bluuebunny/crossref_metadata_2025 huggingface dataset:
Dietary Fat Quality and Lipophilic Index among Mexican‐American Adults with and without Metabolic Syndrome
Authors: Arias‐Gastelum Mayra , Lichtenstein Alice , Matthan Nirupa , Vega‐Lopez Sonia
Published: June 2021

Objective

To assess dietary and red blood cell (RBC) fatty acids (FA) and estimate the Lipophilic Index (LI), a novel index of FA fluidity, among Mexican‐American adults with and without Metabolic Syndrome (MS).

Methods

Fasting blood samples were collected from 75 Mexican‐American adults (65% female; 29% with MS; 38±9 years; BMI=29±5 kg/m2) for the measurement of plasma glucose, serum lipids and RBC FA profiles. Dietary intake was estimated using the Southwest Food Frequency Questionnaire and MS status was determined using NCEP ATP III criteria. LI was calculated as the sum of the products of the proportion of FA and melting points (°C) of each FA, divided by the sum of all FA.

Results

The LI was positively correlated with serum triglyceride concentrations in all subjects, regardless of MS status (r=0.263, p=0.023). Participants reported consuming 29.8±19.2 g/d, 35.2±22.2 g/d, 17.5±11.6 g/d and 1.5±1.2 g/d of saturated, monounsaturated, polyunsaturated, and trans FA, respectively. In contrast, FA levels in RBC were 42.5±2.3 mol %; 18.7±2.3 mol %; 38.0±2.6 mol % and 1.4±0.4 mol % for saturated, monounsaturated, polyunsaturated, and trans FA, respectively. The LI was significantly higher among MS participants relative to those without MS (25.5±1.2 vs 24.7±1.0, p=0.007). There were no significant differences in dietary or RBC FA by MS status.

Conclusion

Participants with MS had a greater LI but further studies with larger samples are required to assess whether LI is a better biomarker than either dietary or RBC FA, for CVD risk prediction in this population.
Long‐term Health Related Quality of Life Outcomes of Patients who have undergone Bariatric Surgery with the Biliary Pancreatic Diversion with Duodenal Switch
Authors: Nirmalraj Maya , Khandalavala Birgit
Published: June 2021

Background

Of the different procedures used for bariatric surgery, BPD/DS is the most uncommon and has been associated with the longest durable weight loss with significant improvement in obesity‐related comorbidities. Improvement in health‐related quality of life (HRQoL) is of paramount importance and an essential metric to evaluate additional outcomes. Previous studies have shown significant improvement following BPD/DS for up to 5 years after surgery.

Methods: We studied HRQoL in 27 patients who have had BPD/DS at an academic medical center over a period of 19 years. This study had utilized the validated SF‐36 survey and concluded that for most patients, surgery improved their HRQoL.

Results: Of the 27 respondents who had BPD/DS surgery, 77.8% were female, and 96.3% were White.

Sex
#
%

Female
21
77.8

Male
6
22.2

Race

White
26
96.3

Black
1
3.7

Mean
SD

Age at Surgery
44.33
8.9

Pre‐Operative BMI
48.89
11.5

Time Lapse Since Surgery (Years)
12.3
0.6

Weight Loss (Kilograms)
41.90
30.4

Their average age at the time of surgery was 44.3 years (SD=8.9), their average pre‐surgery BMI was 48.9 (SD=11.5) and their average time since surgery was 12.3 years (SD=0.6). Mean SF‐36 scores are shown in Table 2.

Table 2: SF‐36 Scores
Mean
SD

Physical Component Score (PCS)
47.677
12.5408

Mental Component Scores (MCS)
48.308
12.0977

General Health
63.52
27.694

Physical Functioning
71.67
28.723

Role limitations due to physical health
62.04
43.506

Energy/Fatigue
53.08
28.812

Pain
72.5
24.789

Emotional well‐being
72.31
20.612

Role limitations due to emotional health
70.37
42.701

Social Functioning
79.81
20.334

Conclusion

In summary, our study was comprised of patients in the US who had undergone BPD/DS. Both, their physical and mental scores were close to the population norm.
Pregnancy Body Weight and Neonate Gut Microbiota
Authors: Mueller Noel , Goldani Helena , Pizoni Aline , Matte Ursula , Werlang Isabel , Goldani Marcelo , Shin Hakdong , Dominguez‐Bello Maria
Published: June 2021

Background

Babies born to overweight/obese (OWOB) moms have elevated obesity risk. A study using culture‐based methods suggested this may be due to neonates' acquisition of obese bacteria from their moms.

Aims

We used high‐throughput sequencing to examine if maternal pregravid BMI and pregnancy weight gain are associated with neonate gut microbiota.

Methods

We obtained the 1st (meconium) and 2nd stool from 62 elective C‐section (CS; n=26 OWOB) and 21 vaginally delivered (VD; n=5 OWOB) babies. We consented mothers at a hospital in Porto Alegre, Brazil. Bacterial V4 16S rRNA region was sequenced with the Illumina platform. We performed metagenomic analyses using QIIME and used the linear discriminant analysis (LDA) effect size method, which integrates statistical significance and biologic relevance, to compare microbiota of neonates from moms with pregravid OWOB vs normal BMI, and excess vs normal pregnancy weight gain, by delivery mode.

Results

In the meconium of CS and VD babies, microbial composition did not vary by pregravid BMI or pregnancy weight gain. The 2nd stool of VD babies from OWOB moms, however, had greater abundance of 11 phylotypes, including the Bacteroides genus, and lower abundance of 23 phylotypes compared to the 2nd stool of VD babies from normal BMI moms.

image

In contrast, only 3 phylotypes differed in the 2nd stool of CS babies from OWOB vs normal BMI moms. Results were similar for pregnancy weight gain and neonate gut microbiota according to delivery mode.

