bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Caloric Restriction on Cardiometabolic Risk and Bone Biomarkers in Ovariectomized Rats
Authors: Ahn HyeJin , Choue Ryowon
Published: June 2021

The purpose of the study was to investigate the effects of caloric restriction on cardiometabolic risk and bone biomarkers in ovariectomized (OVX) rats. Seven–week–old female SD‐rats were randomized to sham‐operated control fed ad libitum diet (SHAM‐AL), sham‐operated fed caloric restriction diet (SHAM‐CR), ovariectomy‐operated control fed ad libitum diet (OVX‐AL), or ovariectomy‐operated fed caloric restriction diet (OVX‐CR). For 8 weeks, the OVX‐AD group was pair‐fed with the SHAM‐AD group and the CR groups were fed diet with 50% fewer calories than their counterparts. Cardiovascular risk biomarkers (glucose, insulin, TG, TC, LDL‐C, HDL‐C, NOS, endothelin‐1) and bone biomarkers (cytokines, osteoclast and osteoblast scores, ALP, OC, DPD, NTx, and other bone‐related parameters) were analyzed at the end of the experiment. As results, glucose, TG, TC, LDL‐C, NOS and endothelin‐1 were the highest in the OVX‐AL group, but HDL‐C was high in the CR groups (p<0.05). IL‐1α, IL‐6, RANKL/OPG ratio, osteoblast scores, and OC were low and DPD and NTx were high in the CR groups (p<0.05). IL‐1α, IL‐6, RANKL/OPG ratio, osteoclast scores, ALP, OC, and DPD were high in the OVX groups (p<0.05). As a conclusion, the caloric restriction produced positive effects on cardiometabolic risk biomarkers, especially in OVX groups, however it produced negative influences on bone mass in both SHAM and OVX groups.
Activity of GSHpx and toenail Se contents in Korean young adults
Authors: Lee Okhee , Chung Yongsam , Moon Jongwha
Published: June 2021

Selenium, is a key constituent of enzyme, glutathione peroxidase, so effective in decreasing various oxidative stress. Toenail selenium level is a long term indicator for selenium nutritional status. The low activity of glutathione peroxidase in RBC is known to indicate a state of selenium deficiency. This study is aimed to measure the toenail selenium levels in Korean young adult and GSHpx activity in young adults. For the measurement of toenail selenium content, the instrumental activation analysis (=INAA) method was employed. Serums were subsequently used to analyze glucose and insulin level. The RBC was used to analyze the GSHpx activity. Toenail were washed in ultra‐sonicator and freeze‐dried at temperature of −60 C for 12~24 h. The toenail selenium contents were measured by INAA method using HANARO‐reactor. Analytical quality control was conducted using NIST SRMs. The toenail Se level and RBC GSHpx did not show significant difference by gender in our study participants. The toenail selenium status and GSHpx activity did not show significant correlation with insulin and glucose, but with c‐reactive protein level. This result suggests that the selenium status would be associated with inflammation in young adult and showed a vast research on the selenium status in Korean people is necessary for the evaluation of selenium association with risk factors for health.

Grant Funding Source: National Research Foundation of Korea
Dietary alkali‐extracted rice protein improves some diabetes‐related markers in obese type 2 diabetic rats
Authors: Watanabe Reiko , Kubota Masatoshi , Yamaguchi Miki , Saito Akihiko , Fujii Mikio , Fujimura Shinobu , Kadowaki Motoni
Published: June 2021

The explosive increase in patients of type 2 diabetes mellitus (T2DM) is becoming a serious problem all over the world. T2DM is a metabolic disease caused by lifestyle factors, and may lead to many complications such as retinopathy, neuropathy and chronic renal failure. The control of blood glucose levels by the diet therapy is the most important treatment to delay the progression of T2DM. Rice is a staple food as a major source of not only energy but also protein in many Asian Countries including Japan. Therefore, we focused on effects of alkali‐extracted rice protein (AE‐RP) on T2DM. The AE‐RP used included a high crude protein content (>;90%). Male obese type 2 diabetic Zucker Diabetic Fatty (ZDF) rats and non‐diabetic Zucker lean rats were fed 20%AE‐RP or casein (C) diets, for 8 or 15wk. The diabetes‐related markers in the blood and liver were determined. AE‐RP had suppressive effects on fasting blood glucose levels (175 vs. 377 mg/dL) at 6 wk, and on hemoglobin A1c (5.9 vs. 7.3%) at 8 wk in ZDF rats compared to C group. In addition, the concentrations of plasma alkali phosphatase and total cholesterol in AE‐RP‐fed ZDF rats were significantly lower than those of C‐fed group. These findings suggest that AE‐RP has beneficial effects on the progression of diabetes in obese type 2 diabetic rats. This study was supported in part by a Grant‐in‐Aid for Scientific Research to RW from the Japan Society for the Promotion of Science.
Astragalin inhibits allergic asthma through modulating oxidative stress‐responsive MAPK signaling
Authors: Cho In‐Hee , Kang Young‐Hee
Published: June 2021

Allergic asthma is airway obstruction and chronic inflammation and is triggered by inhaled allergens such as dust mite allergen and pollen. Since inflammation is often associated with an increased generation of reactive oxygen species (ROS), and the biochemical environment in the asthmatic airways is favorable for ROS‐mediated reactions, it can be assumed that oxidative stress is a mechanistically‐imperative factor in asthma. Astragalin (kampferol‐3‐O‐glucopyranoside), a flavonoid with anti‐cancer, antimicrobial properties, has been isolated from tea seeds. This study elucidated whether oxidative stress activated allergic asthmatic responses in airway epithelial BEAS‐2B cells, which was suppressed by treating 1–20 μM astragalin. When airway epithelial cells were exposed to 2 μg/ml LPS, nontoxic astragalin at 1–20 μM suppressed LPS‐induced ROS production, evidenced by fluorescent DCFH‐DA staining. Astragalin attenuated LPS‐enhanced expression of PKCβ2 and p22phox and H2O2‐upregulated phosphorylation of JNK and p38 rapidly within 30 min. In contrast, the phosphorylation of Akt and ERK increased due to astragalin treatment. Additionally, astragalin attenuated LPS‐ or H2O2‐induced eotaxin‐1 protein expression of epithelial cells. Therefore, dietary astragalin can be effective in ameliorating allergic airway diseases through modulating oxidative stress‐responsive MAPK signaling pathway.
Inhibitory effects of anthocyanin‐rich purple corn extract on diabetes‐associated glomerular angiogenesis
Authors: Kang Min‐Kyung , Kang Young‐Hee
Published: June 2021

