bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship between amount of food intake and bone mineral density in Korean 20‐30 years‐old women
Authors: Ann Jiyoung , Kim Minji , Shin Yoonjin , Woon Youngjun , Kim Yangha
Published: June 2021

The aim of this study was to investigate the relationship of food intake to bone mineral density (BMD) in Korean 20‐39 y women. In accordance with the amount of food intake, those who had food intake of the upper 50% were put into the control group (Control), while those who had food intake lower 50% were put into the low intake group (Low Intake: LI). The amounts of food intake were 1583 g in Control and 943 g in LI, respectively. The BMD of lumbar spine in control was significantly higher compared to that of LI. Nutrient intakes, such as energy, protein, P, Fe, Zn, Vitamin A, Vitamin E were higher in Control, but lower in LI compared to Korean Dietary Reference Intakes (KDRI). The intake of Ca was below KDRI in all subjects. Correlation analysis revealed that BMD was positively associated with intake of energy, protein, Ca, P, Fe, Zn, Vitamin A, Vitamin E. In conclusion it is confirmed that one of the most effective ways to maximize bone mineral density in young women may be to maintain an adequate food intake.
Analysis of Branched‐Chain α‐Keto Acid Dehydrogenase Complex (BCKDC) Activity and Activity State in Rat Skeletal Muscle by Isotope Ratio Mass Spectrometry (IRMS) using 13C‐Labeled Stable Isotope Substrate
Authors: Matsumoto Hideki , Akita Keiichi , Sakai Ryosei , Shimomura Yoshiharu
Published: June 2021

The BCKDC is the rate‐limiting enzyme in the catabolism of branched‐chain amino acids (BCAA). The radiochemical method has been used to measure the low level of the BCKDC activity in tissues such as skeletal muscle. In the present report, we introduced a non‐radioisotope method, using the α‐keto[1‐13C]‐isocaproate ([1‐13C]‐KIC) as stable isotope substrate. 13CO2 was released from [1‐13C]‐KIC in the reaction catalyzed by BCKDC. The ratio of 13CO2/12CO2 was measured by isotope ratio mass spectrometry. The detection limit of 13CO2 was estimated a few times lower than that of the radiochemical method. Using the purified BCKDC from bovine kidney, the production of 13CO2 was found to be linear with respect to reaction time (r2 = 0.9978) and quantity (r2 = 0.9920) of the enzyme. The muscle BCKDC activity measured by the new method was almost the same as that by the radiochemical method reported previously. Thus, we concluded that the novel assay method for the BCKDC activity is applicable to tissues containing the low level of the enzyme activity.
Quantitative Gas Chromatographic Method for C16:1t9, C18:1t9, C18:1t11, and C18:2tt9,12 trans Isomers Using Negative Chemical Ionization and Stable Isotope Internal Standards
Authors: Watson Amy D , Ribera Ashley , Vesper Hubert
Published: June 2021

Trans fatty acids have long been associated with risk factors for cardiovascular disease, thus making their consumption a major public health concern. Since the individual trans isomers of fatty acids typically make up less than 5% the total fatty acid content in a given biological matrix, epidemiological studies on the subject have been difficult. In addition, the absolute concentrations of these analytes were oftentimes subject to high variability in earlier reports. As part of the CDC's ongoing biomonitoring activities, we report an improved, highly sensitive GC/MS method that enables the measurement of 31 fatty acids including the trans fatty acids C16:1t9, C18:1t9, C18:1t11, and C18:2tt 9,12 in human plasma. At present, no other methods reported in the literature have been able to detect all of the aforementioned trans isomers in a single chromatographic separation.

In a set of 35 anonymous individuals, the median concentrations and ranges for the trans fatty acids were 0.0062 (0.0039‐0.0281), 0.0387 (0.0147‐0.2056), 0.0557 (0.0196‐0.5546), and 0.0016 (0.0000‐0.0074) µM for C16:1t9, C18:1t9, C18:1t11, and C18:2tt 9,12, respectively. The sum total concentration of the fatty acids analyzed was 12.13 µM. Therefore, the aforementioned trans fatty acids comprised only 0.73% of the fatty acids quantitated for this assay. The method reported herein will allow for more detailed studies on trans fatty acid intake and health effects associated with this intake.
Children's body mass index percentile (BMI%), social desirability score (SDS) and dietary recall accuracy for energy
Authors: Baxter Suzanne D , Guinn Caroline H , Royer Julie A , Hardin James W , Mackelprang Alyssa J , Smith Albert F
Published: June 2021

We examined whether 4th‐grade children's dietary recall accuracy for energy varied by BMI% and SDS. Each of 374 children (96% Black) was observed eating 2 school meals (breakfast, lunch) and interviewed to obtain a 24‐hour recall that included the 2 observed school meals. At the end of the interview (after the recall), children responded to 14 social desirability questions. We weighed and measured children, and calculated BMI%. We quantified accuracy of recalling energy intake at school meals by calculating conventional report rates (total reported energy/total observed energy) and the 2 components of report rate ‐ correspondence rate (total correctly reported energy/total observed energy) and inflation ratio (total intruded energy/total observed energy). For each measure, ANOVA was conducted with sex, BMI% x sex, SDS x sex, and BMI% x SDS x sex in the model; results were adjusted for other covariables. Report rates increased (indicating less underreporting) as BMI% and SDS decreased (2 p values = 0.001). Correspondence rates increased (indicating better accuracy) as SDS decreased (p = 0.013). Inflation ratios decreased (indicating better accuracy) as BMI% increased (p < 0.001). Report rates alone do not provide a complete picture of the relationship of children's dietary recall accuracy for energy with BMI% and SDS; BMI% and SDS appear to be related to separate aspects of recall accuracy. Funded by R01 HL074358.
Analysis of Zn contents in children's foods and assessment of Zn nutritional status in elementary school children by INAA
Authors: Lee Okhee , Chung Yonginsam , Moon Jongwha
Published: June 2021

