bluuebunny/crossref_metadata_2025 huggingface dataset:
Polyphenols suppress oxidative stress in bovine articular chondrocytes
Authors: Yagi Haruyo , Ulici Veronica , Tuan Rocky S
Published: June 2021

Reactive oxygen species (ROS) and nitric oxide (NO) have been implicated in chondrocyte senescence and cartilage aging, pathogenesis of osteoarthritis (OA) and rheumatoid arthritis. Naturally occurring polyphenolic compounds (polyphenols), such as curcumin (turmeric), resveratrol (grape), and epigallocatechin‐3‐gallate (EGCG) (green tea) have been known for their anti‐inflammatory and chondroprotective effects. However, the potential protective effects of these polyphenols against oxidative stress in chondrocytes are unclear. To investigate this, bovine articular chondrocytes were pre‐treated with polyphenols at varying concentrations, and then exposed to hydrogen peroxide (H2O2) as an ROS inducer or S‐nitroso‐N‐acetylpenicillamine (SNAP) as a nitric oxide (NO) donor. Alternatively, chondrocytes were co‐treated with polyphenols and H2O2. Intracellular ROS/NO were measured using a fluorescent dye technique (H2DCFDA for ROS; DAF‐FM for NO). Direct interaction between polyphenols and H2O2 was analyzed by quantitative peroxide assay. Our results showed that polyphenol pre‐treatment inhibited both H2O2‐induced ROS and SNAP‐induced NO in a dose dependent‐manner in chondrocytes. Polyphenol co‐treatment also inhibited both ROS and NO production in chondrocytes. Interestingly, our results indicate that all three polyphenols rapidly and completely blocked H2O2‐induced ROS without significant direct effect on H2O2 itself. Further, we found that only EGCG rapidly suppressed inducible NO production. These findings demonstrate that polyphenols are capable of suppressing inducible oxidative stress in chondrocytes, and suggest their potential therapeutic value for clinical application. Support: PA Commonwealth Dept of Health
Anthocyanins affect glutathione S‐transferase activity in human HepG2 cells
Authors: Galambos Alexis G , Fischer Joan G
Published: June 2021

Anthocyanins (ANTH) may act as antioxidants either directly or by increasing the activities of antioxidant enzymes and raising glutathione (GSH) concentration. Previous studies in our lab showed that rats consuming blueberry ANTH fractions had increased liver glutathione S‐transferase (GST) activity. Subsequent studies in HT‐29 colon cells showed that ANTH increase GST activity at low doses (< 10 μmol/L). The aim of this study was to examine possible protective mechanism(s) of ANTH by determining the effects of malvidin (MAL) and delphinidin (DEL) on antioxidant enzyme activity, cell viability and oxidative stress in human hepatocellular carcinoma (HepG2) cells. HepG2 cells were incubated in 75 cm2 flasks with 0, 5, or 10 μmol/L MAL or DEL for 24 h at 37°C, 95% CO2, 5% air, followed by incubation with 0–200 μM tert‐butyl hydroperoxide (t‐BOOH) for 2 h. Activity of GST was then assessed. t‐BOOH significantly (p = .049) increased GST activity above the control. ANTH increased GST activity (p=.058) above the control, with MAL having a greater effect (54%–84%) than DEL (29%). ANTH did not oppose the effects of t‐BOOH on GST activity. While t‐BOOH reduced cell viability, pre‐incubation with ANTH returned viability to that of the control. Further studies will examine the effects of MAL and DEL on glutathione peroxidase and glutathione reductase activities, GSH concentration and lipid peroxidation. Funding: GA‐AES.
Involvement of AMPK on the synergistic upregulation of phase II detoxifying enzyme activities by 1′‐acetoxychavicol acetate and sodium butyrate
Authors: Yaku Keisuke , Matsui-Yuasa Isao , Kojima-Yuasa Akiko
Published: June 2021

Introduction

1′‐acetoxychavicol acetate (ACA) naturally obtained from the rhizomes and the seeds of the Zingiberaceae plant such as Languas galangal and Alpinia galangal. Butyric acid is one of major products of gut fermentation. Though ACA and sodium butyrate respectively upregulated phase II enzymes in intestinal epithelial cells (IEC 6), the interaction between these compounds is not clear. The present study was conducted to examine the interaction between ACA and sodium butyrate on regulation of phase II enzyme activities in IEC 6.

