bluuebunny/crossref_metadata_2025 huggingface dataset:
Process evaluation of the HOT project (Healthy Outcomes for Teens) website in 3 venues
Authors: Chapman‐Novakofski Karen , Castelli Darla , Peterson Sharon , Goss David , Lloyd Les
Published: June 2021

The purpose of this study was to identify the most effective access point for interactive web materials in providing diabetes prevention instruction to middle school students. Website use and content knowledge were compared across three different venues: (a) part of health/physical education curriculum, (b) after school and (c) voluntary weekend access. Participants viewed the materials as part of an after‐school computer program, pull out from physical education classes, or as part of the health education curriculum in middle school (n=182). The weekend participants, 5th‐8th graders, (n=46) were introduced to the website and provided with a copy of the website home page and their access code. Each participant was reminded at least twice by phone or email to visit the website. The structure and accountability of the school curriculum resulted in the greatest access and gains in content knowledge both within and beyond the school day, as free time access was highest in this group. Social marketing techniques used in the voluntary access group had the lowest engagement. Benefits to participants within the school structure included completion of class assignments, MP3 players; no access barriers existed. Weekend participants received no compensation or benefit and may have had access difficulties. Each venue has implementation possibility but more structure, benefit and fewer barriers have most promise.
Mode of delivery and early nutrition modulate microbial colonization and fermentation products in the neonatal piglet
Authors: Wang Mei , Radlowski Emily C , Monaco Marcia H. , Chen Long , Miller Michael J. , Fahey George C. , Gaskins H Rex , Donovan Sharon M.
Published: June 2021

Herein, the hypothesis that delivery route and nutrition would interact to modulate the composition of the microbiota and short chain fatty acids (SCFAs) production was tested. Piglets derived by vaginal or Cesarean delivery were sow‐reared (SR) or fed formula (F) or with prebiotics (F+P). On d 7 and 14, intestinal contents were collected. Bacterial density in ileum and ascending colon (AC) on d 14 was quantified by real‐time PCR. Delivery mode and nutrition each affected microbial populations, but no significant interactions were observed. Bacteroides density was higher in ileum and Clostridium cluster XIVa was lower in the AC of VD than CD piglets. Total Bacteria differed in the AC, with SR > F+P > F piglets. Lactobacillus and Bifidobacterium were higher in the ileum and AC of SR than F or F+P piglets and ileal Bacteroides density of SR > F > F+P piglets. Clostridium cluster XIVa density was higher in ileum of SR than F or F+P piglets. In addition, butyrate, isobutyrate, isovalerate and valerate were higher in VD than CD and SR than F or F+P.F+P piglets had higher acetate than SR and F, whereas propionate of SR > F and F+P was intermediate. An interaction between delivery and nutrition was observed for butyrate, isobutyrate and isovalerate, wherein their concentrations were greatest in VD/SR, intermediate in CD/SR and lowest in F and F+P piglets, independent of delivery route. (BMS Freedom to Discover Award)
Atherogenic Diet Promotes Atherosclerotic Lesion Formation by Enhancing Inflammatory Response in the LDL Receptor Null Mouse
Authors: Wang Shu , Wu Dayong , Matthan Nirupa , Lamon‐Fava Stefania , Lecker Jaime , Wu Yi Chieh , Lichtenstein Alice
Published: June 2021

The effect of an atherogenic diet on inflammatory response in elicited peritoneal macrophages and livers in relation to aortic lesion formation was assessed in LDL receptor null (LDLr‐/‐) mice. Mice were fed an atherogenic (20% fat, 0.2% cholesterol, w/w) or control (4% fat, 0.02% cholesterol, w/w) diet for 32 weeks. The atherogenic diet, relative to the control diet, resulted in significantly higher plasma concentrations of monocyte chemoattractant protein‐1 (MCP‐1, P<0.01), tumor necrosis factor alpha (TNF‐alpha, P<0.01) and interleukin‐6 (IL‐6, P<0.001), more aortic wall macrophage deposition, and greater accumulation of both aortic free cholesterol (4.9‐fold, P<0.001) and esterified cholesterol (6.4‐fold, P<0.001). Elicited peritoneal macrophages isolated from the atherogenic diet, relative to the control diet, fed mice accumulated more esterified cholesterol (P<0.001), had higher mRNA levels (P<0.05) and secretion (P<0.01) of MCP‐1. The atherogenic diet, relative to the control diet, significantly increased the production of IL‐6 (P<0.05) and TNF‐alpha (P<0.05) from the livers. These data suggest that the atherogenic diet resulted in higher plasma inflammatory factor concentrations, higher elicited peritoneal macrophage secretion and liver production of inflammatory factors, which in turn were associated with greater aortic cholesterol accumulation.
Electrophysiological Characterization of a TRPC‐like Current in Cerebral Arterial Smooth Muscle
Authors: Anfinogenova Yana , Welsh Donald G.
Published: June 2021

The goal of this investigation was to isolate and characterize a TRPC‐like current in cerebral arterial smooth muscle cells (VSMCs). Briefly, VSMCs were enzymatically isolated from rat cerebral arteries and whole cell currents monitored using patch clamp electrophysiology. Our initial experiments revealed two subpopulations of VSMCs that displayed either an outwardly rectifying or a doubly rectifying current. The reversal potential of both currents shifted with the Na(+) equilibrium potential and were potently blocked by micromolar concentrations of Gd(3+). A pharmacological characterization subsequently revealed that the doubly rectifying TRPC‐like current was insensitive to Cl(‐) channels inhibitors (DIDS and niflumic acid), amiloride and flufemamate. In contrast, tamoxifen elicited a dual effect both attenuating and activating this current at submicromolar and micromolar concentrations, respectively. Similar to findings from cultured smooth muscle cells, hyposmotic challenge and vasoconstrictor agonists (UTP) activated this TRPC‐like non selective cation current. To summarize, this study is the first to characterize the electrical and pharmacological properties of a doubly‐rectifying TRPC‐like cation current in cerebral VSMCs. Ongoing investigations continue to define the molecular composition of this TRPC‐like current along with it's mechanisms of physiological regulation.
Down‐regulation of exclusively testis‐expressed microRNA‐469 by gonadotropin‐regulated testicular helicase (GRTH/DDX25) controls male germ cells development
Authors: Dai Lisheng , Tsai‐Morris Chon‐Hwa , Villar Joaquin , Zhang Jiabao , Dufau Maria L.
Published: June 2021

