bluuebunny/crossref_metadata_2025 huggingface dataset:
Qualitative Assessment of Nutrition Education and Health Needs of University Mid‐Age Female Staff
Authors: Yen Wan‐ju Jennifer , Lewis Nancy M.
Published: June 2021

Researchers have described women as facing a dramatic increase in the risk of heart disease, osteoporosis, stroke, and Alzheimer's, the onset of these as the result of the impact of changing hormone levels, particularly the decline in estrogen. As large numbers of women reach midlife, understanding their perceptions of health stability, opportunities for change in dietary attitudes and practices and the influences on them will become increasingly important to nutrition educators. The objective of this qualitative study was to identify mid‐age female's beliefs and interests around the topic of nutrition. Data were collected using audiotaped semi‐structured individual interviews. Eight female staff (aged 45 to 65) in a Midwestern university were recruited using snowball sampling. Interviews were transcribed verbatim. Data were coded and themes developed by sorting and summarizing coded transcript segments. Three themes emerged: “Health” related to their individual conditions, such as avoiding illnesses and weight management, with aging; “Life style” described what occasions lead to participants’ current dietary action; “Availability” described the factors that influenced participants’ healthy eating behaviors. The results of this study provide a context to develop a nutrition intervention for this target population.
Alpha linolenic acid increases lipid efflux through inhibition of stearoyl CoA desaturase 1 during foam cell formation
Authors: Zhang Jun , Heuvel John Vanden , Kris‐Etherton Penny
Published: June 2021

Epidemiologic studies indicate that higher alpha linolenic acid (ALA) intake is associated with reduced cardiovascular disease risk. Stearoly CoA desaturase (SCD) is a rate‐limiting enzyme in triglyceride production by converting saturated fatty acids to monounsaturated fatty acids. Increased SCD activity is related with obesity, metabolic syndrome and atherosclerosis. There is some evidence that SCD1 also is involved in the cholesterol efflux from macrophage foam cells, important mediators of the atherogenic plaque. The current study was conducted to investigate the regulation of SCD1 by ALA and its contribution to cholesterol efflux. To induce foam cell formation, RAW 264.7 macrpohages were treated with oxidized low‐density lipoprotein (oxLDL). ALA treatment with and without oxLDL significantly decreased SCD1 expression by 88% and 82%, respectively, in macrophages. In a newly developed fluorescence‐based lipid efflux experiment, lipid efflux increased about 2 fold following ALA treatment (100 μM) compared with control. This effect was independent of reduced expression of ATP binding cassette transporter A1 and G1 (ABCA1, ABCG1). ALA inhibited SCD1 expression and would be beneficial to atherosclerosis regression via increasing efflux from lipid ladened foam cell.
Soluble cellulose reduces expression of lipogenic and inflammatory genes in fat fed hamsters
Authors: Hong Yun‐Jeong , Bartley Glenn , Turowski Maciej , Anderson Kerr , Yokoyama Wallace
Published: June 2021

The underlying cause of the metabolic differences between normal and insulin resistant hamsters fed identical high fat diets except for dietary fiber solubility were investigated by determining relative expression of genes for fat metabolism and inflammation. Male Syrian hamsters switched from standard laboratory chow to a high fat diet become insulin resistant and hyperlipidemic. If the same high fat diet contains hydroxypropylmethylcellulose, as the dietary fiber source, most of the adverse physiological effects of the high fat diet are ameliorated. Relative expression of genes related to fat metabolism, nuclear receptors, and inflammation were determined in order to understand the molecular basis for the normalization of blood and liver lipids and adipose mass. Primers for hamster leptin, adiponectin, CYP51, FXRalpha, CD36, F4/80, MAC‐1, GLUT4, SCD1, TNF alpha, PPAR alpha and gamma, SREBP‐1c and others in liver and/or adipose were developed. Dietary soluble cellulose consistently lowered SCD1 and increased CYP51 expression relative to insoluble microcrystalline fiber feeding. Adipose GLUT4 and inflammatory marker expression was decreased. PPARalpha expression but not PPARgamma increased in some studies.
Randomized controlled trial comparing the bioavailability of vitamin D3 from fortified Cheddar cheese, fortified low‐fat cheese, and supplement
Authors: Wagner Dennis , Sidhom Gloria , Whiting Susan J , Rousseau Derick , Vieth Reinhold
Published: June 2021

Objective: We compared the bioavailability of vitamin D3 from novel fortified hard cheeses and from vitamin D3 supplement.

Methods: 60 adults were randomly given weekly servings of fortified Cheddar cheese (34 g), fortified low‐fat cheese (41 g), or vitamin D3 supplement. These contained 28,000 IU vitamin D3, equivalent to 4,000 IU/d. 20 adults were also randomized to placebo. Bioavailability of vitamin D3 was assessed by comparing the serum 25‐hydroxyvitamin D [25(OH)D] response over 8 weeks.

Results: In the placebo group, initial 25(OH)D, 55.0 Â± 25.3 nmol/L (mean Â± SD), declined over the 8‐wk winter protocol, to 50.7 Â± 24.2 nmol/L (paired t test, p=0.046). In the vitamin D3‐treated groups, the mean increases in 25(OH)D from baseline to 8 wk were: 65.3 Â± 24.1 nmol/L (Cheddar cheese), 69.4 Â± 21.7 (low‐fat cheese), 59.3 Â± 23.3 (supplement with food), and 59.3 Â± 19.6 (supplement without food); these changes were significantly different from the placebo group (ANOVA, p<0.0001) but not from one another (ANOVA, p=0.62). Compared with baseline, serum parathyroid hormone decreased significantly with both the fortification (paired t test, p=0.003) and the supplementation (paired t test, p=0.012) strategies.