Diabetic nephropathy (DN) is one of the major diabetic complications and the leading cause of end‐stage renal disease. Abnormal angiogenesis results in new vessels that are often immature and play a pathological role in DN, contributing go renal fibrosis and disrupting glomerular failure. Purple corn has been utilized as a daily food and exerts disease‐preventive activities. This study was designed to investigate whether the anthocyanin‐rich purple corn extract (PCE) prevented glomerular angiogenesis under hyperglycemic condition. Human endothelial cells were treated in conditioned media of mesangial cells exposed to 33 mM high glucose (HG‐HRMC). PCE decreased endothelial expression of the angiogenesis markers VEGF and HIF‐1α induced by HG‐HRMC‐ conditioned media. PCE attenuated angiogenic endothelial marker PECAM‐1 and integrin β3 enhanced in HG‐HRMCconditioned media. In the in vivo study, db/db mice were treated with 10 mg/kg PCE for 8 weeks. It was revealed that PCE alleviated glomerular angiogenesis of diabetic kidney by attenuating the induction of VEGF and HIF‐1α. In addition, PCE diminished plasma level of soluble VEGF. These results demonstrate that PCE antagonized glomerular angiogenesis due to chronic hyperglycemia. Therefore, PCE may be a potent therapeutic agent targeting abnormal angiogenesis in DN leading to kidney failure.
Lepidium meyenii (maca) consumption prevent benign prostatic hyperplasia
Authors: Noratto Giuliana , Condezo‐Hoyos Luis , Gasco Manuel , Gonzales Gustavo
Published: June 2021

Lepidium meyenii (maca) is a Peruvian Andean with effects against benign prostatic hyperplasia (BPH); a disease associated with male sexual dysfunction.

BPH was induced in mice with two doses of testosterone enanthate (TE) (83mg/kg, day 1 and 7), a negative control received the vehicle. Experimental groups received TE alone or TE in combination with Finasteride (FN) (0.6 mg/Kg/day), a drug used to treat BPH, red maca (RM)( 167 mg/Kg/day), or black maca (BM)( 167 mg/Kg/day) extracts during 21 days. At the end of the study, weights of body and reproductive organs (prostate, seminal vesicles (SV), testicles, preputial glands (PG), and epididymis) were recorded and compared to negative controls.

Results showed that TE induced BPH increasing weights of prostate and SV to 225% and 149% of negative controls, respectively. Treatments with FN, RM and BM prevented these effects and maintained prostate weights at 71%, 44%, and 82% of TE, respectively. The SV weights were maintained at 73%, 80% and 81% of TE, respectively. Weights of testicles, and PG were also increased by TE, and treatments with RM were more effective than BM or FN in maintaining weights closer to negative controls. Overall, RM protected against changes in weights of male reproductive organs that accompanied HPB at higher extent than FN or BM. The underlying mechanisms to explain the RM protective effects are being investigated.
Onion peel extracts and exercise decrease cardiovascular risk in rats fed a high fat diet
Authors: Chyun Jong Hee , Yang Yuan , Yim Jung‐Eun
Published: June 2021

Quercetin derivatives in onion peel extracts (OPE) have been regarded as the most important flavonoids to improve lipid profiles. The present study was aimed to examine the effects of OPE and exercise on fat deposition and blood lipid profiles in a diet‐induced obese Sprague‐Dawley rats. Twenty eight male Sprague‐Dawley rats were randomly assigned into four groups: High‐fat diet (HFD, 15% beef tallow + 1% cholesterol + 5% corn oil), HFD with exercise (HFD‐Ex), HFD with 0.15% OPE (HFD‐OPE), and HFD with both exercise and 0.15% OPE (HFD‐Ex‐OPE). After feeding experimental diets for 4 weeks, we weighed epididymal fat and retroperitoneal fat, and examined blood lipid profiles and liver lipid peroxide (TBARS). Epididymal fat and retroperioneal fat weights were remarkably lower in the HFD‐Ex and HFD‐Ex‐OPE group than the HFD group. The liver TBARS was significantly reduced and serum HDL cholesterol was significantly increased in HFD‐Ex, HFD‐OPE, and HFD‐Ex‐OPE. Atherogenic index and cardiovascular risk factor(CRF) were lower in HFD‐Ex, HFD‐OPE, and HFD‐Ex‐OPE than those in HFD group. Our results suggest that OPE and exercise might improve cardiovascular (CVD) risk with reduction of epididymal and retroperioneal fat. These findings provide a basis for the use of OPE with exercise to reduce CVD risk.
The effect of black chokeberry (Aronia melanocarpa) on the prevention of obesity in C57BL/6J mice
Authors: Baum Jamie Ilene , Shouse Stephanie A , Gilbert William , Prior Ronald L , Howard Luke R
Published: June 2021