Gonadotropin‐regulated testicular RNA helicase (GRTH/DDX25), a testis‐specific member of the DEAD‐box family, is an essential post‐transcriptional regulator of spermatogenesis. Failure of expression of Transition protein 2 (TP2) and Protamine 2 (Prm2) proteins (chromatin remodelers, essential for spermatid elongation and completion of spermatogenesis) with preservation of their mRNAs expression was observed in GRTH‐null mice (azoospermic due to failure of spermatids to elongate). These were indentified as target genes for the testis‐specific miR‐469, which is increased in the GRTH‐null mice. Further analysis demonstrated that miR‐469 repressed TP2 and Prm2 protein expression at the translation level with no effect on mRNA expression, through binding to the coding regions of TP2 and Prm2 mRNAs. This indicates miR‐469 silencing function at the translational level. The corresponding primary‐miRNAs (pri‐miRNAs) and the expression levels of Drosha and DGCR8 (both mRNA and protein) were increased significantly in the GRTH‐null mice, suggesting that GRTH regulates the processing pathway of miRNAs at the transcriptional level. Collectively, these studies indicate that GRTH, a multifunctional RNA helicase, acts as a negative regulator of miRNA biogenesis at the transcriptional level by regulating miRNAs expression and consequently their function during spermatogenesis.
Structure‐activity relationships of diacetylene‐based LpxC inhibitors
Authors: Lee Chul-Jin , Liang Xiaofei , Park Eugene , Zeng Daina , Swanson Shauna , Chen Xin , Nicholas Robert A. , Raetz Christian R.H. , Toone Eric J. , Zhou Pei
Published: June 2021

The LpxC enzyme in the lipid A biosynthesis pathway is one of most promising and yet clinically unexploited antibiotic targets for the treatment of multidrug‐resistant Gram‐negative infections. Unfortunately, extant LpxC inhibitors typically display large variations in potency against different Gram‐negative pathogens. Recently, structural studies in our laboratory revealed that significant structural variations exist among diverse LpxC orthologs. When LpxC orthologs bind to LPC‐009 inhibitor, the vicinity of the active site displays large conformational variations, creating different morphologies of the substrate‐binding hydrophobic passage. The LPC‐009 inhibitor whose pliable diacetylene scaffold is essential for its broad‐spectrum inhibition overcomes structural variations in the hydrophobic passages of LpxC orthologs. To further improve potency and antibiotic profiles of LPC‐009, a series of diacetylene‐based derivatives were designed from the lead compound LPC‐009, synthesized and evaluated for antibiotic profiles against diverse Gram‐negative pathogens. In addition, the crystal structures of LpxC in complex with newly designed LpxC inhibitors provide insights for structure‐activity relationships and future structure‐based optimization of LpxC inhibitors.

This research was supported by National Institutes of Health Grants AI055588 (to P.Z.) and GM‐51310 (to C.RH.R).
Effects of Isoflavones Supplements and Exercise on Biomarkers of CVD in Korean Postmenopausal Women
Authors: Lee HanSongYi , Kim Jinkyung , Lee Okhwa , Choue Ryowon
Published: June 2021

The objective of this study was to assess the combined effects of isoflavones supplements (70 mg/d of isoflavones) and exercise (81 min/day) in Korean postmenopausal women. We recruited postmenopausal women who were free of diagnosed cardiovascular disease (KMC IRB 1002‐02). In this double‐blind randomized controlled trial, 67 postmenopausal women were assigned to four groups: 1. Placebo (PLA, n=9) 2. Isoflavone supplements (ISO, n=11), 3. Exercise and placebo (Ex+PLA, n=25), 4. Exercise and isoflavone supplements (Ex+ISO, n=22). Dietary isoflavones intakes were analyzed by a 3‐day food records. Blood levels of lipids, insulin, glucose, hormones, hs‐CRP, sVCAM, and sI‐CAM were analyzed at baseline and 12 weeks. The average age and menopausal period were not different among the groups. The levels of total and LDL‐chol in the PLA were significantly higher than in the ISO, Ex+PLA, and Ex+ISO groups (p<.05). Levels of hs‐CRP, sICAM‐1 and sVCAM‐1 were not different among the groups. However, sICAM‐1 levels increased after supplementation, in the Ex+ISO group and sVCAM‐1 levels reduced after supplementation, in the ISO group (p<.05). In conclusion, isoflavone supplements and exercise for 12 weeks prevented the increased levels of blood total‐ and LDL‐cholesterol, however further study is to confirm the beneficial effects on cardiovascular disease in postmenopausal women.
An estimate of nutritional status of students in Astana
Authors: Slivkina Nataliya , Dalenov Erbolat , lderbayev Oralbek
Published: June 2021

