bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of Annexin A2 in Glutamate‐induced Extracellular matrix (ECM) Changes and Retinal Ganglion Cell Death
Authors: Valapala Mallika , Vishwanatha Jamboor K
Published: June 2021

Glutamate excitotoxicity mediated increase in intracellular Ca2+ levels has been implicated in the pathogenesis of glaucoma. The downstream events leading to RGC damage after excitotoxic injury are poorly understood. It is suggested that retinal ganglion cells (RGCs) respond to excitotoxic injury by activating protease‐mediated remodeling of the extracellular matrix (ECM) through the plasminogen activator (PA)‐plasminogen (PG) system which results in detachment induced apoptosis of RGCs. Annexin A2 is a Ca2+ and phospholipid binding protein, it serves as a proteolytic center at the cell surface by recruiting matrix remodeling proteases and their substrates. Our goal is to determine the role of the annexin A2 and its associated proteases in glutamate excitotoxicity. Here we characterize the expression of annexin A2 in rat retinal sections, primary RGCs and in a transformed cell line; RGC‐5. Live cell Ca2+ imaging and time lapse confocal microscopy demonstrate glutamate‐mediated Ca2+ influx in RGC‐5 cells. A23187, a Ca2+ ionophore increases the phospholipid binding affinity of annexin A2. We demonstrate that annexin A2 translocates to the membrane of RGC‐5 cells upon glutamate treatment in a process that is independent of new protein synthesis and regulated by Ca2+.These results suggest that annexin A2 acts as a facilitator in glutamate induced changes in the ECM and subsequent RGC death.

(Supported in part by grants from the National Center for Minority Health and Health Disparities, NIH (P20 MD001633), and Alcon Research, Inc.)
A role for protein phosphorylation in CYP3A4 ubiquitin‐dependent proteasomal degradation (UPD)
Authors: Wang YongQiang , Liao M. , Deng C‐H. , Krutchinsky A. , Correia M. A.
Published: June 2021

Cytochromes P450 (P450s) are known to incur phosphorylation. The precise role of this posttranslational modification is however unclear. Various roles have been ascribed, including marking P450s for degradation. Indeed, we have previously found that CYP3A4, the major human liver P450, and its rat orthologs not only are phosphorylated in the course of their UPD, but also this phosphorylation is enhanced after their structural inactivation. Peptide mapping coupled with mass spectrometric analyses of in vitro phosphorylated CYP3A4 have identified three major target sites: T264, S420 and S478. To determine whether phosphorylation of these residues is relevant to in vivo CYP3A4 degradation, we heterologously expressed wild type and CYP3A4 with single, double or triple mutations of these residues to Ala in S. cerevisiae pep4Δ‐strains with a functional UPD but nonfunctional vacuolar degradation, after verifying that such mutations did not affect CYP3A4 structure and/or function. Our findings revealed that relative to CYP3A4, its S478A mutant was significantly stabilized in these yeast, and this stability was greatly to markedly enhanced for its S478A/T264A, S478A/S420A and S478A/T264A/S420A double and triple mutants. Together these findings indicate that phosphorylation of a buried, otherwise concealed SRS‐6 S478 residue by cytosolic kinases is important for CYP3A4 UPD. Supported by NIH GM44037.
RGS13 restricts G‐protein‐coupled receptor‐induced responses in mast cells
Authors: Bansal Geetanjali , Rao Sudhir , Nocka Karl , Druey Kirk
Published: June 2021

Mast cell degranulation and release of vasoactive mediators and cytokines promotes allergic reactions and innate immunity. Regulator of G protein signaling (RGS) proteins attenuate G‐protein‐coupled receptor (GPCR)‐evoked signaling pathways by binding heterotrimeric G protein alpha subunits and acting as GTPase accelerating proteins (GAPs). Several RGS proteins were expressed in mast cells including RGS13. We found previously that RGS13 suppressed IgE‐mediated degranulation of mouse bone marrow‐derived mast cells (BMMCs) and anaphylaxis (Bansal et al Nat Immunol in press). To determine how RGS13 affected GPCR‐mediated signal transduction in human mast cells, we examine responses of the human mastocytoma cell line (HMC‐1) overexpressing RGS13 or expressing an RGS13‐specific short, hairpin siRNA (shRNA) to GPCR stimulation. Compared to control cells, HMC‐1 cells with reduced RGS13 amounts exhibited heightened Ca2+ influx in response to adenosine, C5a, sphingosine‐1‐phosphate (S‐1P), and CXCL12. Chemotaxis and cytokine (interleukin 8, IL‐8) secretion were increased in shRGS13‐HMC‐1 cells while overexpression of RGS13 in HMC‐1 cells inhibited CXCL12‐induced Ca2+ mobilization and chemotaxis. These results suggest that RGS13 may exert control over GPCR‐evoked biological responses in human mast cells.
Effect of bioflavonoids on the proliferation of DU145 human prostate cancer cells
Authors: Jean‐Pierre Ninochka , Faldu Gaurav , Patel Suketu , DePass Anthony , Joseph Cecil K.
Published: June 2021

Flavonoids are ubiquitously occurring and widely consumed secondary metabolites of plants and are therefore an integral part of the human diet. Apigenin, luteolin, Myricetin and Rutin are flavonoids that have various pharmacological activities including inhibitory effects on the growth and proliferation of tumor cells. We have investigated the effects of these flavonoids alone and in combination on the proliferation of androgen‐independent human prostate (DU145) cancer cells in culture. Our results indicate that apigenin, luteolin, myricetin and rutin inhibited cell proliferation and decreased the viability of DU145 cells after 96 hours of treatment. To determine whether the decreased viability and cell proliferation was due to cell death by apoptosis or necrosis, we performed western blot analysis using antibodies raised against the pro‐apoptotic protein Bad and the anti‐apoptotic proteins Bcl2 and BclxL, We observed a dose‐dependent and time dependent increase in the pro‐apoptotic Bad gene and a dose dependent and time dependent decrease in BcL2 and BclxL when cells were treated with flavonoids for 72 hours. DNA fragmentation studies and expression of matrix metalloproteinase (MMP) – 2 using identical doses and time points confirmed the pro‐apoptotic effect of flavonoids on the proliferation of DU145 cells.
Coordinate suppression of B cell lymphoma by PTEN and SHIP
Authors: Miletic Ana V. , Anzelon‐Mills Amy N. , Mills David M. , Omori Sidne A. , Pedersen Irene M. , Ravetch Jeffrey V. , Bolland Silvia , Rickert Robert C.
Published: June 2021

