bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of structural and functional determinants of Ca2+‐dependent kinetics of voltage‐gated calcium channel regulation by CaM
Authors: Taiakina Valentina , Spafford David , Guillemette Joseph Guy
Published: June 2021

Calmodulin (CaM), a widely expressed and highly conserved Ca2+‐sensing regulatory protein with at least 300 known targets, has been recently implicated in the Ca2+‐dependent inactivation and facilitation of voltage‐gated calcium channels. Due to its bi‐lobed structure, CaM is able to serve as a bi‐functional sensor and regulator of channel activity. The particular contribution of each of the two lobes and the associated forms of regulation are not universally understood or generalized for each channel type, and even less so are the particular channel regions responsible for the association and regulation by CaM. In this study we examined unique regions outside the canonical channel CaM‐binding domains for their ability to dictate the electrophysiological profile of any channel type. Channel chimaeras were constructed and patch‐clamp recordings were performed to determine the Ca2+‐dependent kinetics profiles. Furthermore, the peptides corresponding to putative CaM‐binding regions of the channel were synthesized and the binding properties were analyzed. The implications for the channel regulatory model and some unifying principles are discussed. This work received NSERC funding.
Inhibition of lytic function of human NK cells by Tributyltin: Role of Protein Kinase C
Authors: abraha abraham berhanemeskel , Whalen Margaret M
Published: June 2021

Protein kinase C (PKC) catalyzes the phosphorylation of serine/threonine residues and is involved in signal transduction associated with cell proliferation, differentiation, and apoptosis. The environmental pollutant, tributyltin chloride (TBT) has been reported to decrease human Natural killer (NK) cell lytic function. We previously showed that TBT induces activation of MAPK in NK cells. In the present study, we examined the role of PKC in TBT‐ induced inhibition of NK lysis of tumor cells and TBT‐induced activation of MAPKs by using a PKC inhibitor. A 1 h treatment with Bisindolylmaleimide I caused 85% decrease in the ability of NK cells to lyse K562 tumor cells in a 2‐h 51Cr‐release assay. Western blot analysis using an antibody to phospho‐MAPKs showed that Bisindolylmaleimide I was able to completely block TBT‐induced activation of p44/42 whereas activation of p38 and JNK were not blocked. However, when the activation of PKC was blocked by the presence of Bisindolylmaleimide I, subsequent exposure to TBT produced even greater inhibition of lytic function of NK cells than was seen with TBT alone. These results indicate the pivotal role of PKC in the TBT‐induced activation of p44/42 in NK cells, possibly by activating Raf ‐> MEK ‐> MAPKs and suggest the negative effect of TBT in NK lytic function was not prevented even when the activation of PKC was blocked.
ANTIBACTERIAL PROPERTIES OF SOME NEW PYRIDO – 1,2,4 – TRIAZOLE‐ 3 – THIONES
Authors: Govori Sevdije R , Rifati Arleta , Leci Osman , Haziri Arben
Published: June 2021

8‐amino‐[1,2,4] triazolo [4,3‐a] pyridine‐ 3(2H) ‐ thione (7), 6‐amino‐[1,2,4] triazolo [4,3‐a] pyridine ‐ 3(2H) ‐ thione (8) and 5‐amino‐1,2‐dihydro‐3H‐pyrazolo [4,3‐b] pyridine ‐ 3 ‐ thione (9) ,are synthesized through corresponding hydrazine derivatives (4 ‐6) with CS2 in very good yields.

The identity and structure of these compounds was proven on the basis of spectral data and elemental microanalysis.

The new compounds (7‐9) showed strong activity in reducing the microbial growth. The inhibitory effects of compounds were affected by their structure. All compounds showed the most effective antibacterial activity against Escherichia coli , Staphylococcus aureus , and Klebsiella pneumonie .
Gambogic acid covalently interacts with IKK‐2 to inhibit the NF‐kB signaling pathway
Authors: Gandhi Ujjawal H , Kalantari Parisa , Palempalli Uma D , Vunta Hema , Arner Ryan J , Prabhu K. Sandeep
Published: June 2021

Gambogic acid (GA) is a polyprenylated xanthone abundant in the resin of Garcinia morella and G. hanburyi with a long history of use as a complementary and alternative medicine. The anti‐tumor activity of GA has been well demonstrated and is thought to arise partly due to the anti‐inflammatory activity associated with GA. Here we demonstrate that GA inhibits the LPS‐dependent expression of nuclear factor‐kB (NF‐kB) target pro‐inflammatory genes in murine macrophages. Western immunoblot, NF‐kB luciferase reporter, and gel shift analyses revealed that GA strongly blocked the activation of NF‐kB induced by LPS; while 9,10‐dihydroGA that lacks the reactive alpha,beta‐unsaturated carbonyl group was ineffective. Moreover, GA was able to decrease nuclear p65 levels in RAW264.7 macrophages, where the expression of TfR1 was down‐regulated by RNA interference. In‐vitro kinase assays coupled with interaction studies using biotinylated GA as well as proteomic analysis demonstrated that IKK2, a key kinase of the NF‐ ?ÛB signaling axis, was covalently modified by GA at Cys179 causing significant inhibition of its kinase activity. Taken together, these data demonstrate that covalent modification of IKK2 as an underlying mechanism of the potent anti‐inflammatory activity of GA. NIH AT004350 (KSP)
Flavonoids can be cytotoxic to breast tumor cells despite inhibiting caspase‐3
Authors: Beckford Jeremy , Yadegarynia Sina , White J Brandon , d'Alarcao Marc
Published: June 2021

Flavonoids are polyphenolic plant natural products found extensively in the human diet. This class of compounds has been of considerable medicinal interest because of numerous reports of anti‐inflammatory and anti‐cancer activities. However, a detailed understanding of the biochemical mechanism of these activities is lacking because the compounds have been reported to be antioxidants and also affect multiple cell signaling pathways, including those mediated by protein kinase C, phosphoinositide 3‐kinase, Akt/protein kinase B, tyrosine kinases, and mitogen activated protein kinase.