Conclusions

Pregnancy weight is associated with microbiota from the 2nd (but not 1st) stool of VD (but not CS) neonates, indicating maternal‐offspring transfer of obese bacteria may occur at birth.
Effects of Obesity on Serum Cations using Obese Zucker Rat Model
Authors: Hakkak Reza , Korourian Soheila , Jagetia Saagar , Romani Andrea
Published: June 2021

Obesity has been epidemic in the United States for more than two decades. Obesity is associated with type 2 diabetes, cardiovascular disease and certain cancers. We investigated the effect of obesity on serum cations content. Five‐week‐old female lean and obese Zucker rats were used. After one week of acclimation, Lean and Obese female rats were maintained on AIN‐93 G diet for 155 days. At the end of the experiment, serums were collected, and different cations were measured by atomic absorbance spectrophotometry in addition to proteins. Obese rats gained significantly (P<0.001) more weight than lean rats. Obese rats showed increased serum protein (P<0.05) and Ca2+ levels (p<0.01), and decreased serum K+ (P<0.05) and Mg2+ (p<0.05) levels compared to lean rats. Consequently, Mg2+/Ca2+ ratio decreased as a result of obesity. No significant differences in Na+ levels were observed. Our results indicate the presence of detectable changes in total protein and cation content in obese animals vs. lean animals. Whether similar changes occur in adipose tissue and other organs affected by obesity remains to be determine [Supported by NIAAA11593 to AMPR and ABI to RH]
Monounsaturated Fatty Acid High‐Fat Diets Impede Adipose IL‐1β Secretion and Insulin Resistance
Authors: Roche H , Finucane O , Lyons C , Murphy A , Reynolds C , Healy N , Tierney A , Morine M , Alcala‐Diaz J , Lopez‐Miranda J , O'Neill L , McGillicuddy F
Published: June 2021

Mice were placed on chow, MUFA or SFA‐HFD or switched to a MUFA‐HFD following SFA‐induced insulin resistance (IR). Glucose homeostasis was assessed. Stromal vascular fraction (SVF) was cultured ±ATP. Macrophages were LPS (10ng/ml:3h) primed, cultured ±oleic acid (OA;250µm)/palmitic acid (PA;250µm:24h), ±ATP(5mM:1h). Media was harvested to measure IL‐1β secretion. Protein AKT, AMPKα1, pro‐IL‐β were determined by immunoblot analysis. Implications of dietary fat on insulin sensitivity were examined in a sub‐cohort from the CORDIOPREV and LIPGENE studies.

MUFA‐HFD fed mice displayed improved IR coincident with reduced pro and active IL‐1β, with sustained adipose pAMPK activation. We further demonstrate that monounsaturated fatty acid; oleic acid, can impede ATP‐induced IL‐1β secretion from LPS primed cells in an AMPK dependent manner, in vitro. MUFA‐HFD mice display hyperplastic adipose, with enhanced adipogenic potential of SVF and improved insulin sensitivity. Switching from SFA to MUFA‐HFD failed to reverse IR but improved hyperinsulinemia. This effect is recapitulated in man wherein high‐SFA consumers, but not high‐MUFA, display reduced insulin sensitivity according to habitual SFA:MUFA intake.

These novel findings suggest that dietary MUFA can attenuate IL‐1β mediated IR and adipose dysfunction despite obesity via preservation of AMPK activity.
Lycopene Supplementation and Oxidation: a Metaregression Analysis of Lag Time to Absorption
Authors: Minto Clara , Vecchio Maria Gabriella , Gregori Dario
Published: June 2021

Background

LDL oxidation plays an important role in the pathogenesis of CVD and carotenoids could be related to decreased susceptibility of this oxidation.Several authors have suggested that Lycopene supplementation may have beneficial effects on LDL oxidation.This study evaluates the association between Lycopene addition and mean of lag time oxidation through a meta‐analysis of 3 intervention trials

Methods

Relevant articles published until September 2014 were collected using electronic database PubMed.The search string was “vitamins”, limited to “clinical trials” and “humans”.18091 records were retrieved.Out of them, studies with detailed information of dose of Lycopene, Lag Time and Duration were retrained for the analysis.A metaregression model has been estimated using Generalized Least Squares, accounting for within‐study correlation (assuming a Continuous AutoRegressive of first order structure) and weghts proportional to study variance and study size. Effect of dose has been estimated as Cumulative Effect (Dose per day over the weeks of study duration)

Results

90 patients were included in the analysis,receiving different doses of Lycopene.

Study
Population
N
Duration
Dose
Lag Time (min) mean ± SD

Hininger:2013
Males non smokers
51
12 weeks
15 mg/d LP
Week 0
Week 12

healthy

106± 7,37
95,9±7,53

Bub:2000
Males non smokers
23
2 weeks
40 mg/d LP
0‐2 week
2‐4 week

healthy

106±13,4
124±9,9

Carrol:2000
males and females
16
12 weeks
13,3 mg/d LP
baseline
after 12 weeks

healthy

54±20
62±22

Susceptibility of LDL to metal ions‐mediated oxidation in vitro was assessed by measuring conjugated diene formation at 234 nm.A significant dose‐response relationship has been found between cumulative dosage of Lycopene and Lag Time(effect 0.01085,S.E. 0.000908,p<0.001)

Conclusions

Data suggest that a high‐dose of Lycopene decreases lag time.This result support the assumption that Lycopene supplementation could be protective from diseases related to LDL oxidation
Evaluating Extraction Protocols for Quantifying Hepatic Retinol and Retinyl Esters
Authors: You Hong , White Wendy
Published: June 2021