Conclusions: Vitamin D3 is equally bioavailable from fortified hard cheeses and from supplement. Therefore, hard cheese is a viable food for vitamin D fortification, and can help to enhance vitamin D status.
Glycosaminoglycans isolated from fish muscle enhance iron uptake by Caco‐2 cells
Authors: Llopis José Moisés Laparra , Tako Elad , Glahn Raymond P , Miller Dennis D
Published: June 2021

Numerous attempts have been made to identify the factor in muscle tissue that enhances Fe absorption but the nature of the promoter remains controversial. Previous research suggested that sulfated glycosaminoglycans (GAGs) from fish muscle promote Fe uptake in Caco‐2 cells (1). This study continues that research to confirm that GAGs enhance Fe uptake by Caco‐2 cells.

Fish haddock filets were cooked and lyophilized. GAGs were isolated and separated into low and high sulfur fractions (2). Enzymatic digestion with chondroitinase ABC (50 mU/mg, 1h, 37ºC) and reverse phase chromatography (RP‐HPLC) were used to characterize the GAGs. Nonheme Fe was added to the isolated fractions and to whole cooked fish and an in vitro digestion/Caco‐2 cell model was used to assess their effect on Fe uptake (1).

RP‐HPLC analyses showed that retention times of the main components in the high sulfur fraction were consistent with chondroitin/dermatan sulfate‐related structures. This fraction increased Fe uptake up to 185% relative to controls exposed to nonheme Fe. Treatment of the isolated fraction with chondroitinase eliminated the enhancing effect. Unfractioned fish caused a 40% higher Fe uptake than caused by GAG isolates.

These results indicate that specific GAGs contribute to the enhancing effect of fish on Fe uptake by Caco‐2 cells but that other factors are also involved.
Copper induces, and the copper chelator tetrathiomolybdate inhibits, activation of human aortic endothelial cells
Authors: Wei Hao , Zhang Weijian , Frei Balz
Published: June 2021

Copper may play an important role in enhancing oxidative stress and vascular inflammation. We investigated whether copper can induce activation of human aortic endothelial cells (HAECs), and whether copper chelation by tetrathiomolybdate (TTM) can inhibit this process. We found that incubating HAECs with 6.25 to 100 μM copper for 4 to 16 hrs dose‐ and time‐dependently up‐regulated vascular cell adhesion molecule‐1 (VCAM‐1), intercellular adhesion molecule‐1 (ICAM‐1), and monocyte chemotactic protein‐1 (MCP‐1) mRNA and cell surface protein levels. Maximum induction of VCAM‐1, ICAM‐1, and MCP‐1 mRNA levels (3.0±0.2, 4.2±0.8, and 5.8±1.4‐fold increase, respectively) was observed following incubation with 100 μM copper for 4 hrs. Maximum induction of cell surface protein levels of VCAM‐1 and ICAM‐1 was observed after 8 hrs of incubation with 100 μM copper (4.6±0.5 and 3.6±0.3‐fold increase, respectively). TTM in 10 μM significantly inhibited copper (100 μM) induced increases of surface protein levels of VCAM‐1 and ICAM‐1 by 67.0±4.9% and 78.2±4.8%, respectively. Furthermore, copper in 100 μM caused activation of activator protein 1 (AP‐1) and nuclear factor kappa B (NFκB) by 1.5±0.02 and 2.4±0.05 fold, respectively, which was significantly inhibited by TTM (10 μM). In conclusion, our data indicate that excessive copper may cause endothelial activation by up‐regulating expression of adhesion molecules and chemokines, and that TTM may be helpful in ameliorating copper‐induced vascular inflammation. This research was supported by NIH grant CERCAT, NIH P01 AT‐002034.
Redox regulation of HIV‐1 transcription by selenoprotein thioredoxin reductase
Authors: Prabhu K. Sandeep , Kalantari Parisa , Reddy C. Channa , Henderson Andrew J
Published: June 2021

Selenium slows HIV disease progression, increases CD4(+) T cell counts, and reduces mortality. However, the underlying mechanism of selenium is still unclear. Here we demonstrate that selenium, in the form of the host disulfide oxidoreductase, thioredoxin reductase (TR), regulates the transcription of HIV. Biochemical and genetic experiments indicate that TR reduces the disulfide bonds in the HIV transactivating protein Tat and, thus, regulates HIV‐1 transcription. These studies shed light on a novel redox mechanism that host cells use to repress viral transcription. In addition, these studies suggest that selenium may provide a simple and inexpensive adjunct therapy in conjunction with current anti‐retroviral therapeutics to control HIV replication.
The effects of excessive selenium metabolism on gene expression of HepG2 cells
Authors: Chen Yu Hsin , Yang Feili Lo
Published: June 2021

High level of selenium (Se) in culture medium is lethal to cells. This study utilized cDNA microarray technique to explore the influence of excessive Se on the gene expression of HepG2 cells. Exposure of HepG2 cells to 20 μM of sodium selenite for 12 hr resulted in a greater than 5‐fold changes in the expression of 96 genes. These genes are involved in a widespread of cellular events such as cell growth and/or maintenance (46), cell proliferation (10), cell differentiation (11), cell death (46), apoptosis regulator activity (9), morphogenesis (5), regulation of gene expression, epigenetic (24), transcription and transcription regulator activity (11,13), translation factor activity (43), cell‐cell signaling (12), nucleotide and nucleic acid metabolism (11), protein phosphatase activity (3), protein tagging activity (7), protein metabolism (1), carbohydrate metabolism (3), energy pathways (6), mitochondrion organization and biogenesis(6), electron transport (9), ion channel activity (8), calcium ion binding (6), cell adhesion molecule activity (4) and immune functions (1). Therefore changes in gene expression during the early phase in response to excessive Se may result in changes in functions and metabolism of hepatocytes leading to their decreased survival. Further analyses on the correlation among the genes in clusters of cellular activities upon the influence of excessive Se metabolism are pending. (Sponsored by Office of Research and Development, Fu Jen Catholic University, Project 409531060134)
Calculating Estimated Average Requirements (EARs) for zinc (Zn) According to Dietary Phytate
Authors: Hambidge K Michael , Miller Leland V , Westcott Jamie E , Krebs Nancy F
Published: June 2021

Background: Estimation of the DRIs (dietary reference intakes) for Zn by the Institute of Medicine (IOM, 2001) specifically included calculations only for phytate‐free diets.

