bluuebunny/crossref_metadata_2025 huggingface dataset:
Ceramide 1‐phosphate transfer protein depletion in the cell drives autophagic response: potential implications of this newly identified protein in cell death and survival (737.6)
Authors: Mishra Shrawan , Karanjeet Kul , Zou Xianqiong , Hinchcliffe Edward , Brown Rhoderick
Published: June 2021

Ceramide 1‐Phosphate (C1P) is a biologically active sphingolipid that stimulates cell proliferation, macrophage migration, and inflammatory events. We recently characterized a novel human lipid transfer protein encoded by GLTPD1 on chromosome 1 (locus 1p36.33). We named the protein, ceramide 1‐phosphate transfer protein (CPTP) due to its lipid specificity and discovered that CPTP transports C1P to the plasma membrane from TGN after synthesis by ceramide kinase. CPTP depletion by RNAi leads to C1P accumulation in TGN (~6‐fold), stimulating cPLA2α and inducing pro‐inflammatory eicosanoid production (Simanshu et al., Nature 500: 463‐67, 2013). Here, we investigated whether CPTP depletion leads to programmed cell death and discovered an autophagic response in human cervical cancer cell lines (HeLa) and human lung adenocarcinoma epithelial cells (A549). Transfection with ShRNA (pSuper.GFP.ShRNA.CPTP vector) resulted in GFP‐LC3 puncta and acidophilic acridine orange staining by epifluorescence microscopy and beclin 1(BECN1) overexpression by RT‐PCR. CPTP mutants deficient in C1P binding/transfer (K60A & R106L) also exhibited autophagy. We conclude that normal levels of viable CPTP are needed for cells to suppress autophagy.

Grant Funding Source: [Support: NIGMS‐45928 & 72754; Hormel Fnd]
Regulation of Hsf4b nuclear translocation and transcription activity by phosphorylation at threonine 472 (739.2)
Authors: Hu Yanzhong
Published: June 2021

Hsf4b, a key regulator of postnatal lens development, is subjected to posttranslational modifications including phosphorylation. However, the phosphorylation sites in Hsf4b and their biological effects on the transcription activity of Hsf4b are poorly understood. Here we examined 17 potential phosphorylation residues in Hsf4b with alanine‐scanning assays and found that a T472A mutation diminished Hsf4b‐mediated expression of Hsp25 and αB‐crystallin. In contrast, the phosphomimetic mutation of T472D enhanced their expression. Further investigation demonstrated that Hsf4b could interact with nuclear‐transporter importin β‐1 and Hsc70 via amino acids 246‐320 and 320‐493, respectively. T472A mutation reduced Hsf4b’s interaction with importin β‐1, while enhancing its interaction with Hsc70, resulting in Hsf4b cytosolic re‐localization, protein instability and transcription activity attenuation. At the upstream, MEK6 was found to interact with Hsf4b and enhance Hsf4b’s nuclear translocation and transcription activity, probably by phosphorylation at sites such as T472. Taken together, our results suggest that phosphorylation of Hsf4b at T472 by protein kinases such as MEK6 regulates Hsf4b interaction with the importin β‐1‐Hsc70 complex, resulting in blockade of it nuclear translocation and transcriptional activity of Hsf4b.

Grant Funding Source: national nature sciences foundation of china
Variations in phytochemicals, proximate and nutrient constituents of Trichosanthes cucumerina L. plant (804.32)
Authors: Adeyemo Oyenike , Olaitan Olanrewaju , Smith Oshinowo
Published: June 2021

Trichosanthes cucumerina L. is used for food and medicine. In this study, the young and old T. Cucumerina seeds, leaves, fruits and roots (dried powder) were analysed for phytochemicals, proximate and nutrient constituents using chemical tests and standard spectrophotometric methods. Phytochemical qualitative analysis revealed the absence of cardiac glycosides, and saponin in all plant parts while tannins and phenols were present only in old leaves and terpenoids in young and old seeds and roots. Phlobatanin is not contained in young fruit and seeds but present in other parts. Steroids, flavonoids, alkaloids and reducing sugar were found to be major phytochemicals in all plant parts and their levels increased significantly (P < 0.05) in T. cucumerina old parts. The young and old parts of T. cucumerina variant contain varying concentration of nutrients. Calcium ranged from 13.05 to 152.41 mg/100 g with a mean of 88.26 mg/100 g, potassium (13.20‐26.60, mean = 23.32 mg/100 g), magnesium (30.28‐50.10, mean = 39 mg/100 g), sodium (3.45‐52.25, mean = 22.9 mg/100 g and iron (10.40‐65.50, mean = 32.96 mg/100 g) were the most abundant. Proximate of all parts showed a high moisture content (mean =71.8 %) except in roots and old seeds contained high crude fats (mean = 15.75 %). The study has revealed the T. Cucumerina variant has useful nutrients and flavonoids and can be utilized as sources of minerals and antioxidants respectively.
Associations of breakfast consumption patterns with body weight across 10 years: results from NHANES 2001‐10 (810.22)
Authors: Albertson Ann , Thompson Douglas
Published: June 2021

The association of breakfast consumption patterns with body weight has been documented in several studies. This study expanded the existing literature in two ways: 1) the association of breakfast consumption patterns (Ready‐to‐Eat Cereal vs. Other Breakfast vs. Skipped Breakfast) with body weight measures was examined in a large, diverse sample spanning 10 years (NHANES 2001‐10); and 2) changes in breakfast consumption patterns across 10 years were described within the same sample. The results were broken out by age (children and adolescents vs. adults). Results generally replicated past studies indicating that Ready‐to‐Eat Cereal breakfast is associated with the most favorable body weight measures and skipping breakfast is associated with the worst body weight measures, even after statistically adjusting for relevant covariates. However, specific patterns differed somewhat by age and body weight measure (Body Mass Index, Waist/Height Ratio, Overweight/Obese vs. Not). The types of food consumed at breakfast differed markedly between age groups; for example, coffee was most frequent among adults, whereas pancakes/waffles/french toast were much more frequent among children and adolescents. There were also noticeable shifts in breakfast foods consumed across the 10 years. For example, children and adolescents (not adults) showed an increasing trend in consumption of pancakes/waffles/french toast, whereas adults (not children and adolescents) showed a strongly increasing trend in oatmeal consumption.
Anti‐adipogenic properties of selected phenolics in 3T3‐L1 cell model (811.31)
Authors: Tan Huiyuan , Tse Iris M.Y. , Li Edmund T.S. , Wang Mingfu
Published: June 2021