Everyone knows that nutrition is an important factor that determines the physical condition and health of the body. At the same time, among students at the high workload of the educational process, financial insolvency and other reasons are common disorders of nutrition. The aim was to evaluate some indicators nutritional status of students in Astana. Calculation of the energy of students was based on estimates of basal metabolism (BM) according to the equation of Harris‐Benedict and the rate of physical activity (RFA), established by the timing. Mean values of BM were 1561,34±48.17 for men and 1360,66±42.14 for girls. RFA was 1.87±0.08 for men and 1.75 for girls. Diurnal energy consumption were 2864,59±61,30 for men and 2382,36±59,47 for girls. Analysis of the diets of students showed that the average daily caloric foods was 2726,0±58,76 kcal for men and 2396±52,44 kcal for girls. Ketle index's mean values ranged from 20,02±3,34 for men and 21,71±3,47 for girls. Underweight were 23.14% men and 19.1% girls. Daily caloric foods were lower than the daily consumption of energy among men. Estimate of nutritional status revealed a low intake of protein (69 g/day), including animal (51.3%), nutrition imbalance, due mainly starchy food. Ratio of protein, fat and carbohydrate was 1:1,2:6,4.
Feeding practices correlated with authoritative parenting style and responsive feeding style scores
Authors: Dickin Katherine L , Hubbs-Tait Laura , Sigman-Grant Madeleine , Jahns Lisa , Mobley Amy
Published: June 2021

Study objective was to identify correlations of authoritative parenting and responsive feeding styles with parental practices and child behaviors previously found to protect children from or increase risk of child obesity. Participants were 144 low‐income mothers of 3‐ to 5‐year‐old children (71 girls, 73 boys). Feeding practices measures were Healthy Children Healthy Families Behavior Checklist and Parental Overt and Covert Control. Caregiver's Feeding Style Questionnaire and Preschool Behavior Questionnaire–Head Start measured feeding and parenting styles. Authoritative parenting style continuous scores were correlated with child active play, r = .252, p =.002, low frequency of child drinking soda, r=.388, p<.001, and maternal covert control items. Responsive feeding style scores were correlated with frequencies of child eating fruit, r=.268, p = .004, vegetables, r=.215, p=.01, and eating together with mother, r=.178, p=.03. Both authoritative parenting and responsive feeding style scores were correlated with low child screen time and fast food intake, p<.05. Results show parenting and feeding styles are linked not only to similar but also to unique risk and protective behaviors and practices.
CVT‐10216 selectively suppresses binge eating of palatable foods and attenuates dopamine release in the accumbens of rats
Authors: Bocarsly Miriam E , Avena Nicole M , Hoebel Bartley G , Paredes Daniel , Arolfo Maria P , Yao Lina , Fan4 Peidong , Diamond Ivan
Published: June 2021

CVT‐10216 is a highly selective, reversible inhibitor of ALDH‐2 that reduces alcohol and cocaine intake in rats. Given overlaps in behavior and neurochemistry seen with binge eating and drug use, we examined the effects of CVT‐10216 on binge eating of palatable foods and subsequent dopamine (DA) release in the nucleus accumbens (NAc). Male Sprague‐Dawley rats (n=12/group) were maintained on 1) a binge schedule (12‐h deprivation then 12‐h access to chow and a 10% sucrose solution or vegetable shortening), 2) ad libitum chow with sucrose or shortening, or 3) only chow. After 25 days, CVT‐10216 (0, 7.5, or 15 mg/kg, i.p.) was administered. The 15 mg/kg dose decreased 1 h shortening and chow intake in binge eating rats (p<0.05) and decreased 24 h shortening intake in both the binge and ad libitum groups (p<0.05). This dose selectively decreased 1 h and 24 h intake of sugar in the sugar binge‐eating group (p<0.05). There was a slight decrease in 1 h chow intake for the ad libitum chow control group (p<0.05), but no effect on 24 h intake. In vivo microdialysis was performed in a new set of rats (n=8/group) fed binge sucrose or ad libitum sucrose and chow. CVT‐10216 (15 mg/kg) attenuated the rise in extracellular DA levels in rats binge eating sucrose (p<0.05). These preclinical findings suggest that selective, reversible ALDH‐2 inhibitors may have therapeutic potential to reduce binge eating behavior in clinical populations.

Grant Funding Source : Gilead Sciences
Leptin and adiponectin are both suppressed upon activation of Nucleotide Oligomerization Domain Containing Protein 1 during adipocyte differentiation
Authors: Purohit Jaanki S , Hu Pan , Chen Jiangang , Zhao Ling
Published: June 2021

Obesity is associated with chronic inflammation. Toll‐like receptors (TLR) and NOD‐like receptors (NLR) are two families of pattern recognition receptors that play important roles in inflammation in adipocytes. It has been reported that TLR4 and TLR2 activation induce proinflammatory changes that impair adipocyte differentiation. However, the effects of activation of NOD1 and NOD2, the two prominent members of NLR, on adipocyte differentiation have not been studied. Here we report that activation of NOD1, but not NOD2, by the synthetic ligand dose‐dependently suppresses 3T3‐L1 adipocyte differentiation as revealed by oil red O stained cell morphology, attenuated genes expression of adipocyte markers and lipid accumulation. However, both NOD1 and NOD2 activation significantly suppress the differentiation of primary preadipocytes derived from human subcutaneous fat. In contrast to TLR4 activation, NOD1 activation suppresses both adiponectin and leptin in adipocytes, demonstrating differential effects of NOD1 activation on adipocytes. Overall, our results support that NOD1 represents a novel target for obesity prevention and/or treatment. The work is supported by The University of Tennessee faculty start‐up funds to LZ.
Nitrogen metabolisms of dietary glutamate and the other nonessential amino acids in rats
Authors: Nakamura Hidehiro , Kawamata Yasuko , Kuwahara Tomomi , Torii Kunio , Sakai Ryosei
Published: June 2021

Previous reports have shown that nitrogen (N) supply as nonessential amino acids (NEAAs) is required for normal growth in rats and that glutamate (Glu) is one of the most potent growth promoters among NEAAs. Our recent N‐balance study has also indicated that dietary Glu‐N was retained more than glycine‐N or glutamine‐N, suggesting differences in N‐metabolisms of these NEAAs. Thus, the present study aimed to compare N‐metabolism of dietary Glu with those of the other NEAAs using stable isotope techniques.

