosculation with the host vasculature by day 5‐7, with functional vessels by day 2. Vascular density was measured over a 7 day period; day 3, 455 ± 186 cm/cm2; day 5, 546 ± 305 cm/cm2 and day 7, 707 ± 370 cm/cm2. Following this fluorescently labelled prostate (PC3‐GFP) or oral (SCC4‐GFP) cancer cells (1 x 105) are injected via the heart to simulate metastatic spread. Cells initially adhere to the microvascular endothelium and/or to metatarsal matrix 5 days after injection (10 ± 7 cells/metatarsal). Bone viability has been assessed throughout the 4 week study duration and demonstrates viable tissue within the DSC. Histology of the metatarsal confirms the presence of tumour cells, viability of the metatarsal, active growth plate and new vessels formation. This model will allow homing, interactions and extravasation between the tumour and bone microenvironment to be studied in detail. Funded by Sixth European Framework Programme (PROMET).
ANP dependent regulation of vascular permeability and plasma volume in mice
Authors: Samardzic Haris , Lenz Joyce F , Adamson Roger H , Lin Yueh‐Chen , Karlsen Tine V , Rygh Cecilie B , Reed Rolf K , Kuhn Michaela , Curry Fitz‐Roy E
Published: June 2021

Mice with endothelial‐restricted deletion of the guanylyl cyclase‐A (GC‐A) receptor for ANP (EC GC‐A KO) have expanded intravascular volumes, and reduced tissue albumin content with acute increases in plasma ANP (Sabrane et al. J Clin Invest 115:1666, 2005). To further test the hypothesis that ANP regulates plasma volume by modulating vascular permeability, we used pharmacological methods to modulate permeability, and measured blood to tissue albumin clearance in mouse models. Pretreatment of isoflurane‐anaesthetized mice with rolipram (8 mg/kg IP, 30 min) was used to stabilize the endothelial barrier in C57 black mice. Albumin clearances, measured using a dual label fluorescence method (FASEB J 20:A708), were: 24.6±5 and 13.8 ±1.3 (μl/g dry/30 min x104, SEM) in skin and skeletal muscle respectively (n=18) in the presence of ANP alone (500 ng/kg/min), and 9.7±1.3 and 9.2±0.7 in skin and muscle in the presence of ANP with rolipram pretreatment (n=17). Fractional reductions in ANP/rolipram clearances were similar to those measured in preliminary results in EC GC‐A KO mice (dual isotope method). In KO as compared to control littermates, ANP‐stimulated albumin clearance was significantly reduced in skin (53%) (n=6) and there was a trend for reduced clearance in muscle (by 80%), trachea (75%) and heart (75%). Is albumin clearance attenuated in both KO and rolipram treated mice after acute plasma volume expansion?
Effects of aging on the cutaneous microcirculation evaluated by a novel neurovascular testing in humans
Authors: FROMY Berengere , SIGAUDO‐ROUSSEL Dominique , SAUMET Jean Louis
Published: June 2021

Aging alters vascular and nervous systems. Skin pressure‐induced vasodilation (PIV) relying on intact neurovascular interaction tests the capacity of the skin to resist to pressure‐induced ischemia. We hypothesized that PIV would be altered in older subjects because of the endothelial dysfunction and would be aggravated by a neuropathy. Subjects were divided according to their age (20‐35 or 60‐75 yrs) and the presence of peripheral neuropathy. In the skin microcirculation, PIV was reduced in non neuropathic older subjects (12±7%) compared to young subjects (62±4%) and was totally abolished in neuropathic older subjects (‐31±10%). The skin endothelium‐dependent responses to acetylcholine and local heating were likewise reduced in both groups of older subjects, showing a comparable endothelial dysfunction. The quantitative sensory tests showed an early C‐fiber dysfunction in non neuropathic older subjects, which was emphasized in neuropathic older subjects. Therefore, for a given endothelial dysfunction, the alteration of the neurovascular interaction was mild to severe according to the degree of the severity of the C fiber dysfunction in older subjects. The skin neurovascular interaction testing could discriminate subjects into subgroups presenting different capacities to resist against tissue ischemia with comparable endothelial dysfunction. Supported by PHRC 2005‐541 and MEDD 0000375.
Differences among exercisers and non‐exercisers during pregnancy
Authors: May Linda E , Gustafson Kathleen M
Published: June 2021