Many phenolics are known to possess anti‐obesity properties through mechanisms that inhibit adipogenesis, stimulate lipolysis or induce apoptosis. This study aimed to examine the anti‐adipogenic effects of two phenolics, oxyresveratrol and cyanomaclurin in 3T3‐L1 preadipocytes. At doses that did not induce cytotoxicity (based on the lactate dehydrogenase assay), the exposure of oxyresveratrol (0‐100μM) or cyanomaclurin (0‐600μM) in the first three days of differentiation led to a dose‐dependent decrease in triglyceride accumulation on day 9 post‐confluent. Results of cell proliferation [3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐Diphenyltetrazolium Bromide (MTT) assay] and cell cycle [Fluorescence‐activated cell sorting (FACS) analysis] studies demonstrated that both compounds inhibited differentiation through inducing cell cycle arrest by retaining the preadipocytes in the G1 phase during the first two days post‐confluent. By studying the gene expression of the major transcriptional factors (Peroxisome proliferator‐activated receptor γ, CCAAT/ Enhancer‐binding proteins α), as well as their effects on related proteins expression in cell cycle modulation (cyclins and cyclin dependent kinase inhibitors), the regulatory pathways are identified. This research helps to identify novel agents with anti‐obesity properties by provide preliminary data on their mechanism of actions.
The c‐RAF/MEK/ERK MAPK pathway regulates a class of GCN2‐independent amino acid response genes (818.6)
Authors: Donelan Will , Shan Jixiu , Kilberg Michael
Published: June 2021

During adaptation to protein/amino acid (AA) limitation, the general control non‐derepressible 2 (GCN2) serine/threonine protein kinase initiates the amino acid response (AAR) by binding uncharged tRNAs, which activates its kinase activity. Subsequently, GCN2 phosphorylates eukaryotic initiation factor 2 (eIF2) leading to suppression of general protein synthesis, but induction of translation for specific mRNA species, including that for the transcription factor ATF4. ATF4 is a major effector of AAR‐induced transcription for many genes involved in restoring homeostasis or inducing apoptosis. GCN2 is currently the only well characterized sensor for AA deficiency in mammalian cells and the GCN2‐eIF2‐ATF4 pathway is considered the predominant AAR signaling mechanism. However, evidence from several laboratories has been published suggesting that GCN2‐independent AAR signaling exists. In this study, HepG2 human hepatoma clonal cell lines, expressing shRNA against GCN2, were generated to investigate the possibility of alternative AAR signaling mechanisms. Based on previous research, the MAPK pathways were likely candidates and therefore, were screened using chemical inhibitors. Using IL‐8 as a model MAPK‐responsive and AA‐responsive gene, it was established that c‐RAF/MEK/ERK signaling contributes to GCN2‐independent regulation of a wide spectrum of genes following AA limitation.

Grant Funding Source: Supported by NIH, DK092062 and DK094729
High dose C‐3 alpha epimer of 25‐hydroxyvitamin D over 8 weeks reduces cortical mineral density of lumbar vertebra in weanling rats (828.2)
Authors: Djekic‐Ivankovic Marija , Mak Ivy , Wang Krystyna , Lavery Paula , Agellon Sherry , Weiler Hope
Published: June 2021

C‐3 alpha epimer (EPI) of 25‐hydroxyvitamin D (25(OH)D) is elevated in infancy. Short‐term treatment with the EPI of calcitriol reduces parathyroid hormone (PTH) in rats. This study tested the effect of EPI on growth, bone biomarkers and volumetric bone mineral density (vBMD). Weanling rats (4 wk; n=6 male; n=6 female/diet) were randomized to receive AIN‐93G diets modified in vitamin D for 8 wk: control AIN‐93G (C‐1: 25 µg vitamin D3/g diet), control 25(OH)D (C‐2: 12.5 µg 25(OH)D/g diet), level one EPI (EPI‐1: 12.5 µg EPI/g diet) or level two EPI (EPI‐2: 25 µg EPI/g diet). Measures were weight and biomarkers at 0, 4 and 8 wk and in vivo lumbar vertebra 3 vBMD using microcomputed tomography (LCT‐200, Aloka) at 0 and 8 wk. Differences among groups were tested using ANOVA, Tukey’s post hoc test and P<0.05. No differences in vitamin D status as measured using LC‐MS/MS were observed at baseline. Average serum EPI (range ng/ml) at 8 wk was C‐1: 4 (0‐12), C‐2: 2 (0‐7), EPI‐1: 49 (27‐78) and EPI‐2: 98 (41‐173). Weight, ionized Ca and C‐telopeptides of Type 1 collagen were not affected by diet. PTH was lowest in EPI‐2 vs all groups at wk 4; and lower in EPI‐1 and EPI‐2 vs C‐2 at wk 8. Cortical vBMD was lowest in EPI‐2 vs all other groups at wk 8; but not trabecular or total vBMD. These data suggest that pharmacological concentrations of EPI reduce PTH and cortical density during rapid growth.