Fischer strain male rats were given hourly meals containing [15N]Glu or other [15N]NEAAs. Concentrations and 15N‐enrichments of each amino acid (AA) in portal and arterial plasma were measured to estimate their first‐pass N‐metabolism and their N‐contribution for systemic AA.

Virtually all of the dietary Glu‐N and aspartate‐N were metabolized in the gut for production of various AA such as alanine and citrulline. Meanwhile, substantial amount of the other dietary NEAAs were released into the portal vein directly without any metabolism in the gut. The incorporations of dietary Glu‐N in circulating AA were similar to those of aspartate‐N, alanine‐N and amino‐N of glutamine but greater than those of amide‐N of glutamine, glycine‐N, serine‐N. These results suggest efficient utilization of dietary Glu‐N for the intestinal synthesis of the other NEAAs and hence its significance as N‐source for whole body.

Grant Funding Source: Ajinomoto Co., Inc.
High fasting ghrelin concentrations are related to low fatty acid flux in overweight subjects
Authors: Hovhannisyan Yelena , Lee Joseph , Ramos-Roman Maria , Bradford Dora , Valdez Maressa , Parks Elizabeth
Published: June 2021

Ghrelin plays a role in glucose metabolism. Given the competitive interaction between free fatty acids (FFA) and glucose as fuel sources, the present study determined relationships between acylghrelin and postprandial metabolism in overweight, nondiabetic Hispanic and African American men and women (n=19, BMI 34.8 ± 6.7 kg/m2, 47.1 ± 9.3 yrs). Subjects were admitted for 24 h and consumed a 40g‐fat meal test. Blood metabolite and hormones profiles were determined biochemically, substrate oxidation by indirect calorimetry, insulin sensitivity by IVGTT, and FFA flux by stable isotope infusion and GC/MS. Fasting ghrelin was negatively correlated with fasting insulin (r = −0.47, P = 0.04) and FFA flux (r = −0.55, P = 0.03); it tended to be positively related to glucose oxidation (r = +0.46, P = 0.06). After the meal, the absolute fall in ghrelin correlated with fasting ghrelin (r = +0.88, P < 0.001) suggesting a biological limit of activity. The % decrease in ghrelin correlated positively with insulin sensitivity (r = +0.47, P = 0.04) and fasting TG (r = +0.53, P = 0.02). These data suggest that in overweight subjects, high fasting ghrelin signals efficient glucose use, lower FFA flux, and allows for greater postprandial ghrelin reduction. The connection between ghrelin and fasting FFA flux and TG suggests its role in body weight regulation is mediated through FFA metabolism, as well as though glucose metabolism.
Effects of retinoid catabolism on the gene expression in primary rat hepatocytes and hepatoma cells
Authors: Li Yang , Chen Guoxun
Published: June 2021

Vitamin A (retinol, ROL) is reversibly oxidized into retinal (RAL) by retinol dehydrogenases; retinal is irreversibly oxidized by retinal dehydrogenases into retinoic acid (RA) in cells. RA regulates gene expression via activation of retinoic acid receptor (RAR) and retinoid X receptor (RXR). We have shown that retinoids synergize with insulin to induce Srebp‐1c expression in primary rat hepatocytes. Therefore, we hypothesize that RA production controls the regulation of hepatic gene expression.

The expression of Cyp26a1 (an RA responsive gene) was examined using real‐time PCR and used as a marker for RA production from ROL and RAL in primary hepatocytes and hepatoma cells. Here, we show that retinol and retinal dose‐dependently induced Cyp26a1 expression via activation of RAR and RXR in those cells. The expression levels of Rdh2, Rdh10, Raldh1, and Raldh3 (major retinoid catabolic enzymes) were examined in hepatocytes from Zucker lean (ZL) and fatty (ZF) rats. Only Rdh2 and Raldh1 were expressed at comparable level as that of 36B4 gene (the invariable control gene) in hepatocytes and hepatoma cells. In addition, the hepatocytes from ZF rats had higher expression level of Raldh1 than that from ZL rats.

We conclude that hepatocytes actively metabolize retinol into RA, which directly regulate gene expression. The elevated Raldh1 expression in ZF hepatocytes may contribute to the altered lipid metabolism in ZF liver.

Sponsor: Guoxun Chen, 1215 W. Cumberland Ave. 229 Jessie Harris Building, Knoxville, TN 37996

Grant Funding Source : AHA
Polymorphisms in FADS2 correlated with altered desaturase activity in a type‐2 diabetic cohort
Authors: Huang Pei-Chi , Jhang Huei-Ru , Hsu Chih-Cheng , Kothapalli Kumar , Brenna Tom , Huang Meng-Chuan
Published: June 2021

Recent evidence reveals that single nucleotide polymorphisms (SNP) in fatty acid desaturase 2 (FADS2) is correlated with alteration in blood fatty acid profile in some ethnic groups. We aimed to examine effects of FADS2 (intronic SNP, rs174589) on Δ6 and Δ5 desaturase activities based on ratios of 18:3n‐6/18:2n‐6 and 20:4n‐6/20:3n‐6 respectively. In 2008, we cross‐sectionally analyzed erythrocyte fatty acids in type 2 diabetic patients (n=702) recruited from the Diabetes Management through Integrated Delivery System (DMIDS) cohort established in Taiwan. Frequencies of rs174589 genotypes GG, GC and CC were 0.739, 0.234 and 0.027 respectively. Levels of hemoglobin A1c in CC genotypes were significantly different from those of GC and GG genotypes. Δ6 desaturase activity (18:3n‐6/18:2n‐6) was significantly reduced in GC (0.005±0.003, p=0.003) and CC (0.004±0.003, p=0.025) than GG genotypes (0.006±0.003). Similarly, Δ5 desaturase activity (20:4n‐6/20:3n‐6) was significantly diminished in C allele carriers than the GG homozygotes (p<0.05). In conclusion, rs174589 in FADS2 is associated with Δ6 and Δ5 desaturase activities in type 2 diabetes patients and impact of these alterations in fatty acids profiles on clinical parameters warrants further investigation.
SURVEY OF HUMAN MILK FATTY ACID LEVELS OF U.S. WOMEN WHO PROVIDE EXPRESSED MILK FOR FEEDING THEIR PRETERM INFANTS
Authors: Berseth Carol Lynn , Mitmesser Susan Hazels , Harris Cheryl , Hoffman Dennis R , Diersen-Schade Deborah
Published: June 2021