We have undertaken a study to determine which of the various cellular activities are necessary and sufficient to induce cell death in a variety of breast cancer cell lines. As part of this study, we have evaluated the ability of various flavonoids to induce apoptosis by evaluating their effect on various markers, including caspase 3/7 activation, and cytochrome C release from the mitochondria. We have found that several flavonoids, in addition to inducing cell death are inhibitors of caspase 3. This poster will report the details of our studies on the inhibition of caspases by flavonoids.

Support of research was through the National Institutes of Health RISE Program at San Jose State University to JB.
A methanol extract from Collinsonia canadensis root decreases lipopolysaccharide‐induced inflammatory response in murine macrophages
Authors: Hanlon Paul Robert , Williams Melissa , Barnes David
Published: June 2021

Collinsonia canadensis, or Stone Root, a perennial plant in the mint family has its origins as a medicinal herb in Native American culture. HPLC analysis identified rosmarinic acid as a major component in Collinsonia shoots and the roots at 20.1 and 15.8 mg/g, respectively. A 80% methanol extract of the Collinsonia root also contained two other major compounds with spectra typical of phenolic compounds. The anti‐inflammatory properties of Collinsonia extract were then assessed using a lipopolysaccharide (LPS)‐stimulated macrophage model (RAW 264.7 cells). The 80% methanol extract from Collinsonia inhibited LPS‐stimulated nitrite production at a concentration corresponding to 5 mg of dry Collinsonia root per mL of media without producing toxicity. The three major phenolics from Collinsonia root were then purified using preparatory HPLC. Treatment of RAW cells with rosmarinic acid at an equimolar concentration as was present in the 5 mg/mL Collinsonia extract did not significantly inhibit LPS‐stimulated nitrite production. However, a combination of all three major phenolic compounds equimolar to the 5 mg/mL Collinsonia extract treatment did result in significant inhibition of LPS‐stimulated nitrite production. This initial characterization of a methanol extract from Collinsonia demonstrated high concentrations of phenolic compounds with anti‐inflammatory activity in macrophage cells.
Mango improves glucose and lipid parameters in mice fed high fat diet.
Authors: Li Wenjia , Peterson Sandra , Perkins‐Veazie Penny , Clarke Stephen , Kuvibidila Solo , Smith Brenda Jane , Lucas Edralin
Published: June 2021

Mangoes contain high amounts of beta carotene, vitamin C, and phenolic compounds that may confer cardiovascular health benefits. The objective of this study is to compare the effects of freeze dried mango with a known PPARγ agonist, rosiglitazone (rosi), and a known PPARα agonist, fenofibrate (feno), in modulating glucose and lipid parameters using a mouse model of diet‐induced obesity. Eight wk old male C57BL/6 mice were randomized into 6 treatment groups (n=25/group): control diet, high fat (HF) diet, HF + 1% mango (w/w), HF + 10% mango (w/w), HF + rosi (50mg/kg diet), and HF + feno (500mg/kg diet). Dietary treatment lasted for 8 or 16 wks. At both time points, HF fed mice had the highest fat mass and the two doses of mango were comparable to rosiglitazone in attenuating the increase in fat mass due to HF diet. Similar to rosiglitazone, mango modulated the rise in plasma glucose, cholesterol, and NEFA associated with HF diet for 8 wks but no treatment effects were observed after 16 wks. Mice on the low dose mango diet for 8 wks had the best response after a glucose challenge. Plasma leptin was significantly elevated in the HF group after 8 wks of dietary treatment which was modulated by rosiglitazone, fenofibrate, and the two doses of mango. Fenofibrate caused an enlargement of liver which was not observed with the other treatments. These findings suggest that mango may be beneficial in lowering risk factors of cardiovascular disease. (Supported by the Mango Board and College of Human Environmental Sciences, Oklahoma State University)
Ginsenosides and their role in modulating TNF‐alpha signaling to enhance apoptosis in MDA‐MB 231 breast cancer cells
Authors: Hantak Alison M , Murphy Laura L
Published: June 2021

The aim of the present study was to determine if ginsenosides enhance tumor necrosis factor‐ (TNF‐α) induced apoptosis through modulation of TNF‐α signaling pathways. MDA‐MB 231 breast cancer cells were treated with 0.5 mg/ml of ginsenoside fraction (GF), 12.5 ug/ml of the ginsenoside metabolite Rh2, or 100 ng/ml of TNF‐α, alone or in combination, for 24 hours. The inclusion of GF or Rh2 together with TNF‐α increased percentage of cell death, but all three in combination further enhanced this effect. Flow cytometric analysis showed that GF or Rh2 with TNF‐α induced a G0/G1 arrest, while GF+TNF+Rh2 produced a robust increase in the pre‐G0/G1 cell population relative to other treatments. Further co‐treatment with a pan caspase inhibitor (Z‐VAD‐FMK) indicated that treatment‐induced cell death is caspase dependent. To analyze the role of TNF‐α in this system, proteins were collected at 15, 30, and 60 minutes after treatment and analyzed for phosphorylated levels of IkB and JNK. TNF‐α stimulated these downstream targets, but GF and Rh2 together with TNF‐α diminished this effect. Cells were then pretreated with inhibitors of NFκB (PDTC) or JNK (SP60012), resulting in further cell death. Together, these results suggest that GF and Rh2, alone or in combination, can enhance the apoptotic activities of TNF‐α by modulating the activities of downstream pro‐survival factors. (Supported by CA121074)
Luteolin attenuated pro‐inflammatory conditions induced by activated microglia and protected against neuronal cell death
Authors: Jang Saebyeol , Johnson Rodney W
Published: June 2021

Excessive production of proinflammatory mediators by activated brain microglia plays an important role in sickness behavior and neurodegenerative disorders. Luteolin, a flavonoid found in high concentrations in celery, green pepper, and perilla leaf and seeds, has been shown to reduce proinflammatory molecules produced by lipopolysaccharide (LPS)‐stimulated macrophages and microglia. In the present study, the anti‐inflammatory and subsequent neuroprotective effects of luteolin were investigated. Pretreatment of primary microglia and BV2 murine microglial cells with luteolin inhibited LPS‐stimulated production of proinflammatory cytokines such as TNF‐α and IL‐1β at both the gene and protein levels. Luteolin also suppressed LPS‐induced nitric oxide release in BV2 cells by regulating the expression of inducible nitric oxide synthase. In addition, the release of prostaglandin E2 and cyclooxygenase‐2 mRNA expression were inhibited by luteolin. In addition, treating Neuro‐2a cells with conditioned media from LPS‐stimulated microglia resulted in neuronal cell death. However, treating microglia with luteolin prior to LPS reduced neuronal cell death in a dose‐dependent manner. These results suggest that luteolin may be a useful agent counteracting neurotoxicity associated with activated microglia. This work was supported by NIH grants AG16710 and MH069148.