The objective of this investigation was to apply a mathematical model (J Nutr, 2007) of daily total absorption of Zn (TAZ) as a function of daily total dietary intake of Zn (TDZ) and of phytate (TDP) to derive EARs for Zn in adults consuming various quantities of phytate.

Methods: The IOM estimated physiologic requirements for Zn for men (3.8 mg Zn/d) and women (3.3 mg Zn/d) were used for this purpose. An updated version of the model was used to calculate the TDZ and TDP values which produce TAZs equal to these physiological requirements.

Results: The relationship between TDZ and TDP to yield a constant TAZ is linear and described by the following equation for the case of TAZ = 3.8 mg/d: TDZ = 0.0093 × TDP + 8.5, where TDZ and TDP are in mg/d. For a TDP of 1000 mg/d, the model predicts an EAR that is approximately double, and for a TDP of 2000 mg/d an EAR approximately triple, the ‘no phytate’ EAR of the IOM.

Conclusion: As these and greater intakes of phytate are typical in developing countries, these calculations have major implications for many populations, not only for EARs, but also for RDAs and Upper Limits.

Supported by IAEA and MRDDR Center Grant P30 HD004024.
Supplementation of Fucoxanthin Rich‐Seaweed Extract Improves Lipid Profiles and Suppresses Body Fat in Mice
Authors: Woo Myoung‐Nam , Jeon Seon‐Min , Kim Hye‐Jin , Yeo Jiyoung , Shin Young Chul , Choi Myung‐Sook
Published: June 2021

This study investigated the hypolipidemic effect of fucoxanthin from seaweed ethanol extract in mice fed a high‐fat diet. Male C57BL/6J mice were divided into 3 groups; control (HF; 20% fat, w/w), positive control (PC; HF diet + 0.2% conjugated linoleic acid, wt/wt), and fucoxanthin (HF‐Fx; HF diet + 0.2% Fx, wt/wt) and fed for 6 weeks. Fucoxanthin supplementation significantly lowered body weight and relative weights of total white adipose tissue in the HF‐Fx group compared to the HF group. HF‐Fx supplementation significantly lowered plasma and hepatic lipid levels compared to the PC and HF groups. The accumulation of hepatic lipid droplets and the epididymal white adipocyte size were less in the HF‐Fx and PC groups than in the HF group. Hepatic phosphatidate phosphohydrolase, malic enzyme, fatty acid synthase, and glucose‐6‐phosphate dehydrogenase activities were significantly lower in the HF‐Fx group compared to the HF group. Consequently, the fucoxathin supplementation with a high‐fat diet seemed to be effective for suppressing body weight gain and for improving plasma and hepatic lipid metabolism in mice.
The Effect of Spice Extracts on Caspase‐3 Activity as an Indicator of Apoptosis of Human HL‐60 Cells
Authors: Meadors Joanna L. , Nantz Meri P. , Percival Susan S.
Published: June 2021

Spices and herbs contain antioxidant compounds which are hypothesized to reduce the effects of oxidative stress and may, in some cases, contribute to the prevention of chronic diseases. The Oxygen Radical Absorbance Capacity assay was used to measure the antioxidant capacity of twelve common spices and results confirmed that cloves have a high antioxidant capacity while black pepper is relatively low. We asked if the antioxidant capacity was related to the ability to cause apoptosis. Caspase‐3 is the committed step in apoptosis. Human HL‐60 cells were treated with different concentrations of spice extracts for various times. Caspase‐3 activity was measured with a synthetic substrate and cell death was measured using MTT. The assays revealed that both black pepper and cloves induce caspase‐3 activity in HL‐60 cells and that cell death occurs with both spices. However, the amount of cell death does not appear to be correlated with the antioxidant capacity of the spice. These data suggest that non‐antioxidant compounds in spices may contribute to cell death or that mechanisms other than activation of caspase‐3 may be involved. Funding in part by University Scholars Program at UF.
Isoliquiritigenin inhibits JNK/AP‐1 signaling in DU145 human prostate cancer cells
Authors: Kwon Gyoo Taik , Cho Han Jin , Chung Won Yoon , Moon Aree , Park Jung Han Yoon
Published: June 2021

Isoliquiritigenin (ISL), 4,2′,4′ ‐trihydroxychalcone, found in licorice, shallot, and bean sprouts, is a potent antioxidant with anti‐carcinogenic effects. We previously observed that ISL inhibited cell migration, invasion and adhesion and both basal and EGF‐induced secretion of uroplasminogen activator (uPA), vascular endothelial growth factor (VEGF), MMP‐9 and TIMP‐1 in DU145 cells. The present study examined whether ISL inhibits phosphorylation of Akt and mitogen‐activated protein kinase (MAPK) in DU145 cells. EGF stimulated phosphorylation of extracellular signal‐regulated kinase (ERK)1/2, p38 MAPK, Jun N‐terminal kinase (JNK), c‐Jun, and Akt. Treatment of cells with ISL led to significant decreases in basal and EGF‐induced phosphorylation of JNK, c‐Jun, and Akt. However, phosphorylation of neither ERK1/2 nor p38 MAPK was altered. The JNK inhibitor SP600125 inhibited basal and EGF‐induced secretion of uPA, VEGF, MMP‐9 and TIMP‐1 as well as cell migration. Both ISL and SP600125 inhibited AP‐1 DNA binding activity. These results suggest that inhibition of JNK/AP‐1 signaling may be one of mechanisms by which ISL inhibits cell migration, invasion, and adhesion in DU145 cells.
The novel antioxidant ergothioneine found in dietary mushrooms inhibits monocyte binding to human aortic endothelial cells
Authors: Martin Keith R
Published: June 2021