Products derived from black chokeberry are claimed to be beneficial in diseases associated with oxidative stress due to high concentrations of anthocyanins and procyanidins, which possess strong antioxidative potential. Extracts from black chokeberries have been demonstrated to have anticancer, cardioprotective, and anti‐diabetic effects. The purpose of this study was to determine if supplementation with Aronia (black chokeberry) juice concentrate (AJ) has anti‐obesity properties. Male C57BL/6J mice were placed on either a low‐fat (LF; 5% fat), LF+AJ (113 g/kg diet), higher‐fat (HF; 30%), or HF+AJ for 12‐weeks. Mice on the LF+AJ diet had lower body weights compared to the other dietary treatments (p<0.05). Mice on the LF and LF+AJ had less total weight gain compared to those receiving high‐fat diets (p<0.05). As expected, mice receiving the LF diet had less epididymal fat (p<0.05), however mice on the LF+AJ had significantly lower epididymal fat than LF controls. There was no difference in fasting plasma insulin, glucose or triglycerides between groups. Mice receiving AJ supplementation had higher cecum content weights (p<0.05). The results from this study demonstrate that supplementation with AJ has the potential to reduce body fat under normal dietary conditions, however further research is needed to determine mechanisms of action. Funded by Arkansas Biosciences Institute.
Effects of Konjac glucomannan, inulin and cellulose on colonocyte DNA Integrity and anti‐genotoxic responses against azoxymethane in mice fed a high‐fat diet
Authors: Wu Wen‐Tzu , Chen Hsiao‐Ling
Published: June 2021

The aim of study was to compare effects of konjac glucomannan (KGM), inulin, and cellulose on the DNA integrity and gene expressions of enzymes in colonocytes of C57BL/6J mice before and within 24 h of a single dose of azoxymethane (AOM). Six‐week‐old mice were fed a control diet with 20% (w/w) corn oil and 1% (w/w) cellulose or that supplemented with KGM, inulin or cellulose (5%, w/w) for 3 weeks. Fresh feces were collected 3 days prior the AOM (10 mg/kg BW, i. p.) injection on day 22. Each mouse was killed before (0 h), or 6, 24 h after the injection. The colonocytes were immediately isolated for further analyses of DNA damage (denoted by tail moment) and gene expressions. The DNA damage of colonocytes was the lowest in the inulin group at 0 h. The increased AOM‐induced DNA damage was ameliorated by KGM and inulin. The initial (0 h) gene expressions of glutathione peroxidase 2 (GPX2) in the colonocytes were lower in the control and cellulose groups, which may lead to AOM‐induced up‐regulation (to ≥ 2 folds of initial level) within 24 h. Addition of KGM or inulin to control diet beneficially promoted the concentration and daily fecal excretion of bifidobacteria and lactobacilli. The present study suggests that KGM and inulin protect the colonocytes from DNA damage induced by AOM, which may be related to their prebiotic effects. This study was supported by the National Science Council Grant NSC‐101–2320‐B‐040–018 ‐MY2, Taiwan.
Chemical properties and antioxidant activity of beverage with ethanol extracts from the roasted Maegmundong (Liriope platyphylla tubers)
Authors: Lee Kyongae , Kim Boram
Published: June 2021

The antioxidant activity of beverage with ethanol‐extract from the roasted Maegmundong (Liriope platyphylla tubers) were studies along with free amino acid and mineral composition analysis, and saponin content determination. Antioxidant activity determined by electron‐donating ability and ABTS radical scavenging activity. Five major free amino acids of the Maegmundong beverage were proline (14.00mg%), methionine (13.95 mg%), alanine (11.46 mg%), aspartic acid (10.10 mg%) and arginine (7.53 mg%). The mineral compositions of the Maegmundong beverage were potassium(244.32mg%), magnesium(10.14mg%), sodium(7.68mg%), calcium(7.54mg%) and iron(0.55mg%). Crude saponin in the Maegmundong beverage was 6.9%. Electron donating ability and ABTS radical scavenging activity of the Maegmundong beverage were 97.5 and 984.7%, respectively, of control L‐ascorbic acid. Contents of total flavonoids and polyphenols were 33.2 and 812.5 mg/mL, respectively. High electron donating ability and ABTS radical scavenging activity of the Maegmundong beverage were possibly related with amino acid composition as well as high contents of polyphenols and flavonoids in beverage
Effects of feeding cranberry extract enriched in A‐type proanthocyanidins on weight gain and gene expression in the liver and brain of pigs
Authors: Jang Saebyeol , Monagas Maria , Molokin Aleksey , Lakshman Sukla , Bruna Gonzalo , Urban Joseph , Solano‐Aguilar Gloria
Published: June 2021

Cranberry products have purported beneficial effects on human health, including antimicrobial, antioxidant and anti‐inflammatory activities. These effects have mainly been attributed to A‐type proanthocyanidins. We studied the effects of cranberry extract containing ~7% proanthocyanidins (CEPA) on metabolic parameters and gene expression in pigs. Four‐week‐old pigs were fed 14 mg of CEPA/Kg of body weight/day added to a basal or 40% high fat diet for 12 weeks and compared to pigs fed similar diets without added CEPA. Two‐way ANOVA revealed a significant main effect of fat and a trend towards a CEPA × fat interaction on body weight gain showing decreased body weight gain only in pigs fed CEPA with a basal diet. Supplementing CEPA in the basal diet also decrease the expression of hepatic genes SCD‐1 and FASN. There was no significant effect of CEPA on high fat‐ induced increase of liver inflammatory gene such as IL‐1â and TLR4. Notably, brain‐derived neurotrophic factor (BDNF) and insulin‐like growth factor (IGF)‐1 gene expression in the hippocampus was increased by CEPA independent of diet suggesting an effect on neuropeptides linked to cognitive function. Overall, this study showed that daily feeding of CEPA led to body weight loss in basal diet associated with decreased lipid metabolism gene expression. Furthermore, CEPA supplement enhance BDNF and IGF‐1 gene expression in the hippocampus.
Sulforaphane activates a protective Nrf2 response and reduces inflammatory markers in microglia cells
Authors: Townsend Brigitte , Jang Saebyeol , Johnson Rodney
Published: June 2021