Grant Funding Source

NIH
Role of grape powder extract (GPE), quercetin (QUE), and resveratrol (RSV) in attenuating markers of inflammation in primary cultures of human (pre)adipocytes
Authors: Chuang Chia‐Chi , Bumrungpert A. , Kennedy A. , Martinez K. , Walker‐West T. , McIntosh M.
Published: June 2021

Obesity‐associated inflammation in white adipose tissue (WAT) is strongly implicated in the development of insulin resistance. GPE is rich in phenolic phytochemicals like QUE and RSV that possess anti‐inflammatory properties. Thus, we examined the ability of GPE, QUE, and RSV to attenuate tumor necrosis factor (TNF)‐α ‐mediated activation of mitogen‐activated protein kinases (MAPK), nuclear factor kappa B (NFκB) and activator protein (AP)‐1 and induction of inflammatory genes in primary cultures of human (pre)adipocytes. We found that GPE, QUE, and RSV attenuated TNF‐α ‐induced expression of inflammatory adipocytokines [i.e., interleukin (IL)‐6, IL‐1β, IL‐8], markers of macrophage recruitment into WAT [i.e., monocyte chemoattractant protein (MCP)‐1], and negative regulators of insulin sensitivity [i.e., protein tyrosine phosphatase (PTP)‐1B]. Furthermore, we found that GPE and QUE attenuated TNF‐α ‐activated MAPK (i.e., ERK1/2, JNK1/2), NFκB, and AP‐1 (i.e., c‐Jun). RSV, however, only modestly attenuated TNFα‐activation of NFκB and c‐Jun, and had no effect on TNF‐α ‐activation of ERK1/2 and JNK1/2. Taken together, these data suggest that GPE, QUE, and RSV attenuate TNF‐α ‐mediated inflammation in primary cultures of human (pre)adipocytes by different upstream mechanisms. (Funded by the WB Kellogg Institute for Food and Nutrition Research; Grape Powder supplied by the CA Table Grape Com.)

Grant Funding Source

WB Kellogg Institute for Food and Nutrition Research
Bioavailability of Herbs and Spices
Authors: Nieves CJ , Montero Cindy , Percival Susan S.
Published: June 2021

The purpose of this study was to examine the bioavailability of 11 herbs and spices as a prelude to studying other health benefits. We hypothesized that changes in antioxidant activities in blood would be a sensitive measure of absorption. After an overnight fast, volunteers (n=10‐12 each spice) had a baseline blood draw, consumed capsules for 7 d, and returned for a second blood draw. Serum antioxidant capacity and DNA strand breaks in lymphocytes were measured. Serum antioxidant capacity was not significantly different between baseline and 7 d due to large individual variation. Intrinsic DNA strand breaks were remarkably similar between and among subjects. Strand breaks induced by H2O2 were well controlled. A RM 2‐way ANOVA measured significant differences in time and treatment for the number of strand breaks per cell or the percent of cells with strand breaks. There was no appreciable cell death during the oxidative stress phase of the experiment. Herbs and spices that did not have a significant effect were clove, black pepper, cayenne, cinnamon, and oregano. Herbs and spices that protected lymphocytes against DNA strand breaks were paprika, rosemary, ginger, heat‐treated turmeric, sage, and cumin. We conclude that compounds from these herbs and spices are absorbed. Support from UF/IFAS Ag Exp Station & McCormick Science Institute
Characterization of curry leaf polyphenolics and their antioxidant activity
Authors: Wilson Ted , Singh Ajay P , Freeman Michelle M , Singh Vartika , Olson Rachel M , Vorsa Nicholi , Somasundaram Siva , Luthria Dave
Published: June 2021

Curry leaf is commonly used in Indian cooking as a flavoring agent whose popularity in Western countries is increasing, however little is known about its phenolic profile. This study characterized the curry leaf phenolic profile and antioxidant capacity. Curry leaves were extracted using ethanol, methanol or acetone to facilitate isolatation of phenolic compounds using liquid chromatography coupled to electrospray mass spectrometry in tandem mode (LC‐MS‐MS) with negative ion detection. Several known and unknown phenolics and flavonols were identified and quantified based available standards, retention time, mass spectra in full ion scan mode(MS) and MS‐MS modes(product ion, precursor ion and neutral loss scans). Major constituent phenolics included myricetin‐3‐galactoside, quercetin‐3‐rutinoside, quercetin‐3‐glucoside, kaempferol‐3‐O‐caffeoylate, isorhamnetin‐3‐O‐glucoside and 5‐caffeoyl quinic acids. Lag‐time and TBARS tests demonstrated that curry leaf phenolics are associated with a dose‐dependent ability to prevent cupric‐ion induced oxidation of LDL. Leaf samples extracted with 80% acetone yielded higher concentration of phenolics the best antioxidant capacity. Curry leaf is rich in phenolics that have antioxidant activity in vitro, further studies are warranted in regards to the potential health benefits and to identify its novel phenolics.
Improved weight and body composition following a structured diet plan incorporating healthy convenient prepared meals
Authors: Summers Amber , Nguyen Von , Brosnahan Jen , Zukley Linda , Lowndes Joshua , Pohl Rob , Reimers Kristin , Angelopoulos Theodore J , Rippe James M
Published: June 2021

Excess body fat is a risk factor for the development of many chronic diseases. Lifestyle measures such as decreased caloric intake and increased physical activity constitute first line treatments to reduce excess body fat.