Background

Preterm infants have reduced in utero accretion of docosahexaenoic acid (DHA) and arachidonic acid (ARA) due to early birth. Their postnatal DHA status is in part dependent upon the DHA content of their mother's milk, but DHA content of US human milk is among the lowest in the world, typically ≤0.2% of total fatty acids.

Objective

To characterize DHA, ARA, and eicosapentaenoic acid (EPA) levels in milk expressed by mothers of preterm infants.

Methods

Mothers of infants with a gestational age ≤30 3/7 weeks provided single milk samples during the first or second postnatal month. Fatty acids were determined by capillary column gas chromatography. Maternal consumption of fish and/or DHA supplements during the last 12 weeks of pregnancy was recorded.

Results

Sixteen mothers consumed fish, 25 consumed a DHA supplement, 11 consumed both supplement and fish, and 76 did not consume either. DHA was higher among mothers consuming fish or supplement, and greatest for those consuming both (Table). Milk DHA trended higher than previously seen for U.S. mothers. There was little difference in ARA among the groups; EPA was highest among those using supplements and surprisingly, lowest for those consuming fish.

Conclusion

Milk levels of DHA are lowest among those mothers who did not consume fish or DHA supplements during pregnancy. Dietary counseling for mothers providing milk for their preterm infants may increase their infant's DHA intake.
Impact of normal or low organic mineral (Cu, Zn, Mn) diets on gene expression patterns in the intestine of broiler chicks
Authors: Brennan Kristen M , Samuel Ryan S , Ao Tuoying , Ford Michael J , Pescatore Anthony J , Cantor Austin H
Published: June 2021

We assessed the impact of early life mineral (Zn, Cu, Mn) nutrition on gene expression in the jejunum. Broiler chicks were randomly assigned to 1 of 3 treatments. Corn‐soy diets had organic Zn, Cu, and Mn (BioPlex®, Alltech Inc.) at 100% (normal, N) or 20% (low, L) of NRC requirements. Diets were fed for 5 d, followed by a second diet (d 6–21), creating 3 groups: N‐N, N‐L, or L‐L. On d 21, birds were euthanized and gene expression was analyzed using the Affymetrix Chicken Genome Array. Compared to N‐N, the number of genes significantly changed was 303 for N‐L and 530 for L‐L, with 154 commonly affected. Both N‐L and L‐L led to down‐regulation of genes metallothionein 3 and 4 compared to N‐N. Gene ontology (GO) classifications showed that L‐L had a greater impact than N‐L on genes associated with metal ion transport, Zn ion binding and carbohydrate metabolic processes. IPA analysis showed the top biologic functions affected by N‐L were cell signaling, cellular function and maintenance, and cell morphology; for L‐L they were protein synthesis, cell death and post‐translational modification. Although both groups were fed the same diet d 6 – 21, IL‐17A pathways were enriched with L‐L only. Thus, mineral levels up to 5 d of life altered gene expression in birds on d 21, suggesting that early nutrition has a significant, long term impact on gene response in the intestine of young chickens.
Adequate vitamin B‐6 is associated with increased antioxidant enzyme activity in critically ill surgical patients
Authors: Huang Yi-Chia , Cheng Chien-Hsiang , Chen Fang-Pei , Chiang Ting-Yu
Published: June 2021

Inadequate vitamin B‐6 status may increase oxidative stress and compromise antioxidant capacity. The purpose of this study was to understand the association between vitamin B‐6 status and oxidative stress, antioxidant enzyme capacities in critically ill surgical patients. Patients were allocated into either adequate B‐6 (n = 11) or deficient B‐6 (n = 10) group based on their fasting plasma pyridoxal 5′‐phosphate (PLP) concentration at admission to the surgical intensive care unit (SICU). Plasma and erythrocyte PLP, serum malondialdehyde (MDA), total antioxidant capacity (TAC), superoxide dismutase (SOD), glutathione S‐transferase (GST) and glutathione peroxidase (GPx) were determined on the 1st, 4th, 7th and 10th d of admission. There was no significant difference in TAC and MDA levels, GST and GPx activities between the 2 groups on the 1st, 4th, 7th and 10th in the SICU. The mean SOD activity level was significantly higher in the adequate B‐6 group when compared to the deficient B‐6 group on the 1st and 4th after admission. The change of plasma PLP (rs = 0.68, p < 0.01) significantly correlated with the change of SOD activities after adjusting for potential factors. Higher plasma PLP at admission would be an important contributing factor to increased antioxidant enzyme activity in SICU patients. This study was supported by Taichung Veterans General Hospital (TCVGH‐994101B), Taiwan.
Effect modifiers for the safety and efficacy of iron intervention in the context of malaria: A review of data for an individual patient based meta‐analysis
Authors: Namaste Sorrel , Ashour Fayrouz , Porter Alexandra , Raghavan Ramkripa , Pilch Susan , Raiten Daniel
Published: June 2021

Objective

To evaluate the safety and efficacy of iron interventions in regions with malaria, a review was undertaken to determine the extent of data that could be utilized in an individual patient data (IPD) meta‐analysis. IPDs are particularly useful in assessing the heterogeneity of treatment effects and thus lend themselves to examining the influence of risk modifiers.