Cells exposed to oxidative stress require intrinsic mechanisms to abrogate harmful effects of un‐neutralized reactive oxygen species. Nrf2 is a primary transcription factor involved in detoxification. Induction of antioxidant response element (ARE) genes heme oxygenase (HO1), glutathione‐S‐transferase (GSTA), and NAD(P)H quinine oxidoreductase (NQO1) is directly mediated through Nrf2 activation. While Nrf2 has been broadly studied, data characterizing its role in the central nervous system, particularly in microglia, is limited. In this study, BV2 cells were treated with sulforaphane (SFN) to test its effect on activation of Nrf2. SFN treatment increased mRNA expression of HO1, GSTA, and NQO1. In order to determine if the protective effects of Nrf2 activation could reverse inflammatory characteristics of toxin‐activated microglia, BV2 cells were treated with LPS. Cells responded to LPS with significantly increased expression of inflammatory genes. Pre‐treatment with SFN upregulated ARE genes while down‐regulating the increase in inflammatory cytokines IL‐6, IL‐1β, TNFα, and iNOS caused by LPS. Additionally, SFN negated increased nitric oxide production induced by LPS. Changes in the Nrf2 signaling pathway in microglia may be implicated in cognitive decline associated with an aging inflammatory state. This work paves the way for future studies in vivo. 538 NIH 2 R01AG016710C
Anti‐oxidative and Anti‐inflammatory effect of ethanol precipitate of dandelion (Taraxacum officiniale ) in LPS stimulated SD rats.
Authors: Seo Bo‐young , Song Young‐Sun
Published: June 2021

This study was carried out to investigate the anti‐oxidative and anti‐inflammatory effect ethanol precipitate of dandelion (DEP) SD rats injected lopopolysaccharide (LPS). Thirty five rats were divided into 5 groups; NC (normal control), DEPC (DEP control 300mg/kg BW), LC (LPS control), DEP100+LPS (DEP 100mg/kg BW) DEP300+LPS (DEP 300mg/kg BW ). DEP was administered orally for 2 weeks and endotoxin stimulation was induced by intraperitoneal injection of LPS (6 mg/kg BW) 4 h before sacrifice. NC and DEPC groups were injected with saline solution. Histopathological observation further revealed that LPS‐stimulated moderate levels of inflammatory cell infiltration, centrilobular fatty change. But DEP administration for 2weeks attenuated these trends. Hepatic protein expression of iNOS was significantly (p<0.05) lower in DEP administration groups than LC group. Hepatic TBARS concentration was lower in DEP 300+LPS group than LC group. Expressions of superoxide dismutase and catalase were higher in the DEP groups than in the LC group. From the above results, it is concluded that DEP supplementation may ameliorate oxidative stress and inflammatory processes by LPS. Supported from National Research Foundation of Korea (NRF‐2010–0005599)
Curcumin does not add to the quercetin‐induced decrease in TNF‐α production from lipopolysaccharide‐stimulated peripheral blood mononuclear cells
Authors: Meaney Mary Pat , Neiman David C.
Published: June 2021

Quercetin (QUER) and curcumin (CUR) have potent anti‐inflammatory effects; however, it is unclear whether these two plant‐derived phenolics can act synergistically to reduce inflammation in human cells. The purpose of this study was to determine whether a patented blend of QUER, vitamin C, and vitamin B3 (ratio 50:50:2) (QCB3), with or without CUR, could decrease the in vitro release of TNF‐α from lipopolysaccharide (LPS)‐stimulated human peripheral blood mononuclear cells (PBMCs) from four donors. To test this, isolated PBMCs were incubated in cRPMI media containing 25 μM QUER (in QCB3) and/or 15 μM CUR. After 1 h, PBMCs were stimulated with 0.1 μg/ml LPS for 23 h, and then the media was collected for TNF‐α determination by ELISA. The concentration of TNF‐α adjusted on a per cell basis was lower when compared to control wells for QCB3 (20.1±2.9%, P<0.0001), CUR (46.7±11.4%, P=0.0004), and QCB3+CUR (19.8±5.1%, P<0.0001) treated wells. In addition, the concentration of TNF‐α/cell was lower in QCB3+CUR treated wells than in wells treated with CUR alone (19.8±5.1% v. 46.7±11.4%, P=0.0494). These findings indicate that treatment with QCB3 and CUR decreases LPS‐stimulated TNF‐α production from PBMCs, but that the addition of 15 μM CUR to 25 μM QUER in QCB3 does not enhance the anti‐inflammatory effect of QUER.

Supported by Quercegen Pharma.
Protective effect of Ecklonia cava polyphenol against ethanol‐induced liver injury in rat hepatocytes
Authors: Yamashita Haruka , Tamura Akiko , Sasaki Mio , Matsui‐Yuasa Isao , Terada Masaki , Kojima‐Yuasa Akiko
Published: June 2021

Introduction

Alcoholic liver disease is characterized by a pathological process with progressive liver damage leading to steatosis, steatohepatitis, fibrosis and finally cirrhosis. Ethanol metabolism‐associated oxidative stress contributes to be the pathogenesis of alcoholic liver disease. Here, we examined the effect of the extract of Ecklonia cava, brown algae, on ethanol‐induced liver injury in rat primary hepatocytes.

Methods

Isolated hepatocytes were cultured with or without 100 mM ethanol. Ecklonia cava polyphenol (ECP) was dissolved in dimethylsulfoxide. ECP (0~25 μg/ml) was added to culture medium with ethanol simultaneously. The cells were cultured for 0~24 hrs.

Results

In the cultured hepatocytes, ECP treatment suppressed the ethanol‐induced increase in cell death by inhibiting cytochrome p450 2E1(CYP2E1) expression and activity, which is related to the production of reactive oxygen species (ROS) when large quantities of ethanol are oxidized. On the other hand, ECP treatment increased the activity of alcohol dehydrogenase.