Grant Funding Source: Supported by NSERC and Dairy Farmers of Canada
Feeding patterns and dietary diversity scores of 6 to 36 month old children from Retalhuleu, Guatemala (828.5)
Authors: de Ponce Silvia , Alfonso Víctor , Reinhart Gregory , Villanueva Lisa
Published: June 2021

Feeding pattern and dietary diversity was evaluated in 339 stunted children (HAZ < ‐1) 6‐36 mo of age from Retalhuleu using a food frequency questionnaire (Food and Nutrition Technical Assistance protocol). Dietary assessment was utilized to determine the frequency of food consumption for 7 consecutive days and was considered adequate if the food was included a minimum of 3x/wk in 75% of the population. Results indicated breast milk consumption was not adequate; 73% of children 0‐6 mo were breastfed at least 3x/wk and consumption decreased to 61% in children 19‐24 mo. The primary source of vitamin A was fortified sugar resulting in 88‐96% of the age groups having adequate vitamin A consumption patterns; vegetables rich in vitamin A were not consumed (0%). Meat and poultry had adequate consumption in the 25‐30 mo age group (80%), while younger children had intakes less than 75%. Consumption of cereals and grains increased with age and adequate consumption averaged 84% for infants and 86% for children 25‐36 mo. Hen eggs were the most frequently consumed food item (97%) as were instant soups, bean broths and other liquids (84%). In conclusion, diet diversity is poor in infants and young children from Retalhuleu, Guatemala. Fortified sugar and hen eggs were the only items consumed adequately across all age groups. Opportunities exist to increase breast feeding and improve complimentary feeding practices.
Anti‐aging and neuroprotective effects of Jambolan (Eugenia jambolana) L. via modulation of stress response signaling in C. elegans (829.26)
Authors: Bezerra Maria , Al‐Ani Maraim , Correia Roberta , Vattem Dhiraj
Published: June 2021

The effect of Jambolan (Eugenia jambolana) L. spray dried and freeze dried powders (EJP) on modulating stress response signaling was evaluated in transgenic C. elegans carrying promoter‐GFP constructs for several genes in the IGFR/PI3K/AKT, ARE/NRF2 and Sir2.1/AMPK signaling pathways. Relative fold changes in gene expression in response to EJP treatments were quantified flurometrically. Effect of EJP on longevity was evaluated in C. elegans lifespan assays. Anti‐neurodegenerative effects of EJP were evaluated in C. elegans models for Alzheimer’s disease (AD). Where, time to thermally induced Aβ1‐42 aggregation mediated paralysis was evaluated in transgenic C. elegans (CL4176). Results suggest that the processing technology used to produce EJP powders significantly impacted bioactivity. An upregulation in ARE and Sir2.1 stress response genes was noted in response to EJP treatment. A 10‐13% increase in lifespan was observed with EJP treatment. Supplementation with polar acidic, polar basic and polar neutral fractions delayed Aβ1‐42 induced paralysis by 8‐14%.

Grant Funding Source: Supported by: Vrdür Foundation (USA) & CAPES (Brazil)
Additon of atorvastatin and lycopene produce hypolipidemic and morphological changes in liver of a NAFLD rat model (829.34)
Authors: Piña Rosa , Ramos‐Gómez Minerva , García Olga , Rosado Jorge
Published: June 2021

The present study evaluated the effect of atorvastatin (AT) and/or lycopene (LYC) on fatty liver induced with a high‐fat and high‐cholesterol diet (HFHC). Eighty two male Sprague Dawley rats (200 ± 10g) were randomly assigned to 8 groups: normal 4‐weeks (N4), HFHC diet for 4 weeks (model), normal 8 weeks (N8), HFHC diet for 4 weeks followed by 4 weeks of standard diet (DC), HFHC diet for 4 weeks followed by 4 weeks of standard diet plus 50 mg/kg AT (DC+AT50), HFHC diet for 4 weeks followed by 4 weeks of standard diet plus 20 mg/kg LYC(DC+LYC20), HFHC diet for 4 weeks followed by 4 weeks of standard diet plus 50 mg/kg AT and 20 mg/kg LYC (DC + AT50 + LYC20) and HFHC diet for 4 weeks followed by 4 weeks of standard diet plus 20 mg/kg AT and LYC respectively (DC + AT20 + LYC20). Treatments were given daily by oral intubation. Blood samples and livers were taken for the determination of serum lipid profile and histopathological examination. The model group showed a significant elevation of serum TC, TG, HDL and LDL concentrations in comparison with the normal N4 group. Interestingly, DC group showed similar lipid profile than N8 group and we observed no further reduction by the AT and/or LYC treatments. The model group showed steatosis grade II in 25% and grade III in 75% of the livers. In all the treated groups steatosis decreased significantly. In conclusion, AT and LYC improved liver morphology thus suggesting that these treatments have additional benefits than just the change of diet.

Grant Funding Source: Partially supported by CONACYT
Organic and conventional farming: differences in polyphenol content and antioxidant activity of lettuce cultivated on developed nature‐friendly compost (830.6)
Authors: Park Jeong‐Eun , Kim Min‐Ah , Kwon Dae Young , Jeong Su‐Jin , Noh Sun‐Ok , So Byung‐Ok , Chae Soo‐Wan , Cha Youn‐Soo
Published: June 2021

The purpose of this study was to evaluate antioxidant and cancer cell growth inhibition activity by extract nature‐friendly compost, organically and conventionally cultivated lettuce. We determined the anthocyanin, phenol and flavonoid contents and examined antioxidant assay, such as DPPH, ABTS and reducing power. Also, we performed HepG2 cancer cell growth inhibition assay of lettuce extract. The lettuces of nature‐friendly compost and organic exhibited the highest values of antioxidant activity, total polyphenol and flavonoid contents were the highest in nature‐friendly compost lettuce and organic lettuce. In cancer cell growth inhibition activity in a dose‐dependent manner, nature‐friendly compost lettuce extract had growth inhibition activities greater than 90% on HepG2 cell. The results of this study suggest that the antioxidant and anticancer activities in lettuce cultivars grown under nature‐friendly compost and organic conditions are a promising source of functional food ingredients. (This study was supported by a grant (CUHBRI‐2012‐0 2‐007) of the CNUH‐BRI)