Downstream of the B cell receptor, activation of PI3K results in the generation of phosphatidylinositol‐3,4,5‐trisphosphate (PIP3) and promotes survival and proliferation of B lymphocytes through the activation of proteins including PDK, Akt, TOR, cyclins D and E, and FOXO1. The actions of PI3K are antagonized by the inositol phosphatases, PTEN and SHIP. Whereas deletion of PTEN in T cells causes aggressive T cell lymphomas, loss of PTEN in B lymphocytes does not result in malignancy. Thus, we hypothesized that SHIP compensates for the lack of PTEN in B lymphocytes thereby preventing lymphoma development. To this end, we generated mice lacking both PTEN and SHIP specifically in B lymphocytes (bPTEN/SHIP−/−) and show that bPTEN/SHIP−/− mice develop lethal B‐Non‐Hodgkin's Lymphoma (B‐NHL). Mechanistically, PTEN/SHIP−/− B cells are hyperresponsive to mitogenic stimulation with increased proliferation and mitogen‐activated protein kinase (MAPK) activation. Interestingly, PTEN/SHIP‐deficient B cells also display an atypical mitogenic response to the survival factor BAFF. Taken together, these studies reveal that PTEN and SHIP act cooperatively to suppress B‐NHL and provide the first direct evidence that SHIP can function as a tumor suppressor. This research is supported by a postdoctoral fellowship from the Cancer Research Institute.
Reduced proliferative capacity of highly purified T cells from patients with Autism Spectrum Disorders (ASD)
Authors: Goldberg Jodi Elisabeth , Manske Jill
Published: June 2021

A pilot study assessing the proliferation of peripheral blood T cells from ASD patients as compared to controls was performed. Peripheral blood mononuclear cells (PBMC) were purified via ficoll. T cells were isolated from PBMC by negative selection (>95% CD3+), labeled with cytoplasmic dye, activated in vitro, and assessed for proliferation by flow cytometry on day 5. Results indicate that the percent of T cells entering the cell cycle upon activation with anti‐CD3/anti‐CD28 coated beads from ASD patients (73%) and controls (76.4%) were not statistically different. Likewise the division index, defined as the number of divisions on average that every T cell in the culture undergoes, was also not statistically different between ASD patients (1.8) and controls (2.1) activated with the antibody beads. However, the average number of divisions that a T cell that has divided undergoes, the proliferative index, was significantly less in ASD patients (2.45) activated with the beads than in controls (2.67), p‐value = 0.03. Proliferation of T cells to lectins were not different between ASD patients and controls as measured by the percent of cells entering the cell cycle, division index, or proliferative index. These results indicate that while ASD patient T cells are not profoundly deficient in proliferative capacity, they may demonstrate subtle defects in activation via antibody‐coated beads.
BCR Signaling in Transitional versus Mature B cells
Authors: Andrews Sarah F , Rawlings David J
Published: June 2021

Early transitional (T1) splenic B cells, in contrast to follicular mature (FM) B cells, apoptose and fail to proliferate in response to BCR engagement. Consistent with these distinct responses, NFκB target genes, essential for proliferation and survival, are rapidly upregulated in FM but not in T1 B cells upon BCR engagement. Previous studies have implied that proximal signals required to mediate IKK activation are dysfunctional in immature B cells. We directly evaluated this hypothesis using highly purified T1 vs. FM splenic B cells closely matched for surface IgM expression. Surprisingly, key proximal signals required for NFκB activation, including PI3K, PLCγ2, and PKCβ activation, are intact in early transitional B cells. When we measured the BCR‐triggered, canonical NFκB signaling cascade in T1 vs. FM B cells we saw equivalent levels of IKKα/β phosphorylation, IκBα degradation, and nuclear translocation of c‐Rel and RelA. Taken together, our data strongly indicate that proximal NFκB signaling is fully activated in T1 B cells. In addition, studies using chromatin IPs suggest that the failure to upregulate NFκB target genes is due to an altered capacity to mediate NFκB transcription at the nuclear level. These studies provide important insight into what controls cell fate upon BCR engagement in peripheral B cell subsets. Funded by NIH 5R01HD37091.
Regulation of the Tumor‐Suppressor Ikaros Transcription Factor by Vasoactive Intestinal Peptide Receptor‐1 in HuT 78 Cells
Authors: Van der Steen Travis , Meinhardt Steven , Dovat Sinisa , Dorsam Glenn
Published: June 2021