We examined the effects of a DASH‐Style (DS; n=70) diet and exercise or exercise only (EO, n=58) program on cardiovascular risk factors in 128 participants (BMI 22‐40) for 24 weeks. Both groups received an exercise prescription recommending a minimum of 15 minutes of moderate physical activity, 3 days per week and progressing to 45 minutes, 4 days per week by week 12. They also met with an exercise physiologist once weekly for 12 weeks and biweekly thereafter. The DS group followed a reduced calorie eating pattern and incorporated one commercially prepared healthy frozen meal and one other selected grocery item each day for 24 weeks and received dietary counseling from a dietician once weekly for 12 weeks and biweekly thereafter. .

After 24 weeks, the DS group reduced body weight (‐18.12 ± 10.66 vs ‐6.33 ± 8.24 lbs, p<0.001), BMI (‐2.90 ± 1.64 vs ‐1.00 ± 1.30 kg/m2, p<0.001), waist circumference (‐7.03 ± 4.53 vs ‐3.08 ± 3.79 cm, p<0.001), body fat percentage (‐5.93 ± 3.93 vs ‐1.64 ± 2.88%, p<0.001) and fat mass (‐18.59 ± 11.31 vs ‐6.01 ± 9.01 lbs, p<0.001) to a greater extent than the EO group.

These data suggest that a structured diet plan that combines healthy prepared meals with regular walking exercise can successfully reduce weight and improve measures of adiposity. The convenience of ready to eat meals may improve compliance to a structured program of risk factor reduction
Intestine specific over‐expression of DGAT1 in mice alters triacylglycerol storage in enterocytes, but not body weight in response to a high fat diet
Authors: Fast Angela M. , Lee Bonggi , Zhu Jiabin , Cheng Ji‐Xin , Buhman Kimberly K.
Published: June 2021

Mice deficient in DGAT1 (DGAT1−/−), a key enzyme in triacylglycerol (TG) biosynthesis, are resistant to high fat (HF) diet‐induced obesity (DIO). DGAT1−/− mice have no defect in quantitative absorption of dietary fat; however, their enterocytes store abnormal amounts of TG in cytoplasmic lipid droplets (CLDs). We hypothesized that increased intestine specific‐DGAT1 expression would decrease enterocyte TG storage in CLDs and ultimately result in increased weight gain in response to a HF diet. To test this hypothesis we generated mice with five or twenty fold increased intestine specific DGAT1 over‐expression using the villin promoter (DGAT1IntTG5X and DGAT1IntTG20X). Increasing expression of DGAT1 in the intestine reduced TG storage in CLDs in enterocytes, however, we did not find that DGAT1IntTG5X or DGAT1IntTG20X mice are more susceptible to HF DIO compared to wild‐type mice. These results suggest that intestine specific DGAT1 over‐expression does not affect whole animal energy balance possibly due to high endogenous DGAT1expression in the intestine. This work was supported in part by the PRF and AHA SDG to KKB.
Green tea (GT) extract enhances the biliary secretion of benzo[a]pyrene (BaP) in rats
Authors: Kim Juyeon , Noh Sang K. , Seo Yunjung , Koo Sung I.
Published: June 2021

Recently, we have shown that GT extract (GTE) lowers significantly the lymphatic absorption of BaP in rats. The present study was conducted to examine the effect of GTE on the biliary secretion of BaP in rats. Female rats were fed an AIN‐93G diet with or without (control) GTE at 5 g/kg diet for 4 wk. At 4 wk, rats with bile cannulae were infused at 3.0 mL/h for 8 h via a duodenal catheter with a lipid emulsion containing 1.0 mg BaP labeled with 14C (14C‐BaP), 452 μmol triolein, 20.7 μmol cholesterol, and 3.1 μmol α‐tocopherol (αTP), and 396.0 μmol Na‐taurocholate in PBS buffer (pH, 6.4). Bile was collected via a bile cannula hourly for 8 h. No significant difference was observed in bile flow for 8 h between the groups. However, the biliary secretion of 14C‐BaP was significantly increased in rats fed GTE, compared with controls. The rates of 14C‐BaP secretion for 8 h were 24.24 ± 0.43 %dose in rats without GTE and 31.51 ± 0.83 %dose in rats with GTE. The results indicate that GTE has a stimulatory effect on the biliary secretion of BaP in rats. The mechanism(s) by which GTE increases BaP secretion remains to be determined. Supported by the Korea Research Foundation Grant funded by Korean Government (MOEHRD, Basic Research Promotion Fund) (KRF‐2006‐003‐C00312)
Dietary Fat Intake, Liver Function and Disease Progression in HIV+ Adults in Botswana, Africa
Authors: Baum Marianna K , Le Katherine , Moyo Sikhulile , Makhema Joe , Burns Patricia J , Sales Sabrina , Tsalaile Lesedi , Bussmann Hermann , Li Yinghui , Campa Adriana , Marlink Richard
Published: June 2021

Objective

Although increased energy intake is associated with benefits in HIV+ patients, the effect of fat intake on HIV progression and liver function needs to be established.

Methods

After consenting 714 HIV+ asymptomatic adults, blood was drawn for chemistries, CD4 cell count and viral load. 24‐Hour Recalls were collected and analyzed by Nutribase 7.0.