Most of Korean children snacks are junk food and known to be high in calorie but deficient in micro‐minerals. Zn is associated with children growth and immunity. However, there are little reports on the Zn nutritional status of Korean children because of deficiency in Zn database of children foods. Furthermore, accuracy of existing data is questionable because they were analysed by AAS or ICP methods; these methods are known to be inaccurate due to large losses of minerals during pre‐treatment process. This study was aimed to evaluate the Zn nutrition state using Zn contents in children foods analysed by INAA.The homogenized food samples were freeze‐dried at temperature of ‐60 C°for 48~96 h and analyzed by INAA using with NIST SRM for analytical quality control. The mean Zn contents of one serving size were 0.85mg, 0.61mg, 0.52mg, 0.44mg, in the order of instant noodles and Korean buns> breads> ice creams> cookies> milk and milk products > chocolates > candy and sugars, respectively. The Zn intakes of children was not different by gender. Their zinc intakes were 7.5mg/d and 7.7mg/d and NAR of zn intakes were below 1 in children. Conclusively, Zn contents in children foods are quite varied according to food group and Zn intake was not enough in elementary children.
Abnormal methyl group metabolism and epigenetic regulation in the Zucker (type 2) diabetic fatty rat
Authors: Williams Kelly T. , Schalinske Kevin L.
Published: June 2021

Methyl group metabolism and DNA methylation have been shown to be altered in a streptozotocin‐induced model of type 1 diabetes. The Zucker diabetic fatty (ZDF) rat exhibits similar perturbations of methyl group metabolism, but DNA methylation status and the expression of epigenetic regulatory proteins have not been characterized in this model of type 2 diabetes. Because epigenetic regulation could be linked to secondary complications associated with type 2 diabetes, we chose to examine methyl group metabolism, including DNA methylation, in the liver and pancreas, as well as the heart and kidney. At 12 wk of age, 6 ZDF and 6 lean rats were sacrificed for blood and tissue analysis. As expected, ZDF rats were diabetic (2X increase in glucose and insulin) and exhibited marked hypohomocysteinemia. Glycine N‐methyltransferase (GNMT), a regulator of transmethylation, was divergently regulated in different tissues. In ZDF rats, hepatic GNMT activity and abundance were increased, whereas pancreatic activity was decreased. No change was noted in renal GNMT activity and abundance, or cardiac GNMT abundance, despite a lack of activity. In contrast to type 1 diabetes, hepatic DNA was hypermethylated in ZDF rats. Mechanistically, it appears that expression of epigenetic regulatory proteins (i.e. DNA methyltransferases and methyl CpG binding proteins) was abnormal in diabetic animals as well. (Support: Amer. Heart Assoc.)

Grant Funding Source

American Heart Association
Nutritional Epigenetics: Genome‐wide H3K9 Trimethylation Variations of HepG2 Cells in Hyperlipidemic Conditions
Authors: Cho Sungyun , Jun Hee‐Jin , Lee Sung‐Joon
Published: June 2021

Chromatin modification by aberrant histone lysine methylation can promote dysregulated gene transcription and disease progression. Nutrients may be involved in the epigenetic mechanism, however, their impact on cellular histone lysine methylation is poorly understood. Therefore, we investigated changes of histone 3 lysine 9 trimethylation (H3K9me3) at human gene coding and CpG island regions in hyperlipidemic conditions such as lipid‐loading mimicking high‐fat feeding within HepG2 cells. Chromatin immunoprecipitation linked to microarrays was adopted to get genome‐wide information on histone modifications. We found that H3K9 at coding and promoter regions of 1,235 and 1,188 target genes (= 10 fold change) were strongly hyper‐ or hypotrimethylated in hyperlipidemic conditions, respectively. Pathway analysis data showed that regulation of gene expression including chromatin modification, lipid metabolism including FXR/RXR activation and nutritional disease were significantly affected by changes in H3K9me3 of the corresponding genes. To our knowledge, this is the first study investigating histone methylation variations under hyperlipidemic conditions at genome‐wide level.
The dose dependent effect of chronic folate administration on S‐adenosylmethionine and S‐adenosylhomocysteine in human plasma
Authors: Bailey Steven W. , Alverson Patrica B. , Cohen Michael V. , Ayling June E.
Published: June 2021

A study was undertaken to examine the effect of treatment with folic acid (FA, 2.5 mg/d or 5 mg/d) or 5‐methyl‐6S‐tetrahydrofolate (MTHF, 5 mg/d) for ten weeks on the levels of S‐adenosylmethionine (SAM) and S‐adenosylhomocysteine (SAH) in plasma of predominately healthy subjects with mild hyperhomocysteinemia (>10 μM at screening). SAM and SAH were converted to their 1,N(6)‐etheno derivatives for analysis by HPLC using bromoacetaldehyde which permitted complete reaction within 2.5 hrs. Advantage was taken of the different pH optima of the two stages of this reaction to achieve a high level of reproducibility. An HPLC system was developed so that even without solid phase extraction the fluorescence of both etheno‐SAM and etheno‐SAH could be detected with high specificity and sensitivity (L.O.D. < 0.2 nM in plasma) within a single run. Plasma SAM increased significantly above baseline in comparison to placebo controls in the 2.5 mg/d FA and 5 mg/d MTHF, but not the 5 mg/d FA, treated groups. Previous pharmacokinetic studies have demonstrated that a 5 mg dose of FA remains largely unmetabolized in the blood for many hours after oral administration. It remains to be determined if the effect of high intake of FA on plasma levels of SAM is related to an interference in one‐carbon metabolism or possibly SAM and/or folate exchange between blood and tissues. Supported by NIH grant HL068165, Merck KGaA, & PamLab, LLC.
Demographic and anthropometric determinants of insulin like growth factor‐1; preliminary evidence from the third national health and nutrition examination survey (NHANES III)
Authors: Parekh Niyati , Lu‐Yao Grace
Published: June 2021