Methods

Glutathione S‐transferase (GST) activity and NAD(P)H:quinine oxidoreductase 1 (NQO1) activity were measured by spectrophotometer. Nrf2 and p53 mRNA levels were estimated by RT‐PCR. Nrf2, p53, AMPK protein levels were examined by Western blotting.

Results

ACA and sodium butyrate synergistically induced GST and NQO1 activities. These synergistic effects were inhibited with AMPK inhibitor, Compound C. Sodium butyrate upregulated the expression of Nrf2 mRNA and downregulated that of p53 mRNA, but ACA didn't affect those mRNA levels. The protein levels of intranuclear Nrf2 were increased by ACA or sodium butyrate. However, the combined treatment of ACA and sodium butyrate did not caused a synergic effect on level of intranuclar Nrf2. Intranuclar p53 protein level was increased by ACA, but It was decreased by sodium butyrate alone or by combined treatment of ACA and sodium butyrate. pAMPK levels were increased by combined treatment of ACA and sodium butyrate.

Conclusion

AMPK plays an important role on synergistic upregulation of phase II enzyme activities by ACA and sodium butyrate.

Grant Funding Source : ASN
Hepatoprotective effects of Rubus coreanus Miquel concentrates on liver injuries induced by carbon tetrachloride in rats
Authors: Chyun Jong-Hee , Chae Hyun-Jung , Yim Jung-Eun
Published: June 2021

As well‐being foods pursuing healthy life are getting popular, and interests for Rubus coreanus Miquel (RCM) fruit, kind of Korean blackberry, are heightening as its medical actions, which are protecting liver, brightening eyes, and in curing diabetes, are known. This study was carried out to evaluate hepatoprotective effects of RCM concentrates on liver injuries induced by carbon tetrachloride (CCl4) in rats.

RCM, produced in June~July 2008 at Chunbook, Gochang, mashed fine. The seeds of RCM were removed and the juices were condensed. 32 Sprague‐Dawley rats were divided into normal group, CCl4 group (8 rats), 1% RCM group, and 2% RCM group. Experimental diets were provided to experimental animals for four weeks. We measure blood total cholesterol, HDL‐cholesterol, Glutamic oxaloacetic transaminase (GOT), Glutamic pyruvic transaminase (GPT), and Alkaline phosphatase (ALP). Part of the livers was separated for hitopathological observation, and measured lipid peroxide (TBARS) and liver protein.

The activities of serum GOT, GPT and ALP were elevated by CCl4 administration and levels of hepatic TBARS were also significantly increased in CCl4 groups. However, the hepatic TBARS levels and activities of serum enzymes were markedly reduced by supplementation of the RCM concentrates (p<0.05). In histopathological observation, massive necrosis in centrilobular area and degenerative changes by CCl4 were recovered by dietary supplementation of RCM concentrates.

These findings indicate that RCM concentrates have hepatoprotective effects and may improve symptoms of liver injuries.
Effect of oregano essential oils on lipid oxidation in storage eggs enriched with dietary omega‐3 fatty acids
Authors: Ariza-Nieto Claudia , Ortiz Ronnal , Vasquez Diana , Mayorga Olga , Afanador German
Published: June 2021

Oregano plants produce secondary metabolites, which they are utilized by the plant for growth and reproduction, but they are useful as functional additives in animal feeding systems because their intrinsic bioactivities properties. Oregano antioxidant property is attributed to thymol and carvacrol that react with lipid and hydroxyl radicals converting them into stable products. The aim of this study was to evaluate the effect of oregano essential oils (OEO) on lipid oxidation of n‐3 PUFA‐enriched fresh eggs and storage at 4 ºC y −10 ºC during six months. A total of 160 laying hens 40 wk old were placed in 80 cages and were randomly assigned to one of the 8 experimental diets: palm oil and fish oil (n‐3 PUFA source), with and without supplementation of OEO at 4 qualitative thymol levels. Results showed that lipid oxidation was greater in eggs storage at 4ºC (3.09 mg MDA/g). In n‐3 PUFA‐enriched fresh eggs the lowest lipid oxidation was shown by the group supplemented with OEO low thymol (1.27 mg MDA/g). Lipid oxidation increased in n‐3 PUFA‐enriched eggs stored at 4ºC and −10ºC compared to fresh eggs (P<0.001). At 4ºC greater lipid oxidation was shown in enriched eggs without OEO (5.6 mg MDA/g), but OEO supplementation lowered lipid oxidation around 48%. In the present study, dietary OEO supplementation positively influenced by retarding lipid oxidation in eggs enriched with dietary omega‐3 fatty acids during storage.
Anthocyanins‐rich purple corn extract alleviated diabetesassociated inflammatory glomerulosclerosis and monocyte macrophage infiltration: Role of interleukin‐8
Authors: Kang Min-Kyung , Kang Young-Hee
Published: June 2021