Conclusion

These results suggest that ECP protects against ethanol‐induced liver injury via regulating alcoholic metabolizing enzymes and then suppressing ROS production.
Genistein, a phytoestrogen from soybeans, and estradiol modulate cell growth in normal colonocytes and suppresses ACF formation induced by AOM and TNBS.
Authors: YOO GYHYE , Billimek Autumn , Weige Charles , Allred Clinton
Published: June 2021

Studies indicate estrogens and estrogen signaling can reduce colorectal cancer incidence by altering cell homeostasis. Genistein (GEN) is a phytoestrogen that binds to estrogen receptors (ERs) and may influence the development of colon cancer. Our laboratory has studied the effect of GEN on cell growth in young adult mouse colonocytes (YAMC), a non‐malignant colon cell line, and on tumor formation in mice. GEN (1 or 10 μM) as well as estradiol (E2, 1 nM) suppressed cell growth while no change on cell proliferation in YAMC cells. GEN or E2 resulted in increased Caspase‐3 activity and p53 trascriptional activity in YAMC cells. Ovariectomized (OVX) mice were utilized to determine the effect of GEN on healthy colon mucosa. Mice were treated with either a subcutaneous E2 pellet, 1000 ppm GEN diet, or a phytoestrogen free diet (Control). E2 animals showed an increase in apoptosis with GEN animals showing a similar trend in apoptosis. In a second animal study, mice were OVXed, treated with an E2 pellet, GEN or Control diet, and exposed to azoxymethane, a colon specific carcinogen, and 2, 4, 6‐trinitro benzene sulfonic acid (TNBS), a proinflammatory agent, to examine the effects of GEN on inflammation associated colon carcinogenesis. Treatment with E2 and GEN suppressed aberrant crypt foci (ACF) formation compared to control. Future analyses of these tissues will investigate the effects of GEN and E2 on apoptosis and proliferation.

Grant Funding Source: AICR and ACS
Impact of Tart Cherry Anthocyanins and Dietary Carbohydrate Sources on Intestinal Tumorigenesis in APCMin/+ mice
Authors: Walker Ryan , Bourquin Lesie
Published: June 2021

Consumption of tart cherry anthocyanins (TCA) has been demonstrated to reduce the development of intestinal tumors in APCMin/+ mice, whereas high‐sucrose diets have been shown to increase intestinal tumor development in this model. The aim of this study was to determine the efficacy of TCA to reduce intestinal tumorigenesis in APCMin/+ mice consuming diets rich in sucrose and other dietary carbohydrates. Mice were assigned to six different modified AIN‐93G diets, which contained one carbohydrate source (starch, sucrose, or glucose) and TCA (0 or 800 mg/kg). To enhance tumor development, mice were injected for three consecutive weeks with azoxymethane (8 mg/kg) prior to treatment. Mice consuming the sucrose diet without TCA had the greatest adenoma burden in the colon compared to mice consuming sucrose‐TCA, glucose‐TCA, glucose, and starch (P<0.05). No statistically significant differences between other diets were observed in the colon. Mean tumor burden in mice consuming sucrose without TCA was 1.9 to 5.2 times higher when compared to all other diets. These results demonstrate that feeding TCA reduces the additional intestinal adenoma development observed in APCMin/+ mice consuming sucrose.

Grant Funding Source: MSU Internal Funding
GSTM1 null‐allele polymorphism, oxidative stress and HIV disease progression in an HIV‐infected population
Authors: Parsons Mary , Baum Marianna K , Shin Dong‐Ho , Martinez Janet Diaz , Williams Shawn , Greer Pedro , Martinez Sabrina Sales , Campa Adriana
Published: June 2021

Objective

To examine the effects GSTM1 null‐allele polymorphism on oxidative stress (OS) and disease progression in HIV+ adults.

Methods

GSTM1 genotype was determined by quantitative PCR. OS (mitochondrial 8‐oxo‐2′‐deoxyguanosine, and Complex I and IV) and HIV disease (CD4 count and viral load) markers were measured in 129 HIV+ adults. Those with BMI >;28kg/m2 were excluded. Gene copies were not quantified, thus the Hardy‐Weinberg formula was not applicable.

Results

Functional GSTM1 occurred in 66% (62/94) Blacks, and 33% (11/33) of the Caucasians. Those with functional GSTM1 had higher CD4 count (β=5.55, p=0.04), and lower viral load (β=−1.11, p=0.001), and mitochondrial 8‐oxo‐2′‐deoxyguanosine (β−0.28, p=0.03). Complex I activity (β=7.53. p=0.04) was increased over time compared to those with null‐allele GSTM1, controlling for other variables affecting OS.

Conclusion

The presence of a functional GSTM1 genotype is associated with improved antioxidant activity over time. GSTM1 null‐allele polymorphism appears to have a detrimental effect on immune function and viral load control, suggesting a potential benefit from antioxidants in HIV infected patients with the defective gene.

Supported by NIDA
Smooth muscle nitric oxide responsiveness predicts clinical maturation of hemodialysis fistulae
Authors: Hou Xiuyun , Dember Laura M , Wason Chris , Cohen Richard A , Tong Xiaoyong , Hemodialysis Fistula Maturation Study Group
Published: June 2021

The arteriovenous fistula is the preferred type of hemodialysis vascular access for patients with end‐stage renal disease, but 50% of newly created fistulas fail to mature adequately for use. Stenosis due to neointimal hyperplasia is present in a large proportion of fistulae with maturation failure suggesting that local mechanisms controlling vascular smooth muscle cell (SMC) migration are important contributors to maturation failure. Our objective is to manipulate local factors present in the vein at the time of fistula surgery in order to prevent maturation failure. Using SMC explants from vein tissue obtained at the time of fistula creation from patients enrolled in the NIH Hemodialysis Fistula Maturation Study, we find that among patients with subsequent fistula maturation failure, •NO inhibits SMC migration significantly less. Using strategies initially developed in animal models, we find that impaired •NO inhibition is associated with oxidation of the calcium regulatory protein, sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA), and can be reversed by overexpressing SERCA or by down‐regulating Nox4 NADPH oxidase. Our data suggest that SMC •NO responsiveness may be predictive of fistula maturation outcome and that therapeutic restoration of •NO responsiveness through manipulation of local factors may prevent fistula maturation failure.