Cardiovascular disease (CVD) is a leading cause of mortality in the United States. An accumulating body of evidence suggests that dietary consumption of fruits and vegetables can reduce CVD risk via a plethora of bioactive agents functioning, in part, as antioxidants since oxidative stress is considered an early feature of atherogenesis. In this study, we focused on the naturally occurring metabolite ergothioneine (ERT) found in dietary mushrooms because of its demonstrated antioxidant capacity in mammalian cells. ERT is present in edible mushrooms and is not synthesized, but is accumulated, by humans through diet. In this study, human aortic endothelial cells (HAEC) were incubated overnight (20 h) with increasing concentrations of ERT (0.01–10.0 mM) followed by incubation, after washing, with medium alone or with the pro‐inflammatory cytokine IL‐1beta (5 ng/mL) for 6 h. ERT at concentrations of 0.1–0.3 mM significantly (p < 0.05) reduced VCAM‐1, ICAM‐1, and E‐selectin adhesion molecule (AM) expression maximally by 41%. Binding of U937 human monocytes to HAEC was significantly reduced both in unstimulated monolayers and by 12‐27% in IL‐1beta‐stimulated HAEC pre‐incubated with 1 and 3 mM ERT, respectively. Viability was not different between treatments. These data provide evidence that ERT found in dietary mushrooms can protect against atherogenesis by reducing AM expression and subsequent monocyte binding.
Protective Effect of Luteolin on Inflammatory Factor‐Mediated Insulin Resistance in 3T3‐L1 Adipocytes
Authors: Ding Li , Chen Xiaoli
Published: June 2021

Obesity and insulin resistance have been recently linked to a low‐grade chronic inflammatory response characterized by increased macrophage infiltration, altered cytokine production, and activation of inflammatory signaling pathway in adipose tissue. Pharmacological agents and natural products capable of reducing inflammatory activity possess anti‐diabetic properties. Luteolin, a naturally occurring flavonoid, has been demonstrated to inhibit LPS‐induced TNFα release and activation of NF‐κB pathway in macrophages. However, little is known about the role and mechanism of luteolin on anti‐insulin resistance and diabetes. In this study, we investigated the role of luteolin in the regulation of inflammatory factor‐mediated insulin resistance in 3T3‐L1 adipocytes. Here we show that luteolin treatment for 24 hrs increased insulin‐stimulated glucose uptake in a dose‐dependent manner in 3T3‐L1 adipocytes. Our results also demonstrated that 24 hour treatment of luteolin enhanced insulin‐stimulated PPARγ expression in 3T3‐L1 adipocytes. Interestingly, time‐dependent serine phosphorylation of insulin receptor substrate 1 (IRS‐1) by TNFα was blocked in 3T3‐L1 adipocytes with pretreatment of luteolin for 30 min. Furthermore, luteolin treatment for 24 hrs attenuated LPS‐stimulated TNF‐α secretion from both 3T3‐L1 preadipocytes and adipocytes. When added to 3T3‐L1 cells together with LPS during the entire differentiation process, luteolin antagonized the effect of LPS on the induction of IL‐6 gene expression in 3T3‐L1 adipocytes. Thus, our data demonstrate that luteolin plays a protective role in inflammation‐mediated adipocyte dysfunction and insulin resistance.
Induction of the tumor suppressor protein p53 contributes to fisetin‐induced Bax translocation to mitochondria and apoptosis of HCT‐116 colon cancer cells
Authors: Lim Doyoung , Park Jung Han Yoon
Published: June 2021

Fisetin, a flavonol, has been reported to have anti‐cancer properties. In the previous study, we have demonstrated that fisetin induces apoptosis of human colon cancer cells via activation of caspase‐8 and alteration of Bcl‐2 family proteins. In the present study, we examined the role of p53 tumor suppressor protein in fisetin‐induced apoptosis of HCT‐116 cells. Treatment of HCT‐116 cells with fisetin led to a dose‐dependent increase in p53 and decrease in mdm‐2 protein levels. Reporter gene analysis using a luciferase plasmid containing direct repeats of the transcription recognition sequences for p53 indicated that p53 reporter activity was significantly increased in fisetin‐treated cells. Subcellular fractionation and Western blot analysis, and immunocytochemistry revealed that inhibition of p53 expression by small interference of RNA decreased fisetin‐induced Bax translocation to mitochondria. In addition, inhibition of p53 expression significantly reduced fisetin‐induced chromatin condensation and cleavage of poly(ADP‐ribose) polymerase. These results indicate that induction of p53 led to Bax translocation to mitochondria, which contributes to apoptosis in fisetin‐treated HCT‐116 cells.
Conjugated nonadecadienoic acid (CNA) shares similar molecular mechanisms with conjugated linoleic acid (CLA)
Authors: Park Yeonhwa , Park Yooheon , Rhee Siyeon
Published: June 2021