The hypothesis that mothers who exercise during pregnancy have less time than non‐exercisers for other activities was tested by having pregnant mothers complete a reliable and valid Modifiable Activity Questionnaire. Exercisers participated in moderate or vigorous aerobic exercise at least 30 minutes 3 times per week throughout the pregnancy whereas control mothers did not. Questions pertained to descriptives, daily activities, exercise, and amount of sleep. The data from 27 mothers with uncomplicated pregnancies (17 exercisers and 10 non‐exercisers) have been analyzed. Mothers who exercised during pregnancy slept more each night than non‐exercising moms. Although, all mothers were similar age, education levels, and height, there were significant differences between groups for weight and BMI. Control and exercising mothers with male fetuses were less active than moms with female fetuses and, had higher BMIs. Stay‐at‐home moms exercised less than those who worked out of the home, but stay‐at‐home control mothers were more active than control moms who worked outside of the home. We unexpectedly found differences in BMI based on fetal gender. Additionally, exercising moms do not "steal" time to exercise. These results suggest exercising during pregnancy does not require "stealing" time, but being more time efficient. These data will hopefully encourage pregnant women to evaluate their use of time each day.

Grant Funding Source

Intramural
Combined effects of chronic exercise and estrogen on vascular function in ovariectomized rats
Authors: Chao Jung‐I , Su Chia‐Ting , Yang Ai‐Lun
Published: June 2021

Postmenopause, characterized by estrogen deficiency, contributes to the risk of cardiovascular dysfunction. The purpose of this study was to investigate the combined effects of chronic exercise and estrogen on vascular function in ovariectomized rats. Fifteen‐week‐old female Sprague‐Dawley rats were randomly assigned into 3 groups: sham‐operated (sham), ovariectomized (OVX), and OVX with given exercise and 17 beta‐estradiol injection (OVX+Ex+E2) groups. The thoracic aortas were dissected for acetylcholine (ACh)‐ and insulin‐like growth factor‐1 (IGF‐1)‐mediated vasodilation analysis. To explore the roles of phosphatidylinositol‐3 kinase (PI3K) and NO synthase (NOS), selective inhibitors (i.e., wortmannin and Nω‐nitro‐L‐arginine methyl ester) were used. Sodium nitroprusside (SNP, a NO donor)‐induced vasodilation was also examined. We found that, 1) compared with sham rats, the vasodilation evoked by ACh and IGF‐1 was significantly deteriorated in OVX rats, but the effects were disappeared after the removal of endothelium; 2) the combined intervention significantly improved ACh‐ and IGF‐1‐mediated vasodilation in OVX rats; 3) these altered vascular responses were mainly mediated by the release of PI3K and NOS; 4) no significant difference in SNP‐mediated vasodilation was found among three groups. According to our results, we suggested that chronic exercise and estrogen may cooperatively reverse postmenopause‐altered vascular function.
Restriction of dietary NaCl decreases urinary output in diabetic rats
Authors: O'Neill Heidi , Kwon Tae‐Hwan , Ring Troels , Dimke Henrik , Lebeck Janne , Frøkiær Jørgen , Collins Patrick B , Nielsen Søren , Frische Sebastian
Published: June 2021

One week after induction of uncontrolled diabetes mellitus (DM) in female rats by streptozotocin injection, the effect of dietary NaCl restriction on urinary volume (UV) was investigated. In the first experiment (n=6 in each group), rats were offered a synthetic NaCl‐deficient diet (C1036) for 2 days. In DM rats, UV increased dramatically (111±2 vs 211±7 ml/day; p<0.001), whereas non‐diabetic controls showed no change in UV (13.1±1.9 vs 11.7±1.5 ml/day; p=0.6). In the second experiment (n=5 in each group), DM rats receiving C1036 for 2 days again showed a dramatic increase in UV, however the increase and degree of polyuria were similar in DM rats receiving C1036 supplemented with NaCl to normal dietary levels (170±10 vs 174±7 ml/day; p=0.61). No changes were evident in non‐diabetic control rats receiving either diet. In the third experiment (n=8 in each group) standard rat chow (1320) or a custom‐made standard rat chow with reduced (32%) Na+ content (1320‐Na+32%), were given for 7 days. UV was lower in DM rats given 1320‐Na+32% than in DM rats given standard 1320 chow (114±11 vs 151±7 ml/day; p=0.007). No difference was seen between non‐diabetic control groups receiving 1320‐Na+32% or 1320. We conclude 1) administration of the synthetic chow, C1036, to DM rats increased UV dramatically irrespective of the NaCl content, and 2) without a concurrent change in the dietary background a reduction in dietary NaCl leads to decreased UV in DM rats.
Black walnut extract inhibits secretory pathways in equine cecum and colon via mechanisms that overlap with cyclooxygenase signaling
Authors: Rivier Lauraine H , Blikslager Anthony T
Published: June 2021