Purple corn has been utilized as a daily food and exerts disease‐preventive activities. This study elucidated that anthocyanins‐rich purple corn extract (PCE) prevented glomeruloscleosis. Human endothelial cells and THP‐1 monocytes were treated in conditioned media of mesangial cells experienced to 33 mM high glucose (HG). PCE decreased expression of VCAM‐1 and E‐selectin and monocyte integrins of β1 and β2 induced in HG‐conditioned media. PCE demoted HG‐induced mesangial secretion of interleukin (IL)‐8, demonstrating its inhibition of monocyte adhesion. PCE attenuated monocyte adhesion and infiltration through blocking pro‐inflammatory signaling of Tyk2 and STAT1/STAT3 activated by IL‐8. db/db mice were treated with 10 mg/kg PCE for 8 weeks. PCE alleviated mesangial expansion of diabetic kidney. It was revealed that in kidney glomerulus PCE attenuated induction of the leukocyte adhesion key factors of intracellular cell adhesion molecule‐1 and CD11b. In addition, PCE diminished the levels of the macrophage makers of CD68 and F4/80 in the mesangial interstitum. These results demonstrate that PCE antagonized monocyte infiltration and accumulation possibly through mesangial inflammation of IL‐8 signaling due to chronic hyperglycemia. Therefore, PCE may be potent therapeutic agent for the prevention and treatment of diabetes‐associated glomerulosclerosis accompanying kidney dysfunction.
Anti‐Inflammatory Action of Black Garlic through downregulation of NF‐κB activation and MAP kinase phosphorylation
Authors: Oh Hye Lim , Kim Min Jee , You Bo Ram , Kim Mee Ree
Published: June 2021

In this study we demonstrated that garlic inhibits the inflammatory mediators and the active compounds are different from those isolated from garlic or aged garlic. Lipopolysaccharide‐induced nitric oxide (NO) production was determined by Griess method. The DNA binding activity of NF‐κB was investigated by electrophoretic mobility shift assay. NF‐κB and inducible nitric oxide synthetase (iNOS) transcriptional activity was determined by luciferase assay. Expression of iNOS and cyclooxygenase‐2(COX‐ 2) was determined by western blot. The anti‐inflammatory effects of garlic or aged garlic in in vivo were investigated in Lethality studies inflammatory models. These previous observations, the LPS‐stimulated RAW cells caused phosphorylation of the ERK, JNK and p38 phosphorylation in LPS‐stimulated RAW 264.7 cells, suggesting that MAP kinases are involved in the inhibition of LPS‐stimulated NF‐κB binding by garlic in RAW 264.7 cells. The results discussed above demonstrated that garlic and aged garlic exhibited antagonistic activity in a cellular system. We examined the effects of garlic and aged garlic LPS‐induced serum IL‐1β, IL‐ 6 and TNF‐α production. In conclusion, the results of the present study provide the first evidence that garlic inhibits LPS‐induced NO, PGE2 and cytokine (TNF‐α) production. These inhibitory effects of garlic and aged garlic were found to be associated with NF‐κB and MAP kinase signaling, suggesting a possible approach to the treatment of inflammatory diseases.
Suppressive effect of Petasites japonicas extract on TPA‐induced ear edema in mouse model
Authors: Seo Eun Young , Kim Hyunae , Kim Mee Ree
Published: June 2021