Accurate quantification of hepatic retinoids is a key step in determination of vitamin A status in animal models. Our objective was to compare published non‐saponification extraction protocols for HPLC analysis of hepatic retinol and retinyl esters. Based on this comparison, we then investigated the effects of: 1) homogenizing rat liver in ethanol versus aqueous buffer; and 2) adding water to facilitate phase separation. The extraction protocol of Tan et al. (J Lipid Res 2014;55:1077‐86) resulted in higher hepatic retinoid concentrations (96.9 ± 1.8 μg retinol equivalents/g) than the methods of Poulaert et al. (Food Chem 2014;159:477‐85) (28.0 ± 2.5 μg/g) and Kim et al. (Methods Mol Biol 2010;652:263‐75) (16.2 ± 1.7 μg/g) (P < 0.0001). The corresponding recovery of spiked retinyl acetate was also higher: 97.9 ± 1.8% versus 66.7 ± 2.1% and 53.8 ± 3.5%, respectively (P < 0.0001). Using the method of Tan et al., lower hepatic retinol equivalent concentrations were obtained when: 1) liver was homogenized in phosphate buffered saline (PBS) (37.5 ± 4.3 μg/g) instead of ethanol (91.6 ± 1.3 μg/g) (P < 0.0001); or 2) water was added in an effort to facilitate phase separation (66.3 ± 0.7 μg/g) (P < 0.0001). In conclusion, homogenizing liver tissue in ethanol instead of aqueous buffers and avoiding the subsequent addition of water result in substantially increased extraction efficiency.
Production of Uniformly Labeled 13C‐Lutein and 13C‐α‐Tocopherol in vitro Using Carrot Cell Suspension Culture
Authors: Smith Joshua , Rogers Randy , Jeon Sookyoung , Rubakhin Stanislav , Wang Lin , Sweedler Jonathan , Kuchan Matthew , Erdman John
Published: June 2021

The xanthophyll lutein (Lut) is present in high concentrations in primate retinae, protecting against photooxidation and enhancing visual performance. Interestingly, in the elderly, diets high in Lut slow cognitive decline and circulating levels of both Lut and the lipid‐soluble antioxidant α‐tocopherol (αT) are inversely associated with cognitive impairment. However, Lut deposition, metabolism, and function in the brain are poorly understood, and cost‐effective isotopically labeled Lut tracers are not commercially available. The goal of this work, therefore, is to establish a system for efficient bioproduction of uniformly labeled 13C‐Lut and 13C‐αT. Seven carrot (Daucus carota) cultivars were germinated, separated into tissue subtypes, dedifferentiated into solid callus, and initiated in liquid suspension culture. Conditions for production of Lut and αT in suspension culture were optimized and uniformly labeled 13C‐glucose was provided as the sole media carbon source for four serial growth cycles. Lut and αT yields were 2.6 and 3.3 μg/g cells, respectively, as measured by reverse‐phase HPLC. Cell extracts were fractionated by preparatory reverse‐phase HPLC and analyzed for 13C enrichment by mass spectrometry. 58% of Lut and 50% of αT in carrot extract fractions were uniformly labeled with 13C. These tracers can be utilized in studies of Lut and αT in the brain, enhancing our understanding of their function in cognition.
A modified isotope dilution equation predicts vitamin A total body stores in individuals
Authors: Lietz Georg , Oxley Anthony , Ford Jennifer , Green Michael
Published: June 2021

Isotope dilution is used to successfully predict vitamin A (VA) total body stores (TBS) in populations but does not typically work for individuals. Using plasma kinetic data from 33 healthy young adults dosed with [13C10]retinyl acetate, we determined TBS using model‐based compartmental analysis (Simulation, Analysis and Modelling software [SAAM]) vs the equation TBS = F x S x (1/SA), where F= fraction of the orally administered stable isotope dose in stores, S= ratio of specific activity of retinol in plasma to that in stores, and SA = VA specific activity in plasma at 3 d post dosing. A high correlation (R=0.92) was found for the population. We found that the fractional catabolic rate (FCR) was a proxy for F and developed a mono‐exponential equation using 7 and 14 d data to predict F in individual subjects (R=0.999). Since S is ~1 at 3 d (when VA SA in stores crosses over plasma), the equation for d 3 simplifies to TBS = F x (1/SA). That is, by collecting blood samples at 3, 7 and 14 d and then calculating FCR based on 7 and 14 d data, we can accurately estimate an individual's TBS as individual F x (1/SA) at 3 d (R=0.94).

This study was supported by BBSRC.
Simulation of the Contribution of Beta‐carotene Biofortified Rice on Vitamin A Intake among Women and Children in Bangladesh, Indonesia and Philippines
Authors: De Moura Fabiana , Angeles‐Agdeppa Imelda , Muslimatun Siti , Atmarita A , Gironella Glen , Moursi Mourad , Carriquiry Alicia
Published: June 2021

Vitamin A deficiency continues to be a major public health problem affecting developing countries where people eat mostly staple foods. In Asia, rice is the main staple food providing up to 80% of the total daily energy intake. We used existing datasets from Bangladesh, Indonesia and Philippines where dietary intakes have been quantified at the individual level to 1) determine the rice and vitamin A intake among children 6‐59 months and women; 2) simulate the level of change in prevalence of inadequate intake of vitamin A based on a range of beta‐carotene (4‐20 ppm) and adoption (10‐70%) levels of the biofortified rice. PC‐Side software was used to perform the simulation analysis. Rice intake (mean ± SD, as raw fresh weight) among women and children were, respectively, 422 ± 83 and 138 ± 46 g/d in Bangladesh, 276 ± 60 and 100 ± 20 g/d in Philippines and 160 ± 125 and 95 ± 45 g/d in Indonesia. Depending on the age and physiological group the inadequate intake of vitamin A was 80‐100% in Bangladesh, 20‐80% in Indonesia, and 50‐90% in Philippines. As an example, introduction of biofortified rice at 10 ppm of beta‐carotene reduced the inadequacy of vitamin A intake by 10‐15% when 10% of the rice consumed was the biofortified rice and up to a 40‐50% decrease when a 70% adoption level was considered. Biofortification of rice with beta‐carotene could significantly improve the vitamin A intake in these countries.
Evaluation of the Pathways Involved in the in vivo Biotransformation of Tyrosol into Hydroxytyrosol
Authors: Rodríguez‐Morató Jose , Robledo Patricia , Farré Magí , De la Torre Rafael
Published: June 2021