Previously, conjugated nonadecadienoic acid (CNA) has been shown to reduce body fat dose‐dependently and it was more effective than CLA in mice. In this study, we hypothesized that the effects of dietary CNA on expression of genes involving fat and energy metabolism are similar to CLA's. Fifty female 129Sv/J retired breeders were housed in individual wire‐bottomed cages in a windowless room with a 12‐h light‐dark cycle and fed a semi‐purified powdered diet based on the AIN‐93 diet with 20% total fat. After a week adaptation period, mice were randomly assigned into control and 4 groups of treatment (0.5% of CLA, 0.01%, 0.05%, or 0.1% of CNA as weight of diet). Peroxisome proliferator‐activated receptor‐γ (PPAR‐γ), acetyl‐CoA carboxylase (ACAC), fatty acid synthase (FAS), lipoprotein lipase (LPL), hormone‐sensitive lipase (HSL), glucose transporter 4 (GLUT4), and uncoupling protein‐2 (UCP‐2) from white adipose tissue, and PPAR‐α from the liver were quantified by real‐time quantitative PCR. The mRNA expressions of FAS, ACAC, UCP‐2, and PPAR‐γ in the adipose tissue were decreased by CLA as well as CNA. Meanwhile HSL expression was enhanced by CNA but not by CLA. Hepatic PPAR‐α mRNA expression was increased by both CNA and CLA. Overall, the effects of 0.1% dietary CNA on expression of genes involving fat and energy metabolism were similar to that of 0.5% dietary CLA. This supports our hypothesis that dietary CNA has an effect similar to CLA on lipid metabolism at the molecular level, however, differences on lipolysis may help explain the improved efficacy of CNA on body fat reduction compared to CLA. Supported by AHA SDG and ILSI North America
Effect of whey protein on glucose tolerance and insulin sensitivity in Zucker Diabetic rats
Authors: Janle Elsa Marie , Lachcik Pamela
Published: June 2021

There is some evidence that whey protein, may increase insulin sensitivity and decrease the risk of, or modulate the severity of type 2 diabetes. To test the hypothesis that a high whey protein diet would be more beneficial for type 2 diabetics than a high soy protein diet, Zucker Diabetic rats were placed on test diets at 5 weeks of age. Rats were fed test diets of either high soy protein or high whey protein for 8 weeks. There were no significant differences in food consumption or weight gain between groups. Body composition was analyzed by DXA and whey fed rats had a greater percent body fat than soy fed rats (p = 0.035). At the end of 8 weeks on test diets the rats were implanted with ultrafiltrate probes and jugular catheters. The average daily glucose measured in interstitial fluid samples obtained with the ultrafiltrate probe showed at trend toward lower glucose in the whey fed rats (p = 0.07). Glucose tolerance tests were performed and glucose and insulin were measured. Glucose values measured in blood were significantly lower (p < .05) at baseline, Cmax and at 3 hours. The area under the curve (AUC) showed a trend toward lower glucose for whey fed rats (p = 0.059). The AUC for ISF was significantly lower for whey fed rats than soy fed rats. There were no significant differences in insulin concentrations or insulin sensitivity. Conclusion: Whey protein provided some improvement in glucose tolerance. Support: National Dairy Council.
Hypoglycemic effect of chlorella in Goto‐Kakizaki and Wistar rats
Authors: Kim You Jin , Jeong Hyejin , Kim Mi Kyung
Published: June 2021

The aim of this study was to examine the hypoglycemic effect of chlorella in 6 week‐old type 2 diabetic Goto‐Kakizaki (GK, n=30) and 6 week‐old normal Wistar (n=30) rats. Animals were fed different three experimental diets containing 0%, 3% or 5% (w/w) chlorella for 8 wks. Blood HbA1c and fasting plasma glucagon concentrations were measured. And HOMA‐indexes were calculated using fasting blood glucose and plasma insulin concentrations.

In GK rats, fasting plasma glucagon levels in 3% and 5% chlorella groups were significantly lower than control group, and levels of blood HbA1C, fasting glucose, plasma insulin and HOMA‐indexes in 3% and 5% chlorella groups were tended to be lower than control group.

In Wistar rats, blood HbA1C, plasma insulin levels and HOMA‐indexes of 5% chlorella group were tend to be higher, and fasting plasma glucagon levels of 5% chlorella group were significantly higher, than control and 3% chlorella groups.

In conclusion, the feeding of chlorella was beneficial to improve insulin resistance in type 2 diabetic GK rats and to maintain blood glucose level in normal Wistar rats.

*This work was supported by the grants from Daesang.Co.Ltd. and the second stage of Brain Korea 21 project in 2007.
Strawberry modulates oxidative stress markers in response to high fat meal in overweight men and women
Authors: Linares Amanda , Hyson Dianne , Kappagoda Tissa , Burton‐Freeman Britt
Published: June 2021

Our previous work indicates that consuming a highly purified polyphenolic‐rich grape seed extract decreases LDL oxidation. Oxidative stress markers have been reported to increase after consumption of high fat meals. We hypothesized that strawberry; a polyphenolic rich fruit, would protect LDL from oxidation after a high fat meal (HFM). A randomized, double‐blind, placebo‐controlled, 12‐wk cross‐over trial was conducted in overweight men and women (n=24). After a 10 day run‐in, subjects consumed either a Strawberry (Str) or placebo (Pbo) beverage matched in energy and macronutrient composition for 6 wks. At defined intervals for 6 h, postprandial oxidation of LDL, plasma lipids and F2 isoprostanes were measured before and after HFM challenge with Str or Pbo during run‐in (i.e. low Str intake) and again after 6 wks of daily Str or Pbo consumption. During run‐in, the single Str exposure significantly reduced LDL oxidation (p<0.001). After 6 wks Str or Pbo and HFM re‐challenge, Str attenuated the oxidative response to the high fat meal compared to Pbo. The postprandial lipid profile was generally not affected by treatment, except for a modest reduction in postprandial triglyceride concentrations (p=0.06). The increasing relevance of oxidized LDL to vascular injury suggests a potentially important protective role of strawberry in reducing CVD risk. Funding: CA Strawberry Commission
Fermented soybeans by Monascus starter change fatty acid composition of blood and liver in diet‐induced obese rats
Authors: Kim Sun‐Hee , Chang Moon‐Jeong , Yoo Hyun‐Wook
Published: June 2021