Mutations to the germline encoding Ikaros (IK), a master regulator of lymphopoesis, result in a hyperproliferative phenotype leading to T cell leukemia. There are three identified ser/thr phosphorylation sites termed p1, p2 and p3. Hyperphosphorylation of IK at the p1 site is regulated via casein kinase II (CKII), which reduces its DNA binding affinity and mediates G1/S transition. However, other kinase consensus sites have been identified including protein kinase A (PKA). The G protein coupled receptor (GPCR) called vasoactive intestinal peptide receptor 1 (VPACR‐1) negatively regulates TCR signal transduction by invoking cAMP/PKA activity and suppresses G1/S transition. We hypothesize that VPACR‐1 induced PKA activity phosphorylates IK and impedes CKII phosphorylation, maintaining a quiescent naïve phenotype. To test our hypothesis, naïve and activated HuT 78 cells will be utilized in the presence and absence of VPACR‐1 ligand, vasoactive intestinal peptide, to determine whether different isoelectric (pI) patterns of IK are observed. Presently, 2D gel electrophoresis followed by anti‐IK immunoblot detects three isoforms exhibiting multiple pI's. VPACR‐1 induced PKA‐dependent phosphorylation of IK, which impedes cell cycle entry, will lead to a better understanding of a poorly understood anti‐proliferative GPCR regulate T cell activation. Funding: COBRE‐2P20RR015566 and 1KO1 DK064828.
A critical role for PKC‐theta‐mediated T cell survival in cardiac allograft rejection
Authors: Sun Zuoming , Manicassamy Santhkumar , Yin Dengping , Molinero Luciana L , Alegre Marisa‐Luisa Alegre
Published: June 2021

PKC‐theta mediates the critical TCR signals required for T cell activation. Previously we have shown that in response to TCR stimulation, PKC‐theta−/− T cells undergo apoptosis due to greatly reduced levels of the anti‐apoptotic molecule, Bcl‐xL. Here we demonstrate that PKC‐theta‐regulated expression of Bcl‐xL is essential for T cell‐mediated cardiac allograft rejection. Rag1−/− mice reconstituted with wild type (WT) T cells readily rejected fully mismatched cardiac allografts, whereas, Rag1−/− mice reconstituted with PKC‐theta−/− T cells failed to promote rejection. Transgenic expression of Bcl‐xL in PKC‐theta−/− T cells was sufficient to restore cardiac allograft rejection, suggesting that PKC‐theta‐regulated survival is required for T cell‐mediated cardiac allograft rejection in this adoptive transfer model. In contrast to adoptive transfer experiments, intact PKC‐theta−/− mice displayed delayed, but successful cardiac allograft rejection, suggesting the potential compensation for PKC‐theta function. Finally, a sub‐therapeutic dose of anti‐CD154 antibody, that was not sufficient to prevent cardiac allograft rejection in the WT mice, prevented heart rejection in the PKC‐theta−/− mice. Thus, in combination with other treatments, inhibition of PKC‐theta may facilitate achieving long‐term survival of allografts.
APRIL (TNFSF13) Promotes Collagen Induced Arthritis and IL‐17 Production and Inhibits IFN‐gamma and IL‐4 Production
Authors: Xiao Yanping , Motomura Seiichi , Podack Eckhard R.
Published: June 2021

APRIL or TNFSF13 is a member of TNF super family. Analysis of APRIL knock out (APRIL−/−) mice gave conflicting results regarding B and T cell immunity. Therefore, we generated and characterized APRIL−/− mice and studied collagen induced arthritis (CIA). We confirmed that APRIL−/− mice have low basal IgA levels in serum. Splenic T cell proliferation was found increased in APRIL−/− mice. In the CIA model, the incidence of arthritis was decreased in APRIL−/− mice by about 35%, while disease severity and onset were unaffected. In mice with arthritis, chick and mouse anti‐collagen II specific IgG2a isotype antibody titers were significantly lower in APRIL−/− mice than in w.t. mice. Restimulated lymph node cells of w.t. CIA mice had elevated IL‐17 production which was diminished in APRIL−/− mice. The difference in IL‐17 production was disease specific and not observed upon restimulation of CD4 cells or in other disease models. However, APRIL−/− CD4 cells produced increased levels of IL‐4 and IFN‐? upon restimulation. Our data indicate that APRIL deficiency is responsible for diminished IgG2a levels and for diminished IL‐17 production which may be causally related to the pathogenesis of CIA. The combined effects of APRIL on antibody and IL‐17 may explain the decreased incidence of clinical arthritis in APRIL−/− mice. Our studies reveal an important role for APRIL in the regulation of Th1, Th2 and Th17 cytokines.

This study is supported by AI 061807 and AI 068515.
Licensing of naïve T cells to produce TNF
Authors: Priyadharshini Bhavana , Welsh Raymond M. , Greiner Dale L. , Brehm Michael A.
Published: June 2021

TNF is a pro‐inflammatory cytokine that regulates numerous aspects of immune responses, including the generation of virus‐specific CD8 T cells during an infection. We have shown that naïve T cells in peripheral lymphoid organs produce TNF rapidly after TCR engagement (within 4 hrs) and prior to the acquisition of other effector functions. Given the immunoregulatory role of TNF and the ability of peripheral naïve T cells to produce this cytokine, we questioned at what stage of development do T cells become competent to produce TNF. Our findings indicate that, in contrast to peripheral naïve T cells, single positive thymocytes have a poor ability to produce TNF on ex‐vivo stimulation. Despite this inability to produce TNF, thymocytes express mature TNF message, confirming that TNF production can be post‐transcriptionally regulated. Thymocytes that are adoptively transferred into the periphery of congenic hosts become competent to produce TNF as early as 6 hrs after transfer and before undergoing division. Moreover, stimulation of thymocytes in the presence of splenocytes for 4 hours in vitro is sufficient to partially enable thymic lymphocytes to produce TNF. Together, these findings indicate that thymic lymphocytes require post‐thymic maturation in the periphery to become licensed to produce TNF.
Acceptability of three supplements with identical micronutrient content in Mexican children
Authors: Flores Lourdes , Théodore Florence , Bonvecchio Anabelle , Blanco Ilian , Neufeld Lynnette M
Published: June 2021