Hydroxytyrosol [HT, 2‐(3,4‐dihydroxyphenyl)ethanol] is the principle phenolic compound found in olives and olive products and is also a product of dopamine metabolism. Preliminary data in humans suggest a higher recovery of HT after wine than expected (de la Torre et al. Eur J Nutr 2006;45:307). Further studies in healthy adults designed to assess the postprandial effects of moderate doses of red wine, showed that urinary recovery of HT was 4‐fold the expected based on the amount present in wine. The aims of this study were to: (1) assess the contribution of wine components to urinary HT and (2) identify the enzymes involved in this pathway. Male Wistar rats (n=96) were distributed in 14 groups (n=6‐7). The effect of tyrosine, tyrosol and tyramine on the urinary excretion (0‐4 h) of HT was assessed. We found that tyrosol was converted in vivo to HT. An additional experiment with rats (n=72) employed α‐methyl‐p‐tyrosine examine the involvement of tyrosine hydroxylase in this pathway. Experiments employing human liver microsomes showed that CYP450 catalyzes the conversion of tyrosol into HT. These results suggest potential mechanisms involved in the apparent excess urinary recovery of HT and contribute to the elucidation of the health beneficial effects associated with moderate red wine intake. Funded by FIS‐ISCIII‐FEDER (FIS PI14/00072), CIBEROBN (CB06/03/0028), Generalitat de Catalunya (AGAUR 2014SGR680, and Predoctoral Fellowship (FI‐DGR2012 for JRM).
Molecular Analysis of Cholestyramine Treatment in Hamster Model Identified CYP51 as Cholestyramine‐Responsive Gene
Authors: Huang Haiqiu , Xie Zhuohong , Yokoyama Wallace , Yu Liangli , Wang Thomas
Published: June 2021

Cholestyramine (CA) is a bile acid sequestrant widely used as a cholesterol‐lowering drug to treat hypercholesterolemia, one of the major risk factors for cardiovascular disease. Despite the wide usage of CA, its effect on cholesterol and lipid metabolism at molecular level and over long term remains unclear. The current study aimed to further understand CA's molecular effect on cholesterol and lipid metabolism. Male golden Syrian hamsters were fed diets containing 36 kcal% fat with or without 1% CA, or chow for 35 days. As expected, CA supplementation inhibited the increases in liver weight, VLDL, LDL, hepatic cholesterol and bile acids induced by high‐fat diet. CA significantly increased excretion of bile acids, while cholesterol excretion was unchanged. Interestingly, gene expression assay identified that cholesterol synthesis‐related Cytochrome p450, Family 51 (CYP51) but not HMG‐CoA reductase (HMG‐CoAR) mRNA expression was significantly increased in CA treated group. Cholesterol, bile acids and lipid metabolism associated gene (LDL receptor, CYP7A1, LXRα, and ABCG5/8) expressions were also significantly altered by CA. In summary, CYP51 was identified as a novel enzyme regulated by CA ingestion in hamster model. CYP51 and not HMG‐CoAR up‐regulation was employed to maintain plasma cholesterol in hamster model. CYP51 may warrant further consideration as potential target for management of hypercholesterolemia in human.
Formononetin enhances the expression of genes related to adipocyte differentiation and insulin sensitivity
Authors: Yu Seok‐Yeong , Choi Youngmin , Kwon Young‐In , Wood Richard , Kim Young‐Cheul
Published: June 2021

Astragalus membranaceus (AM) is a traditionally used medicinal herb in Asia for the treatment of diabetes. Recent studies have shown that the extracts from AM roots exert anti‐diabetic effects in animals and humans, however the major bioactive constituents and the potential mechanism by which AM improves insulin sensitivity still remains unclear. Here, we investigated the effect of formononetin, a major bioactive compound isolated from AM, on adipocyte differentiation and insulin sensitivity in C3H10T1/2 pluripotent mesenchymal stem cells. Formononetin at 1 to 20µM concentrations dose‐dependently enhanced adipocyte differentiation, consistent with increased expression of PPARγ and C/EBPα proteins. Formononetin also markedly induced the expression of PPARγ target genes such as fatty acid binding protein 4 (FABP4) and GLUT4 proteins. In addition, formononetin treatment significantly increased glucose uptake as well as the expression of insulin receptor substrate‐1 (IRS‐1), a key molecule for insulin signaling and action. Moreover, the effects of formononetin on differentiation and PPARγ expression were blunted in the presence of PPARγ antagonist (BADGE). These findings suggest that the insulin‐sensitizing effect of AM is mediated by formononetin through activation of PPARγ and insulin‐mediated signaling pathway, resulting in increased insulin sensitivity.
Microbial Metabolite Butyrate Suppresses Murine Mast Cell Activation
Authors: Zhang Hanying , Du Min , Xue Yansong , Yang Qiyuan , Zhu Meijun
Published: June 2021

Background

Mast cell is known for its roles in mediating food allergy and regulating gut inflammation. Butyrate, a major short chain fatty acid produced by gut microbiota, exerts beneficial effects on intestinal homeostasis. However, its role in modulating mast cell function remain undefined. Methods: Murine mastocytoma P815 cells (P815) or murine bone marrow derived mast cells (BMMC) were treated with sodium butyrate (NaB). The proliferation, cell cycle, apoptosis, degranulation and cytokine secretion were assessed.

Results

In P815 cells, NaB inhibited cell proliferation, increased acetylation of both histone 3 and α‐tubulin. The growth inhibition effect of NaB is probably through reducing c‐kit expression and inducing cell cycle arrest and apoptosis, which is indicated by enhanced annexin V/PI staining, caspase‐3 and PARP cleavage. In BMMC, NaB treatment dramatically suppressed FcεRI dependent TNFα and IL‐6 release without affecting degranulation. Furthermore, NaB enhanced histone 3 acetylation but diminished mRNA expression of Tnf ‐α and Il‐6, possibly through inhibiting the transcription initiation as revealed by reduced RNA polymerase II binding to the promoters of Tnf‐α and Il‐6.