Disturbances in the hind‐gut have been linked to the induction of equine laminitis, presumably by initiating enteric signaling pathways that result in release of mediators that affect distant fragile vascular beds such as that in the hoof. Laminitis has been experimentally induced by gavaging with black walnut extract (BWE) toxin, leading to changes in both the gut and laminae. Cecal and colonic tissue were mounted on Ussing chambers and treated with escalating doses of BWE (0.6, 1.25, 2.5 and 5%). Changes in short circuit current (Isc), transepithelial electrical resistance, and 3H‐mannitol fluxes were measured. BWE treatment did not affect gut barrier function as there were no significant changes in resistance or mannitol flux. However, BWE caused significant decreases in Isc when compared with controls, most likely due to inhibition of basal Cl− secretion. Pre‐treatment with the cyclooxygenase inhibitor flunixin (2.7μM) resulted in a similar reduction in Isc, which has previously been shown to inhibit Cl− secretion. Treatment with both BWE and flunixin had no additive effect, further suggesting that BWE and flunixin have overlapping signaling pathways. The precise mechanism by which BWE reduces Isc in the equine gut will require further study, but early evidence is suggestive of an inhibitory effect on Cl− secretion. This in turn may affect colonic luminal contents, with systemic effects such as laminitis.
Probiotics restore ion transport in infected intestinal epithelial cells by modulating glycosphingolipids signaling.
Authors: Resta‐Lenert Silvia C , Maruggi Marco
Published: June 2021

Glycosphingolipids (GSL) are part of intestinal epithelial cell (IEC) membranes and are composed by a sugar moiety attached to ceramide. GSL contributes to many cellular functions, including protein sorting, cell adhesion, cell‐cell signaling and apoptosis.

Probiotics are commensal bacteria with the ability to modify cellular processes in a beneficial way. We have shown that a probiotic preparation, S. thermophilus (ST) and L. acidophilus (LA), has the ability in vitro and in vivo to decrease chloride secretion and diarrhea by regulating kinase signaling and levels of ion transport proteins in IEC. Our ST strain expresses bacterial sphyngomyelinase , whereas LA does not. We hypothesized that ST/LA may regulate chloride secretion by altering GSL signaling in IEC. T84 and HT29/cl.19A cells were infected with enteroinvasive E.coli (EIEC) and S.dublin (SD), and were treated with wild type probiotics or mutated ST and/or LA. While wild type ST/LA, LA and ST alone were able to reverse the effects of EIEC and SD on chloride secretion in IEC, a ST strain carrying a mutation of the SMase gene and SMase‐negative commensal Streptococci had no effect on the dysregulated ion transport after pathogens infection. This effect was mediated by ERK1,2 and JNK, as shown by kinases inhibitors treatment. Moreover, we observed significantly altered CFTR and NKCC1 function and cellular distribution after treatment with ST/LA and LA and ST alone after EIEC and SD infection. Our findings provide insight on mechanisms of probiotics effects and on GSL cell signaling in the gut.
Genetic difference between A/J and C57BL/6J mice response to chronic alcohol consumption.
Authors: Croniger Colleen Marie , DeSantis David , Pisano Sorana , Millward Carrie , Hsieh Chang‐Wen , Nagy Laura
Published: June 2021

Genetics contributes to susceptibility and development of disease. In the present study, the response to chronic alcohol was studied in two inbred strains, C57BL/6J and A/J. We report that the C57BL/6J mice developed steatosis with an increase expression of CCAAT/enhancer‐binding protein β (C/EBPβ) gene. The A/J mice were resistant to the development of steatosis due to a decreased C/EBPβ gene expression. However the A/J mice develop liver injury with increased levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and tumor necrosis factor α (TNFα) that is decoupled from hepatic lipid accumulation, suggesting that other mechanisms contribute to alcohol induced liver injury. These data demonstrate that genetics contribute to the pathophysiology of alcohol‐induced liver injury.
Hyperglycemia triggers oxidative stress, Bad O‐GlcNAcylation and increased myocardial apoptosis
Authors: Rajamani Uthra , Essop M.Faadiel
Published: June 2021

Hyperglycemia causes oxidative stress and cell death that may contribute to onset of the diabetic cardiomyopathy. Oxidative stress may increase flux through the hexosamine biosynthetic pathway (HBP) resulting in greater O‐GlcNAcylation of proteins. We hypothesized that higher oxidative stress under hyperglycemic conditions results in greater O‐GlcNAcylation of apoptotic proteins and myocardial apoptosis. H9c2 cardiomyoblasts were exposed to high glucose with/without HBP modulators and an antioxidant. Apoptosis was evaluated by Hoechst staining, Annexin‐V staining and caspase activity assays. Reactive oxygen species (ROS) levels were quantified by H2DCFDA staining using fluorescence microscopy and flow cytometry. Intracellular O‐GlcNAcylation was assessed by Texas Red fluorescence intensity. O‐GlcNAcylation of Bad was determined by co‐localization of O‐GlcNAc (Texas Red) and Bad (FITC) fluorescence staining. High glucose treatment significantly increased apoptosis while intracellular O‐GlcNAcylation, Bad O‐GlcNAcylation and ROS levels were increased in parallel. HBP inhibition attenuated these effects while activation exacerbated it. All detrimental hyperglycemia‐mediated effects were blunted by antioxidant treatment. Our study identified a novel pathway whereby hyperglycemia results in greater oxidative stress, enhanced Bad O‐GlcNAcylation and increased HBP‐mediated myocardial apoptosis. This maladaptive pathway may be triggered in prediabetic/diabetic patients thereby potentially contributing to the onset of the diabetic cardiomyopathy. Research work was funded by the South African NRF and Stellenbosch University.
Hexosamine signaling reduces ER stress‐induced cardiomyocyte death
Authors: Ngoh Gladys Afor , Hamid Tariq , Prabhu Sumanth D , Jones Steven P
Published: June 2021