Soybeans that are consumed a lot in Asia contain essential fatty acid, linoleic acid, and have a preventive effect of cardiovascular disease. Isoflavones, main bioactive materials of soybean, are found in glycosidic form but fermented isoflavones are converted to aglycone form and increased bioavailability. The Monascus fermented products forms the red mycelium and bioactive monacolin K. This study, therefore, was performed to investigate the effects of Monascus starter on fatty acid composition of blood and liver. Sprague‐Dawley weanling male rats were provided either control low fat (10% fat, LF) or high fat (45% fat, HF) for inducing obesity for 9 weeks and, then were provided three different diets containing casein, steamed soybeans, and Monascus starter treated soybeans for 5 weeks. Monascus starter significantly decreased body weight gain and increased fecal fat excretion. Monascus showed a significant increase of plasma unsaturated fatty acids in HF, and not in LF. There was no difference in fatty acid composition of liver in LF groups. However, Monascus increased unsaturated fatty acids and showed relatively higher ratio of polyunsaturated fatty acids to saturated fatty acids among HF groups, The above results confirm that Monascus fermented soybean products prevent obesity and cardiovascular disease by improving lipid metabolism. 
Anti‐adipogenic compounds isolated from fermented wheat grains
Authors: Park Kun‐Young , Ahn In‐Sook , Nam Sang‐Hae , Do Myoung‐Sool
Published: June 2021

Previously, we studied that fermentation improved anti‐adipogenic activity of wheat grains. The wheat grains fermented with microorganisms for over 40 days decreased body and adipose tissue weights in high‐fat diet induced rats. It also decreased fat accumulation in 3T3‐L1 adipocytes. In this study, we isolated anti‐adipogenic compounds from the fermented wheat grains and evaluated the anti‐adipogenic activites. To do this, anti‐adipogenic compounds was isolated from ethyl acetate fraction of the fermented wheat grains. Fourteen compounds were isolated by silica‐gel column chromatography, prep TLC, and TLC. Six out of 14 compounds were appeared to have strong anti‐adipogenic effects by measurement of leptin secretion level and OD value of Oil red O stained in 3T3‐L1 adipocytes. Two out of 6 compounds were identified to be 1,2,3‐propanetriol‐1‐acetate (K1: glycerol derivative) and 1,2,3‐propanetriol (K2: glycerol) on the basis of 1H‐NMR and 13C‐NMR analysis. K1 decreased leptin secretion by 30%, 45%, 50%, respectively, when it was treated at the concentrations of 10μg/ml, 50μg/ml, and 100μg/ml. K2 also decreased leptin secretion by 10%, 30%, 40%, respectively. K1 and K2 decreased mRNA expression of adipogenesis‐related genes including PPARγ, SREBP‐1c, FAS, and ACS by real time RT‐PCR. Taken together, 1,2,3‐propanetriol‐1‐acetate and glycerol isolated from fermented wheat grains decrease the fat accumulation in 3T3‐L1 adipocytes by inhibiting adipogenesis.
Declines of proliferation predominance of Hirsutella sinensis associate with unparallel alterations of its DNA copies during maturation of Cordyceps sinensis
Authors: Yao Yisang , Zhou Yanjiao , Zhu Jia‐Shi
Published: June 2021

Cordyceps sinensis (Cs) is a precious tonic herb used in China for centuries. We reported simultaneous detection of 2 live fungi, Hirsutella sinensis (Hs) and Paecilomyces hepiali (Ph), and their DNA in fresh Cs (FASEB J 19:1033, 2005; 21:A1079, 2007). A companion study (#1476) indicated a 50% decline of competitive proliferation capability of Hs during Cs maturation. We further examined an association of such a predominance decline with possible alterations of Hs gene copies. Hs and Ph ITS sequences were extended up‐ and downstream to their partial 18S or 28S DNA for designing and producing internal control probes for Southern analysis, with use of 18S‐ and 28S‐specific primers. With use of Hs and Ph specific probes, Southern blotting was performed to examine the gene copies of Hs and Ph in Cs of 2 different maturation stages, less mature Cs (T1) collected in early May and mature Cs (T2) in late June. Southern blotting demonstrated slightly altered amounts of Hs rDNA in mature Cs (caterpillar bodies and stromata) collected in late June, as compared to the less mature counterparts. Our data suggest paradoxical dissociation of the copies of Hs rDNA and competitive proliferation capability of Hs (50% drop) during Cs maturation, indicating other factors possibly accounting for the decline of proliferation dominance of Hs.
Polyglutamine immunoreactivity is present in FTLD‐U and normal controls
Authors: Herndon Emily Susanne , Hladik Christa , Shang Ping , White Charles L.
Published: June 2021

1C2 is a monoclonal antibody raised against the N‐terminus of the human TATA‐box binding protein that contains a string of 38 glutamines. We have previously demonstrated 1C2 immunoreactivity in the substantia nigra, locus coeruleus, and globus pallidus of 2 FTLD‐Ub cases. To investigate this further, we stained sections of the midbrain, pons, and globus pallidus of 10 FTLD‐Ub cases and 2 normal controls using 1C2. In all cases 1C2 immunoreactivity was present in neuropil and in the cytoplasm of neurons, microglia, or both. 9 out of 10 FTLD‐U cases demonstrated 1C2 immunoreactivity in the substantia nigra, but only one revealed immunopositivity in the locus coeruleus. Chromogen deposition was independent of neuromelanin distribution. Granular 1C2 immunoreactivity was present within the neuropil or cytoplasm of globus pallidus microglia in all cases. This strong, reproducible, but previously undescribed 1C2 immunoreactivity warrants further assessment of its binding specificity. Alternatively, expanded CAG repeats may be present in selected CNS locations in normal individuals and those without known CAG repeat diseases. In either case, caution is indicated in interpreting 1C2 immunostaining results in diagnostic and pathogenetic studies of possible CAG repeat disorder cases.