Conclusions

NaB modulates murine mast cell function possibly through enhancing histone acetylation, which is of importance in developing therapeutics for diseases involving mast cells (NIH R15HD073864).
Fermented Berry Beverage Phenolics Reduced Fat Mass and Fasting Blood Glucose in High‐Fat Fed Mice
Authors: Johnson Michelle , Gonzalez de Mejia Elvira
Published: June 2021

The objective was to determine the potential of phenolic compounds from a fermented 30:70% blueberry‐blackberry beverage to reduce obesity and hyperglycemia in three week old male diet‐induced obese C57BL/6j mice. Mice (n= 12/group) were randomized into six groups to drink: an alcohol‐free fermented beverage (AFFB) containing 65.1 ± 1.6 mg cyanidin‐3‐glucoside (C3G) equivalents/L; three doses of a phenolic extract where AFFB was eluted through Amberlite XAD‐7HP and adjusted to 6.4 (0.1x), 59.6 (1x) and 192.4 (3x) mg C3G eq./L; sitagliptin (30 mg/kg BW/d) or water as positive and negative controls, respectively. After a week, mice started on a 60% fat diet (HFD). Daily consumption of anthocyanins was 8.4, 1.1, 9.0, and 19.0 mg C3G/kg BW/d for AFFB, 0.1x, 1x, and 3x, respectively, with anthocyanins present predominantly C3G (65.8%). The BW/food intake ratio and sucrose consumption were similar (p > 0.05). Mice in the 3x group had the lowest body weight gain (10.3 ± 0.3 g) and the water group the most weight gain (16.7 ± 1.2 g, p < 0.05). The weight increase was attributed to a gain in fat mass (18.0 ± 0.8 % BW for 3x vs. 31.3 ± 1.1 % BW for water) which was correlated with visceral fat (0.8 ± 0.1 g for 3x vs. 2.4 ± 0.2 g for water), while absolute lean mass did not differ (p > 0.05). After 3 weeks of treatment, all groups had fasting blood glucose (FBG) above 126 mg/dL. At the end of the study (week 12), FBG for 1x and 3x groups was significantly lower (168 and 184 mg/dL, respectively) than the water (222 mg/dL), and the sitagliptin group (217 mg/dL, p < 0.05). Overall, phenolic compounds, mainly anthocyanins, from a fermented blueberry‐blackberry beverage reduced BW gain, fat mass accumulation, and FBG in diet‐induced obese mice.

Funded by Hatch grant Division of Nutritional Sciences 20/20
In vitro and In vivo Protective Effect of Artemisia capillaris Extract on Osteoporosis through Regulation of Osteoclast Differentiation and Bone Resorption Activity
Authors: Jang Hae‐Dong
Published: June 2021

Bone is dynamic tissue that is constantly destroyed or resorbed by osteoclasts and then replaced by osteoblasts in physiological process referred to as bone remodeling. In this study, the protective effect of Artemisia capillaris extract (ACE) on osteoporosis was investigated using RANKL‐induced osteoclasts and ovariectomized rats. In animal study, The total of sixty 8 week‐old female Spraque‐Dawley rats were randomly divided into sham‐operated group and four ovariectomized (OVA) groups: OVA, OVA + 17β‐estradiol (E2, 50 μg/kg/day) and OVA + ACE (125 or 250 mg/kg/day). Daily oral administration of E2 or ACE began 3 weeks after surgery and lasted for 12 weeks. ACE inhibited the decrease in total BMD and BMC of the femur induced by OVA, which was accompanied by a significant reduction in bone remodeling. The bone turnover markers such as BALP, PICP, OPG, RANKL, TRAP, and ICTP were regulated by ACE treatment. In addition, the underlying mechanism of anti‐osteoporotic effect of ACE was studied using scoparone, its major bioactive compound. The TRAP and bone resorption activity were dose‐dependently by scoparone in RANKL‐induced osteoclast differentiation. The suppressive effect of scoparone on RANKL‐induced osteoclast differentiation was executed by down‐regulating ROS production through NADPH oxidase 1 and mitochondria and by scavenging generated ROS. In conclusion, the protective of ACE on osteoporosis can be accomplished by attenuating RANKL‐induced osteoclast differentiation and bone resorption activity. These results indicate that ACE may utilized as a therapeutic agent for the prevention of bone metabolism‐related diseases.
A 6‐month chronic toxicity study on green tea polyphenols in middle‐aged ovariectomized rats
Authors: Shen ChwanLi , Brackee Gordon , Mitchell Kelly , Song Xiao , Finckbone VelvetLee , Tomison Michael , Han Jia , Wang Jia‐Sheng
Published: June 2021

The project is to evaluate chronic toxicity and safety in middle‐aged ovariectomized rats supplemented with different dosages of green tea polyphenols (GTP) in drinking water. 182 SD were sham (n=39) or ovariectomized (OVX, n=143). Both sham and OVX‐control animals receiving no GTP were assigned for sample collection at baseline, 3, and 6 months. The remaining OVX animals were randomized into 0.15%, 0.5%, 1%, and 1.5% (wt/vol) GTP for 3 and 6 months. Routinely clinical observation was performed. Ophthalmologic examination (control, 1% and 1.5% GTP groups), blood for hematology and clinical chemistry, urine analysis were assessed at baseline, 3, and 6 months. Histologic examination was performed on all tissues from animals in both control and high dose groups at baseline and the end of study. No mortality or abnormal treatment‐related findings in clinical observations or ophthalmologic examinations were noted. No treatment‐related macroscopic or microscopic findings were noted for animals administered 1.5% GTP. There was no difference in baseline body weight among all OVX‐treated treatment groups. The OVX+1.5% GTP group had the smallest body weight of the OVX‐treated groups. No difference in food intake among all treatment groups. In all OVX‐treated groups, GTP supplementation decreased water consumption of rats in a dose‐dependent pattern. At 6 months, GTP intake did not affect complete hematology, clinical chemistry, and urinalysis, except for PO4 and BUN (increased); total cholesterol, BUN, LDH, and urine pH (decreased).This study reveals that the No‐Observed‐Effect Level (NOEL) of GTP is 1.5%, the highest dose used. Supported by NIH/NCCAM AT006691.
Suppression of Obesity by Black Adzuki Beans (Vigna angularis) in High‐fat Diet Fed Obese Mouse Model
Authors: KIM MINA , SONG SEOK‐BO , CHA YOUN‐SOO
Published: June 2021