The objective of this study was to evaluate the acceptability of three nutritional supplements consumed by children 6–24 m of age in a randomized trial. A fortified food (FF, distributed by the Oportunidades program), syrup (SY) or Sprinkles (SK) were randomly assigned at the community level. Mothers of participating children were interviewed twice, after 6 and 12 m. Interviewers were transcribed, coded and analyzed using discourse analysis. Themes explored related to perceived benefits and risks of consumption, barriers to use and overall acceptability. All three supplements were perceived to have health benefits for the children. The FF was perceived as food, while SY and SK as vitamins of a more medicinal nature. Some mothers reported that SK modified the taste of foods, limiting the range of foods with which it can be served, with a preference for more liquid foods. SY presented difficulties due to objection from the children. All 3 supplements had high acceptance, mainly because of perceived health benefits. A number of barriers for use of each were identified, and related more to practical use than to any perceived potential risk. The perceived medicinal nature of SY and SK may limit sharing; a mayor difficulty documented previously with the FF. The results of this analysis clearly highlight the importance of a strong education campaign for ANY type of supplement distributed in the context of public health programs.
The effect of almonds on plasma lipids in persons with prediabetes
Authors: Wien Michelle Ann , Sabate Joan , Monahan‐Couch Lynn , Oda Keiji
Published: June 2021

Background: Persons with prediabetes are at increased risk of developing cardiovascular disease, yet there is scant research on effective meal planning approaches in this population.

Aim and Methods: To evaluate the effect of daily almond consumption on blood lipids of adults with prediabetes, we performed a 16‐week trial in 66 free‐living adults (48 women and 18 men; mean±SE age 53±10 years; BMI 30±5 kg/m2) instructed to consume a background meal pattern yielding 35% fat, 50% CHO and 15% protein. Participants were randomized to consume 20% of energy from almonds (almond‐enriched) or a meal pattern without nuts (nut‐free). Fasting total cholesterol (TC), LDL‐cholesterol (LDL‐C), HDL‐cholesterol (HDL‐C) and triglycerides (TG) were measured at baseline and weeks 8 and 16. Mixed model analysis was performed adjusting for baseline values with all time points included in the model.

Results: Daily supplementation with almonds was associated with a greater reduction in LDL‐C [−9% vs. −1% (P=0.03)] and LDL‐C:HDL‐C [−13% vs. −1% (P=0.04) over the 16‐week trial. Almond participants had a significant reduction in LDL‐C [−10% (P = 0.01)] and a significant increase in HDL‐C [+5% (P=0.04)] at week 16 compared to baseline levels, resulting in a significant reduction in LDL‐C:HDL‐C (P=0.001).

Conclusion: Daily almond consumption within a defined meal pattern is effective in improving blood lipids in adults with prediabetes.
Incidence and severity of hypoglycemia
Authors: Sutton Leslie , Chapman‐Novakofski Karen
Published: June 2021

Hypoglycemia or fear of hypoglycemia may inhibit achievement of optimal blood glucose goals. A needs assessment was conducted to determine if education pertaining to hypoglycemia treatment/prevention was indicated. Twelve questions pertained to experience with low blood glucose; 5 determined the participant's locus of control [LOC]; and 10 identified the participant's stages of change. The survey was evaluated by an “expert panel” of four and then administered to participants of the Illinois Extension's Dining with Diabetes program [n=64]. Most had been diagnosed with diabetes by a physician [n=47], with the remaining taking care of someone with diabetes [n=17]. Most take oral agents [n=42]; few insulin [n=6]. Thirty‐five have had hypoglycemia, with 23 having values < 70 mg/dl. Most have been advised by their doctor (n=50) about hypoglycemia, but many still experience episodes (16 have episodes a few times/year; 13 have episodes several to many times/year). Almost all (92%) have an internal LOC regarding their health, but only about half (55%) do so regarding sickness prevention, indicating an interpretation difference between control over health and over sickness. Preliminary data support the need for additional education concerning the prevention and treatment of hypoglycemia.

This research is supported by Illinois AMVETS Diabetes Education Research Program and Illinois Extension.
Dietary habits and eating disorder observed from Korean obese adults visited obesity clinic
Authors: Lee Bog‐Hieu , Kim Mi‐Hyun , Cho Kyung‐Dong , Han Chan‐Kyu , Choi Changsun
Published: June 2021

The study was undertaken to identify the characteristics and respective factors related to obesity in 156 obese adults visiting the obesity clinic in Seoul. The subjects were classified as moderately obese(25≤BMI<30, n = 81) and severely obese(30≤BMI, n = 75) and asked to complete the anthropometric measurements and dietary surveys including eating disorder along with general characteristics of the subjects. The average BMI of moderately and severely obese subjects were 28.1 kg/m2 and 34.0 kg/m2, respectively. Both suffered problems of constipation, low back pain. Moderately obese and severely obese subjects exercised more than normal adults, but were physically inactive in nature. Severely obese subjects had more drinking and smoking comparable to moderately obese subjects. Also, severely obese subjects did not eat breakfast regularly, eat fast, overeat often times comparable to moderately obese subjects. There were high incidence rates of binge eating disorder and night eating syndrome in severely obese subjects, and high incidence rates of bulimia nervosa in moderately obese subjects. From the findings, the obese adults had improper dietary habits and outcomes of obesity seemed to be influenced by dietary pattern, lifestyle and physical activity. Furthermore, there were diverse and various differences in patterns like dietary habits and lifestyles among obese adults depending on obesity rate. Therefore, it is absolutely necessary to customized approach to treat the obese adults considering obesity rate.
A comparison of diets, exercise patterns and postpartum weight retention based on levels of acculturation in Mexican immigrants and Mexican American women
Authors: Kurtz‐Reyes Myra , Tam Chick F.
Published: June 2021