Funding: R21DK084390 and R01 HL31607
Opioid growth factor (OGF) prevents relapses in mice with relapsing‐remitting EAE, a model for multiple sclerosis
Authors: Hammer Leslie A. , Zagon Ian S. , McLaughlin Patricia J.
Published: June 2021

Relapse‐remitting multiple sclerosis (MS) is a life‐long, inflammatory disease of the CNS that affects about 350,000 people in the U.S. [Met5]‐enkephalin, termed opioid growth factor (OGF) administered daily to C57BL/6 mice with established chronic progressive experimental autoimmune encephalomyelitis (EAE) reversed the progression of behavioral disease within 6 days of treatment. To determine whether administration of OGF modulates the course of relapsing‐remitting EAE (RR‐EAE) when OGF is administered at the time of induction, female SJL/J mice received proteolipid protein 139–151 emulsified in complete Freund's adjuvant, and were injected daily (i.p.) with OGF (10 mg/kg) or sterile saline. OGF treated RR‐EAE mice had a 40% reduction in peak disease score and significantly fewer relapses over the course of 55 days relative to controls. The duration of relapse for OGF‐treated mice was approximately 1 day in comparison to 13 days for controls. Evaluation of overall disease scores revealed that only 15% of RR‐EAE mice receiving OGF ever expressed disease scores ≥ 4 in comparison to 40% of controls. Histological examination of spinal cords revealed that OGF‐treated mice had markedly less demyelination and fewer activated astrocytes than controls. The results indicate that OGF treatment reduces the progression of RR‐EAE, and may be a novel biological therapy for the treatment of MS. Supported by the Shockey Family Foundation.
Hyperactivity and Cognitive Flexibility in animals with Temporal Lobe Epilepsy
Authors: Ramos Fabiane Ochai , Cysneiros Roberta Monterazzo , Scorza Fulvio Alexandre
Published: June 2021

The temporal lobe epilepsy (TLE) can have a substantial impact on cognitive process. The rate of prevalence of Attention Deficit Hyperactivity Disorder (ADHD) in children with epilepsy is estimated from 12 to 39%, but the relationship between these two diseases is poorly understood and no validated animal models is available to exlplore this issue. This study investigated the cognitive flexibility and hyperactivity of animals with TLE. Male Wistar rats at 25 days post‐natal were submitted to epilepsy model by systemic administration of pilocarpine and controls received saline 0.9%. Diazepam (7,5 mg/kg. sc) was administered 3h after the onset of status epilepticus to stop or reduce behavioral seizures. Behavioral tests began at 60 days postnatal. The animals were divided into two groups: control and experimental. Operant conditioning box was used to evaluate executive functions: hyperactivity and cognitive flexibilility. Experimental group showed higher activity (total of lever press) [F[1,24] =48,26, p=0,0001] and reduced percentage of correct response [F[1,24] = 20.15, p=0,0020] than control group. In summary, the results suggest that epileptic animals exhibit hyperactive behavior, reduced performance on task and preservated cognitive flexibility.

Supported by CNPq.
The Effects of Glyphosate Based Herbicides on Chick Embryo Morphology during Development
Authors: Winnick Blake , Dzialowski Edward M
Published: June 2021

Glyphosate based herbicides are among of the most widely used herbicides in the world. The purpose of this study was to determine developmental toxicity of glyphosate, the active ingredient in the common herbicide Roundup, on developing chicken embryos. Few studies have examined toxic effects of glyphosate alone versus the full compound formulations of Roundup, which include adjuvants and surfactants. Adjutants and surfactants are added to aid in solubility and absorption of glyphosate, possibly by making it less water soluble or lipophilic. In this study embryos were exposed on embryonic day 6 to 2% and 1% Roundup and 1% and 2% glyphosate soulutions. Injections were made directly into the air cell of the egg. On embryonic day 18 the embryos were sacrificed for morphometric measurements including: wet embryo mass, dry embryo mass, yolk mass, liver mass, heart mass, tibiotarsus length and beak length. Internal and external pipping was documented in eggs after embryonic day 18. Embryos treated with 2% Glyphosate and 1% Roundup showed significant reductions in wet and dry embryo mass, heart mass, liver mass, tibiotarsus length and beak length. Yolk mass was also significantly smaller in the Roundup animals. Mortality was also documented up to the time of external pipping. Embryos treated with 2% and 1% Roundup showed increased rates of mortality as compared with controls and active ingredient treatments.
Pathology of the Nile rat developing type 2 diabetes
Authors: Lyons Jeremiah , Brown Frances , Remillard Darryn E , Bolsinger Julia , Hayes KC
Published: June 2021

The Nile rat (NR) develops metabolic syndrome and type 2 diabetes (T2DM) in a manner similar to humans (NutrMetab 7: 29, 2010). To characterize the pathology of developing T2DM, relevant organs (pancreas, liver, kidney) were examined histopathologically. Immunohistochemical analyses of the pancreatic β cells were performed, as well. Fatty infiltration of the pancreas and hepatic steatosis were the most prevalent early lesions. Rats with advanced T2DM revealed a loss of pancreatic islet cells and insulin immunoreactivity. Hepatomegaly and discoloration were present macroscopically. This was confirmed as mid‐zonal to diffuse steatosis with microscopic foci of steatitis, accompanied by increased triglycerides and cholesterol esters. Hepatic steatosis was correlated with advancing T2DM, but was more pronounced in rats fed a modCHO/modFat diet compared to hiCHO/loFat rats, where clinical T2DM developed most rapidly. Selected chow‐fed diabetic rats >;1y of age with chronic hyperinsulinemia were found to have nodular hepatomas, not unlike non‐alcoholic steatohepatosis in humans. Histopathology of the kidney revealed the 4 stages of diabetic nephropathy described in humans, including early thickening of the glomerular basement membranes followed by thickening and increased interstitial fibrosis with end‐stage glomerular sclerosis.