The primary objective of this study was to investigate the obesity inhibitory effects of Black Adzuki Bean (BAB) extract on high‐fat diet induced obese mice. Four‐week‐old C57BL/6J mice were fed for 12 weeks with control diets containing 10% kcal from fat (CD) or high‐fat diets containing 60% kcal from fat (HD) or high‐fat diet + 1% freeze‐dried ethanolic extract of BAB (BAB‐1) or high‐fat diet + 0.08% kaempferol.After 12 weeks,animals fed with HD along with BAB‐1 showed significantly lower final body weight compared with animals fed with HD alone. Compared to HD fed mice, dietary BAB‐1 supplemented mice showed significantly reduced epididymal and mesenteric fat weight along with fat cell size. BAB‐1 dramatically decreased signs of liver pathology. BAB‐1 mice had significantly decreased liver weight and number of hepatic lipid droplet compared with HD mice. These mice also had a significantly lower liver triglyceride and total cholesterol level, and lower concentrations of serum ALT and AST. The epididymal fat tissue mRNA levels of the lipogenesis‐associated genes (PPARγ, FAS, LPL, CD36) were significantly suppressed, while higher expression of lipolytic genes (ATGL, HSL, PPARα, ACOX, CPT‐1α) were observed in the BAB‐1 supplemented mice than the HD mice. Overall, the black adzuki bean extract demonstrated as a potential dietary supplement capable of attenuating obesity. (This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development Project No. PJ009218” Rural Development Administration, Republic of Korea)
Effect of Recombinant Lubricin on Coagulation Parameters in Human Blood
Authors: Fareed Jawed , Hoppensteadt Debra , Iqbal Omer , Abro Schuharazad , Sweigert Patrick , Truitt Edward , Jay Gregory , Hopkinson William
Published: June 2021

Lubricin (proteoglycan 4; PRG4) is a proteoglycan which acts as a joint lubricant. Lubricin is present in synovial fluid and plays a major role in synovial homeostasis. A recombinant form of lubricin, rhPRG‐4 is developed to treat a variety of inflammatory diseases. (Lubris Pharmaceutical). The purpose of this study is to determine the effect of rhPRG‐4 on blood coagulation utilizing various experimental studies.

rhPRG‐4 was supplemented with citrated whole blood from normal human donors (n = 10) to obtain concenration range of 0 ‐ 200 mg/mL. Activated clotting time (ACT), thrombelastographic analysis (TEG), and whole blood prothrombin time (PT), partial thromboplastin time (aPTT), and thrombin times (TT) were measured. Similarly, citrated plasma was supplemented with rhPRG‐4 at 0‐200 mg/ML to determine PT, aPTT and TT. The effect of this agent was also studied on the generation of proteases. In addition, rhPRG‐4 was also studied for its effect on platelet aggregation.

rhPRG‐4 did not produce any effect on the whole blood clotting assays, such as the PT, aPTT and TT. Similarly, in the citrated plasma, none of the clotting assays were altered. rhPRG‐4 did not produce any inhibition of thrombin generation in plasma. Furthermore, it did not produce any inhibition or augmentation of agonist‐induced platelet aggregation. These studies suggest that rhPRG‐4 does not modify whole blood and plasma coagulation profile and produce and more relation of platelet function.

These studies suggest that because of no potential alteration of the coagulation system, lubricin is not expected to modulate bleeding or thrombotic processes in clinical indications.
Bovine and Porcine Mucosal Heparins Exhibit Similar Biologic Profiles
Authors: Fareed Jawed , Nader Helena , Lima Marcelo , Hoppensteadt Debra , Walenga Jeanine , Jeske Walter , Kumar Emmanuel , Raake Wolfram , Bakhos Mandouh
Published: June 2021

Introduction

In consideration of the limited supply of porcine heparin (PH), which may result in a global shortage, the current discussions on the reintroduction of bovine heparin (BH) are timely and justified. This study compared 5 different batches each of BHs and PHs both of mucosal origin.

Materials and Methods

Commercially available PHs and BH were obtained from various US and Brazilian vendors. USP reference standard heparin (Lot F01187) was used to compare the potency of these two groups of heparin utilizing ACT, aPTT, Heptest, Calcium Thrombin Time, anti‐Xa and anti‐IIa activities, as well as protamine and PF4 neutralization. The interaction with HIT antibodies was studied using a pooled HIT sera preparation. The molecular weight profile was determined by HPLC.

Results

On a gravimetric basis, the anticoagulant activity of PH was 10‐30% higher in comparison to BH. PH exhibited higher anti‐Xa and anti‐IIa activities. The USP cross referenced potency in the anti‐Xa assay was 160+20 U/mg for PH vs 132+16 U/mg for BH. Protamine and PF4 titration profiles were comparable. In the HIT assay, there was no difference in between the two groups. .The molecular weight of BH was 17.1 ± 0.8 kDa, whereas that of PH was 16.6 ± 0.5 k Da (p>0.05).

Conclusion

While PH exhibits relatively stronger anticoagulant and anti‐protease activities, in other assays both BH and PH exhibit comparable compositional and functional profiles. Thus, BH may be considered as biosimilar to PH and can be accordingly defined and developed for clinical use.
The Allosteric Heat Shock Protein Inhibitor YM‐01 Protects Against Alcohol‐Induced Growth Arrest
Authors: Ganapathy Ekambaram , Khachatoorian Ronik , Patel Harsh , Arumugaswami Vaithilingaraja , French Samuel
Published: June 2021

Background

Oxidative stress is implicated in the pathogenesis of alcohol‐induced liver injury. This is, in part, mediated by the cytochrome P450 CYP2E1 which contributes to reactive oxygen species (ROS) production. YM‐01, a derivative of MKT‐077, is an allosteric heat shock protein inhibitor and modulator of the triage function of heat shock cognate protein 70 (HSC70) and may protect cells from oxidative stress. In this study, we investigated the effect of YM01 on ethanol‐induced growth arrest.