Prevalence of overweight Mexican women in the U.S. has increased. Pregnancy‐related weight gain has emerged as a cause of obesity. This study compares dietary intake, physical activity, energy balance, and postpartum weight retention in low and high acculturated postpartum Mexican immigrants and Mexican American women, and determines variables that may predict postpartum weight retention. Dietary and activity recall, a food frequency, and demographic questionnaire were collected from 67 subjects 6–12 months postpartum, at a community clinic. Language determined level of acculturation (Marin and Marin, 1991). Food Processor 8.0, SPSS 14.0, and Minitab 15.0 were used for analysis. There was no significant difference in calorie intake or energy balance between the low (n=33) and high (n=34) acculturation groups. The high acculturation group ate more calories from fat (p=0.000), expended more energy (p= 0.032 for basal metabolic rate; p= 0.000 for physical activity and specific dynamic action of food), and retained less weight postpartum (9.54 pounds). Variables predicting postpartum weight retention were employment, daily physical activity, pre‐pregnancy BMI, and energy expenditure. Results prove that health programs for postpartum Mexican women in the U.S. be designed based on acculturation level. All postpartum women should reduce calorie intake and increase physical activity for weight management.
Examination of Child Feeding Practices of Latino Caregivers in South Carolina
Authors: Kirkpatrick Claire , Haley‐Zitlin Vivian , Cason Katherine , Mayo Rachel , Montenegro Sergio Nieto
Published: June 2021

The Latino population is the fastest growing segment of the U.S population and is the largest minority group in the country. The growth rate of Latinos in South Carolina from 1990–2000 was 211.2%. This study examined current feeding practices used by Latino caregivers of children aged 2 to 6 in South Carolina and common child feeding problems/concerns of parents. Main topics explored (1) child feeding practices (2) child feeding issues (3) nutritional concerns regarding children and (4) nutrition education. Education questions probed opinions and suggestions on nutrition education curricula targeted at this population and how nutrition education should be provided. Eight focus groups, averaging six participants per group with male and female participants, were held at five counties in SC. All focus groups were conducted in Spanish. A convenience sample of 53 participants with a mean age of 30.7 years with mothers representing 98% of the caregivers participated. Mean child age was 3.66 years old. Results from the focus groups included: Common feeding practices used included “short order cooking” and using rewards, bribes, and punishment to coerce a child to eat; and caregivers were found to be more concerned with underweight children than overweight children, so more feeding practices to pressure a child to eat than to control for over‐consumption were used. Conclusion: Health risks of overweight children should be a focus of nutrition education of this population. Education topics should include feeding practices that enable a child to learn self‐regulation as feeding practices which coerce a child to eat may decrease a child's ability to self regulate and ultimately lead to obesity. Supported in part by NCMHD/NIH #1P20MD000539–01
Low vitamin B12 status is negatively associated with global DNA methylation in young Chinese women
Authors: Zhu Jiang‐Hui , Hao Ling , Li Zhu , Quinlivan Eoin , Maneval David , Yang Thomas P , Berry R J , Crider Krista , Rasmussen Sonja , Bailey Lynn B
Published: June 2021

Vitamin B12 is a required coenzyme for homocysteine remethylation, essential for the generation of S‐adenosylmethionine, the source of methyl groups for DNA methylation. The objective of this study was to assess the association of low vitamin B12 status on global DNA methylation in reproductive age Chinese women. Global DNA methylation was assessed in two groups of women whose plasma vitamin B12 concentration was either deficient (<148 pmol/L) (n=24) or normal (>148 pmol/L)(n=128). To correct for the possible effects of RBC folate on DNA methylation, a subset (n= 24) of the vitamin B12 normal subjects were further matched for RBC folate concentration with the vitamin B12 deficient subjects. DNA methylation was expressed as a percent of methylated cytosines (as measured by a novel LC‐MS/MS assay). The mean±SD % methylcytosine for the B12‐deficient group (3.51±0.57) was lower (p<0.001) than the total normal group (4.42± 0.18), and the RBC folate‐matched group (4.40±0.17). These data suggest that impaired vitamin B12 status might reduce global DNA methylation and provide a rationale for future investigations. This work was supported by a collaborative CDC agreement and the GCRC Grant # MO1‐RR00082.
A medicinal plant extract of Scutellaria baicalensis and Acacia catechu reduced LPS stimulated gene expression in immune cells: A comprehensive genomic study using QPCR, ELISA, and microarray
Authors: Tseng‐Crank Julie , Sung Sookyung , Jia Qi , Zhao Yuan , Burnett Bruce , Yimam Mesfin , Park Dae‐Ryoung , Woo Sung‐Sick
Published: June 2021

A standardized flavonoid extracts from Scutellaria baicalensis and Acacia catechu, UP446 (Univestin™), was profiled favorably in its anti‐inflammatory properties. In this study, quantitative polymerase chain reaction (QPCR), enzyme‐linked immunosorbent assay (ELISA), and cDNA microarray were used to study the effect of UP446 on lipopolysaccharide (LPS)‐induced pro‐inflammatory reactions. One consistent result was that the gene expression levels stimulated or suppressed by LPS were normalized by UP446 co‐treatment. This normalization effect was observed for the well‐known pro‐inflammatory genes cyclooxygenase (COX)‐2, tissue necrosis factor (TNF)‐ 1 □, interleukin (IL)‐1β, NFκB, and IL‐6 using QPCR and TNF‐α using ELISA. This was also observed by microarray. In addition to the above genes, microarray results also included cytokines, chemokines, chemokine receptor, immunoglobulin receptors, transcriptional factors and coactivator, caspase, growth factors, and phosphatases. The microarray results supported not only the anti‐inflammatory property of UP446, but also suggested that UP446 may participate in additional signaling pathways of cell proliferation, cell death and lipid metabolism. This research work was supported by Unigen.
Dietary true digestible calcium to phosphorus ratio affects phosphorus utilization and renal sodium and phosphate co‐transporter gene expressions in post‐weaned pigs
Authors: Wang Zirong , Archbold Tania , Yang Chengbo , Fan Ming Z
Published: June 2021