Supported in part by NIH ADC grant AG12300, the Winspear Family Center for Research on the Neuropathology of Alzheimer Disease, and the McCune Foundation.
Effects of FGF‐2 Overexpression in the Dutch/Iowa APP Transgenic Mouse
Authors: Donahue John Edward , Coffin J. Douglas , Johanson Conrad E. , Van Nostrand William E. , Vitek Michael P. , Blay Eddie , Miller Miles C. , Monfils Paul , McMillan Paul N. , Stopa Edward G.
Published: June 2021

Intracerebroventricular‐injected fibroblast growth factor (FGF)‐2 causes cerebrospinal fluid (CSF) stagnation and a normal‐pressure hydrocephalus (NPH)‐like syndrome in rats. NPH may be part of a pathophysiologic continuum culminating in Alzheimer disease (AD), at least in the context of amyloid‐β (Aβ) retention and plaque formation. We have examined transgenic mice with variable degrees of FGF‐2 and Aβ generation within the CNS and have active breeding pairs of TgFGF2xTgAPP (APP = amyloid precursor protein) to provide TgFGF2, TgAPP, and TgFGF2xTgAPP offspring. A coronal section was made through fixed cerebral hemispheres, with the anterior commissure and posterior limit of the optic chiasm used as sectioning landmarks. Serial 10 μm coronal sections were cut from the caudal portion of the hemispheres using a freezing microtome. A striking finding was the presence of flattened and denuded ependyma in the APPxFGF2 overexpressor mice, indicative of early hydrocephalus, and increased Aβ1–42 neuronal expression in these mice. Similar changes were not observed in APP control mice. This strongly suggests that growth factor overexpression will accelerate the development of Aβ production and AD pathology.
Neuropathological characterization of early onset Alzheimer disease associated with the V717L APP mutation
Authors: Spina Salvatore , Murrell Jill R , Hake Ann M , Epperson Francine , Vidal Ruben , Farlow Martin R , Ghetti Bernardino
Published: June 2021

Objective: To report the neuropathological findings associated with the V717L mutation in the Amyloid Precursor Protein (APP) gene. Methods: The proband presented with short‐term memory loss at age 35 and died at age 44. His brain was processed for histology and immunohistochemistry (IHC). Aβ IHC was carried out using antibodies recognizing Aβ3–7, AβN3pE, Aβ10–16, Aβ17–24, AβN‐40 and AβN‐42. Results: Neuritic plaques (CERAD stage C) were abundant in the intermediate neocortical layers. Abundant diffuse Aβ deposits were observed in the remaining cortical layers as well as in the subpial zone. Mild amyloid angiopathy was seen, predominantly in penetrating vessels. Braak VI‐tau pathology was observed, particularly affecting layer V and VI. Neuritic plaques were immunoreactive against the full panel of Aβ antibodies except for the anti‐AβN‐40. Diffuse deposits contained AβN‐42 species but did not react with antibodies recognizing epitopes located before position 11. Conclusions: Our data suggest a heterogeneous involvement of the cortical layers by the deposition of Aβ species with specific immunohistochemical pattern. They also suggest that diffuse deposits, in the upper and lower cortical layers, are composed by N‐terminally truncated Aβ species. Supported by AG10133 and Alzheimer's Association.
Neuromuscular presentation of severe neonatal onset glycogenosis type IV
Authors: Rivera‐Zengotita Marie , Murdock David , Beaudet A , Bhattacharjee Meena B.
Published: June 2021

Glycogen storage disorder type IV (GSD IV) or Andersen disease is a rare autosomal recessive disorder resulting from deficiency of glycogen branching enzyme (GBE). It is very heterogeneous in its clinical features, age of onset, and tissue involvement; classic GSD IV is rapidly progressive and dominated by cirrhosis and hepatic failure. We report a newborn male with severe hypotonia, poor respiratory effort, and elevated CK. The differential included spinal muscular atrophy, congenital muscular dystrophy/myopathy, hypoxia‐ischemia, and chromosomal disorder. Muscle biopsy at 3 weeks of life was remarkable for sarcoplasmic vacuoles with crystalline/granular, diastase‐resistant PAS positive material, and markedly increased acid phosphatase reaction. Ultrastructural examination showed two types of sarcoplasmic inclusions: membrane‐bound with granular and filamentous material, and non‐membrane bound with filamentous to crystalline material. These findings suggested a lysosomal storage disorder other than GSD II (Pompe's disease). Review of literature suggested that clinical and pathological findings might be consistent with the fatal infantile form of GSD IV. Biochemical testing showed markedly decreased muscle GBE activity, consistent with GSD IV. The rare neuromuscular form of GSD IV should be considered in the differential diagnosis of severe congenital, or neonatal/infantile hypotonia.
Evaluation of a Novel Vibration Perception Threshold Instrument
Authors: May Kelsi J. , Latman Neal S.
Published: June 2021

Monitoring peripheral nerve pathology is critical in order to prevent loss of limb or even loss of life. Semmes‐Weinstein (SW) Monofilaments have been the gold standard in non‐invasive peripheral nerve sensitivity monitoring for over forty years. Vibration perception threshold (VPT) instruments have recently been developed as an alternative method of testing peripheral sensitivity, but their measurements exhibit a low correlation with results obtained using the SW monofilaments. One possible reason for this low correlation may be that the current VPT instruments stimulate different receptors than the SW Monofilaments. The purpose of this study was to determine whether a novel VPT instrument, designed to stimulate the same receptors as the SW monofilaments, provides a higher correlation.

The two instruments used in this study were the standard SW monofilaments and the novel VPT instrument. Each subject was tested for peripheral sensitivity on the fingertips of their non‐dominant hand using both instruments. The 98 subjects tested ranged in age from 22 to 88 years, with 20% male and 80% female.

When analyzed, the SW measurements and novel VPT measurements showed a correlation coefficient of 0.67, significantly greater than the current VPT instruments’ typical correlation of 0.39. The reliability of the VPT measurements demonstrated low intra‐user variability, with an average mean deviation of only 0.3 mV.