We have demonstrated that the post‐translational modification of O‐linked beta‐N‐acetylglucosamine (O‐GlcNAc) confers cardioprotection at least partially through mitochondrial dependent mechanisms. Because ischemia/hypoxia causes endoplasmic reticulum (ER) stress, we evaluated a potential link between O‐GlcNAc signaling and ER stress. We pharmacologically induced ER stress to define a potential link between these two processes. Normoxic induction of ER stress (tunicamycin or brefeldin A) augmented O‐GlcNAc signaling in cardiomyocytes (p<0.05). Prior to induction of ER stress, we adenovirally overexpressed or pharmacologically inhibited O‐GlcNAcase, the enzyme responsible for O‐GlcNAc removal. AdO‐GlcNAcase reduced (p<0.05) ER stress‐mediated elevation in O‐GlcNAc levels and aggravated cell death. Conversely, pharmacological inhibition of O‐GlcNAcase resulted in further elevation in O‐GlcNAc levels and significantly (p< 0.05) attenuated ER stress‐induced cell death. We conclude that ER stress acutely recruits O‐GlcNAc signaling, and, such enhanced O‐GlcNAc signaling represents a partially pro‐adaptive response to reduce ER stress‐induced cell death. These results provide new insights into a possible interaction between O‐GlcNAc signaling and ER stress and may partially explain the mechanism of O‐GlcNAc mediated cardioprotection.

Funding: NIH, AHA, and VA
Calcium Transient Evoked by Capsaicin is Enhanced by Tumor Necrosis Factor Alpha (TNFα) in Pulmonary Sensory Neurons
Authors: Hu Youmin , Lin Ruei‐Lung , Gu Qihai , Lee Lu‐Yuan
Published: June 2021

TNFa, a pro‐inflammatory cytokine, is known to be involved in the pathogenesis of allergic asthma. In DRG neurons, the hyperalgesic action of TNFa is mediated in part by upregulation of TRPV1‐mediated responses. This study was carried out to test whether TNFa increases TRPV1 sensitivity in isolated nodose and jugular ganglion neurons innervating the rat lungs. Responses to increasing concentrations of capsaicin (Cap), a selective TRPV1 activator, were determined by measuring the Cap‐evoked calcium transient Δ(Ca2+)i in these neurons. Our results showed: 1) A pretreatment with TNFa (50 ng/ml) for ~20 h increased significantly the peak Δ(Ca2+)i evoked by 100 nM and 300 nM Cap in both nodose and jugular pulmonary neurons; e.g., Δ(Ca2+)i evoked by Cap (300 nM) in TNFα‐treated jugular neurons exceeded that in control neurons by ~133%. 2) The percentages of Cap (300 nM)‐sensitive cells were increased from 58.1% at control to 79.8% after TNFα in nodose neurons, and from 52.8% at control to 73.1% after TNFα in jugular neurons. 3) A pretreatment with the same concentration of TNFα for a shorter duration (1 h) did not significantly enhance the Cap sensitivity. In conclusion, a long‐term application of TNFα induces a potentiating effect on the pulmonary sensory neurons to Cap, suggesting an increase in TRPV1 sensitivity and/or expression in these neurons. (Supported by NIH grant HL67379)
Effect of MPTP and salsolinol on tissue transglutaminase expression in cultured human dopaminergic SH‐SY5Y cells
Authors: Ioudina Marina
Published: June 2021

Tissue transglutaminase (tTG) expression is increased in the brains and cerebrospinal fluid of patients with Parkinson's disease, suggesting a role for the protein in the neurodegenerative process. tTG is a multifunctional protein that catalyzes post‐translational modification of proteins, resulting in the formation of insoluble protein cross linkage or the incorporation of polyamines into protein substrates, and also functions as a G protein hydrolyzing GTP and ATP. The goal of the present study is to determine whether tTG expression is altered in dopaminergic cells following exposure to toxins linked to Parkinson's disease. Immuocytochemistry using a tTG‐specific antibody was used to compare tTG expression in cultured human dopaminergic SH‐SY5Y cells before and after exposure to the environmental toxin 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) and the endogenous dopamine‐derived neurotoxin salsolinol. Both toxins significantly increased tTG expression in SH‐SY5Y cells, suggesting a direct effect of toxins on tTG expression. Further investigations are necessary to determine whether the upregulation of tTG is correlated with increased tTG activity.