Results

Medium CD4 cell count was 424 cells/µL, mean age was 33±8.1 (78.8% < 40 years), and 72.5% were female. Mean dietary fat intake was 38.25±31.2 g; 36% of participants had BMI>25 kg/m2. Total fat intake was positively associated with alkaline phosphatase (β=0.002, p=0.0025) and gamma‐glutamyl transferase levels (β=0.111, p=0.005). Total fat intake (%) was significantly associated with total cholesterol (β=0.007, p=0.027) and LDL levels (β=0.006, p=0.04). Those in the first CD4 quartile (≤399 cells/µL) had higher mean LDL (2.5±0.8 vs. 2.3±0.8 mmol/L, p=0.0273) than the rest. Compared to those in the CD4 third quartile (≥550 cells/µL), those in the first and second quartile had higher alkaline phosphatase (76.44±21.3 vs. 68.2±20.9 U/liter, p=0.015). These relationships remained significant after adjusting for BMI.

Conclusion

These findings support a significant association between increased dietary fat intake, adverse lipid profile, increased liver enzymes and HIV progression that is not affected by body composition.

Funded by NIDA
Vitamin D receptor (VDR) FokI polymorphisms influence body composition in physically active adolescent boys
Authors: Diogenes Maria Eduarda Leão , Bezerra Flávia F. , Oliveira Karla J.F. , Koury Josely C. , Cabello Giselda K. , Cabello Pedro H. , Mendonça Laura M.C. , Oliveira‐Junior Astrogildo V. , Donangelo Carmen M.
Published: June 2021

Body composition, including bone, lean and fat mass, is known to be under complex physiologic and genetic regulation. The aim of the present study was to identify anthropometric, hormonal and genetic determinants of body composition in Brazilian physically active adolescent boys (12‐14y, n=47) by stepwise multiple regression analyze. Dependent variables were total body mineral content (TBMC), total body mineral density (TBMD), lean mass (LM) and percentage of body fat (%BF), assessed by DXA. Independent variables were chronological and bone age, weight, height, plasma testosterone and IGF‐1, serum iPTH, and VDR genotypes (FokI and TaqI). Results were as follows:

*P<0.05, (P<0.01, #P<0.001

Although body weight was the main factor influencing all the outcome variables, FokI polymorphisms appeared as a modest, but significant factor influencing TBMD and %BF. Our results support the complex regulation of body composition during adolescence. VDR‐ related mechanisms are possibly involved and appear to be sensitive to FokI polymorphisms.

Financial support: CNPq and FAPERJ (Brazil).
Dietary selenium (Se) and copper (Cu) affect the activity and expression of the hepatic selenoprotein methionine sulfoxide reductase B (MsrB) in rats
Authors: Uthus Eric O
Published: June 2021

As reported by Jenkinson et al. (J Nutr 1982) and Prohaska et al. (J Nutr Biochem 1992) Cu deficiency (CuD) decreases the activity and mRNA expression of the selenoprotein GPx. Because both Se and Cu are important in oxidative defense, we wanted to determine the effect of a combined deficiency on the methionine (Met) sulfoxide reductase system, which is comprised of MsrA and MsrB. MsrA reduces L‐methionine S‐sulfoxide to Met and MsrB (a selenoprotein) reduces L‐methionine R‐sulfoxide to Met. In a 2 x 2 experiment, male weanling Sprague‐Dawley rats were fed an amino acid‐based diet containing 0 or 0.2 µg Se (as selenite)/g and <1 or 6 µg Cu (as Cu carbonate)/g for 5 wk. Neither Se nor Cu had an effect on MsrA enzyme activity or mRNA expression. Se and Cu interacted to affect MsrB enzyme activity; both Se deficiency and CuD decreased the activity, however, the decrease by CuD was only significant in rats fed 0.2 µg Se. CuD resulted in downregulation of MsrB mRNA. CuD decreased GPx in Se adequate rats. The results show that Cu has a role in regulation of selenoproteins; whether this role is direct or indirect is not known.

1Pooled SD. 2nmol/min/mg protein, N=4/group. 3fold change, N=7‐9/group. 4SD for delta Ct. 5U/mg protein, N=7‐8/group.
EFFECT OF FLAXSEED FEEDING VS OTHER SOURCES OF DIFFERENT PROTEIN QUALITY ON GROWTH'S RATS
Authors: FERNANDEZ INES , VIDUEIROS SILVINA MARIELA , PERRIS PAULA , PALLARO ANABEL
Published: June 2021

Previous studies demonstrated that Flaxseed has a Biological Value of 67%. Weanling Wistar rats (n=24) were divided in groups that received flaxseed (F), wheat flour (W) or casein (C) diets to compare protein quality effects on body weight gain and nitrogen retention. A forth group received a mixture of casein:flaxseeds (70:30) (CL) to analyze the adverse effects of flaxseeds. Protein concentration was 8 % and all essential nutrient were added following AIN93. Food (FI) and protein intake (PI), initial (Bo) and final (Bf) body weight, body weight gain (BWG) and nitrogen retention (NR) were determined. Statistical analysis was performed using Anova‐SNK (a: p<0.01 respect W; b: p<0.01 respect F).

BWG and NR of F were higher than W and lower than C and CL, being C and CL no different. The results are probably due to the different protein quality of the sources and suggest the addition of flaxseed to casein doesn't have an adverse effect on the growth of animals or the ability to retain nitrogen. Supported UBACYT‐B098.
Effect of calcium and dairy products intake on body composition and blood lipids in Puerto Rican obese adults
Authors: Palacios Cristina , Soltero Sandra , Rios Ruth , Bertran Jose Juan
Published: June 2021

Studies have shown that high calcium diets significantly reduce body weight and fat in obese individuals. The purpose of this study was to examine the effects of increased calcium intake (dairy or supplements) on body weight and fat and blood lipids in Puerto Ricans while on isocaloric diets. A 6‐months randomized, double‐blind, placebo‐controlled trial was conducted in obese adults, aged 21‐50 years and BMI >30 kg/m2. A total of 32 subjects were randomly assigned to one of three groups: 1300 mg/d of calcium from dairy products (foods were substituted with dairies) or supplements and placebo group. Subjects were asked not to change their diet (except for the dairy group) and physical activity during the study, which were monitored monthly. Weight, fat and lean body mass (by DEXA), and blood lipids were measured at baseline and at the end of the study. Twenty five subjects completed the study. Mean intakes of calcium during the trial was 1197±184 mg/d in the dairy group, 1171±196 mg/d in the supplemental group and 668±201 mg/d in the control group, which was significantly lower compared to the 2 treatment groups (P<0.001). No significant differences were observed in the mean change in any of the measurements between the three treatment groups. In conclusion, increasing calcium intake from dairy products or supplements did not alter body weight and fat and blood lipids in this group.
Differences in Physical Activity and Fitness by Weight Status in a Sample of Rural School‐Aged Children
Authors: Shriver Lenka Humenikova , Barrett Aimee , Page Melanie , Topham Glade , Hubbs‐Tait Laura , Harrist Amanda
Published: June 2021