This study was conducted with 36 Yorkshire weanling barrows of average initial and final BW of 11.5 and 17.7 kg and the pigs were fed six diets for six blocks according to a completely randomized block design. The six diets were corn and SBM‐based and contained six true fecal digestible Ca to true fecal digestible P ratios (diet 1, 0.56:1; diet 2, 0.68:1; diet 3, 0.96:1; diet 4, 1.35:1, diet 5, 1.46:1, and diet 6, 1.94:1) formulated by supplementing gradient levels of limestone. Each experimental period consisted of 15 d with 10‐day adaptation and 4‐day collection of representative fecal and urinary samples as well as total volumes of urinary excretions. Changes in the dietary Ca to P ratio had quadratic effects (P = 0.02) on average daily feed intake and average daily gain with optimal responses occurred in diets 3 and 4. Increases in the dietary Ca to P ratio had quadratic effects (P < 0.05) on the efficiency of digestive and post‐absorptive utilization of dietary P with optimal responses observed for diets 3 and 4. Renal sodium‐phosphate co‐transporter gene IIa (NaPi‐IIa) and III (NaPi‐III) mRNA expressions were linearly affected (P < 0.05), while the jejunal NaPi‐IIc and NaPi‐III mRNA abundances were not affected (P > 0.05) by changes in the dietary Ca to P ratio. In conclusion, changes in the dietary Ca to P ratio have significant effects on growth performance, and efficiency of P utilization. Dietary true digestible Ca to true digestible P ratio of 0.96:1 to 1.35:1 should be used to guide the formulation of diets for the post‐weaned pig. Supported by OMAFRA and NSERC.
Carnosic acid attenuates the expression of adhesion molecules by IL‐1β in HUVECs
Authors: Yu Ya‐Mei , Chang Weng‐Cheng
Published: June 2021

Expression of cell adhesion molecules by the endothelium and the attachment of monocytes to endothelium may play a major role in the early atherogenic process. Carnosic acid, an antioxidant polyphenolderived from Sage (Salvia officinalis) and Rosemary (Rosmarinus officinalis), is a lipophilicantioxidant that scavenges singlet oxygen, hydroxyl radicals, and lipid peroxyl radicals, thus prevents lipid peroxidationand the disruption of biological membranes. In this study, we investigated the effects of carnosic acid on the adhesion of U937 cells to IL‐1 β‐treated human umbilical vein endothelial cells (HUVECs) and the expression of adhesion molecules. In addition, we observed the effect of carnosic acid on NF‐κB p50, p65 and the formation of intracellular ROS induced by IL‐1 β in HUVECs. Our data showed that carnosic acid inhibited the adhesion of U937 cells, abolished the expression of ICAM‐1, VCAM‐1 and E‐selectin, attenuated or block nuclear translocation of NF‐κB p50, p65 and decreased the production of ROS in IL‐1 β treated HUVECs. In conclusion, carnosic acid inhibits IL‐1 β‐induced nuclear translocation of p65 and p50, thereby suppressing the expression of ICAM‐1, VCAM‐1 and E‐selectin, resulting in decreased monocyte adhesion. Thus, carnosic have anti‐inflammatory properties and may play an important role in the prevention of atherosclerosis.
Effect of Almonds on Insulin Secretion and Insulin Resistance: A Randomized Controlled Cross‐over Trial
Authors: Kendall Cyril WC , Josse Andrea R. , Marchie Augustine , Nguyen Tri H. , Lapsley Karen G. , Jenkins David JA
Published: June 2021

Background: Cohort studies have shown that nut consumption is strongly associated with a reduced risk of coronary heart disease (CHD). Intervention studies demonstrate that almonds reduce serum LDL‐C. However, their documented reduction in LDL‐C can only explain a proportion of the reduced CHD risk.

Methods: Whole almonds, taken as snacks, were compared to low‐saturated fat (<5% energy) whole‐wheat muffins (control) in the therapeutic diets of subjects with hyperlipidemia. Twenty seven hyperlipidemic men and women each consumed three iso‐energetic (mean 423kcal/d) supplements for 1 month. Supplements consisted of full‐dose almonds (73±3g/d), half‐dose almonds plus half‐dose muffins, and full‐dose muffins. In each phase, subjects were assessed at weeks 0, 2 and 4.

Results: No baseline or treatment differences were seen in fasting serum glucose, insulin, C‐peptide or calculated insulin resistance (HOMA‐IR). However, creatinine corrected, 24‐hour urinary C‐peptide output, a marker of daily insulin secretion, was significantly reduced on both the half‐ and full‐almond doses compared to the control (P=0.002 and P=0.004, respectively).

Conclusion: Almond consumption reduced 24‐hour insulin secretion. This may be a further metabolic advantage of nuts that in the long‐term may help to explain the strong association between nuts and CHD risk reduction.

Funding support: Almond Board of California
Pharmacological Preconditioning With Tyrosol, A White Wine Component, Depends On The Activation Of PI‐3 Kinase Pathway In Ischemic Reperfused Rat Myocardium
Authors: Thirunavukkarasu Mahesh , Samuel Samson Mathews , Penumathsa SureshVarma , Zhan Lijun , Bertelli A.A.E. , Maulik Nilanjana
Published: June 2021