Methods

HepG2 cells overexpressing CYP2E1 (E47) were pretreated with either vehicle or YM‐01 then treated with arachidonic acid (60 µM for 18 hrs), followed by iron nitrilotriacetic acid (Fe‐NAT, 25 µM for 5 hrs), and finally with ethanol (100 mM for 48 hrs). MTT and western analysis was then performed.

Results

In the CYP2E1‐expressing ethanol‐treated cells, cell growth was arrested as determined by cell viability (MTT assay). This arrest was relieved by treatment with YM‐01. Further, western analysis showed that pretreatment with YM‐01 prevented 4‐hydroxynonenal (4‐HNE) adduct formation induced by ethanol (the most abundant lipid peroxidation end product). In addition, we determined ubiquitination levels by Western analysis to determine role of the ubiquitin‐proteasome pathway.

Conclusion

This study indicates that YM‐01 decreases 4‐HNE adduct formation in CYP2E1‐expressing cells and protects against alcohol induced cell growth arrest.
Increased activity of the complement system in the liver of patients with alcoholic hepatitis
Authors: Shen Hong , French Barbara , Liu Hui , Tillman Brittany , French Samuel
Published: June 2021

Inflammation has been suggested as a mechanism underlying the development of alcoholic hepatitis (AH). The activation of the complement system plays an important role in inflammation. Although it has been shown that ethanol‐induced activation of the complement system contributes to the pathophysiology of ethanol‐induced liver injury in mice, whether ethanol consumption activates the complement system in the human liver has not been investigated. Using antibodies against C1q, C3, and C5, the immunoreactivity of the complement system in patients with AH was examined by immunohistochemistry and quantified by morphometric image analysis. The immunoreactivity intensity of C1q, C3, and C5 in patients with AH was significantly higher than that seen in normal controls. Further, the gene expression of C1q, C3, and C5 was examined using real‐time PCR. There were increases in the levels of C1q and C5, but not C3 mRNA in AH. Moreover, the immunoreactivity of C5a receptor (C5aR) also increased in AH. To explore the functional implication of the activation of the complement system in AH, we examined the colocalization of C5aR in Mallory‐Denk bodies (MDBs) forming balloon hepatocytes. C5aR was focally overexpressed in the MDB forming cells. Collectively, our study suggests that alcohol consumption increases the activity of the complement system in the liver cells, which contributes to the inflammation‐associated pathogenesis of AH.
GPC3‐CD81 axis in the HCV mediated liver carcinogenesis
Authors: Xue Yuhua , Bowen Bill , Orr Anne , Koral Kelly , Haynes Meagan , Bell Aaron , Paranjpe Shirish , Mars Wendy , Michalopoulos George
Published: June 2021

Glypican‐3 (GPC3) is highly over‐expressed in human hepatocellular carcinomas (HCC). GPC3 decreases liver regeneration by interacting with CD81. Other groups have shown that activation of CD81 increases phosphorylation of Ezrin (p‐Ezrin), which inhibits Hippo pathway. Hepatitis C virus (HCV) interacts with human CD81 by E2 protein to enter into hepatocytes. We investigated the mechanism of HCV and GPC3‐CD81 axis in liver carcinogenesis. HepaRG human hepatoma cell line was treated with E2 protein to evaluate the Hippo activity. Rat hepatocytes were treated with CD81 agonist antibody to determine if CD81 related Hippo activity depends on p‐Ezrin. Partial hepatectomy (PHx) was used to evaluate the dynamic changes of GPC3‐CD81 axis with Hippo activity. Human and rat HCC cell lines were used to define GPC3‐CD81 interactions. HCC tissue array was used to assess the expression characteristic of CD81‐GPC3. After E2 treatment, Hepa RG cells had higher activity of Hippo. Treatment of rat hepatocytes with CD81 agonist antibody led to lower Hippo activity and higher p‐Ezrin. PHx shows that GPC3 increases at the early stage of liver regeneration while CD81 decreases, while Yap activity increases. Both human and rat HCC cell lines have dramatically decreased CD81 expression levels. HCC tissue array also shows absence of CD81 in most HCC.

Conclusions

Dependent on CD81, GPC3 may function as tumor‐suppressor gene by modulating YAP expression. Persistent E2 HCV protein interacts with CD81 to decrease Yap via Hippo. Hepatocytes with no expression of CD81 would have a growth advantage to undergo clonal expansion, facilitating formation of HCC. After absence of CD81, GPC3 may be unable to control the Hippo, and this maybe the reason for its high expression in HCC.
Regulation of Actin Interacting Protein Drebrin by Mutations in HSPB1
Authors: Mata Marina , Sun Xiankui , Fink David
Published: June 2021

Heat shock protein B1 (HspB1) is an ATP‐independent molecular chaperone with a conserved α‐crystalline domain in the C‐termini region. A number of mutations in HspB1 have been identified in families with Charcot Marie Tooth type IIF (CMT‐IIF) disease or with distal hereditary motor neuropathy II (dHMN II). Mutations outside the crystalline domain (P39L, G84L, P128S, P182L) associate with dHMN. Mutations S135F, R136W and R127W associate with a CMT‐IIF phenotype.

Wild type (wt) HspB1 reduces RhoA activation by translational inhibition of PDZ‐RhoGEF, a RhoA specific GEF. Decreased expression of PDZ‐RhoGEF by HspB1 was accomplished by enhancing expression of specific microRNAs (miR20a and miR128) that inhibit the translation of PDZ‐RhoGEF and resulted in an increased neurite growth response. The regulatory effect of HspB1 on RhoA activity is blocked by mutation in S135F of HspB1 but not by R136W and R127W. This suggested either that these HspB1 mutations alter independent cellular mechanisms or that their effects may converge downstream of RhoA GTPase signaling to cause similar disease phenotype.