Evidence suggests that insulin‐like growth factor (IGF‐1) is associated with cancer progression. The purpose of this study is to describe blood IGF‐1 concentrations in relation with various demographic, anthropometric and lifestyle attributes. We used data from the nationally representative Third National Health and Nutrition Examination (NHANES III; 1988‐1994) survey, which used a cross‐sectional study design (n=6,029). Mean IGF‐1 concentrations were 260ng/ml in the whole population. IGF‐1 concentrations were highest among non‐Hispanic blacks (268ng/ml) and lowest in Mexican‐Americans (247ng/ml); they were higher among men as compared to women (278 versus 241ng/ml), decreased with age (p<0.001), and with body mass index (BMI <25; 274; BMI 35+; 209ng/ml). IGF‐1 concentrations were significantly and inversely correlated with BMI, waist circumference, serum leptin and insulin. IGF‐1 concentrations were lower among participants with self‐reported cancer status (261 vs. 207ng/ml) and did not differ among individuals having the metabolic syndrome or its components. In conclusion, the preliminary results from NHANES III suggest that the IGF‐1 has a complex relationship with body weight, age and sex.
Adiponectin and Leptin Affect Cell Cycle Regulation and Adiponectin Receptor Function
Authors: Connor Michael , Walker O'Llenecia
Published: June 2021

To establish a molecular link between obesity and cancer we examined the effects of the adipocyte produced peptides leptin (LEP) and adiponectin (ADIPO) on MCF7 cell cycle regulation. p27 levels decreased and increased with LEP and ADIPO, respectively, suggesting adipokine‐mediated MCF7 cell cycle regulation. LEP overcame the effects of ADIPO on p27 protein levels. LEP increased adiponectin receptor (ADIPOR) levels despite inhibiting ADIPO action, suggesting that LEP is stabilizing and inactivating ADIPOR. Inhibition of AKT enhanced ADIPOR protein levels, increased 14‐3‐3/ADIPOR binding and inhibited AMPK phosphorylation. Thus, phosphorylation of ADIPOR by AKT disrupts ADIPOR/14‐3‐3 interactions and promotes ADIPOR degradation. ADIPO induces AMPK‐mediated phosphorylation of p27 at T198 and stabilizes p27 protein, effects similar to those seen after AMPK activation by AICAR. ADIPO and AICAR effects on p27 phosphorylation were inhibited by the AMPK inhibitor compound C. ADIPO also induced decreases in ADIPOR protein levels. Our data suggest that ADIPO and LEP exert antagonistic effects on mammary cell cycle regulation, in part by regulating the function and expression of ADIPOR.
Longer‐term Effects of a Low Glycemic Index Diet on Glycemic Control in Type 2 Diabetes
Authors: Kendall Cyril , Esfahani Amin , Parker Tina , Banach Monica , Mitchell Sandra , Jenkins David
Published: June 2021

Background

Nut consumption, including peanuts, has been associated with a reduced risk of coronary heart disease (CHD). More recently, interest has grown in the potential value of including nuts in diets of individuals with diabetes.

Objective

To determine if tree nuts and peanuts improve glycemic control in non‐insulin dependent diabetes, as assessed by HbA1c and to assess whether these outcomes relate to improvements in CHD risk (serum lipids, blood pressure and oxidative stress and inflammatory biomarkers).

Methods

Approximately 120 NIDDM subjects (BMI ≤32kg/m2) treated with oral hypoglycemic agents (HbA1c 6.5‐8.0%) were recruited to a 3 month parallel design study. Subjects were randomized to one of three treatments: 1) Test (Full Dose Nut Diet): Raw nuts were added as supplements to the subject's usual diet based on required energy intake (≥2,400kcal/d received 100g/d nuts, ≈600kcal; 1,600‐2,400kcal/d received 75g/d nuts, ≈450kcal; ≤1,600kcal/d received 50g/d, ≈300kcal); 2) Test (Half Dose Nut Diet): Subjects received half dose of nuts and half dose of control muffin according to calorie needs; and 3) Control: whole wheat muffins were matched with energy content of nut supplements. One‐week weighed diet histories were obtained and fasting blood samples collected at baseline and weeks 2, 4, 8, 10 and 12 for markers of glycemic control and CHD risk factors.

Results

Final data to be presented.
Whole and fractionated yellow pea flour consumption alters post‐ prandial glucose response and insulin parameters, but not lipids or glucose levels, in overweight or obese, hypercholesterolemic humans
Authors: Marinangeli Christopher , Jones Peter J.H.
Published: June 2021

Yellow pea flour is a low fat, high fiber food; however, underlying mechanisms regarding health benefits of whole (WPF) and fractionated (FPF) yellow pea flours remain understudied. In a controlled feeding crossover study, 23 hypercholesterolemic, overweight and obese men and women received for 28 d two muffins/d containing WPF, FPF or white wheat flour (WF) as control. Treatments were separated by a 28 d washout. Two WPF muffins contained 50 g of whole yellow peas, with the amount of FPF in FPF treatments equaling the amount of pea‐derived fiber in WPF treatments. WPF and FPF consumption failed to affect fasting total cholesterol, LDL‐C, HDL‐C, and glucose. Subjects with a BMI between 25 and 34.9 receiving FPF tended (p=0.075) to have reduced post‐prandial glucose responses compared to WF. Insulin levels for WPF (31.8 ±6.5 pmol/ml, p=0.02) and FPF (32.0±6.5 pmol/ml, p=0.026) were lower compared to WF (37.9±6.5 pmol/ml). Insulin homeostasis model‐assessment showed that consumption of WPF (p=0.019) and FPF (p=0.012) increased insulin sensitivity 20% compared to WF. Results suggest that at energy balance, WPF and FPF do not alter fasting lipid and glucose levels and exert minimal effects on post‐prandial glucose response in hypercholesterolemic men and woman with elevated BMIs. Conversely, 50 g/d WPF and FPF consumption improves fasting insulin levels and sensitivity and thus provide dietary health benefits. Supported by Pulse Innovation Project, Canada
Bioaccessibility, biotransformation and transport of [alpha]‐mangostin from Garcinia magostana (Mangosteen)
Authors: Bumrungpert A. , Kalpravidh R. W. , Suksamrarn S. , Chaivisuthangkura A. , Jaratrungtawee A. , Chitchumroonchokchai C. , Failla M. L.
Published: June 2021