Tyrosol (2‐(4‐hydroxyphenyl) ethanol) (Tsl) a mono‐phenolic compound, in white wine possesses antioxidant activity & is able to reduce the release of pro‐inflammatory cytokines. The present study sought to determine the mechanism of Tsl mediated cardioprotection from ischemia reperfusion injury. Isolated perfused rat hearts were divided into five groups: ‐ (1) control heart (2.5h of perfusion) (C) (2) hearts subjected to 30min ischemia [I] & 2h reperfusion [R] (IR) (3) perfused with Tsl (1 μM, 20 min) followed by 30 min I & 2h R (TIR) (4) LY294002 (5μM) + Tsl followed by IR (LYTIR) (5) SN50 (18μM) + Tsl followed by IR (SNTIR). We have observed improved post‐ischemic ventricular functional recovery during the 2 hrs of R & the reduced infarct size & cardiomyocyte apoptosis in TIR compared to IR. Increased phosphorylation of pro‐survival proteins Akt & eNOS was observed in TIR treated compared to the IR. The gel‐shift analysis showed significant upregulation of DNA binding activity of NF‐κB in the Tsl treated TIR group compared to non‐treated IR. The use of PI‐3kinase inhibitor, LY294002 & NF‐κB inhibitor SN50 completely blocked Tsl mediated cardioprotection. We therefore demonstrate for the first time that Tsl mediated cardioprotection is by activation of the survival pathway through PI‐3kinase/NF‐κB & maybe used as a novel therapeutic agent against ischemia‐reperfusion injury.
Immunohistochemical expression of PTTG in brain tumors
Authors: Salehi Fateme , Scheithauer Bernd W , Kovacs Kalman , Cusimano Michael , Sharma Soniya , Munoz David G
Published: June 2021

Pituitary tumor transforming gene (PTTG) plays a role in many cellular processes. PTTG overexpression is seen in several tumor types and correlates with survival time/recurrence. We evaluated PTTG expression in various types of brain tumors (n=94). Immunohistochemistry was performed using a monoclonal PTTG antibody (DCS‐280, Abcam;Cambridge, MA) and the streptavidin‐biotin‐peroxidase complex method. The intensity of PTTG immunoreactivity was evaluated semi‐quantitatively on a 0‐4 scale. PTTG expression was evident in most tumor cells and was predominantly nuclear. In glial tumors, PTTG immunoreactivity was higher in glioblastomas (IV), anaplastic oligoastrocytomas (III), anaplastic oligodendrogliomas (III), oligoastrocytomas (II), oligodendrogliomas (II), and pilocytic astrocytomas (I) (range: 3.1–3.5), whereas notably lower PTTG was seen in myxopapillary ependymomas (I) and ependymomas (II) (1.5 & 1.6). In non‐glial tumors, hemangiopericytomas and schwannomas had higher PTTG score (3.4), than meningiomas (2.3). Thus, it appears that PTTG expression is not associated with tumor grade, but rather with tumor type, the most striking difference being between ependymomas and other glial tumors. PTTG may be a valuable therapy target in some brain tumors. Acknowledgments Authors thank the Jarislowsky Foundation and the Lloyd‐Carr‐Harris Foundation for their generous support.
Whole body Bi‐214 and bedroom radon concentration in Multiple Sclerosis
Authors: Lykken Glenn Irven , Magness Andrew T. , Momcilovic Berislav
Published: June 2021

Environmental radon may be an induction factor of multiple sclerosis (MS) due to its high fat solubility and emission of high energy alpha particles. In this study we analyzed the bedroom home ambient air radon (222Rn) and whole body (WB) retention of a radon daughter (214Bi) from each subject's bedroom. The 15 MS subjects came through the Grand Forks MS Support Group, and 15 subjects with no apparent health problems (H) were recruited locally. The WB 214Bi (Bq/m3) was assessed by WB counting and the 222Rn was measured with charcoal canisters. The 222Rn (Bq/m3) range (mean ± SD) was 307 ± 411 in the bedrooms of MS subject vs. 126 ± 122 of the H subjects, respectively; the 214Bi (Bq) was 333 ± 312 in the MS subjects, vs. 243 ± 144 in the H subjects, respectively. Our preliminary data indicate that the bedroom 222Rn exposure and WB214Bi retention are higher in the MS than H subjects. The study was approved by the local IRB and conducted according to the principles of the Declaration of Helsinki. The work was sponsored in part by the USDA ARS Grand Forks Human Nutrition Research Center and the University of North Dakota Public Scholarship Fund.
Nicotine Self‐administration in Combination with the Tobacco Constituent, Norharmane
Authors: Arnold Monica M , Belluzzi James B , Leslie Frances M
Published: June 2021

Tobacco is one of the most abused drugs in today's society. Despite this, nicotine, the best characterized psychoactive component of tobacco is only weakly reinforcing in animals. A reason for this discrepancy may be that nicotine might act in concert with other components found in tobacco to create its addictive potential. There are over 4000 constituents found in cigarette smoke and of interest to the present study are those that inhibit the enzyme, monoamine oxidase (MAO). MAO activity has been shown to be decreased in the brains of smokers and it has been suggested that MAO inhibition may play a role in the reinforcing properties of cigarettes. In light of this, we have investigated the potential rewarding properties of nicotine in combination with norharmane (NH), a component of cigarette smoke known to inhibit MAO‐A and B reversibly. Preliminary data indicate that addition of NH (0.025 – 2.5 μg/kg/inj) to the i.v. solution supports acquisition and maintenance of self‐administration of a low dose of nicotine (7.5 μg/kg/inj) at an FR‐1 schedule without prior food training. An intermediate dose of NH (0.25 mg/kg/inj) supports continued self‐administration of nicotine as the task is made progressively harder through progressive ratio. Following extinction of self‐administration of nicotine/NH, drug‐seeking behavior can be reinstated by both cues and priming drug injection. These findings suggest that the addictive properties of cigarettes may result from the combined actions of nicotine and other tobacco constituents.
Chronic Blockade of mGluR signaling rescues Fragile‐X Synaptic Phenotype
Authors: Nagarajan Naveen , Bear Mark F
Published: June 2021