This work was supported by Touro University Nevada Research Grant Award.
Flavonoids from Inula Britannica L. inhibits injury‐induced neointimal hyperplasia through suppressing oxidative stress generation
Authors: Zhang Hong‐Bing , Wen Jin‐Kun , Wang Yan‐Yan , Zheng Bin , Han Mei
Published: June 2021

The total flavonoid extracts (TFE) were prepared from Inula Britannica L., containing 45.5% quercetin, 25.85% luteolin, 12.96% 6‐methoxyluteolin, 4.33% spinacetin and 1.85% isorhamnetin. To explore the effect of TFE on neoinimal hyperplasia induced by balloon injury, the rats administered orally TFE at doses of 12.5, 25, and 50mg/kg/d by gastric gavage from 3 days before balloon injury to 14 days after the injury, respectively. We found that the rats treated with high dose of TFE (50 mg) showed a reduced neointimal hyperplasia, and the ratio of intima (I) to media (M) area (I/M) of balloon injured‐carotid arteries was significantly reduced by over 70% after TFE treatment, compared with model group. The inhibitory effect of TFE (50 mg) on neointimal hyperplasia was almost consistent with Atorvastatin, as a positive control. The plasma SOD activity was obviously increased in TFE (50 mg)‐treated group (P < 0.01), meanwhile, plasma MDA production was markedly decreased (P < 0.05). At 14 days after balloon injury, the neointimal and medial layer of the carotid arteries showed an increase in O2? production. TFE (50 mg) significantly prevented the injury‐induced O2? production, and inhibited the injury‐induced p47phox expression in carotid arteries. These results suggest that the TFE inhibits the neointimal hyperplasia induced by balloon injury through suppressing oxidative stress generation.
A transcriptomic approach to the survival signaling enhanced by Neuroprotectin D1 in response to oxidative stress
Authors: Calandria Jorgelina M , Bazan Nicolas G
Published: June 2021

Oxidative stress triggers apoptotic damage on RPE cells, enhancing the enzymatic production of neuroprotectin D1 (NPD1) that down‐regulates the expression of pro‐inflammatory enhancing the expression of the anti‐apoptotic proteins. We hypothesize that NPD1 signaling promotes the modulation of a group of genes that contributes to the homeostatic survival response of RPE cells. 15LOX1 knockdown cells were used to study the NPD1‐dependent transcriptional control. The oxidative stress‐induced expression in the silenced cells and controls with and without NPD1 was assessed by microarray technology. To construct the NPD1‐transcriptome, enrichment analysis was used to identify the over‐represented genes for common, similar and unique sets. Functional ontologies, Gene Go and GO cellular processes were applied. More than 300 genes were shown to be modulated in response to NPD1. Chemokines such as pro‐mitotic CXCL1 were down‐regulated by 6 hs of oxidative stress and up‐regulated by NPD1, and CXCL3 was down‐regulated by NPD1. The expression of transcriptional regulators was observed at early times of oxidative stress treatment. MAPK pathway components also were modulated by NPD1. The transcription factor network analysis showed the involvement of NF‐kB, c‐Jun and EGR1 among others. Different cellular processes were affected by NPD1 treatment involving stress and immune response signaling via MAPK. The data suggests that NPD1 induce the modification of gene expression, enhancing the intrinsic pathways related with RPE cell survival regulation. (NIH/NEI Grant EY005121).
Ethanol Inhibition of Methionine Synthase in Rat Liver and Cortex: Role of Glutathione Depletion and Different Cobalamin Cofactors
Authors: Waly Mostafa Ibrahim‐Ahmed , Kharbanda Kusum , Deth Richard
Published: June 2021