We constructed lentiviral vectors carrying full length human HspB1 cDNA of wt or mutant S135F, R136W or R127W. Cortical neurons derived from SD rat pups E15 were transfected to expressed wt or mutant HspB1transgene to determine their effect on actin cytoskeleton.

The studies showed S135F, R16W and R127W mutations cause HspB1 to partition more readily with the cytoskeletal fraction and enhance the binding of HspB1 to F‐actin leading to the loss of Drebrin, an F‐actin side‐binding protein, from the F‐actin domain. These findings highlight the diverse mechanisms by which HspB1 functions as an important regulatory protein of the actin cytoskeleton.
Convulsant effects of high efficacy synthetic cannabinoid JWH‐018 in mice
Authors: Camper Kiristen , Caperton Caitlin , Tai Sherrica , Fantegrossi William
Published: June 2021

Medical cannabis has apparent efficacy in controlling seizure in treatment‐resistant epilepsy, particularly in children. In contrast, abuse of high efficacy synthetic CB1 agonists (SCBs) can result in significant toxicity, including morbidity not typically associated with marijuana, such as seizure. Here, the convulsant effects of the aminoalkylindole SCB JWH‐018 were investigated in mice, and compared with those of the classical chemical convulsant pentylenetetrazole (PTZ) using an observational rating scale. JWH‐018 and PTZ dose‐dependently elicited convulsant effects in mice. The CB1 antagonist / inverse agonist rimonabant blocked convulsant effects of JWH‐018, but not those of PTZ. Pretreatment with Δ9‐THC dose‐dependently blunted the convulsant effects of JWH‐018, but did not alter PTZ‐induced convulsions. Daily administration of JWH‐018 for five consecutive days resulted in tolerance to its convulsant effecs, but daily treatment with PTZ increased its convulsant effects. Finally, no cross‐tolerance was observed when PTZ was administered to mice previously made tolerant to JWH‐018‐elicited convulsions. These data suggest that CB1 agonists can elicit convulsant effects in the mouse, although intrinsic efficacy is likely to be an important factor. These studies supported, in part, by an NIGMS IDeA Program award (GM110702), by the UAMS Translational Research Institute (RR029884), and by T32 DA022981. KDC and COC received Summer Undergraduate Research Fellowships from the American Society of Pharmacology and Experimental Therapeutics.
Use of Daniovision™ to Measure Activity in 3rd Instar Drosophila melanogaster Larvae
Authors: Graham Stephanie , Rogers Ryan , Alper Richard
Published: June 2021

The noncompetitive GABAA receptor antagonist picrotoxin (PTX) decreases activity in 3rd instar Drosophila larvae (Eur J Neurosci, 2006). The purpose of these studies was to replicate the data obtained by Stilwell et al. using an automated data acquisition/data reduction system, Daniovision™ by Noldus.

Larvae were extracted from food (Carolina Biologic Supplies 4‐24) in 20% sucrose at the 3rd instar stage. The larvae were randomly placed into vials containing vehicle (20% sucrose + 0.3% DMSO) or PTX from 0.03‐3 mM: n = 20‐30 larvae/treatment. After incubation for 2‐24 hr, individual larvae were placed in separate wells of a 6‐well culture plate previously coated with 2% agar, then placed in the Daniovision™ apparatus. The activity of each larva was digitized and recorded for 5 minutes using Ethovision software.

PTX reduces 3rd instar larvae activity in a concentration‐dependent manner, consistent with findings reported by Stilwell et al. Incubation in 1 mM PTX for 4 or 24 hours decreased activity, with consistent reduction produced by 3 mM, variably by 0.3 mM, and never by 0.03 mM. PTX, 0.03‐3 mM, did not alter activity in a mutant strain, Rdl, known to have a point mutation in the PTX binding site on the GABAA receptor conferring a lower affinity for PTX.

We have shown that an automated system can be used to quantify activity in 3rd instar Drosophila melanogaster larvae. As human observation and review of videos is not required, and multi‐well plates can be used, we have refined an in vivo high‐throughput screen for anti‐seizure compounds.

Supported by funds from the University of Saint Joseph.
Knockdown of the D2 Autoreceptor from the Ventral Tegmental Area Increases Temporal Impulsivity
Authors: Bernosky‐Smith Kimberly , Qui Yanyan , Li Jun‐Xu , Bass Caroline
Published: June 2021

Impulsivity associated with abnormal dopamine function occurs in a variety of disorders, including addiction. Temporal impulsivity is characterized by the preference for smaller rewards sooner over larger rewards after a delay, caused by excessive discounting of future rewards. Addicts have abnormally high discounts rates and prefer the smaller rewards sooner. While impulsivity has been inversely correlated with D2 receptor availability in the midbrain, it is difficult to mechanistically link the two due to the diverse neuroanatomical localization of the receptors, which are found in multiple brain regions, neuronal subtypes, and compartments within the neurons. To test the hypothesis that Ventral Tegmental Area (VTA) D2 autoreceptor hypofunction is linked to impulsivity, we have developed a method to knockdown D2 autoreceptors from the VTA. By injecting a viral vector to deliver short interfering RNAs (shRNA) targeting the D2 receptor in rats, D2 receptor knockdown is restricted to the VTA. This leaves postsynaptic D2 intact in the nucleus accumbens, prefrontal cortex, and other corticomesolimbic structures, all which have some influence in impulsive action. Rats were trained in a delayed discounting (DD) task to assess choice impulsivity until a stable discounting curve was obtained. Rats then received bilateral VTA infusions of the D2 shRNA or a scrambled control virus. Over time the discounting curve of the VTA D2 knockdown rats shifted to the left, indicating a preference for the smaller, immediate reward, whereas the curve for control rats remained stable. Together these results demonstrate that a decrease in D2 autoreceptors enhances temporal impulsivity.