Fragile X syndrome (FXS) is caused by transcriptional silencing of the FMR1 gene, which encodes the fragile X mental retardation protein (FMRP). The “mGluR theory” of FXS suggests that key aspects of the syndrome are caused by exaggerated Group 1 mGluR signaling. A prediction of the theory is that some rescue could be achieved by down‐regulating mGluRs. We studied the maturation of hippocampal synapses from Fmr1 knockout (KO) and wild‐type (WT) mice to test this prediction. Our results indicate that dendrites of neurons lacking FMRP have a significantly higher density of GluR2 and synapsin puncta than controls. GluR2 puncta measured 7.0 ± 1.4 in WT (n = 4) and 11.1± 0.8 in KO (n = 7) neurons (P = 0.04). Synapsin puncta measured 7.2 ± 1.1 in WT (n = 5) and 11.7 ± 0.9 in KO (n = 10) neurons (P = 0.017). Synaptic GluR2 expression was differentially affected by transient activation of mGluRs with DHPG (100 μM) in WT and KO neurons. DHPG treatment produces significant loss of synaptic GluR2 in cultures from KO (43 ± 6 % of untreated controls), but not WT animals (100 ± 20 % of untreated controls). Interestingly the exaggerated effect of GluR2 receptors, synapses and spines under basal conditions were rescued by chronically treating hippocampal cultures with MPEP, an mGluR5 antagonist.
BDNF protects against phencyclidine neurotoxicity in neonatal brain via activation of the PI‐3K/Ak/GSK‐3β and MEK/ERK pathways
Authors: Xia Yan , Wang Cheng Z , Anastasio Noelle C , Johnson Kenneth M.
Published: June 2021

NMDA receptor blockade by phencyclidine (PCP) in neonatal rats causes wide‐spread apoptosis and certain schizophrenia‐like behavioral abnormalities later in life. Disruption of BDNF signaling may play a role in the development of schizophrenia and mediate the neurotoxic effects of NMDAR blockers. Therefore, this study was designed to test the potential protective effects of BDNF on PCP‐induced apoptosis in corticostriatal slice culture and to explore the possible mechanisms. We observed that BDNF prevented PCP‐induced capase‐3 activation and DNA fragmentation dose‐dependently. The Trk B receptor inhibitor, K252a, abolished the protective effects of BDNF, indicating that BDNF acts through TrkB receptors. Further investigation of the downstream signaling transduction of TrkB activation revealed that BDNF prevented the dephosphorylation of Akt, GSK‐3β and ERK induced by PCP. In addition, pharmacological blockade of these pathways with either the MEK inhibitor PD98059 or the PI‐3K/Akt pathway inhibitors, LY294002 and Akt inhibitor V, completely abolished the protective effects of BDNF. Finally, the observation that the inhibitors of either the PI‐3K/Akt or the MEK/ERK pathway selectively prevented the stimulating effects of BDNF on the two pathways suggests that the two pathways are independently involved in the neuroprotection against PCP toxicity afforded by BDNF. Supported by DA‐02073.
Neuroprotection Against Ethanol Mediated Oxidative Stress and Apoptosis via Nrf2/ARE.
Authors: Stewart Jennifer Lynn , Henderson George I
Published: June 2021

This study characterizes a neuroprotective pathway responsible for coordinated enhancement of astrocyte glutathione (GSH)‐homeostatic machinery. Injuries in fetal rat cortical neurons associated with Fetal Alcohol Syndrome are correlated to ethanol (E) exposure leading to increased oxidative stress, GSH depletion, and ultimately neuroapoptosis. We hypothesize that neuroprotection is regulated by activation of the anti‐oxidant response elements (ARE) of cytoprotective genes, via the master transcription factor Nrf2. Primary cultured rat neonatal astrocytes were treated with 4mg/ml E at various time points. There was increased nuclear Nrf2 expression within 15 minutes after E treatment, and lasted up to 4 hours. Immunohistochemical studies corroborate cytoplasmic to nuclear translocation. Enhanced nuclear Nrf2 expression was further augmented by BSO pre‐treatment. In addition, the transcription factor Maf, which obligatorily dimerizes with Nrf2 to activate gene transcription, was increased by E or BSO. Known ARE inducers and oxidative stress mimicked the cytoplasmic to nuclear translocation seen in E treatment. Next, cells were pre‐incubated with Actinomycin D and subsequently treated with E. Nrf2 expression induced by E was abolished suggesting that E increases Nrf2 by stimulating gene transcription. Transfection of Nrf2 siRNA completely ablated Nrf2 expression, and furthermore blocks the tbHQ induced Nrf2, GGT, and Mrp1 expression. Taken together, this suggests that Nrf2 is required for activation of ARE. The role of E in Nrf2‐ARE control of astrocyte neuroprotection need to be further investigated. These points of regulation may allow for pharmacological augmentation of this crucial neuroprotective mechanism.
Biochemical Characterization of G93A Mutant human Cu, Zn Superoxide Dismutase Purified from Transgenic Rat Spinal cord
Authors: Bhogaraju Venkata K. , Reed Ronald L. , Crow John P.
Published: June 2021

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder with no cure and mutations to Cu‐Zn superoxide dismutase (SOD1) remain the only proven cause of the disease. SOD1, a 32‐kDa homodimer, is a ubiquitously expressed cytosolic antioxidant enzyme with one Cu and one Zn per monomer stabilized by an intra‐subunit disulfide (Cys 57 and Cys 146) bond. It also contains two more cysteines (Cys 6 and Cys 111) thought to be present as reduced thiols. To identify the potential mechanisms by which the mutant SOD1 (mSOD1) mediates neurotoxicity, we purified the mSOD1 from pre‐symptomatic and end stage ALS‐associated G93A mutant rat spinal cords (a model for ALS) using simple reverse‐phase preparative HPLC. The purified mSOD1 was characterized in terms of Cu and Zn levels, SOD activity, and thiol content using chromophoric chelation assay, SOD activity assay and thiol assay respectively. Our results indicate that Zn levels were significantly higher in mSOD1 purified from end stage cord to that of pre‐symptomatic cord, however no significant differences were observed in Cu levels and specific activities. Interestingly, thiol assay revealed that Cys 111 exists primarily in an oxidized state with Cys 6 primarily in the reduced state. These results suggest that biochemical impact of altered Zn affinity and cysteine modifications on SOD1 could have important implications in disease progression.