The objective of this observational study was to determine whether significant differences in physical activity and fitness exist between healthy‐weight, at‐risk‐for‐overweight, and overweight school‐aged children. Twelve rural public schools participating in a multidisciplinary research project were randomly selected; data were collected from 257 third‐graders during PE classes. Variables included BMI‐for‐age, moderate and vigorous physical activity, and fitness performance (FITNESSGRAM battery tests: PACER, Curl‐up, and Sit & reach). Analysis of variance showed overweight children spent significantly less time in moderate (p<0.05) and total moderate & vigorous activity (p<0.01) than other children. Cardiovascular endurance (PACER) of overweight children was significantly lower compared to both children at a healthy weight (p<0.001) and at‐risk‐for‐overweight (p<0.05). Total moderate & vigorous activity was positively associated with cardiovascular endurance (PACER) (p<0.05) but not with other fitness variables. Differences in physical activity and fitness were identified between healthy‐weight, at‐risk‐for‐overweight and overweight children in our sample. Increased moderate activity may not only reduce obesity but also have a positive influence on cardiovascular endurance of overweight children.
Awareness of the birth process among university students: knowledge of risks associated with cesarean deliveries
Authors: Joyce Stephanie , Johnson Deborah , Nolte Joy V , Granger Lauren A , Kavanagh Katie F
Published: June 2021

Traumatic birth experiences may have a direct impact on breastfeeding success. Our purpose was to collect baseline and follow‐up data on university students' knowledge of risks of cesarean deliveries (medically indicated or not), before and after screening of a documentary featuring natural childbirth as an option for those with low risk of complications. The event was free and included a discussion panel, comprised of a Certified Nurse Midwife, a Registered Nurse, and a Medical Doctor to answer audience questions. Pretests and posttests were administered. Chi‐squared tests were performed at the 0.05 level of significance. Of the 225 attendees, 205 completed the pretest and 162 completed the posttest. Participant knowledge regarding risks associated with cesarean delivery significantly increased. This included maternal recovery time (80.5 vs. 85.2%); respiratory distress in infants (28.3 vs. 45.1%); infant lethargy (25.4 vs. 42.6%); maternal mortality (33.2 vs. 67.9%); infant mortality (24.9 vs. 61.1%); and delayed initiation of breastfeeding (46.8 vs. 71%). Results indicate that exposure to the documentary and expert discussion panel did result in increased knowledge of risks of cesarean deliveries, especially those associated negatively impacting breastfeeding success. Funding: The Dept of Nutrition, Dean's Office, and The Center for Public Health at The University of Tennessee, Knoxville.

Grant Funding Source

Internal University funds
Nutritional status of methadone‐treated pregnant women
Authors: Tomedi Laura Elizabeth , Bodnar Lisa M. , Bogen Debra , Hanusa Barbara , Wisner Katherine L.
Published: June 2021

Maternal nutrition has profound effects on a child's long‐term health. There are no known studies of the nutritional status of mothers on methadone, a treatment for opiate addiction. Pilot study objective: evaluate dietary intake in a cohort of 26 methadone‐treated (MT) pregnant women compared to a cohort of 198 pregnant women not treated with methadone (non‐MT). Usual dietary intake in the 2nd trimester of pregnancy was assessed using a self‐administered modified Block98 food frequency questionnaire. Nutrient intakes were adjusted for energy using the nutrient‐density approach, and categorized into tertiles. MT women were significantly leaner than non‐MT women (underweight, 11.5% vs. 3.0; normal 53.8% vs. 52.5%; overweight 11.5% vs. 19.2%; and obese 7.7% vs. 25.3%; (p<0.05)). MANOVA analysis showed that MT women were more likely than non‐MT women to be in the lowest tertile of nutrient density for dietary folate equivalents, vitamins C, E and A, calcium, iron and fiber. MT women were also more likely to be in the lowest tertile for percent of energy from protein and fat. However, MT women were less likely than non‐MT women to be in the lowest tertile for carbohydrates. These pilot data suggest that MT mothers may consume a low nutrient‐dense diet that is high in refined carbohydrates. Project supported by The Gerber Foundation, Children's Hospital of Pittsburgh RAC, NIH grants K01 MH074092, R01 MH060335 & K12 HD043441.
Differences in Dietary Habits, Serum Lipid Profiles, and interest in Health among Korean College Women stratified by BMI
Authors: Shin Kyung Ok , Hwang Hyo Jeong , An Chang Hun , Choi Kyung Soon , Chung Keun Hee
Published: June 2021

The principal objective of this study was to compare the food habits, nutritional status, blood lipids, and health interest among these women, in accordance with their obesity status. The data utilized herein was collected from randomly selected groups in Seoul. The subjects, aged 20‐24 years old (n=419), participated by answering questionnaires. Our analysis showed significant differences among the underweight, normal weight, and obesity groups according to BMI, weight, skeletal muscle mass, body fat mass, fat free mass, percentage of body fat, waist‐hip ratio, and basal metabolism (p<0.05). Cholesterol intake in the obesity group (121.92 ?114.52mg) was lower than that of the underweight group (164.67 ?114.52mg) and normal‐weight group (143.31 ?99.58mg/dl) (p<0.05). Serum cholesterol in the normal group (189.46 ?29.05mg/dl) was significantly higher than in the underweight group (177.04 ?26.36mg/dl ) (p<0.05). Serum triacylglycerol in the obesity group (132.00 ? 64.66 mg/dl ) was higher than in the underweight and normal groups (p<0.05) The ratio of HDL‐C / LDL‐C in the obesity group (0.41 ?0.06) was higher than in the underweight (0.52 ?0.45) and normal groups (0.59 ?0.23). The number of subjects in the normal group that reported taking regular exercise was higher than in the other groups. The reported duration of exercise period ranged from 30 minutes to one hour in all groups. Interest in health was high among all groups.