Methods

Eligible randomized trials were identified using computerized and manual searches. Studies were stratified by iron status, age, malaria patterns, and iron mode. The number of studies and the upper/lower expected pooled sample size range was calculated.

Results

A total of 3,602 trials were screened, 523 underwent text review and 165 included. There were 106 supplementation and 50 fortification trials; mainly in meso (50%) and hyperendemic (29%) regions. Serum ferritin was the predominant iron biomarker (n=27, 12,122–17,671 >2yr; n=47, 17,891–34,305 >5yr; n=9, 3,026 pregnancy) and was rarely accompanied by CRP or AGP. Anthropometry was the most frequent health endpoint (52%), followed by morbidity (44%), and clinical malaria (12%).

Conclusion

There are sufficient studies to warrant an IPD. However, the limitations in the number and nature of studies highlight the need for a more robust research agenda, including well designed protocols to support nutrition programs and policy.

Support: Bill & Melinda Gates Foundation; NIH
Two GC‐biased genotypes of Ophiocordyceps sinensis with distinct maturational patterns in the stroma of Cordyceps sinensis
Authors: Yao Yi-Sang , Gao Ling , Zhao Jiagang , Zhu Jia-Shi
Published: June 2021

We reported simultaneous detections of the rDNA of Paecilomyces hepiali and at least 5 Ophiocordyceps sinensis (Os) genotypes in Cordyceps sinensis (Cs), the precious Chinese herb for anti‐aging. Southern blotting following EcoRI digestion showed 2 groups of genotypes, EcoRI sensitive (GC‐biases) and EcoRI resistant (AT‐biases and other mutants). SNP genotyping revealed multiple mutants in Cs with transition or transversion point mutations, differentially expressed in Cs stroma and caterpillar. Cs maturation associates altered expressions and phenotypes of these fungal species or mutants. In this study, we found that GC‐biased genotypes expressed little in the premature Cs stroma, but predominantly in the premature caterpillar. The differential expression was altered non‐proportionally with Cs maturation. Two GC‐biases were identified having identical sequences: (1) Hirsutella sinensis and (2) a moiety truncated at its 5′ and 3′ ends. The 2 GC‐biases have distinct expression patterns during Cs maturation. In conclusion, Cs stroma contains 2 GC‐biased genotypes with distinct maturation patterns. It remains to be determined if the 2 GC‐biases and all other Os genotypes are intra‐ or inter‐species mutants. Although ITS sequencing has been widely used for fungal species identification, our data indicates that the method may have limited power in distinguishing between fungal mutants.
A low zinc concentration detected in sera under zinc deficiency induces dysregulation of cytokine productions in rats
Authors: Ishizuka Satoshi , Suwendi Erwin , Iwaya Hitoshi , Lee Jae-Sung , Hara Hiroshi
Published: June 2021

Zinc (Zn) deficiency has been observed in patients with ulcerative colitis (UC), although the precise contribution of Zn to UC prevetion has not been elucidated until now. We investigated Zn status during dextran sulfate sodium (DSS) ‐ induced colitis. Rats were fed Zn adequate diet (ZA, 30 mg Zn/kg diet) as a control or Zn deficient diet (ZD, 5 mg Zn/kg diet) for 21 days and then treated with 2% DSS through deionized drinking water for 7 days. The disease activity index (DAI) was recorded daily during the DSS treatment. Serum Zn concentration in the rats fed the ZD diet was significantly lower than in those fed the ZA diet at day‐7 and ‐ 14. DSS treatment considerably reduced the serum Zn to the same extent in both groups. The ZD rats showed exacerbated colitis as assessed by clinical outcomes including weight loss, increased DAI, and shortening of colon length. In vitro study corroborated these results, showing that large amounts of TNFα were produced under ZD medium by rat mesenteric leukocytes in response to stimulants. Interestingly, such increase in the TNFα production was not found under ZA medium. These results indicate that excess TNFα production due to Zn deficiency increases the disease activity in DSS‐induced colitic rats.
Dietary Arctium lappa L. root extracts: Molecular mechanisms of action as anti‐allergic agents
Authors: Sohn Eun-Hwa , Kim Ye-Jin , Kim Taeseong , Namkoong Seung
Published: June 2021

Atopic dermatitis is a chronic, allergic inflammatory skin disease that is accompanied by markedly increased levels of inflammatory cells. Arctium lappa L. is a traditional medicine in Asia. This study examined the effect of ALBE (butanolic extracts of A. lappa) on the release of β‐hexosaminidase in antigen‐stimulated‐RBL‐2H3 cells and the expression of IL‐4, IL‐5, pMAPKs, and NF‐κB activation in ConA‐stimulated splenocytes. We observed significant inhibition of β‐hexosaminidase release, the production of IL‐4 and IL‐5, NF‐κB translocation and the phosphorylation of MAPKs. Additionally, ALBE suppressed the expression of ICAM‐1 and the production of NO‐iNOS induced by TNF‐α in A549 endothelial cells. These results suggest that ALBE may have beneficial effects in the treatment of allergic diseases, including atopic dermatitis. This study was supported by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.
Determining source populations of newly identified cases of chronic wasting disease in white‐tailed deer
Authors: Green Michelle L. , Manjerovic Mary Beth , Mateus-Pinilla Nohra E. , Kelly Amy C. , Shelton Paul , Novakofski Jan
Published: June 2021