Vitamin B12 is an essential cofactor for normal activity of the ubiquitous enzyme methionine synthase (MS), which methylates homocysteine as part of the methionine cycle. Studies have shown that chronic ethanol (ETOH) exposure in rats results in inhibition of MS and depletion of glutathione (GSH), the major intracellular antioxidant. We investigated the ability of either hydroxocobalamin (OHCbl) or methylcobalamin (MeCbl) to support MS activity. OHCbl is converted to MeCbl via a GSH‐dependent mechanism. In liver and cortex of the control animals, MeCbl‐based activity was higher than OHCbl‐based MS activity. ETOH treatment reduced both MeCb1 and OHCb1‐based MS activity in liver; however, in cortex ETOH reduced OHCb1‐based MS activity, but not MeCb1‐based activity. ETOH lowered liver and cortex GSH levels by 30%. Our results indicate that ethanol inhibits MS activity in both liver and cortex in conjunction with depleted intracellular GSH levels, but only in cortex was a unique protective effect for MeCbl observed against the ETOH‐mediated inhibition of MS activity. Thus intracellular oxidative stress may be responsible for the ETOH‐induced decrease in MS activity, and MeCb1 and/or GSH administration could represent a strategy to overcome this inhibitory effect.
Possible protective mechanism for dietary glutamate against Helicobacter pylori‐induced gastric atrophy in rodents
Authors: San Gabriel Ana Maria , Nakamura Eiji , Hasumura Mai , Uneyama Hisayuki , Torii Kunio
Published: June 2021

L‐glutamate is an abundant dietary amino acid, which can be free or bound to other amino acids within proteins. It plays an essential role in transdeamination and gluconeogenesis, and becomes a substrate for the synthesis of the antioxidant glutathione and the polyglutamated folic acid. The intestine extracts most of the dietary glutamate where acts as the main fuel for nutrient absorption. However, glutamate metabolism in the gastric mucosa is poorly understood. Recently, we found that dietary supplementation with monosodium glutamate can ameliorate the inflammation and the atrophy of pepsinogen‐secreting chief cells and acid secreting parietal cells caused by chronic infection with Helicobacter pylori. To uncover the protective mechanism of glutamate, we isolated chief cells from rat stomach by counterflow elutriation and density gradient. When cells became confluent, we exposed them to either buffer or 20 mM glutamate for 2 h. The molecular profile between both treatments was compared by GeneChip analysis, which suggested that L‐glutamate may detoxify ammonia through the synthesis of glutamine and signal cells through glutamate receptors and glutamate transporters.
Women with gestational diabetes mellitus (GDM) have breast milk with lower concentration of docosahexaenoic acid (DHA) compared to women without GDM
Authors: Salguero Julissa Marisel , Fontenot Emily , Page Alicia , Durham Holiday , Servan Olivia , Tang Tianyi , Tulley Richard , Elkind‐Hirsch Karen , Gordon Stewart , Lammi‐Keefe Carol
Published: June 2021

Arachidonic acid (ARA) and DHA are essential for normal infant growth, vision, and neurological development. Breast feeding is encouraged, but data on breast milk composition for women with GDM are lacking. Fatty acid (FA) content of breast milk of women with (n=5, GDM) and without (n=9, C) GDM was assessed. Milk samples were collected at 2, 6, 10 and 12 weeks using a breast pump. Milk FA methyl esters were prepared by direct transesterification and separated, identified and quantitated with gas chromatography. GDM not consuming fish oil/DHA supplement during lactation had lower milk DHA (wt %) compared to C without supplementation at weeks 2, 6, and 10 (2 wks: GDM, 0.28; C, 0.36, p = 0.03; 6 wks: 0.16 vs. 0.28, p = 0.006; 10 wks: 0.17 vs. 0.30, p = 0.004). There was no difference for ARA. Further, GDM women had lower milk n‐3/n‐6 ratios at all weeks (2 wks: 0.22 vs. 0.27, p = 0.08; 6 wks: 0.20 vs. 0.28, p = 0.02; 10 wks: 0.23 vs. 0.32, p = 0.005; 12 wks: 0.22 vs. 0.28, p = 0.06). Mean dietary DHA intake was similar for groups (40 mg/day). ARA mean dietary intake was 95 mg/day for GDM and 130 mg/day for C. Based on these findings and the previous finding of Lammi‐Keefe (Wijendran et al, Lipids, 2000; 35; 927‐931) demonstrating that infants of women with GDM have low concentration of DHA at birth, the importance of DHA supplementation during breast feeding for women with GDM is underscored. (Supported in part by National Fisheries Institute and LSU AgCenter).
A new method for the detection of mitochondrial oxidative stress
Authors: Speicher Danielle M , Hardwick James , Chilian William , Yun June
Published: June 2021