Grant Funding Source: supported by a grant (CUHBRI‐2012‐0 2‐007) of the CNUH‐BRI
Tart cherry juice increases sleep time in older adults with insomnia (830.9)
Authors: Liu Ann , Tipton Russell , Pan Weihong , Finley John , Prudente Alfredo , Karki Namrata , Losso Jack , Greenway Frank
Published: June 2021

Recent studies indicate that tart cherries (Prunus cerasus) may have sleep‐enhancing effects. In a randomized crossover trial, we evaluated the effects of tart cherry juice vs. a placebo juice on various markers of sleep quality in older adults (age 68 ± 9 years). Seven participants with insomnia completed two treatment periods (tart cherry juice and placebo juice), 2 weeks each, separated by a 2 week washout period. Participants consumed 8 ounces of juice twice per day. Overnight polysomnography (PSG) was used to evaluate sleep architecture, such as the distribution of sleep stages, sleep latency and state transitions. Participants also completed questionnaires related to sleep, depression, anxiety, and fatigue. Tart cherry juice consumption increased sleep time by 85 ± 17 minutes compared to placebo as measured by PSG (P=0.02). Expression of indoleamine 2,3 dioxygenase (IDO) and PGE2 decreased after tart cherry juice consumption (P<0.05) suggesting that its effects on sleep can in part be ascribed to the improvement in the expression of IDO and PGE2. This work is registered at ClinicalTrials.gov NCT01669317.

Grant Funding Source: Supported by the Cherry Marketing Institute
Allele frequency of a 22‐bp insertion/deletion polymorphism of FADS2 in a U.S. population (831.6)
Authors: Kothapalli Kumar , Carlson Susan , O'Brien Kimberly , Ojukwu Kinsley , Zou James , Hyon Stephanie , Ran‐Ressler Rinat , Brenna James Thomas
Published: June 2021

Introduction. Fatty acid desaturase 2 (FADS2) is a critical rate limiting enzyme in the biosynthesis of long chain polyunsaturated fatty acids (LCPUFA). Recently, we identified a 22 bp insertion/deletion (INDEL) polymorphism within FADS2 intron 1 near a sterol response element to be associated with desaturase expression depending on levels of SREBP‐1c agonists. Cultured lymphocytes from individuals (n=16 Japanese) homozygous for 22 bp deletion (D/D, n=5) showed stronger upregulation of both FADS1 and FADS2 in response to simvastatin. Little is known of the prevalence of this INDEL apart from these 16 Japanese participants.

Our aim was to investigate the allele frequency of the INDEL in US residents from 311 opportunistic human samples originating from numerous sites.

Methods. A 629 base pair portion of FADS2 intron 1 flanking the sterol regulatory element DNA sequence was amplified by PCR. Amplicons were separated on 2% agarose gels to indentify INDEL genotypes.

Results. We identified three genotypes (D/D), (I/D), (I/I) for the 22 bp INDEL for each individual. Our data shows D/D alleles in 43% (n=134), I/D alleles in 39% (n=120) and I/I alleles in 18% (n=57).

Conclusion. A high frequency (43%) of D/D genotype is found in US population, when compared to Japanese (24%). Our data indicate that the deletion genotype is prevalent and may contribute to individual variability in response to LCPUFA consumption or statins.

Grant Funding Source: NIH R01AT007003
Identification of a conformational epitope induced in the kringle 5‐latent protease domain of Glu‐plasminogen Induced by its interaction with cells (833.2)
Authors: Han Jane , Kim Kee‐Hwan , Parmer Robert , Miles Lindsey
Published: June 2021

Activation of Glu‐plasminogen is markedly enhanced on cell surfaces, arming cells with the broad spectrum proteolytic activity of plasmin that promotes cell migration. This may be due to conformational changes in Glu‐plasminogen when bound to cells. To explore these conformational changes we have developed anti‐plasminogen mAbs that preferentially recognize receptor‐induced binding sites in Glu‐plasminogen. We used specific proteolysis and immunoaffinity chromatography, followed by LC‐MS‐MS to define the epitope(s) recognized by two mAbs. Plasminogen was digested with trypsin and applied to a normal mouse IgG affinity column and the flow‐through applied to either a mAb 109 or mAb 20 immunoaffinity column. The eluate from the mAb 109 column contained two peaks with masses of 1298 AMU and 1030 AMU in LC‐MS‐MS corresponding to a nonlinear epitope contained within H494‐R504 within the kringle 5 domain and L652‐K661 within the latent protease domain, respectively. For mAb 20, MALDI TOF results were consistent with an epitope contained within L650‐I663, overlapping the mAb 109 epitope. Furthermore, mAb 109 and mAb 20 cross‐reacted in pairwise epitope mapping using surface plasmon resonance. The conformational changes detected by mAbs 109 and 20 are likely to reflect changes in Glu‐plasminogen conformation that modulate its ability to be activated on the cell surface.

Grant Funding Source: Supported by HL080146 and CA166473
Pathological changes in the lungs of Cux1 transgenic mice (834.1)
Authors: Harris Ethan , Andres Erica , Jang Inae , Petrie Alex , Doo Caroline , Safavi Sahar , Herndon Betty , Vanden Heuvel Gregory , Molteni Agostino , Baybutt Richard
Published: June 2021