Funding source: Foster Biomed Res Lab and the Malaysian Palm Oil Board.
Lipoprotein metabolism indicators improve cardiovascular risk prediction
Authors: de Graaf Albert A , van Schalkwijk Daniel B , Tsivtsivadze Evgeni , Parnell Laurence D , van der Werff‐van der Vat Bianca J.C. , van Ommen Ben , van der Greef Jan , Ordovas Jose M
Published: June 2021

We investigated whether lipoprotein metabolism indicators derived from a computational model (van Schalkwijk et al J Clin Bioinf 2011) could improve cardiovascular (CVD) risk prediction for 1981 subjects (145 cases, 1836 controls) from the Framingham Heart Study offspring cohort. We applied a statistical learning algorithm using a support vector machine to select conventional risk factors and lipoprotein metabolism indicators that contributed to predicting risk for general CVD. Improvement was quantified by the change in the Area‐Under‐the‐ROC‐Curve (ΔAUC) and by risk reclassification, using low, middle, and high risk categories (Net Reclassification Improvement, NRI), and a category‐independent method (Integrated Discrimination Improvement, IDI).

Two calculated VLDL lipoprotein metabolism indicators improved CVD risk prediction. Added to a multivariate model with the best performing Framingham risk markers, these markers significantly improved CVD prediction (ΔAUC=0·0177 p=0·0055), risk reclassification (NRI=0·090 p=0·014) and the category‐independent method (IDI=0·051 p<0·0001). The novel risk markers could reclassify 25% (p<0·0001) of all subjects, who were inappropriately classified in the ‘intermediate risk’ category by Framingham markers, to the low risk category.

Conclusion

two calculated VLDL metabolism indicators significantly improved CVD risk prediction.
Dimerization of G‐protein coupled Receptors (GPCRs) in Appetite Regulation and Food Reward
Authors: Schellekens Harriet , Dinan Timothy G , Cryan John F
Published: June 2021

Heterodimerization of GPCRs involved in appetite signalling may explain functional crosstalk observed between neuropeptide systems. The ghrelin (GHSR1a) and the serotonin 2C receptor (5‐ HT2C) have been implicated in homeostatic control of appetite and satiety and in neurocircuits involved in the rewarding aspects of food. Interestingly, serotonin has been shown to effectively block ghrelin's orexigenic actions and the 5‐HT2C receptor exerts a tonic inhibitory influence over dopamine neurotransmission. Moreover, ghrelin mediates rewarding effects of palatable foods via neuronal dopamine secretion. Thus, investigations into how such interactions occur at the molecular level are warranted and under investigation in this study. We show, to our knowledge for the first time, compelling evidence for the existence of a novel 5‐HT2C/D1 and 5‐HT2C/GHS‐R1a receptor heterodimer in vitro and ex vivo. Colocalization and agonist‐mediated co‐internalization was demonstrated in lentivirally‐transduced cells and endogenously in hypothalamic rat neurons. Interestingly, dimerization of the unedited 5‐HT2C‐INI, but not the edited 5‐HT2C‐VSV isoform, was shown to attenuate GHS‐R1a‐mediated signalling. Similarly, D1‐ induced cAMP signalling was reduced in cells co‐expressing both isoforms of the 5‐HT2C receptor. The 5‐HT2C‐mediated attenuation of both GHS‐R1a and D1 signalling was fully restored following pharmacological blockade of the 5‐HT2C receptor. These findings may uncover novel mechanisms important for the future pharmacological targeting of the GPCRs in the homeostatic regulation of body weight as well as hedonic appetite signalling, which both play a significant role in the development of obesity. The work was supported by Enterprise Ireland under Grant Number CC20080001.
Role of the Dopamine D3 Receptor Signaling Complex in the receptor Tolerance and Slow Response Termination properties
Authors: Kota Kokila
Published: June 2021

Dopamine is a predominant neurotransmitter and improper regulation of dopaminergic transmission results in disorders such as Parkinson's disease, schizophrenia and addiction. Dopamine receptors are G‐protein coupled receptors (GPCRs) and are classified as D1‐D5. Although the D3 receptor (D3R) shares high homology with D2, it possesses distinct signaling properties when repeatedly stimulated with agonist. The D3R exhibits two unique signaling properties, tolerance and slow response termination (SRT) to Dopamine and classical D2‐like receptor agonists. Tolerance is defined as a reduction in signaling response upon a second exposure to agonist and SRT is seen as a slow termination of the response upon agonist removal. GPCRs interact with a variety of proteins including but not limited to their cognate G‐protein, regulatory and modulatory proteins such as Regulator of G protein Signaling proteins, Kinases and Phosphatases. The molecular mechanisms underlying the D3R tolerance and SRT are unknown. We hypothesized that the D3R and D2R interacts with a different set of proteins and differences in this signaling complex contributes to the phenomena of Tolerance and SRT. Our results suggest that the D2R and D3R couple to different Gαi/o proteins, suggesting a role for the G proteins in the D3R tolerance and SRT properties. Sucrose density gradient experiments indicated that the molecular mass of the D3R is different in the tolerant and non‐tolerant states while Mass Spectrometry analysis identified specific proteins interacting with the D3R in its tolerant state.
Conserved serines of transmembrane 5 in the dopamine D1 receptor interact with non‐catechol D1 ligands
Authors: Lee Sang‐Min , Bhattacharya Supriyo , Nagarajan Vaidehi , Mailman Richard
Published: June 2021

Full dopamine D1 agonists have been shown to have excellent efficacy for Parkinson's disease (PD), equal to levodopa, the current gold standard therapy. Unfortunately, no full D1 agonist has been approved because all current full D1 agonists have poor oral bioavailability caused by a catechol group. We investigated the binding sites of non‐catechol D1 ligands using ergoline and tetrahydronaphthalene scaffold backbones to provide a structural basis for new drug design. Homology modeling suggests that several ergoline derivatives show a docking pose close to the conserved serines (S5.42, S5.43, and S5.46) in transmembrane 5 (TM5). Each of these serines was mutated to alanine (S5.42A, S5.43A, and S5.46A) to remove a potential hydrogen bonding site. As predicted, the S5.46A mutation significantly decreased the affinity of ergoline derivatives, and S5.43A mutation showed an equal or less decrease than S5.46 mutation. Conversely, the S5.42A mutation greatly increased the affinity of ergoline derivatives. The affinity of the tetrahydronaphthalene rotigotine was decreased by S5.42A or S5.43A, but not S5.46A indicating possible hydrogen bonding with S5.42 or S5.43. Together, these data suggest that the conserved serines in TM5 influence the interactions of non‐catechol D1 ligands in site‐ and scaffold‐dependent manners.
Effects of methadone, fentanyl and nalbuphine on intracranial self‐stimulation in rats: modulation by morphine exposure
Authors: Altarifi Ahmad , Negus S. Stevens
Published: June 2021