A consequence of mitochondrial (mt) electron transport and energy production is continual formation of reactive oxygen species (ROS). MtROS production is usually compensated by high activities of antioxidants, e.g., superoxide dismutases 1 and 2. However, under various physiological or pathological states, mtROS production exceeds antioxidant defenses and oxidative (Ox) modification of mt proteins and mtDNA occurs. Current techniques to detect mtOx stress include estimations of ROS, Southern blotting to measure mtDNA fragmentation, or immunostaining and HPLC to measure Ox stress biomarkers such as 8‐hydroxy‐2′‐deoxyguanosine (8OHdG). These techniques are laborious and require specialized procedures (use of radioisotopes) or equipment (HPLC). Our goal was to devise and evaluate a new, simple method of assessing mtOx stress. Total cellular or mtDNA was isolated from liver and heart from Zucker lean and obese rats. Equal amounts of DNA were fixed to a nitrocellulose membrane and an immunoblotting protocol was performed with an antibody against 8OHdG. Levels of 8OHdG in heart mtDNA were less in Zucker lean than those from obese rats, consistent with other measurements from our laboratories showing greater mtROS production in obese compared to lean rats. The proposed method is useful to measure ROS‐induced DNA damage because it is convenient, semi‐quantitative, and accurate even with small amounts of DNA.
Healthy Bodies, Healthy Souls: Design of a church‐based environmental intervention program to prevent diabetes risk behaviors
Authors: Gittelsohn Joel , Summers Amber , Rowan Megan , Goheer Attia , Martins Paula , Graham Karlene , Mahendraratnam Nirosha
Published: June 2021

Churches have great potential to engage communities and sustain health interventions. We are conducting a diabetes risk behavior intervention trial in 12 African American churches in Baltimore city, divided into 4 cohorts (3 churches/cohort) matched by size and denomination. One church in each cohort is randomly selected to receive a workshop intervention (American Diabetes Association's Project POWER), while a second church receives an environmentally‐enhanced intervention plus workshop intervention, and a third church is comparison. The enhanced program includes changes to church food and physical activity environments. The program will be evaluated in 375 non‐diabetic adults, using psychosocial, behavioral (diet, physical activity) and health (BMI, blood pressure, waist circumference) measures. Initial baseline data (n=138) show that 68% are female, mean age 48 years. Over 78% of respondents have a high school degree, and 73% have household incomes >$30,000. 32.4% of the sample is obese and 51.8% are overweight. 44.2% are prehypertensive and 24.6% are hypertensive. Mean waist circumference is 100.8cm (SD: 16.9) for men and 97.2cm (SD: 18.2) for women, which indicate that 40.9% of men and 69.1% of women at high risk for heart disease. This study will test church‐specific interventions to reduce diabetes risk behaviors and improve health measures in a high‐risk minority population.

Grant Funding Source: American Diabetes Association Clinical Research Award
Activity‐Based Probes for Penicillin‐Binding Protein Imaging
Authors: Kocaoglu Ozden , Lanning Bryan , Cozy Loralyn , Calvo Rebecca , Kearns Daniel , Carlson Erin E.
Published: June 2021

We aim to investigate the timing and localization of penicillin‐binding proteins (PBPs) during peptidoglycan (PG) biosynthesis using a combination of fluorescence microscopy, gel electrophoresis, and liquid chromatography‐mass spectrometry (LC‐MS) techniques facilitated by the use of a β‐lactam‐based probe. β‐lactam‐containing compounds are the most widely used class of antibiotics that inhibit PG biosynthesis by binding PBPs. We designed and synthesized fluorescent and biotin derivatives of the antibiotic cephalosporin to selectively label high molecular weight (HMW) PBPs in B. subtilis PY79 in vivo. Our microscopy results indicate that the probe facilitated specific labeling of active HMW PBPs that are localized at newly synthesized septa. The MS analysis identified the labeled proteins as PBP 1a and b, PBP 2a, PBP 2b and PBP 4. Both microscopy and gel‐based analysis indicated that our probe gives cleaner staining and is more selective than BOCILLIN‐FL, a commercially available penicillin analog, which labels both high and low molecular PBPs. Our cephalosporin probe is currently being applied to time‐course studies to visualize the roles of HMW PBPs in B. subtilis.
Urinary sugars (sucrose and fructose) associations with self‐reported sugars intake: the influence of plausibility of reported energy intake
Authors: Sharma Rashmi S , Roberts Susan B. , Polissar Nayak L. , Boushey Carol J. , Maroney Owen H. , Fieselmann Krista N. , Ebner Rachel E. , Fuss Paul J. , McCrory Megan A.
Published: June 2021

Background/Aim

Urinary sucrose and fructose may serve as biomarkers of sugars intake, the latter which are thought to be underreported in dietary assessment. We examined associations of urinary sugars with reported sugars intake in adults recruited for a study on diet and chronic disease risk.

Methods

Healthy, non‐smoking, weight‐stable adults (n=76) performed 3×24h urine collections (UC) and completed 3 multiple pass 24h dietary recalls over 10d. Results are shown for those (n=67; aged 18–58y, BMI 19–36kg/m2) who had ≥1 complete UC (urinary PABA recovery ≥85%). Urinary sugars were measured by an enzymatic kit (Roche) and mean daily amounts calculated. 3d mean intakes of sucrose, fructose and added sugars were computed with Nutrition Data System for Research (v.10/11). Plausible (n=52) and implausible EI reporters (n=15) were determined (Huang et al, 2005).