Cux1 is a transcriptional repressor gene and part of the network controlling G1‐S phase transition. It represses the expression of the cyclin kinase inhibitors (CKI) p21 and p27. To determine the function of Cux1, we generated transgenic mice constitutively expressing Cux1. These mice develop multiorgan hyperplasia resulting from the aberrant repression of p27. We have previously characterized the changes in kidney development, spermatogenesis, and liver development in the Cux1 transgenic mice. In this study we have focused on the changes in the lung resulting from constitutive Cux1 expression. Histological staining of mice lung tissue showed extensive inflammation, localized areas of atelectasis alternated to emphysema, severe bronchiectasis, thickened bronchial basal membrane, and pluristratification of bronchial epithelium interspaced with areas of necrosis. Peribronchial arteries exhibited mild media thickening and enlarged adventitia with significant presence of fibroblasts (trichrome staining). The damage, bronchiectasis, and inflammation in particular were more evident in lungs of Cux1 transgenic mice. In conclusion, our results suggest that increased Cux1 expression results in multiple lung pathologies and may contribute to bronchiectasis.
Targeted mice reveal a role for RAGE in an early inflammatory response to tobacco smoke (834.4)
Authors: Wood Tyler , Winden Duane , Barton David , Betteridge Bryce , Marlor Darek , Wright Alex , Jones Cameron , Chavarria Michael , Rogers Geraldine , Reynolds Paul
Published: June 2021

The receptor for advanced glycation end‐products (RAGE) has increasingly been demonstrated to be an important modulator of inflammation in cases of pulmonary disease. Published reports involving tobacco smoke exposure have demonstrated increased expression of RAGE, its participation in pro‐inflammatory signaling and its role in irreversible pulmonary remodeling. The current research evaluated the in vivo effects of short‐term tobacco smoke exposure in RAGE null and control mice compared to identical animals groups exposed to room air only. Q‐PCR and immunoblotting revealed elevated RAGE expression in controls after four weeks of exposure and an anticipated absence of RAGE expression in RAGE null mice regardless of smoke exposure. Inflammatory cell behaviors were assessed by determining the activation of ras, intracellular signaling kinases, and cytokine synthesis and secretion. Furthermore, bronchoalveolar lavage fluid (BALF) was procured from RAGE null and control animals prior to assessment for inflammatory cells and molecules. As a general theme, inflammation induced by tobacco smoke exposure was influenced by the availability of RAGE. These data reveal captivating information suggesting a role for RAGE signaling in lungs exposed to tobacco smoke and further research is necessary in order to fully explain roles for receptors such as RAGE in cells coping with exposure.

Grant Funding Source: Supported by the Flight Attendant’s Medical Research Institute (FAMRI) and a BYU MEG Award (PRR).
Neural and cell‐cell junction autoantibodies in megacity children: the neurodegenerative impact (835.2)
Authors: Calderon‐Garciduenas Lilian , Vojdani Aristo
Published: June 2021

Millions of children in the USA, Mexico and around the world are chronically exposed to concentrations of air pollutants, including fine particulate matter, above safety standards. Mexico City children exposed to high concentrations of particulate matter (PM) exhibit a breakdown of the nasal, olfactory, and brain‐blood‐barriers (BBB), along a brain gene imbalance in oxidative stress, inflammation, innate and adaptive immune responses, and the accumulation of misfolded proteins observed in the early stages of Alzheimer (AD) and Parkinson’s diseases.

We measure serum autoantibodies associated with epithelial and endothelial barriers, and neural proteins in 95 Mexican children age 11.02 ±3.6 years, with exposures to megacity severe air pollution versus clean air controls. MC children had significantly increased IgG high affinity antibodies for cell‐cell junction and neural proteins including: occludin‐zonulin, actin, S‐100, myelin oligodendrocyte glycoprotein, and myelin basic protein (p<0.001). The major factor determining the impact of brain autoantibodies is the integrity of the BBB, thus, cell‐cell junction and brain autoantibodies might contribute to the white matter integrity alterations, structural/volumetric brain changes and the selective impairment in subscales tapping on attention, short term memory and learning abilities present in highly exposed children. Increasing research supports dysregulated immune responses in the pathophysiology of AD. Early neurodegeneration‐related dysregulated immune responses might indicate pathways that can be targeted for childhood interventions aiming at AD prevention. AD affects 5.2 million Americans today. By 2050, 13.8 million will be affected. Early identification of subjects at risk and effective prevention strategies are urgently needed.
Effect of peripherally restricted PEGylated Neuromedin U on cardiovascular function (837.14)
Authors: Emmerson Paul , Foltz Lisa , Bricker Gerald , Qian Yuewei , Wang He , Suter Todd , Ruble Cara , Flora David , Sundquist Sarah , Statnick Michael , Coskun Tamer , Patel Nita , Leishman Derek
Published: June 2021

Background: Administration of NMU agonists have been shown to regulate smooth muscle contraction, stress, analgesia, and metabolism. Central NMU receptor activation has been shown to increase heart rate in rodent models. To further examine the effect of peripheral NMU receptor activation on cardiovascular function, a long acting pegylated (40Kda) neuromedin U analog (PEGNMU) was designed and its effect on cardiovascular function examined in rats and cynomolgus monkeys.

Methods: cynomolgus monkey NMU receptors were cloned and expressed in HEK293 cells. The activity of NMU peptides was characterized in receptor binding and FLIPR based functional assays. Cardiovascular function was assessed by telemetry in instrumented rats and cynomolgus monkeys.

Results: Cynomolgus monkey NMU (cNMU) receptors were >95% homologous to human receptors. NMU peptides activated both human and cynomolgus monkey receptors with similar potency and selectivity. PEGNMU was found to be a potent full agonist at both cNMU receptors and exhibited a half life in cynomolgus monkeys of 33 hours. Administration to rats and cynomolgus monkeys resulted in increased diastolic and systolic blood pressure with no change or slight decrease in heart rate and cardiac contractility.

Conclusion: The data presented illustrate a role for peripheral neuromedin U receptors in the regulation of cardiovascular function in vivo.
The effects of radiation on schedule‐controlled responding and quinpirole‐induced yawning (839.6)
Authors: Davis Catherine , Hienz Robert
Published: June 2021

A previous report noted that rats receiving head‐only radiation exposure displayed neurobehavioral deficits on the rodent psychomotor vigilance task (rPVT), including changes in accuracy and premature responding. These changes were only apparent in a subset of irradiated rats, such that radiation‐sensitive and radiation‐insensitive groups emerged. Group specific changes in levels of the dopamine D2 receptor and dopamine transporter (DAT) protein were also evident, in addition to differential sensitivity to the rate‐decreasing effects of quinpirole on schedule‐controlled responding.