a‐ and g‐Mangostin (MG) are the most abundant prenylated xanthones present in the fruit of the mangosteen tree. These compounds have been reported to possess numerous bioactivities that have provided the impetus for use of mangosteen products in functional foods and dietary supplements. The health‐promoting benefits of mangosteen are dependent on delivery of the xanthones to target tissues. Here, we used simulated digestion and Caco‐2 cells to investigate the digestive stability, bioaccessibility and intestinal cell transport and metabolism of α‐ and γ‐MG. Transfer of a‐ and g‐MG to the aqueous fraction during simulated digestion was efficient (65‐74%) and dependent on bile salts, suggesting that micellarization is required for optimal bioaccessibility of xanthones. Cell uptake of xanthones from micelles was dose dependent and intracellular concentrations were maximum by 1h. Free and phase II metabolites of α‐MG were present in apical and basolateral media. The distribution of free plus conjugated α‐MG in the apical, cellular and basolateral compartments after 6 h was 79.7 ± 2.5, 11.8 ± 2.2 and 8.5 ± 1.7%, respectively; total recovery of α‐mangostin in wells ranged from 98‐101%. Transepithelial transport of free α‐MG significantly increased during prandial‐like compared to fasted conditions, suggesting fat mediated enhancement of absorption. Supported by NRC of Thailand and OARDC.
Protective effects of bamboo (Sasa borealis) leaf extracts against oxidative stress induced by t‐BHP in HepG2 cells
Authors: Park Yeon Ok , Lim Hyeon‐ Sook
Published: June 2021

Bamboo (Sasa borealis) has been widely utilized as a medicinal plant in Korea. In this study, hepato‐protective effects of bamboo leaf extracts against oxidative damage induced by tert‐butyl hydroperoxide (t‐BHP) were evaluated using HepG2 cells. Two crude extracts and five fractions of the 70% ethanol (EtOH) extract of the bamboo leaves were prepared by extracting using water or 70% ethanol and by fractionating sequentially. The two crude extracts and the five fractions were non toxic up to 150 ¥ì g/mL concentration, as determined by the 3‐(4,5)‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐tetrazolium bromide (MTT) assay. Decreased cell viability (%), increased leakage of lactate dehydrogenase (LDH), increased activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR), and raised concentration of malonedialdehyde (MDA) and decreased reduced glutathione (GSH) by t‐BHP were restored dose‐dependently by adding the crude extract of water or 70% EtOH or each fraction of the 70% EtOH extract. Among the two crude extract and five fractions, chloroform, ethyl acetate, and n‐butanol fractions showed the highest protective potential against oxidative damage. The results indicate that bamboo leaf extracts protect against oxidative damage in HepG2 cells.
Pathogenesis of heparin contaminant mediated vascular and hemostatic responses in animal models
Authors: Losada Angel , Litinas Evangelos , Hoppensteadt Debra , Jeske Walter , Fareed Jawed
Published: June 2021

Earlier this year, unfractionated heparin (UFH) and low molecular weight heparins (LMWH) were found to be contaminated with oversulfated chondroitin sulfate (OSCS) and high molecular weight dermatan sulfate (DS). The main contaminant, OSCS, had a molecular weight (MW) similar to UFH, but exhibited different biologic actions. OSCS does not interact with antithrombin, but activates the complement, kallikrein‐kinin and fibrinolytic systems via contact activation. To investigate the hemodynamic and hemostatic responses, contaminated (CH) and non‐contaminated heparin and isolated OSCS were administered to groups of rats and parameters such as blood pressure, bleeding time (BT) and antithrombotic actions (ATA) were measured. CH produced markedly higher ATA and BT responses relative to UFH. OSCS did not produce a strong hemodynamic effect at doses up to 2 mg/kg IV. However, when mixed with UFH, an augmentation of the hemodynamic, ATA and BT effects was seen. No significant differences were noted between OSCS and LMWH at doses up to 2 mg/kg (either IV or SC). OSCS isolated from heparin or LMWHs exhibited MW differences, but both produced similar effects on the contact system and platelet activation. These studies suggest that OCSC not only produces anticoagulant effects but also exerts pharamcodynamic effects/interactions with UFH. The presence of exogenous DS may further impact on their pharmacologic effects.
Investigating Therapeutic Approach of IBD Using Recombinant Glycoprotein Mucin2
Authors: LI XIAOXIAO , Wei Bo , Goodglick Lee , Wen Tao , Xia Lijun , Braun Jonathan
Published: June 2021