Results

Mean(±SEM) urinary sucrose and fructose were 28±2 and 38±4 mg/d, respectively. Reported intakes were 56±5 (sucrose), 25±2 (fructose), and 84±7 (added) g/d. Urinary sucrose was significantly associated with dietary sucrose in the plausible sample (r=0.36; p=0.008) and non‐significantly in the total sample (r=0.19; p=0.12). A similar pattern was seen for urinary sucrose and added sugars intake. Urinary fructose and sugars intake were not significantly associated.

Conclusion

These data provide strong evidence that foods containing sucrose and added sugars are underreported.

Grant Funding Source: R01DK075862 and Purdue University
Orange juice consumption is associated with enhanced antioxidant intake status in the U.S. population: a cross‐sectional study
Authors: Lee Sang Gil , Lloyd Beate , Wang Ying , Yang Meng , Chung Sang-Jin , Chun Ock K.
Published: June 2021

Antioxidants, including polyphenolic antioxidants, are important to reduce oxidative stress and inflammation, key mechanisms in the development and progression of chronic diseases. The aim of the study was to evaluate the impact of 100% orange juice (OJ) on diet quality and antioxidant intake status of 13,971 U.S. population aged ≥ 4 y in the NHANES 2003–2006. OJ was found to be the largest single contributor to fruit intake and 3rd largest contributor to total antioxidant intake (TAC) in the US, with tea #1 and vitamin C supplements #2. Furthermore, OJ consumers (OJC) had a healthier life style as well as greater intakes of whole fruits and vegetables than non‐OJ consumers (OJNC). OJC had higher intakes of carotenoids, vitamin C, and flavonoids compared to OJNC after adjusting for age, gender, ethnicity, and energy intake (p<0.01). Particularly, the intakes of β‐cryptoxanthin, hesperidins, and naringenins were increased by the amount of OJ consumption (p<0.001). Dietary TAC also showed similar increases relative to OJ consumed compared to NOJC (p<0.001). Overall, OJC have higher antioxidant intakes and greater intakes of fruit than OJNC.
Early life risk factors for obesity in childhood: A cumulative risk model
Authors: Dev Dipti , McBride Brent
Published: June 2021

The objective of this study is to identify multiple levels of influence in predicting weight gain in pre‐school children (2–5 years). A cumulative risk approach is taken to predict overweight (85th percentile ≤BMI <95th percentile) and obesity (95th percentile ≤BMI) based on the following predictors (SES, ethnicity, maternal BMI, family size, breast feeding duration, age of introduction of solid foods, parental feeding practices, parental fruit and vegetable intake, family media use, family encouragement of physical activity, child food consumption, sleep patterns and childcare attendance). Baseline data from the Synergistic Theory and Research on Obesity and Nutrition Group (STRONG) kids were used for data analysis. Primary caregiver‐child dyads (n= 497) were recruited from childcare centers in Eastern Illinois. Caregivers completed survey and child height and weight was measured. Current sample statistics n= 497 (254 males, 293 females), with 34 Hispanics, 131 Blacks, 278 Whites and 54 Asians. 254 children enrolled in Child and Adult Care Food Program (CACFP), 169 Non‐CACFP and 74 in Head Start programs. 25% of children were classified as overweight. Using a cumulative risk model, it was found that the combination of child, family level and childcare risk factors could predict overweight and obesity. The results suggest the need for targeted obesity interventions based on above characteristics.

Grant Funding Source: Administration of Children and Families(ACF), and the Illinois Transdisciplinary Obesity Prevention Program (I‐TOPP)
Don't forget about dad: Exploring paternal influences on children's weight status and eating behaviors
Authors: Vollmer Rachel L. , Sigman-Grant Madeleine , Mobley Amy R.
Published: June 2021

While mothers remain the primary gatekeeper for children's nutrition, fathers have assumed more child care duties over time. The objective of this study was to examine the relationship of paternal feeding style (Caregiver Feeding Style Questionnaire) and eating behaviors (Healthy Children, Healthy Families checklist) on child's weight status and associated behaviors. Low‐income fathers (n=30) were interviewed individually. Each father and child (ages 3–10 years) were weighed and measured. Authoritarian (37%) and indulgent (33%) feeding styles were most common while authoritative (17%) and uninvolved (13%) were less common among fathers. Feeding style was not associated with a child's weight but paternal BMI was associated (r=.377, p=.04) with child's BMI z‐score. High responsiveness feeding scores of fathers were correlated with frequent family meals (rs=.545, p=.002) and less soda consumption (rs=‐.560, p=.001) in children. Father's reported vegetable (rs=.722, p<.001), dairy (rs=.634, p<.001) and soda (rs=.548, p=.002) intake was significantly associated with their child's reported intake. Further research should address paternal influences on children's eating behaviors and weight status.

Grant Funding Source : none