To better understand DA system differences in radiation sensitivity, a cohort of rats was trained to perform the rPVT and then trained on a fixed‐ratio 30 fixed‐interval 2‐min schedule of reinforcement; dopaminergic compounds were administered prior to the FR‐FI session to determine the rate‐altering effects of these drugs in individual subjects. A second cohort of rats was trained to perform the rPVT and was then observed for quinpirole‐induced yawning. Pre‐irradiation rPVT, FR/FI, and yawning data was compared to post‐irradiation data in the same animals, in order to determine how radiation might impact the dopamine system as measured by these behaviors induced by administration of dopaminergic compounds.

Grant Funding Source: This work was funded by NSBRI through NASA NCC 9‐58‐PF02602 and 9‐58‐NBPF02802.
Comparison of antioxidative and antifibrotic effects of Allium sativum in rat model of chronic pancreatitis (840.4)
Authors: Sharma Surendra , Rana Satya , Bhasin Deepak , Rana Surinder , Nada Ritambhra , Malhotra Samir
Published: June 2021

OBJECTIVE: Allium sativum (Garlic) is considered effective antioxidant. This study aimed to compare the antioxidative and antifibrotic effects of Allium sativum in L‐Arginine induced chronic pancreatitis in rats METHODS: Oral administration of freshly homogenized Allium sativum (0.25g/kg per day) started two day before induction of chronic pancreatitis. L‐arginine hydrochloride (250mg/100g bw/day) was administered intraperitoneally (IP) in 2 repeated doses of 1 hour interval on day 1, repeated with 250mg/100 g bw/day, on day 4, 7, 10, 13, 16 & 19 was used to induce CP. Histological examination by masson’s trichrome staining and measurement of reduced glutathione (GSH) and lipid peroxidation (LPO) of pancreatic tissue were performed to evaluate pancreatic damage and fibrosis. IHC staining of α‐SMA was done to evaluate the activated pancreatic stellate cells and mRNA levels (densitometry analysis) of transforming growth factor‐β1 (TGF‐β1), fibronectin 1 and collagen‐α1(I) were performed by reverse transcription polymerase chain reaction.

RESULTS: Allium sativum, supplementation increased the level of GSH and decreased the level of LPO which indicates reduction in oxidative stress. It also reduces inflammation and fibrosis. It also down‐regulates the mRNA expression of TGF‐β1, fibronectin 1 and collagen‐α1(I) in L‐arginine induced CP.

CONCLUSION: Oral administration of Allium sativum improves pancreatic inflammation and fibrosis in L‐arginine induced CP rats. Therefore, Allium sativum can be a novel option for alleviating inflammation and fibrotic process in CP.
M4 muscarinic acetylcholine receptor modulation of associative learning and behavioral flexibility in a novel touchscreen cognitive assessment (845.8)
Authors: Dencker Ditte , Gould Robert , Bubser Michael , Bridges Thomas , Wood Michael , Duggan Mark , Wood Michael , Fink‐Jensen Anders , Wess Jürgen , Conn P , Jones Carrie
Published: June 2021

Recent findings indicate that selective activation of the M4 muscarinic acetylcholine receptor (M4 mAChR) may represent a novel treatment approach for the psychotic symptoms and cognitive disturbances observed in several neuropsychiatric disorders. In the present studies, cognitive performance of M4 mAChR knock‐out mice (M4‐KO mice) was examined across a series of computer touchscreen‐based pairwise discrimination tasks assessing susceptibility to interference from previously learned competing sets of information. While M4‐KO mice acquired the initial discrimination task in a similar number of sessions as wildtype mice, the M4‐KO mice showed impaired acquisition on all subsequent visual discriminations. In addition, the novel M4 positive allosteric modulator VU0467154 reversed disruptions in pairwise discrimination induced by MK‐801, an antagonist of the N‐methyl‐D‐aspartate subtype of glutamate receptors. These data reveal that selective M4 mAChR activation is critical for modulation of normal associative learning and behavioral flexibility in visual discrimination tasks. Moreover, our studies suggest that selective M4 PAMs may have therapeutic potential for the treatment of cognitive deficits observed in patients with schizophrenia.

Grant Funding Source: This work was funded by grants# 2R01 MH73676‐06, 1R01 MH086601‐04, AstraZeneca.
Nicorandil increases Na/Ca exchange current in heart (850.2)
Authors: Watanabe Yasuhide , Kimura Junko
Published: June 2021

Nicorandil is a K(ATP) channel opener, nitric oxide donor and antioxidant, and is used as a drug for the treatment of angina pectoris. This durg induces cardioprotection and vasodilatation after ischemia reperfusion. It was previously reported that nicorandil may exert antiarrhythmic actions by abolishing both spontaneous and triggered activity [early after‐depolarization (EADs), delayed after‐depolarization (DADs)] (Lathrop et al., Br J Pharmacol, 1990). The triggered activity induced by DADs is one of the mechanisms of ventricular arrhythmias associated with intracellular Ca2+ overload.

In this study we examined the effect of nicorandil on Na+/Ca2+ exchange current (INCX) in isolated guinea pig ventricular myocytes using the whole‐cell patch clamp technique.