Inflammatory bowel disease (IBD) is characterized by a chronically inflamed mucosa of the gastrointestinal tract, caused by an underlying immune imbalance and alteration of mucosal barrier function. No effective medical cure has been developed for IBD due to our limited understanding of disease pathogenesis. Previous studies have demonstrated that the patients with Ulcerative colitis (UC)‐like mucosal inflammation exhibit a disrupted mucosal barrier. We speculate that the alteration of mucosal barrier integrity is initiated by the destructive interactions between abnormal enteric microbiota and mucous layer. Consistently, analysis of O‐glycan, a major component of colonic mucin in bowel rinse samples from patients with UC has revealed an alteration of mucin structure. This study further explores a novel therapeutic approach for IBD treatment by using exogenous recombinant glycoprotein to block the harmful interactions driven by the colonic bacteria. We established the CHO‐DG44 cell lines producing a recombinant Mucin2, the most abundant secretary glycoprotein in the colon and characterized the O‐glycans on the recombinant Mucin2 protein. Purified O‐glycosylated Mucin2 are being evaluated for the therapeutic effect in the mice model of mucosal inflammation caused by an impaired mucosal barrier. The study would provide a proof of concept to the therapeutic benefit of treating human IBD with recombinant glycoprotein administration.
RV‐induced type 1 IFNs induce antiviral gene expression and apoptosis in intestinal epithelia
Authors: Frias Amena Husain , Jones Rheinallt , Vijay‐Kumar Matam , Gewirtz Andrew T.
Published: June 2021

Rotaviruses (RV) are the leading cause of diarrhea in children. RV primarily infects intestinal epithelial cells (IEC), which typically clear the infection within 7 days via an innate immune‐mediated mechanism. Type 1 IFNs (α/β) mediate various aspects of the immune response to a number of viruses in a variety of cell types. Thus, we sought to define whether RV induced type I IFN in IEC and determine the role of such IFN in the antiviral immune response. Intestinal epithelial cells (HT29) were infected with RV (MOI 1) alone or in the presence of neutralizing antibodies to IFN‐α/β. Viral protein synthesis, antiviral signaling (IRF7, STAT1, and PKR) and PARP cleavage (an apoptotic event) was detected by western blotting. Cell morphology was assessed by light microscopy. Type 1 IFN and IL‐8 (inflammatory cytokine) induction was measured by qRT‐PCR or ELISA. RV robustly induced IFN‐β, but not IFN‐α. In accordance, blockade of IFN‐β was without effect, while blockade of IFN‐β completely ablated the potent induction of p‐STAT1, p‐PKR and IRF7 induced by RV alone. IFN‐β blockade also resulted in modest suppression of viral protein expression. Most surprisingly, RV alone induced epithelial cytopathology/ apoptosis that was abrogated when IFN‐β activity was blocked. Thus, RV‐induced IFN‐β is essential for IEC antiviral signaling and may also promote viral replication and apoptosis of infected cells.
NADPH oxidase is involved in polychlorinated biphenyl‐mediated alterations of tight junction proteins in cells of the gastrointestinal tract
Authors: Choi Yean Jung , Eum Sung Yong , Son Heejeong , Hennig Bernhard , Toborek Michal
Published: June 2021

Exposure to polychlorinated biphenyls (PCBs) can contribute to numerous disease states. The main route of exposure to PCBs is through the gastrointestinal tract; however, little is known about the effects of PCBs on intestinal epithelial barrier functions. In the present study, we evaluated the effects of several highly‐chlorinated PCBs using human intestinal Caco‐2 cell line as an in vitro epithelial model system. Epithelial cells are comprised of tight junction (TJ) proteins that form an intricate assembly of membrane, cytoplasmic and accessory proteins linked to an actin cytoskeleton and which contribute to limited paracellular diffusion between adjacent epithelium. We demonstrated that exposure to PCBs increased permeability of FITC‐labeled dextran (4 kDa) across Caco‐2 monolayers. Consistent with permeability results, PCB disrupted expression of TJ proteins occludin and ZO‐1. These effects were associated with upregulation of the expressions of NADPH oxidase subunits as detected by immunoblotting. In addition, inhibition of NAPDH oxidase by apocynin significantly blocked PCB‐mediated increase in epithelial permeability and alterations of TJ protein expression. Similar effects were exerted by pretreatment with Rho kinase and Ras inhibitors suggesting that PCBs can disrupt epithelial barrier integrity through induction of redox‐regulated mechanisms. Supported by NIH/NIEHS (P42 ES 07380).
FPR1 does not mediate neutrophil infiltration in experimental colitis
Authors: Stadnyk Andrew William , Farooq Shukkur M.
Published: June 2021

One hallmark of the inflammatory bowel diseases is the infiltration of neutrophils (PMN) into the mucosa. The mechanism behind this infiltration is unclear but bacterial N‐formyl peptides are candidates since these are potent chemoattractants and activating factors for PMN. We investigated this hypothesis by inducing colitis in mice deficient for the formyl peptide receptor, FPR1. FPR1‐/‐ and wild type (WT) mice were administered dextran sodium sulphate (DSS) through their drinking water for 5 days followed by one day of pure water (acute phase) or 4 days of water (recovery phase), or for 2 DSS cycles (chronic phase) with the second cycle commencing on day 22. During acute colitis FPR1‐/‐ mice lost up to 5% of their starting body weight whereas WT mice lost up to 20%. However, the histopathology was similar between the two groups at both the acute and recovery phase points. This included ulcers, PMN infiltration, and loss of crypt architecture. In contrast, during the chronic phase FPR1‐/‐ mice experienced weight loss of up to 20%, greater clinical illness, and 15% shorter colons than WT mice, all indications of a more severe disease. Whether this is reflected in differences in the histopathology remains to be evaluated. We conclude that FPR1 is not necessary for PMN infiltration into the acutely inflamed colon; however, it may play a role in chronic colitis. Funded by the CCFC and an IWK Health Centre Fellowship to SMF.
TNF‐α‐induced cyclooxygenase‐2 expression in human synoviocytes
Authors: Chen Chun‐Hua , Yang Chuen‐Mao
Published: June 2021