Therefore, this study indicated that the novel VPT Instrument provides measurements of peripheral nerve sensitivity that have a stronger correlation with SW monofilaments than currently available VPT instruments.
Coexposure of mice to trovafloxacin and lipopolysaccharide results in a unique gene expression profile and liver injury dependent on early growth response‐1 transcription factor activation
Authors: Shaw Patrick , Ditewig Amy , Waring Jeff , Liguori Michael , Blomme Eric , Copple Bryan , Ganey Patricia , Roth Robert
Published: June 2021

Trovafloxacin (TVX), a fluoroquinolone antibiotic, has been linked to several cases of severe idiosyncratic hepatotoxicity in people. Previously, we showed that a modest inflammatory stress induced by lipopolysaccharide (LPS) renders mice sensitive to nonhepatotoxic doses of TVX, producing midzonal hepatic coagulative necrosis and increased plasma ALT activity. Hierarchical cluster analysis and principal component analysis of hepatic gene expression data from mice taken at 3 h after LPS, a time before the onset of liver injury, revealed treatment‐related clustering. One gene identified from gene expression analysis was the transcription factor early growth response‐1 (egr‐1). Egr‐1 mRNA was selectively increased in TVX/LPS‐treated mice at 3 h. In addition, the nuclear accumulation of egr was selectively increased in the livers of TVX/LPS‐treated mice at 3 h. The selective increase in egr‐1 was corroborated by the selective increase of two specific egr‐1 activation products, nab2 and dscr‐1. In addition, the expression of tumor necrosis factor α and plasminogen activator inhibitor 1, both of which are expressed upon egr‐1 activation and involved in the development of TVX/LPS‐induced liver injury, were selectively increased in TVX/LPS‐treated mice (Supported by NIH grants DK061315 and GM075865.)
The identification of μ opioid receptors on colonic circular smooth muscle cells
Authors: Hawkins Edward G , Ross Gracious R , Gabra Bichoy H , Dewey William L , Akbarali Hamid I
Published: June 2021

Morphine‐induced constipation severely hampers the clinical utility of this excellent pain reliever. Studies of the chronic effects of morphine on gastrointestinal motility have mainly focused on inhibition of both excitatory and inhibitory motor neurons in the myenteric plexus of the ileum. In the present study we examined the direct effects of morphine on colon smooth muscle. Isometric force was measured from colonic circular muscle ring preparations in an organ bath. Morphine (10nM – 1 μM) produced both tonic and rhythmic phasic contractions in the presence of atropine (1 μM). Tolerance did not develop to repeated exposure to morphine. The expression of μreceptor (MOR) was determined by immunohistochemistry using anti‐MOR antibody. In cross‐sections of the colonic wall, MOR was detected in both enteric ganglia and circular muscle. MOR expression was confirmed in enzymatically‐isolated single smooth muscle cells. The specificity of the MOR antibody was further verified in MOR‐expressing CHO cells. These data demonstrate the expression of MOR on smooth muscle cells and suggest that part of morphine's effect on gastrointestinal motility may be due to direct stimulation of circular smooth muscle. In addition to its well known effects on neuronal function, morphine‐induced enhanced non‐propulsive segmental contractions are important in the genesis of constipation. Supported by DK46367, DA01647.
Effects of early life stress on stimulant self‐administration in rhesus monkeys
Authors: Ewing Sarah B. , Howell Leonard L.
Published: June 2021

Early life stress can have effects on behavior and stress reactivity which are linked to sensitivity to stimulants in rodents. The present study investigated whether rhesus monkeys that experienced early life stress (n=4) would show altered sensitivity to the reinforcing effects of stimulants as compared to controls (n=3). Experimental monkeys experienced periodic maternal separations between 3–6 months of age and exhibited a phenotype of altered HPA axis function and hyper‐responsivity to stress, predictive of enhanced propensity to stimulant use. Monkeys were trained initially to self‐administer cocaine (0.1mg/kg/inj) under a fixed ratio (FR) 1 schedule and parameters were increased once all 10 injections were taken for 3 consecutive sessions. Ultimately the schedule was a second order schedule of FR 20 components with drug injection following the first ratio completed after 10 min elapsed (FI 600). Dose‐effect curves were determined for cocaine (0.01–1.0 mg/kg/inj) and amphetamine (0.003–0.3 mg/kg/inj) with each dose tested until response rates were stable over 5 consecutive sessions. As compared to control animals, experimental animals showed reduced behavioral output during the acquisition of self‐administration and in the dose‐response curves for both stimulants. The results do not provide support for early life stress leading to enhanced vulnerability to stimulant use. (Supported by USPHS grants MH58922, DA10344, DA00517 and RR00165.)
N,N‐dialkyltryptamines inhibit plasma membrane and vesicular serotonin transport
Authors: Cozzi Nicholas , Gopalakrishnan Anupama , Anderson Lyndsey , Feih Joel , Ruoho Arnold
Published: June 2021

The plant hallucinogen N,N‐dimethyltryptamine (DMT) has been used for religious and other purposes for many centuries. Over the past few decades, DMT has been reported in human brain, and pharmacological studies have shown that DMT binds to monoamine receptors. Recently, several N,N‐dialkyltryptamines were reported to modify monoamine uptake and release in rat brain synaptosomes, indicating that these compounds also affect monoamine transporters. To further clarify the mechanism of action of hallucinogenic tryptamines, we synthesized DMT and several other N,N‐dialkyltryptamines and tested them for their abilities to inhibit [3H]5‐HT uptake via the plasma membrane serotonin transporter (SERT) and via the vesicle monoamine transporter (VMAT2). SERT transport was assayed in human platelets and VMAT2 uptake was assayed in Sf9 cells infected with a recombinant baculovirus expressing rat VMAT2. The compounds were also tested as inhibitors of [3H]dihydrotetrabenazine (TBZOH) binding. Our results show that DMT and other N,N‐dialkyltryptamines inhibit [3H]5‐HT transport at SERT and VMAT2 at low micromolar concentrations. The Ki for TBZOH binding/Ki 5‐HT uptake ratios suggest that these compounds may act as competitive substrates for VMAT2.