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy caused by prions, and affects deer, elk and moose. CWD was first reported in Illinois in 2002 and continues to spread. Spark deer, or new CWD cases found in disease free locations > 60 miles from infection zones, are worrisome because of their risk of infecting healthy populations. In Illinois, six spark cases have been reported south of the north core area. It is unknown whether spark deer originated from the core and migrated to southern areas or southern individuals contracted CWD from within their locale. The objective of this study was to genetically identify source populations of spark cases. Using microsatellites, randomly selected samples from north and south source populations were genotyped, tested for Hardy‐Weinberg and linkage disequilibrium using CERVUS, and allele frequencies calculated using ARLEQUIN. Assignment tests compared genotypes of each spark case to the north and south to calculate the probability of genotype occurrence in each population. Based on the probability, we assessed likelihoods that these spark deer originated from the disease core or local population. The results of the study provided a better understanding of potential mechanisms of spark case occurrence, the data required to evaluate sparks in Illinois and potential sources of CWD spread. This project was supported by the IDNR and UIUC‐OVCR.
Modulation of mdr‐1 Expression and Activity by Verapamil and Resveratrol Potentiates the Cytotoxicity of Docetaxel and Doxorubicin in Solid Tumor Cell Lines
Authors: El-Moselhy Mohamed Ahmed Mohamed , El-Sherbiny Gamal Ahmed , Abd Abdel-Latif Hekma Abdel-Tawab , Al-Abd Ahmed Mohamed , Nofal Salwa Mohamed , El-Eraky Wafaa Ibraheem
Published: June 2021

P‐glycoprotein (P‐gp) is a member of ATP‐dependent membrane transport protein superfamily encoded by mdr‐1 gene family. In tumor cells, expression of P‐gp results in reduced intracellular drug concentrations, which decreases the cytotoxicity of antitumor drugs. Herein, we evaluated the added value of combining two P‐gp blockers (verapamil and resveratrol) with docetaxel and doxorubicin and the possible underlying mechanism(s) in solid tumor cell lines (MCF‐7, HeLa and HepG2). Verapamil possessed weaker anti‐cancer properties than resveratrol (IC50 of 92.1 to 107.6 μM and 35.0 to 84μM, respectively). Doxorubicin and docetaxel IC50's of 0.48 to 0.72 μM and 25.9 to 77.8 nM, respectively. Verapamil combination with doxorubicin and docetaxel significantly decreased the IC50's of both agents to 0.034 to 0.12 μM and 2.5 to 6.0 nM, respectively. Resveratrol increased the potencies of both chemotherapeutic agents to IC50 of 0.12 to 0.34 μM and 7.2 to 53.02 nM, respectively. The interaction of resveratrol with both agents was additive in HeLa and Hep‐G2 cell lines. Surprisingly, only resveratrol but not verapamil interfered with the expression of mdr‐1 and several apoptotic genes BAX, BcL2 and p53. P‐gp efflux activity was significantly inhibited by resveratrol and verapamil with subsequent accumulation of P‐gp substrate within the intracellular compartment. In conclusion, resveratrol was comparable to verapamil in potentiating the anti‐cancer effects of doxorubicin and docetaxel in solid tumor.
Differential responses to social stress are associated with qualitatively different responses of dorsal raphe nucleus (DRN)‐serotonin (5‐HT) neurons to corticotropin‐releasing factor (CRF)
Authors: Wood Susan K , Zhang Xiao-Yan , Reyes Beverly , Van Bockstaele Elizabeth J , Valentino Rita J
Published: June 2021

Psychosocial stress is linked to psychiatric disorders such as depression. Using a social stressor in rats (resident‐intruder model) we previously identified two phenotypic responses to social stress. One population (45%) exhibited a passive response and assumed a defeat posture within a short latency (SL, <300s). The other phenotype (55%) exhibited proactive behaviors and a longer defeat latency (LL, >300s). Furthermore, SL rats developed endocrine and behavioral characteristics resembling depression. As DRN‐5‐HT neurons are implicated in psychiatric disorders and are regulated by the stress neuropeptide, CRF, their responses to CRF were compared in social stress and control rats. CRF (30 ng, intra‐DRN) decreased 5‐HT neuronal firing in control and SL rats. In contrast, CRF activated 5‐HT neurons of LL rats. Because inhibition and activation of DRN‐5HT neurons has been linked to CRF1 and CRF2 receptors, respectively the effects of a CRF2 antagonist, antisauvagine‐30 (ASV) were examined. ASV (300 ng, icv) prevented CRF‐induced activation of DRN‐5‐HT neurons in LL rats. Consistent with this, electron microscopy revealed that social stress resulted in recruitment of CRF2 to the plasma membrane of DRN neurons selectively in LL rats. These data suggest that differential trafficking of CRF receptors in DRN neurons underlies phenotypic differences in responses to and consequences of social stress. MH05280
Determining the Importance of Changes in Dopamine when Modafinil is Used as a Treatment for Cocaine Dependence
Authors: Brewer Alex Jawann , De La Garza Richard
Published: June 2021

Cocaine (COC) dependence is a significant public health issue. Currently, there is no drug approved by the FDA to treat COC dependence; however, recent data indicates that modafinil (MOD) is effective in reducing the subjective and reinforcing effects produced by COC. How MOD produces these effects is unclear. Dopamine (DA) has been widely speculated to mediate the effects of MOD, and MOD was recently shown to bind and inhibit the DA transporter, which increases DA in brain. In addition, DA has long been implicated as critical to the rewarding and reinforcing effects produced by stimulants such as COC. Based on the preceding information, I hypothesize that MOD reduces the subjective and reinforcing effects produced by COC by increasing DA in the brains of COC‐abusing individuals. This hypothesis will be tested in my dissertation project by enrolling participants who are not currently seeking treatment into a phase I clinical trial. Participants will be administered MOD plus either lisdexamfetamine, which will increase brain DA, haloperidol, which will decrease brain DA, or placebo, then challenged with COC. Changes in DA concentration will be measured several ways, both directly and indirectly. Outcome variables will include the participants’ subjective responses, self‐administration outcomes, and measures of plasma prolactin and DA.
The nicotinic subunit β4 plays a modulatory role on the antiaddictive action of varenicline
Authors: Dang Justin , Arias Hugo , Jackson Gary
Published: June 2021