COX‐2 is implicated in the pathogenesis of rheumatoid arthritis (RA) and has previously been shown to be induced by TNF‐α is implicated in the pathogenesis of rheumatoid arthritis (RA) and has previously been shown to be induced by TNF‐α. However, the molecular mechanisms underlying TNF‐α‐induced cyclooxygenase‐2 (COX‐2) expression in human synoviocytes (HS) which contributes to inflammatory responses in RA were largely unknown. Here, Western blotting and RT‐PCR analyses showed that TNF‐α induced COX‐2 protein and mRNA expression as well as phosphorylation of p42/p44 MAPK, p38 MAPK, and JNK which was attenuated by the inhibitors of MEK1/2 (U0126), p38 MAPK (SB202190), and JNK (SP600125), and transfection with siRNA of ERK1, p38, and JNK2, respectively. Furthermore, TNF‐α‐induced COX‐2 expression was inhibited by a selective NF‐¿B inhibitor (Bay11‐7082) and consistent with that TNF‐α stimulated both I¿B‐α degradation and NF‐¿B translocation into nucleus in these cells. NF‐¿B translocation was blocked by Bay11‐7082, but not by U0126, SB202190, and SP600125. Taken together, these results suggested that in HS, TNF‐α‐induced COX‐2 expression was independently mediated through phosphorylation of MAPKs and NF‐¿B pathways.
Immunological effects of polyphenols contained in Fermented Grape Marc (FGM) on human healthy peripheral blood cells
Authors: Jirillo Emilio , Marzulli G. , Jirillo F. , Tafaro A. , Kaneko M. , Kumazawa Y. , Amati L.
Published: June 2021

The effects of polyphenols extracted from FGM of Negroamaro and Koshu wines were evaluated on human Peripheral Blood Mononuclear Cells (PBMC) in terms of regulatory, inflammatory and anti‐inflammatory cytokine production. Lyophilized FGMs were suspended in water or ethanol (100 mg/ml) and supernatants were added to 1 x 106 PBMC/ml in culture media. After 4 hours incubation at 37°C, cells were marked with anti‐human CD4 and CD14 moAbs, respectively . To study intracellular cytokine profile, cells were permeabilized and marked with anti‐human IL‐10, IL‐12 and TNF‐α moAbs, respectively. The analysis with the flow cytometer revealed that Koshu FGM markedly enhanced intracellular content of IL‐10 by CD4+ T cells when it was suspended in water but not in ethanol. On the other hand, Koshu greatly increased the enhanced intracellular of IL‐12 by CD14+ cells (monocytes) when it was suspended in ethanol but not in water. Negroamaro FGM was able to augment the intracellular content of TNF‐α by CD14+ cells, when it was suspended in water but not in ethanol. On the contrary, Koshu markedly enhanced the intracellular content of TNF‐α when it was suspended in ethanol but not in water. These results may represent a starting point for the use of FGM as modulators of the immune response in different pathologies as well as anti‐inflammatory agents via release of IL‐10 (e.g., Koshu suspended in water).

This work was in part supported by MIUR and MIN. SALUTE (Rome, Italy).
Modulation of reactive oxygen species and MMP9 by Rac1 prevents asbestos‐induced pulmonary fibrosis
Authors: Murthy Shubha , Tephly Linda , Perry Sarah , Adamcakova‐Dodd Andrea , Keller Richard , Metwali Nervana , Wang Yongqiang , Glogauer Michael , Thorne Peter , Carter A. Brent
Published: June 2021

The release of reactive oxygen species (ROS) and cytokines by alveolar macrophages has been demonstrated in asbestos‐induced pulmonary fibrosis, but the mechanism linking alveolar macrophages to the pathogenesis is not known. The GTPase Rac1 is a second messenger that plays an important role in host defense. In this study, we demonstrate that Rac1 null mice are protected from asbestos‐induced pulmonary fibrosis. We hypothesized that Rac1 induced pulmonary fibrosis via generation of ROS. Asbestos increased ROS in a Rac1‐dependent manner with ROS being significantly greater in wild type (WT) mice compared to Rac1 null mice. Extracellular matrix degradation was dependent on Rac1 activity and ROS. Expression of the macrophage specific metalloproteinase, MMP9, was significantly less in WT cells compared to Rac1 null cells. In addition, Rac1 null cells exhibited enhanced gene expression driven by AP1, an important transcription factor in MMP9 expression. Over expression of catalase decreased ROS generation and increased MMP9 expression in vitro. Importantly, administration of catalase to WT mice attenuated the development of fibrosis in vivo. For the first time, these results demonstrate that Rac1 plays a crucial role in asbestos‐induced pulmonary fibrosis. Moreover, it suggests that a simple intervention may be useful to prevent progression of the disease. This work was supported by NIH grant ES‐015981.
Dissecting the Biological Function of NF‐¿B2p100
Authors: Yang Liqun , Ding Han‐Fei
Published: June 2021

Nuclear Factor‐kappaB2 (NF‐¿B2), a member of the NF‐¿B family of transcription factors, has an important role in the development and function of the immune system. It is synthesized as a protein of 100‐kDa (p100). In response to certain cytokines, p100 undergoes proteolytic processing, leading to the generation of p52 that corresponds to the N‐terminal half of p100. p52 functions as a transcription activator with anti‐apoptotic activity; constitutive activation of p52 can lead to autoimmune inflammation. Previous in vitro studies have provided evidence that, in addition to serving as a precursor for p52, p100 functions as an I¿B molecule with the ability to restrain p52 activity. However, the physiological relevance of this I¿B activity of p100 has never been demonstrated. To address the question, I have generated p52 transgenic mouse lines with or without NF‐¿B2 p100. Compared to p52 transgenic mice carrying wild‐type p100 alleles, p52 mice deficient in NF‐¿B2 p100 show a marked increase in the mortality rate with severe inflammation in the lung, characterized by extensive infiltrations of lymphocytes and macrophages. Lymphocytes from these mice contain significantly higher levels of ¿B‐binding activity. These data demonstrate the function of p100 as a mechanism to limit the activity of p52, which has important implications in clinical control of inflammation.