In conclusion, our results show that college‐aged women in Seoul should be encouraged to amend their overall dietary habits, make a dietary plan that fits their individual needs, and maintain an effective exercise schedule.
Tenofovir Toxicity Targets Mitochondria of Renal Proximal Tubules
Authors: Kohler James John , Hosseini Seyed , Hoying‐Brandt Amy , Green Elgin , Tran Dung , Russ Rodney , Santoianni Robert , Lewis William
Published: June 2021

Tenofovir disoproxil fumarate (TDF) is an analog of adenosine monophosphate that inhibits HIV reverse transcriptase in HIV/AIDS. Despite its therapeutic success, renal tubular side effects are reported. Mechanisms and targets of TDF toxicity were determined using "2 by 2" factorial protocols that included HIV transgenic mice (TG) and wild‐type (WT) littermates with or without oral TDF (5 weeks). A parallel study employed didanosine (ddI) instead of TDF. At termination, heart, kidney and liver samples were retrieved. Laser‐capture microdissection (LCM) isolated renal proximal tubules for molecular analyses. Mitochondrial DNA (mtDNA) abundance, histo‐ and ultrastructural pathology were analyzed. TDF increased mtDNA abundance in TG whole‐kidneys, but not hearts or livers. In contrast, ddI decreased mtDNA abundance in livers of WTs and TGs, but had no effect on hearts or kidneys. LCM‐captured renal proximal tubules from TGs exhibited decreased mtDNA abundance with TDF. Histological analyses of kidneys showed no disruption of glomeruli or proximal tubules with TDF or ddI treatments. Ultrastructural features in renal proximal tubules from TDF treated TGs included increased number and irregular shaped mitochondria with sparse fragmented cristae. Results indicate tissue‐specific toxicity from TDF and ddI monotherapies. TDF toxicity targets mitochondrial renal proximal tubules in an AIDS model.
Early Embryonic Lethality of Mice with Disrupted Transcription Cofactor PIMT/NCoA6IP Gene
Authors: Jia Yuzhi , Viswakarma Navin , Matsumoto Kojiro , Pyper Sean , Rao M Sambasiva , Reddy Janardan K
Published: June 2021

PIMT (PIPMT, NCoA6IP), first isolated as a transcription coactivator PRIP (NCoA6)‐interacting protein, binds RNA and possesses RNA methyltransferase activity. PIMT is evolutionarily conserved and is localized predominantly to the nucleus. It is expressed in most adult tissues and in all embryonic stages in the mouse. PIMT interacts with key coactivators such as PRIP, CBP, p300 and MED1. Since the disruption of these PIMT‐interacting coactivator genes causes embryonic lethality in the mouse, we reasoned that PIMT gene knockout might also affect embryonic development. We have used homologous recombination to generate mice with a disrupted PIMT gene in order to understand the biological significance of this gene in mammalian development and growth. Heterozygous (PIMT+/−) mice develop normally and are indistinguishable from those of their wild‐type (PIMT+/+) littermates. Disruption of both PIMT alleles results in early embryonic lethality due to apoptosis and decreased proliferative potential of the blastocyst cells. PIMT deficient embryos die shortly after implantation and then resorbed. Mouse embryonic fibroblasts with conditional knockout of PIMT gene, show defective wound healing and G2 phase arrest of cell cycle. We conclude that PIMT is indispensable for early embryonic development. This work was supported, in part, by grants from the National Institutes of Health Grants CA104578 and GM23750
Hepatocyte retinoid X receptor alpha (RXRalpha) deficiency impairs liver regeneration through multiple pathways
Authors: YANG XIAOXIA , GUO MINGLEI , WAN YU‐JUI YVONNE
Published: June 2021

Many nuclear receptors have been implicated in liver regeneration. RXRα forms heterodimers with these nuclear receptors. In the current study, we characterize the role of hepatocyte RXRα in liver regeneration. RXRα deficiency results in delayed DNA synthesis, mitosis, and hepatocyte proliferation in response to partial hepatectomy. RXRα plays a unique role and controls multiple pathways dictating normal liver regeneration. In regenerating RXRα‐null livers, the cytokine interleukin 6/signal transducer and activator of transcription 3 (IL‐6/Stat3) pathway, the growth factor signaling, and the peroxisome proliferator‐activated receptors alpha (PPARα)‐regulated circadian cell cycle progression are altered; many of these changes are not found in the regenerating liver of other nuclear receptor knockout mice. Our data suggest that hepatocyte RXRα is a critical regulator, through cross‐talk with signaling pathways mediated by other types of liver cells, to ensure normal progression of liver regeneration. This work is supported by grants from National Institutes of Health (CA53596) and COBRE (P20 RR021940).
SMP30/Regucalcin is a direct transcriptional target of Wnt signaling in the liver
Authors: Zeng Gang , Bowen Kari‐Nejak , Monga Satdarshan S.
Published: June 2021