The role of β4‐containing nicotinic receptors (AChRs) in drug addiction and in the anti‐addictive effects of varenicline was studied. A cocaine‐induced locomotor sensitization model was developed to compare the anti‐addictive effect of varenicline between wild‐type (β4+/+) and mutant (β4−/−) mice, male and female. Mice were first habituated for 75 min, injected with saline (control) or cocaine (15 mg/kg), and then the locomotor activity was measured for 75 min. Mice were treated intermittently during days 1–5, 8, 11, 12, 14, 18, 19, 21, 28, 35, 42. Mice were pretreated with varenicline (1 mg/kg) before cocaine injection, or treated with varenicline alone. Two‐way ANOVA analyses indicate that varenicline decreases the cocaine activity in wild‐type and mutant males (p<0.05). The decreased activity produced by varenicline on wild‐type and mutant females was not statistically significant (p>0.36). Pretreated mutant males, but not mutant females, have lower activity compared to wild‐type mice (p<0.068). Varenicline alone had comparable results with saline. The results support the view that the anti‐addictive effect of varenicline is more pronounced in male than in female mice, and that the lack of the β4 subunit increases the anti‐cocaine effect of varenicline. In this regard, β4‐containing AChRs may play a modulatory role in cocaine addiction and could be a possible target for drug addiction treatment.

Grant Funding Source: Science Foundation Arizona
Donepezil and tacrine differentially interact with estradiol in female rats responding under a multiple schedule of repeated acquisition and performance
Authors: Leonard Stuart , Weed Peter F. , Winsauer Peter J.
Published: June 2021

In females, ovarian hormones can facilitate learning and memory processes but such beneficial effects are often lost or diminished following long‐term hormone deprivation. Because the absence of estradiol disrupts learning and memory presumably by reducing the activity of the cholinergic system, acetylcholinesterase (AchE) inhibitors may reverse this disruption by enhancing the functional status of the cholinergic system. This study investigated the interactive effects of estrogen and two AchE inhibitors, donepezil (0.56–5.6 mg/kg) and tacrine (0.56–10 mg/kg), in subjects responding under a multiple schedule of repeated acquisition and performance of response chains. Dose‐effect curves were determined in female rats that were gonadally intact, ovariectomized, or ovariectomized with estradiol replacement. In all three groups, both drugs produced dose‐dependent rate‐decreasing and error‐increasing effects in the acquisition and performance components. However, OVX females were less sensitive to the error‐increasing effects of donepezil in the acquisition component and the error‐increasing effects of tacrine in the performance component. These data extend the current knowledge regarding the effects of estrogen on nonspatial learning and memory tasks, and help determine if estradiol's capacity for mediating responding on such tasks is related to its ability to affect cholinergic neurotransmission. Supported by DA019625 and Contract NSF(2010)‐PFUND‐198 Board of Regents of Louisiana
Alpha1 and alpha2 adrenoceptors in the medial amygdaloid nucleus have different role on cardiovascular responses to acute restraint stress in rats
Authors: Fortaleza Eduardo Trindade , Scopinho América Augusto , Corrêa Fernando Morgan
Published: June 2021

Introduction and Goals

Medial amygdaloid nucleus (MeA) is involved in the modulation of the cardiovascular responses during exposure by restraint stress (RS). We investigate the involvement of the alpha‐adrenoceptors into the MeA therein mediate cardiovascular responses to RS in rats.

Methods

We used Wistar rats. It was done implanting bilateral guide cannula into the MeA to microinjections of drugs or vehicle. The animals received a polyethylene catheter into the femoral artery for blood pressure (BP) recording.

Results

The RS caused BP and heart rate (HR) increases in aCSF treated animals (n=8). RX821002 (alpha2‐ adrenoceptor antagonist) treatment significantly enhanced restraint‐evoked HR increase without significant effect on the BP response, when compared with control animals (ΔMAP: F1,66= 3,496, P= 0,0660, n=5; ΔHR: F1,66= 15,22, P= 0,0002, n=5; twoway ANOVA). In contrast, the treatment with WB4101 (alpha1‐adrenoceptor antagonist) significantly reduced restraint‐evoked HR increase without significant effect on the BP response, when compared with control animals (ΔMAP: F1,66= 0,8556, P= 0,3583, n=5, and ΔHR: F1,66= 3,558, P= 0,0637, n=5; two‐way ANOVA).

Conclusion

Results suggest that adrenoceptors either alpha(1) and alpha(2) have different roles into the MeA on the HR increase evoked by RS, enhance and reducing the cardiac responses.

Financial Support: CNPq, CAPES.
Mass‐spectrometric analysis of changes in brain lipid expression and distribution after brain injury
Authors: Cox Brian , Post Jeremy , Bull Gregory , Gouty Shawn , Woods Amina
Published: June 2021

Mechanical impact injuries to the cortex of rats induce neuronal loss, vascular damage, glial activation, infiltration of circulating immune cells, and edema, followed by partial revascularization, partial neuronal recovery and remodeling of neuronal connectivity. We report here changes in lipid metabolism in the region of injury after a controlled cortical impact (CCI) injury to the cortex of anesthetized Sprague‐Dawley rats. At selected time points after induction of a CCI injury inducing tissue damage and subsequent loss of some cortical tissue but not major functional impairment, brains were removed and processed for imaging mass spectrometry of 16 μm coronal sections through the region of injury. Following injury, significant time‐related changes in numerous lipid species in both gray and white matter around and through the site of injury were observed. Specifically, increases and decreases in the levels of specific sphingomyelin species were noted at 24 hrs, 3 days and 7 days after CCI injury. Phosphatidylcholine 36a:2 was found in greater abundance in both grey and white matter in the area of injury than in the contralateral cortex, while another phosphatidylcholine 38:4 showed no change in distribution between sham and injured animals. Lipid imaging presents new insights into the cellular consequences of injury, (Supported by US Dept of Defense through the CNRM).