Supported by NCI Grant R01 CA106550 (to H.‐F. D.).
Complex Vascular Lesions in the Lungs of Domestic Fowl Selected for Susceptibility to Pulmonary Arterial Hypertension: Incidence and Histology
Authors: Wideman Robert F , Hamal K. , Bayona M. , Lorenzoni A. , Cross D. , Rhoads D. , Erf G. , Anthony N.
Published: June 2021

Humans developing pulmonary arterial hypertension (PAH) have a poor prognosis for survival when plexiform lesions progressively obstruct their terminal pulmonary arterioles. Plexogenic arteriopathy rarely develops in existing animal models of PAH. Rapidly growing chickens spontaneously develop PAH accompanied by the formation of plexiform‐like complex vascular lesions (CVL). We now report the frequency with which at least one CVL per tissue section was observed in the lungs of chickens from a line selected for susceptibility to PAH: 48% of females (n = 128 lungs) and 44% of males (n = 132 lungs) spontaneously developed CVL; 38% of right lungs and 51% of left lungs exhibited CVL; and at 8, 12, 16, 20, 24, and 52 weeks of age the CVL incidences were 52, 50, 51, 40, 36, and 18%, respectively. CVL formed distal to branch points in muscular inter‐parabronchial pulmonary arterioles that typically exhibited medial hypertrophy and intimal proliferation. Perivascular mononuclear cell infiltrates consistently surrounded the affected arteriole branch points. Proliferating intimal cells fully or partially occluded the arteriole lumen adjacent to a mature CVL, and the dilated main body of the CVL encompassed diverse cell types, including foam‐like giant cells and a glomeruloid‐like capillary network. These CVL in the lungs of chickens strikingly resemble published photomicrographs of plexiform lesions in the lungs of human PAH patients, potentially providing biomedical researchers with a much‐needed animal model of spontaneous plexogenic arteriopathy. Supported by NIH grant 1R15HL092517.
The Establishment of Acetic Acid‐Induced Cerebral Infarct Model, and Evaluation of Tissue Plasminogen Activator (t‐PA), Edaravone, Argatroban and Ticlopidine
Authors: Shibata Keita , Hashimoto Terumasa , Nobe Koji , Ohata Hisayuki , Honda Kazuo
Published: June 2021

The aim of the present study is to establish a novel embolic infarct model that was similar to human clinical condition and to evaluate the effects of some anti‐cerebral infarct agents were examined. Thrombotic occlusion was induced by local application of acetic acid to the right common carotid artery in the male Mongolian gerbil. Infarct area was assessed by the 2% TTC staining. Neurological scores were determined by a modified method described by Longa et al. The infarct area and the neurological scores increased in acetic acid concentration‐dependent manner (60, 80, 100%). Infarct area reached the maximum at 24 hr after ischemia and sustained until 5 days later. However, neurological scores decreased following the maximal response. Treatment with t‐PA reduced both values in a dose‐dependent manner (0.01, 0.1, 10 mg/kg). A free radical scavenger, edaravone (3 mg/kg), a thrombin inhibitor, argatroban (10 mg/kg), and an antiplatelet agent, ticlopidine (10 mg/kg) slightly affected. These results indicate that this model may be a useful for evaluating the cerebral infarct therapeutic drugs and provide the information about pathophysiological mechanisms of cerebral infarction.
High molecular weight Fibroblast Growth Factor 2 (FGF2) requires an active Fibroblast Growth Factor Receptor 1 (FGFR1) to depress post‐ischemic cardiac function
Authors: Bodmer Janet R. , Wright Edward , Newman Gilbert , Schultz Jo El J.
Published: June 2021

Fibroblast growth factor 2 (FGF2) is a family of isoforms with potential both as therapeutic molecules, as well as endogenous targets for pharmacological modulation in cardiac ischemia. Our laboratory has previously demonstrated that the secreted low moledular weight (LMW) FGF2 isoform protects the heart against myocardial dysfunction induced by ischemia‐reperfusion (I/R) injury by activating the FGF receptor 1 (FGFR1), while the mechanisms by which the high molecular weight (HMW) FGF2 isoforms produce the opposing and detrimental effects on cardiac function following I/R are less clear. The purpose of this study was to examine whether FGFR1 activation is necessary for the effects of HMW FGF2. Using an isolated work‐perfoming heart model, mice overexpressing the 24kDa isoform of FGF2 were subjected to global low‐flow ischemia in the presence of an inhibitor to FGFR1. The blockade of FGFR1 resulted in higher post‐ischemic contractility in transgenic hearts (p<0.05). We also found differences in the activation of kinases downstream from FGFR1, including protein kinase C (PKC) and mitogen activated protein kinases (MAPKs), indicating that HMW FGF2 alters the targets of FGFR1 signaling cascades. These findings indicate that HMW FGF2 mediates its effects by modulating FGFR1 signaling, to induce post‐ischemic myocardial dysfunction.