Facilitation of intracranial self‐stimulation (ICSS) is often interpreted as an abuse‐related drug effect. Initial exposure to the intermediate‐efficacy mu opioid agonist morphine primarily depresses ICSS; however, repeated morphine exposure reduces expression of ICSS rate‐decreasing effects and increases expression of abuse‐related ICSS facilitation. This study examined effects of morphine exposure on expression of abuse‐related ICSS facilitation produced by other opioids with different efficacies at mu receptors. Male Sprague Dawley rats with electrodes implanted in the medial forebrain bundle responded for a range of brain stimulation frequencies (56–158 Hz) during daily behavioral sessions. Effects of the high‐, intermediate‐ and low‐efficacy mu agonists methadone, fentanyl and nalbuphine were assessed during escalating regimens of morphine exposure (Vehicle, 3.2, 18 mg/kg/day). During vehicle treatment, methadone and fentanyl only decreased ICSS, whereas nalbuphine produced weak facilitation that was not dose dependent. Chronic morphine produced tolerance to rate‐decreasing effects of methadone and fentanyl and increased expression of ICSS facilitation by all three mu agonists. These results suggest that morphine exposure increases expression of abuse‐related facilitation of ICSS by mu agonists with a broad range of efficacies at mu receptors. Supported by R01 NS070715 and JUST
Effect of Phycocyanin on Cell Cultures Under Oxidative Stress
Authors: Cervantes‐Cervantes Martha Patricia , Jimenez‐Rodriguez Stephanie , Muñoz‐Sanchez Jose Luis
Published: June 2021

Cyanobacteria has been used as antioxidant in some toxicological models, as well as fluorescent markers and, but little is known about its role in oxidative stress protection. To analyze the effect of phycocianin (PC) on J774A.1 cell cultures, were exposed to an oxidative media generated by ascorbic acid in the presence of Cu(II), and in a control culture without exposure to ROS. After a period of incubation cell viability was obtained and total protein content was measured by Bradford assay; the electrophoretic pattern was obtained by SDS‐PAGE. Results showed that there are differences in the viability and these correlate to the total protein content found under oxidative stress conditions and different PC concentrations. Electrophoretic patterns suggest that the expression of some proteins depend upon the PC concentration added to the oxidative media; this result is related to the content of total protein. Supported by COFAA and EDD, IPN.
An Increase in Expression of the ER Stress Responsive Protein IRE1α Occurs in Response to the Anti‐HIV Drug Efavirenz in Primary Human Hepatocytes.
Authors: VanAusdall Jennifer Lynn , Bumpus Namandje
Published: June 2021

Efavirenz (EFV), one of the most commonly prescribed drugs for HIV, has been associated with liver toxicity; however the mechanism( s) for this is not well understood. In non‐stressed conditions, BiP (binding immunoglobulin protein) binds IRE1α (inositol‐requiring enzyme 1α), PERK (PKR‐like endoplasmic reticulum kinase), and ATF6 (activating transcription factor 6), rendering them inactive. BiP dissociates when ER stress is sensed, allowing these proteins to become activated. Immunoblotting revealed that primary human hepatocytes treated with 10μM of EFV expressed IRE1α at a 3‐fold increase at 6 hours, while no significant expression changes were observed at 3 and 24 hour time points. A commensurate increase in IRE1α mRNA levels was observed using reverse transcriptase PCR. The chaperone proteins calnexin and grp94, whose expression is upregulated by ER stress, demonstrated modest increases (1.5 fold) after 6 hours of EFV treatment, as did BiP (1.5 fold). No significant change in expression was observed in PERK or ATF6 with EFV treatment. Interestingly, results obtained from human samples were not recapitulated in primary mouse hepatocytes, despite being a conserved pathway, indicating a possible species difference. The results above suggest that EFV may exert its effects through the IRE1α pathway specifically, not by inducing the global unfolded protein response.
Physical interactions among NADPH‐cytochrome P450 reductase, CYP1A2, and CYP2B4 in the endoplasmic reticulum
Authors: Connick John Patrick , Reed James Robert , Backes Wayne L
Published: June 2021

In previous reports, our laboratory has shown that CYP1A2 and CYP2B4 form complexes in mixed reconstituted systems. Formation of the CYP1A2•CYP2B4 complex alters the function of these proteins, particularly in their ability to associate with NADPH‐P450 reductase (CPR). When in mixed reconstituted systems, the affinity of the CYP1A2 moiety of the complex for CPR was dramatically increased, demonstrating not only that CYP1A2 and CYP2B4 form complexes, but also that protein function is affected in the CYP1A2•CYP2B4 complex. To further explore the nature of these P450•P450 interactions, we used bioluminescence resonance energy transfer (BRET) to characterize the complexes in living cells. Transfection of GFP‐containing CYP1A2 and luc‐containing CYP2B4 into HEK293T cells led to a strong BRET signal, indicative of the CYP1A2•CYP2B4 complex. Next, we determined if the CYP1A2•CYP2B4 complex led to an alteration in CPR‐CYP binding by measuring the effect of unlabeled CYP1A2 on BRET for the CYP2B4‐GFP CPR‐luc pair. The results showed that CYP1A2 caused a dramatic disruption of the CYP2B4‐GFP•CPR‐luc complex. Conversely, CYP2B4 was unable to disrupt the CYP1A2‐GFP•CPR‐luc complex. These results are consistent with our model of CPR binding the CYP1A2•CYP2B4 complex and provide further evidence that functional P450•P450 interactions occur in living cells.