Nicorandil enhanced INCX in a concentration‐dependent manner. The EC50 values of nicorandil were 15.0 μM and 8.7 μM for the outward and inward components of INCX, respectively. Eight‐Br‐cGMP, a membrane permeable analog of cGMP also enhanced the INCX in a concentration‐dependent manner. ODQ, a guanylate cyclase inhibitor (10 µM) almost completely abolished the effects of both nicorandil and Eight ‐Br‐cGMP on INCX. DADs induced by electrical stimulation with ouabain disappeared in the presence of 100 µM nicorandil. We conclude that nicorandil enhances the function of Na+/Ca2+ exchanger (NCX) via guanylate cyclase, and this may partially contribute to the cardioprotection of nicorandil by acceleratiing Ca2+ exit via NCX.
In vitro responses of insulin resistant and nonresistant digital vessels of healthy and laminitic horses (851.11)
Authors: Venugopal Changaram , Holmes Earnestine , Beadle Ralph , Kearney Michael , Eades Susan
Published: June 2021

Hyperinsulinemia leads to insulin resistance of blood vessels and interferes with circulation of lamina of the equine foot. The objective of the study was to compare the digital vessel responses of clinically healthy and laminitic horses with and without experimental induction of insulin resistance. Vessel segments (artery and vein) collected after euthanasia, were cut into 3 mm wide rings and were prepared in tissue baths containing Tyrode’s solution for response studies. Two rings from each horse were used as non‐resistant and two were made insulin‐resistant. All rings were contracted with phenylephrine (10‐6.5M). When the contraction reached a plateau, insulin (10‐5M) was added and the response monitored. The response to insulin was calculated as a percentage of the phenylephrine response. The results showed that the responses of the non‐resistant rings differed significantly between the groups. However, when all rings were made insulin‐resistant, the significance between the groups disappeared. In the laminitic group, responses of resistant and non‐resistant rings did not differ, whereas in the healthy group, the vessel responses differed significantly. In all experiments, arterial and venous rings followed the same pattern but the magnitudes were greater in arterial rings. The findings suggest that the vessel responses to insulin are altered in laminitis which could be due to insulin resistance.

Grant Funding Source: Supported by Equine Health Studies Program of LSU‐SVM
Effect of exhaustive exercise on contrast sensitivity and neurovascular coupling (853.11)
Authors: Yamaguchi Yuji , Ikemura Tsukasa , Miyaji Akane , Hayashi Naoyuki
Published: June 2021

Neurovascular coupling (NVC) is assessed as an increase response to a given visual stimulation in the blood flow velocity in the posterior cerebral artery (PCAv). To investigate whether the change in NVC with exhaustive exercise, if any, associates with contrast sensitivity, we measured the PCAv during rest and exercise in 13 healthy males while they performed a leg‐cycle exercise at 75% of maximal heat rate until exhaustion. NVC was estimated as the relative change in PCAv from the mean value obtained during 20 s of eye closing to the peak value obtained during 40 s of visual stimulation involving looking at a reversed checkerboard. The conductance index (CI) of the PCA was calculated by dividing PCAv by the mean arterial pressure. Contrast sensitivity was measured before and after the exercise. At exhaustion, PCAv response to visual stimulation significantly decreased from 16±1 (at rest; mean±SE) to 11±1%, while it significantly increased to 20±2% with a significant increase in CI response to visual stimulation at recovery period. Contrast sensitivity significantly improved after exhaustive exercise, compared to the resting baseline. These results suggest that NVC decreases by exhaustive exercise and overshoot after the exercise, and that contrast sensitivity increases by an exhaustive exercise, implying that NVC associates with a change in contrast sensitivity.

Grant Funding Source: Supported by Kozuki Foundation
A Bayesian statistical model to characterize blood pressure increase in salt‐sensitive hypertension in the rat (856.6)
Authors: Dayton Alex , Ahn Kwang‐Woo , Laud Purushottam , Kurth Terry , Mattson David , Cowley Allen
Published: June 2021

In salt‐sensitive rat models of hypertension, characteristic patterns of increase in blood pressure (BP) are observed when salt intake in the diet is increased. In this study, we used a Bayesian statistical model to quantitate these patterns and draw conclusions about the differences between sets (strains or treatments) of rats.

Time‐course BP data for individual rats was fit to a mixture of two cumulative normal distributions by Gibbs sampling, a process for approximating multivariate probability distributions. The mixture was defined by seven parameters, which describe aspects of the timing and extent of BP increase. Probability distributions for each parameter describe the frequency of a certain value over all Gibbs samples. Sets of rats were assessed on a single parameter level by taking parameter values for each rat as samples from a group parameter representing all the rats, which allowed us to assess differences between experimental sets.

The model precisely fit all observed BP data, and is sufficient to detect significant differences in parameters between experimental sets.

This Bayesian model provides a useful tool for distinguishing sets of rats based on the character of their BP increase. These insights provided by this analysis into the timing and extent of BP increase may provide a framework for the analysis of further data generated with these rats, and the mechanisms underlying these BP increases.
Novel PKC pathway provides insight into the regulation of tone in vascular smooth muscle (865.13)
Authors: Sivilich Sarah , Trappanese Danielle , Moreland Robert
Published: June 2021

Vascular smooth muscle (VSM) tone is the phenomenon of intrinsic force developed in the absence of a stimulus. Although the primary site of VSM regulation is on the thick filament, our research has focused on the potential role of thin filament proteins in signaling events contributing to vascular tone. We hypothesized that protein kinase C (PKC)‐dependent phosphorylation of mitogen‐activated protein kinase phosphatase‐1 (MKP‐1), an upstream mitogen activated protein kinase (MAPK) regulatory protein, would increase MKP‐1 activity and decrease MAPK activity. We used the MKP‐1 inhibitor, sanguinarine, to investigate the effect of PKC on p42/p44 MAPK in swine arterial smooth muscle. Smooth muscle strips were mounted for isometric force measurement and incubated with 3 µM Bis (PKC inhibitor), 10 µM histamine, or both, in the presence or absence of 30 µM sanguinarine. Histamine significantly increased p42/p44 MAPK activity; the p42 isoform was further enhanced by inhibition of PKC. Sanguinarine reversed the histamine and Bis‐dependent increases in p42 MAPK. MKP‐1 phosphorylation was increased by histamine and Bis, and surprisingly was further amplified in the presence of sanguinarine. These findings suggest a novel PKC pathway, which indirectly modulates MAPK activity through phosphorylation of MKP‐1. Our current studies are examining potential regulatory proteins as downstream sites of action by MAPK.