Research supported by NIH grant NIH/NHLBI RO1 HL075633
Potential Role of PP2A in Regulating Ca2+‐Activated Cl− Channels in Pulmonary Artery Smooth Muscle Cells
Authors: Ayon Ramon , Sones William , Forrest Abigail , Perrino Brian , Valencik Maria , Sanguinetti Amy , Greenwood Iain , Leblanc Normand
Published: June 2021

Ca2+‐activated Cl− currents (IClCa) represent a major excitatory mechanism in vascular smooth muscle cells (SMCs). Our group previously showed that CaMKII and Calcineurin (CaN) exert opposed regulatory effects on IClCa in arterial SMCs. In this study, we determined whether other phosphatases also modulate IClCa in rabbit pulmonary artery (PA) myocytes. In a previous study, our group (Ayon et al., Faseb J. 22: 937.16, 2008) detected significant mRNA and protein expression of multiple isoforms for PP1, PP2A and PP2B in rabbit PA tissue homogenates. Recently, we have extended our studies to show significant cytoplasmic immunolabeling of PP1α, PP1β/d, PP1γ, and PP2A in isolated PASMCs. Results from previous patch clamp experiments on freshly isolated PA myocytes (Ayon et al., Faseb J. 22: 937.16, 2008) showed differential responses when manipulating endogenous PP1 and PP2A as opposed to supplying these phosphatases exogenously. To address this issue we examined the effects of Fostriecin (FOS), a highly specific inhibitor of PP2A, on an exogenous application of PP2A in the presence of ATP. Cells dialyzed with 150 nM FOS produced a delayed but pronounced block of exogenous PP2A‐induced recovery of IClCa which contrasted with the inability of FOS to alter the recovery of IClCa evoked by endogenous phosphatase(s). These results suggest that endogenous PP1 and CaN, not PP2A, are involved in IClCa regulation in PASMCs.
PKA Enhances the Sensitivity of IP3 Receptors in Swine Airway Submucosal Gland Cells.
Authors: LU SILU , FARLEY JERRY M
Published: June 2021

Increased sensitivity of airway submucosal gland cells (SMGC) to ACh by PKA activation results in increased transepithelial ion movement. The mechanism of the increased sensitivity induced by PKA is unclear. Since ACh‐induced transepithelial ion movement is dependent on the IP3/Ca2+ signaling pathway, we hypothesized that PKA enhances sensitivity of IP3 receptors in airway SMGC, resulting in increased [Ca2+]i. Patch clamp of swine airway SMGC was performed in these studies. We showed that forskolin decreased the EC50 for IP3 about 4‐fold measured as the peak KCa current induced upon whole cell formation with IP3 in the pipette. VIP (10 nM), a co‐neurotransmitter present in cholinergic airway nerve fibers, enhanced the response to 0.3 µM IP3. Neither VIP nor IP3 (0.3 µM) induced KCa current when applied alone. VIP induced a significant KCa current in SMGC only when the internal solution contained IP3. Removing external Ca2+ did not abolish the VIP‐induced sensitization of the peak IP3 response although the plateau response was lost. This suggests that Ca2+ release is sensitized as well as Ca2+ influx. Elevating external Ca2+ to 5 mM revealed a KCa current induced only in the presence of IP3 (0.3 µM). Thus Ca2+ entry may also be increased by IP3 (0.3 µM). These results suggest that PKA increases the sensitivity of IP3 receptors by phosphorylation, resulting in the increased intracellular Ca2+ release and Ca2+ entry.
Calcium‐Calmodulin Dependent Kinase II (CaMKII) N‐methyl D‐aspartate Receptors (NMDAR) Interactions in Development and Disease
Authors: Gustin Richard , Baucum Anthony , Colbran Roger
Published: June 2021

Postsynaptic CaMKII translates dynamic changes in local calcium concentrations into diverse functional responses. Proper CaMKII localization, phosphorylation, and activity are crucial for long‐term potentiation as well as normal learning and memory, and are disrupted in animal models of multiple neurological diseases (van Woerden et al., 2007). To determine how changes in CaMKII expression and phosphorylation over a developmental timeline (rodent postnatal day: 7, 25, 90) affects its interactions with CaMKII‐associated proteins (CaMKAPs), we have developed a fractionation protocol which allows us to probe interactions in cytosolic, membrane‐associated, and post‐synaptic density (PSD)‐associated pools of proteins. Both CaMKII and NMDAR subunits are present in multiple subcellular compartments, but a complex of these proteins can only be co‐immunoprecipitated from the PSD pool. In addition, CaMKII differentially interacts with NMDAR subunits across the developmental timeline. Other CaMKAPs that modulate CaMKII localization and activation are also being explored. Ultimately, we will determine how these interactions are disrupted in mouse disease models that exhibit hyperphosphorylation of CaMKII (i.e. Angelman Syndrome, Parkinson's Disease) as well as other models of altered CaMKII phosphorylation (T286A CaMKII knockin mice).
Modulation of lysine acetylation‐stimulated repressive activity by Erk2‐mediated phosphorylation of RIP140 in adipocyte differentiation
Authors: Ho Ping‐Chih , Gupta Pawan , Tsui Yao‐Chen , Ha Sung gil , Huq MD Mastaqul , Wei Li‐Na
Published: June 2021

Receptor‐interacting protein 140 is a co‐regulator for transcription factors. Previous studies showed that either phosphorylation or lysine acetylation of RIP140 directly enhanced its trans‐repressive activity. Here, we identified p300 as a specific lysine acetyltransferase, and ERK2 as a specific kinase for threonine phosphorylation, of RIP140 in vivo. We further determined two specific acetylated lysine residues (Lys158/Lys287) that were critical for its gene repressive activity. We then delineated signal transduction from Erk2‐mediated phosphorylation of RIP140 that enhanced its recruiting p300 for subsequent lysine acetylation, and demonstrated the kinetics of activation of this signal transduction pathway in differentiating adipocytes. The physiological significance of this cell signal transduction pathway was illustrated in rescuing experiments where the defect in fat accumulation of RIP140‐null cultures was rescued by re‐expressing the wild type RIP140 or its phospho‐mimetic mutant, but not its acetylation deficient mutant. These results demonstrate ERK2‐meidated threonine phosphorylation, followed by p300 recruitment for lysine acetylation, which ultimately enhances the gene‐repressive activity of RIP140 and its role in fat accumulation in differentiated adipoctyes. This work was supported by DK 54733, DK 60521, K02‐DA13926 and DA11190 to L.‐N. Wei.