TheWnt/β ‐catenin pathway plays an essential role in liver biology. It is critical to identify novel transcriptional targets of this pathway to enable us to understand the basis of some of the functions of this pathway. By analyzing livers from β‐catenin conditional knockout mice (KO), we identified regucalcin as a downstream target of β‐catenin‐TCF signaling in the liver. Regucalcin is a Ca2+ binding protein that regulates Ca2+ transport, oxidative stress, and vitamin C metabolism. Western blotting and immunohistochemistry demonstrated the presence of regucalcin protein in the cytosol and nucleus of wild‐type mouse livers at 1, 3, and 6 months; however, regucalcin expression was markedly diminished in KO. Also, higher regucalcin levels were observed in HepG2 (active β‐catenin) as compared to Hep3B (normal β‐catenin) hepatoma cells. Next, regucalcin promoter was analyzed and several TCF‐4/beta‐catenin binding sites were identified. Luciferase reporter assays using deletion mutants of regucalcin promoter identified the TCF4 binding consensus sequence at ‐163 to ‐157 site to be the important for full transcriptional activation. Also, ChIP assays demonstrate a direct interaction of β‐catenin and TCF4 with this area of regucalcin promoter. In conclusion, we have identified regucalcin as a direct target of β‐catenin in the liver and provide evidence of direct regulation of its gene expression by β‐catenin/TCF interactions.
MALLORY DENK BODY FORMATION IS ASSOCIATED WITH AN INCREASE OF THE IMMUNOPROTEASOME AND A DECREASE OF THE 26S PROTEASOME
Authors: Oliva Joan , Bardag‐Gorce Fawzia , Li Jun , French Barbara A , French Samuel W
Published: June 2021

Mallory Denk body (MDB) formation is seen in the liver cells of alcoholic patients and in other non‐alcoholic diseases. Proteasome inhibition is hypothesized to be the one of causes of MDB formation. Mice fed the drug diethyl‐1, 4‐dihydro‐2, 4, 6‐trimethyl‐3, 5‐pyridinedicarboxylate (DDC) for 10 weeks is the experimental model used to study MDB formation. 20S proteasome chymotrypsin‐like activity was not changed in the liver of mice refed DDC. However, ATP‐stimulated 26S proteasome chymotrypsin‐like activity was decreased by DDC feeding. These results confirm the reduction of 26S‐dependent ubiquitin‐proteasome proteolysis, which is reflected in the accumulation of ubiquitinated cytokeratins. Three constitutively expressed subunits of the 20S proteasome are known to be replaced by the inducible subunits of the immunoproteasome LMP2, LMP7, and MECL‐1 when stimulated with IFNg. There was an up regulation of LMP2, LMP7 and MECL‐1 protein levels and gene expression in the liver of mice fed DDC. Gene expression and protein levels of FAT10, a member of the ubiquitin‐like modifier family, which is also induced by IFNg and TNFa, were induced by DDC feeding. The marked up regulation of FAT10 could account for the block of cytokeratin turnover by the 26S proteasome. These results support the concept that the proteasome population is switched from the 26S proteasome to the immunoproteasome. The lack of 26S proteasome formation causes an accumulation of altered cytokeratin proteins. Protein aggresomes such as MDB then form. Supported by NIH/NIAAA Grants 8116 and P50‐011999 Alcohol Center Grant, Liver/Pancreas.
Muscarinic receptor regulation of osmosensitive taurine transport in human SH‐SY5Y neuroblastoma cells
Authors: Foster Daniel J , Vitvitsky Victor M , Banerjee Ruma , Heacock Anne M , Fisher Stephen K
Published: June 2021

The ability of G‐protein‐coupled receptors to regulate osmosensitive uptake of the organic osmolyte, taurine, into human SH‐SY5Y neuroblastoma cells has been examined. When monitored under isotonic conditions taurine influx was mediated exclusively by a Na+‐dependent, high‐affinity (Km=2.5 μM) saturable transport mechanism (Vmax=0.087 nmol/mg protein/min). Reductions in osmolarity of >20% (under conditions of constant NaCl) resulted in an inhibition of taurine influx (>30%) that could be attributed to a reduction in Vmax, whereas the Km for uptake remained unchanged. Inclusion of the muscarinic cholinergic agonist, oxotremorine‐M (Oxo‐M), also resulted in an attenuation of taurine influx (EC50~0.7 μM). Although Oxo‐M‐mediated inhibition of taurine uptake could be observed under isotonic conditions (~25‐30%), the magnitude of inhibition was significantly enhanced by hypotonicity (~55‐60%), a result that also reflected a reduction in the Vmax, but not the Km, for taurine transport. Oxo‐M‐mediated inhibition of taurine uptake was dependent upon the availability of extracellular Ca2+ but was independent of protein kinase C activity. These results suggest that taurine uptake is reduced upon muscarinic receptor activation in a volume‐dependent fashion, thus facilitating taurine depletion from cells upon exposure to hypotonicity. Supported by NS23831 (SKF) GM007767 (DJF) and DK64959 (RB).
The glucuronidation of native and oxidized estrogens can be effectively inhibited by compounds structurally related to UDP‐glucuronic acid in human recombinant UGT1A10
Authors: Mazur Anna , Bratton Stacie Moseley , Miller Grover P , Finel Moshe , Radominska‐Pandya Anna
Published: June 2021

Human UGT1A10 glucuronidates physiologically important substrates such as native and hydroxylated estrogens and drugs. Modulation of UGT1A10 activity can alter estrogen metabolism and disposition resulting in alterations of its status in the body. Here, the effect of bi‐dentate inhibitors containing a lipophilic and uridine moiety on UGT1A10 estradiol (E2) and 4‐hydroxyestradiol (4‐OH‐E2) glucuronidation was investigated. Comparisons of detailed inhibition analyses (IC50 and Ki values) for the best inhibitor in the series, PP55B, were made, and for each substrate these data were found not to follow classical inhibition models. Increasing concentrations of PP55B affected the glucuronidation of E2 and 4‐OH‐E2 very differently. For E2, 2‐25 µM of inhibitor caused an activation of activity (~50%) and followed by a gradual decrease in activity at higher concentrations with 100% inhibition seen at ~600 µM. A multiple binding site model may explain this observation. For 4‐OH‐E2, no effect was seen from 2‐25 µM PP55B, followed by a rapid decline in activity was seen with 100% inhibition seen at ~300 µM. This data indicates that UGT1A10 can be effectively inhibited by PP55B. These findings are undergoing further analysis to elucidate the mechanism of action, which could have significant clinical and pharmacological applications for designing UGT inhibitors. NIH‐DK60109, GM075893 (AR‐P)