Grant Funding Source: NIH DK 85734
Activation of AMP‐activated protein kinase attenuates arterial stiffening and hypertension in klotho‐deficient mice (867.1)
Authors: Lin Yi , Velury Divya , Sun Zhongjie
Published: June 2021

Background: Arterial stiffening is a progressive aging‐related disorder. Klotho is an anti‐aging protein but its role in regulation of arterial stiffening is unknown.

Methods and Results: Heterozygous Klotho deficiency (KL+/‐) mice and WT littermate mice were fed on high fat diet (HFD) or normal diet (ND). Pulse wave velocity (PWV), an index of arterial stiffening, was increased in KL+/‐ mice fed on HFD for 5 weeks. Systolic blood pressure and blood glucose levels increased earlier with greater magnitudes in KL+/‐ mice fed on HFD. Notably, protein expression of collagen I, Runx2, and TGFβ1 were increased but protein expression of phosphorylated AMPKα (pAMPKα), phosphorylated eNOS (peNOS), and Mn‐SOD were decreased in aortas of KL+/‐ mice fed on HFD for 5 weeks. Interestingly, daily injection of AICAR, an activator of AMPK, abolished the increases in PWV, blood pressure, and blood glucose in KL+/‐ mice fed on HFD. AICAR not only abolished the downregulation of pAMPKα, peNOS, and Mn‐SOD levels but also attenuated the increased levels of collagen I, Runx2, TGFβ1 and superoxide, elastic lamellae breaks, and calcification in aortas in KL+/‐ mice fed on HFD.

Conclusions: Klohto deficiency promotes HFD‐induced arterial stiffening and hypertension. The promoting effects of klotho deficiency on arterial stiffening may be due to downregulation of AMPKα activity.

Grant Funding Source: Supported by HL105302 and HL 102074.
Direct visualisation of smooth muscle cell phenotypic plasticity and migration by long‐term imaging (867.12)
Authors: Sandison Mairi , Chalmers Susan , Dempster John , McCarron John
Published: June 2021

An abnormal increase in the number and distribution of SMCs is a key feature of the vascular remodelling that underlies cardiovascular disease. The predominant explanation for these changes is that medial SMCs undergo phenotypic modulation followed by migration to the intima and subsequent proliferation. However, there has been no direct demonstration of this dedifferentiation process by tracking the fate of contractile SMCs and the very existence of phenotypic plasticity has been questioned. We therefore employed high‐resolution, multi‐wavelength fluorescence and simultaneous phase contrast, time‐lapse microscopy to monitor the fate of unambiguously identified, freshly isolated, mature SMCs. Phenotypic modulation of the initially contractile SMCs was clearly observed in response to serum‐containing culture media. Extensive cellular remodelling occurred, though a consistent sequence of events was observed in which spindle‐shaped SMCs initially rounded up following exposure to media. The cells remained round for varying lengths of time (1‐3 days) before spreading outwards and, at this stage, repetitive contractions occurred (measured via bursts of high speed imaging). Finally, the cells flattened further and migration began. Surprisingly, after the onset of migration, the extrusion of subcellular structures (e.g. mitochondria) from one cell and their subsequent uptake by another cell was observed. The phenotypically modulated SMCs also formed direct connections with other cells (tunnelling nanotubes). These results highlight the complexity of the remodelling process, the highly dynamic cellular interactions involved and the significant cell‐cell variation in response and will enable a detailed picture of SMC remodelling to be built.

Grant Funding Source: Supported by the Wellcome Trust (092292/Z/10/Z) and the British Heart Foundation (PG/11/70/29086)
The role of adventitial resident Sca‐1+ progenitor cells in angiotensin II‐induced aortic stiffening (867.3)
Authors: Wu Jing , Hoglund Virginia , Dong Xiu , Chen Wei , Majesky Mark , Harrison David
Published: June 2021

The aortic adventitia contains progenitor cells that express the surface marker stem cell antigen‐1 (Sca‐1). Sca‐1+ cells have been reported to transform into smooth muscle cells and promote neo‐intimal formation. We have previously found a striking deposition of adventitial collagen in mice with experimental hypertension. This fibrotic layer of collagen contributes to aortic stiffening and likely promotes systolic hypertension. Because this collagen deposition occurs at the site of Sca1+ cells, we hypothesized that Sca‐1+ progenitor cells serve as a source of adventitial collagen. Using confocal microscopy we found that chronic angiotensin II infusion causes a dramatic increase of Sca‐1+ cells in the aortic adventitial and peri‐adventitial region. Flow cytometry confirmed that aortic Sca‐1+CD31‐ cells are tripled in angiotensin II‐induced hypertension (5.6±1.6 ×103 vs. 15.9±2.4 ×103 cells/per aorta). These Sca‐1+ cells represent approximately one fourth of total adventitial cells. Angiotensin II infusion induces proliferation of these cells as evidenced by positive ki‐67 staining. We then isolated aortic Sca‐1+ cells in sham and angiotensin II‐treated mice using magnetic‐activated cell sorting and found collagen 1a1 (1.0±0.2 vs. 5.4±0.9), 3a1 (1.0±0.4 vs. 4.7±0.8), 5a1 (1.0±0.4 vs. 3.5±0.8) and fibronectin‐1 (1.0±0.3 vs. 2.7±0.4) genes were significantly upregulated in these cells. These fibrotic genes were also upregulated by 2‐3 fold in CD31+ cells suggesting endothelial to mesenchymal transition. Immunofluorescence staining demonstrated colocalization of Sca‐1 and vimentin, a marker of fibroblasts, indicating that these progenitor cells differentiate into fibroblasts during the onset of hypertension. In conclusion, resident aortic Sca‐1+ progenitor cells likely differentiate into fibroblasts in hypertension and contribute to adventitial collagen deposition and promote angiotensin II‐induced aortic stiffening.

Grant Funding Source: Supported by NIH R01 grant HL105294