Inflammation is a beneficial host response to foreign pathogens or tissue injury, and it eventually leads to the restoration of normal tissue structure and function. Petasites japonicas (Pj) is known as an herb for treating asthma, oxidant stress and gastric ulcer in traditional Oriental medicine. In this study, we evaluated the anti‐inflammatory effects of Pj extract on 12‐O‐tetradecanoylphorbol 13‐acetate (TPA)‐induced ear edema in mouse model. The Pj extract (10, 30, 50 mg) was acquired from whole plants of Petasites japonicas using butanol. After the PJ extract was administered to mice via gavage consecutively for 5 days, an edema was induced on the ear by topical application of 2.5 μg/ear of TPA dissolved in 10 μl of acetone. Ear thickness was measured using a Digimatic Micrometer (Mitsutoyo Co., Tokyo, Japan) before TPA application and 4 h after. ELISA was performed to determine the expression levels of cytokines. The levels of ear thickness were not significant between normal group and experimental groups, However, the ear weight in group Pj 10 was lower than that in TPA‐only group, and similar to that in normal group. TPA increased the expression of pro‐ inflammatory cytokines. The Pj extract significantly inhibited the release of interleukin (IL)‐ 1β, IL‐6 and tumor necrosis factor (TNF)‐α, when compared to TPA only. Especially, Pj at relatively low doses, reduced the level of pro‐inflammatory cytokines dose‐dependently. From present results, it was suggested that the extract of Petasites japonicas showed anti‐inflammatory action in TPA‐ induced ear edema mouse model.
Effects of aromatic alcohols on inflammatory response in RAW 264.7 cells
Authors: Jordan Chad , Harris Gabriel Keith
Published: June 2021

The aromatic alcohols (AOH) tyrosol (TY), tryptophol (TRY), and 2‐phenylethanol (2PE) are signaling molecules secreted by yeast and bacteria and consumed in fermented products. Since the majority of the immune system resides in the intestinal tract, AOH may influence immune signaling. This study examined effects of AOH on lipopolysaccharide (LPS) and interferon‐γ (IFN‐γ) induced inflammation 18 and 30 hours post induction in RAW 264.7 mouse macrophage cells. Cell viability, Prostaglandin E2 (PGE2), and Nitric Oxide (NO) were assessed using MTT assay, ELISA, and Griess reagent, respectively. At 18 hours post induction, TY and TRY inhibited PGE2 production by 86% and 92%, while 2PE increased PGE2 production by 32%. At 30 hours post induction, PGE2 production was inhibited by 83% and 92% by TY and TRY, while PGE2 production was increased 37% by 2PE. PGE2 comparisons were highly significant (p < 0.001). At 18 hours post induction, TRY inhibited NO production by 16% (p<0.05), while TY and 2PE showed no effect. At 30 hours post induction, NO production was inhibited 13% by TRY (p < 0.001) and increased 16% by 2PE (p < 0.001) while TY showed no effect. TRY also resulted in more typical cell morphology compared to TY and 2PE. TY and TRY showed anti‐inflammatory activity by inhibiting PGE2 production, but only TRY reduced NO production. 2PE showed immune stimulatory activity by increasing NO and PGE2 production.
Liuwei Dihuang Aqueous Extract Reduces Weight Gain in Obese‐Prone Rats through Multi‐Mechanisms
Authors: Wang Yanwen , Nair Sandhya , Zhang Junzeng , Sun Changhao
Published: June 2021

Liuwei Dihuang (LWDH), a traditional Chinese herbal formulation, has been demonstrated to reduce weight gain in obese rats; however, the active components and mechanisms are unknown. In the present study, water extract of LWDH was obtained and determined for its effect and mechanism of action on weight gain in male obese‐prone CD rats. The rats were divided into three groups and fed a high‐fat diet (60 kcal% from fat). Two treatment groups received 600 (WE600) or 1200 (WE1200) mg/kg/d LWDH water extract dissolved in water once a day via gavage feeding for 10 wk. The control rats were gavaged with the control vehicle. It was found that WE1200 lowered body weights after 3 weeks of treatment and the effect was retained throughout the remaining study period. WE1200 also lowered visceral fat mass, serum free fatty acids, serum leptin, as well as blood cholesterol and triacylglycerides. Oxygen consumption, carbon dioxide production and fat oxidation were increased in both light and dark periods, while carbon dioxide oxidation increased in the light period in the WE1200 group. Energy expenditure was increased in the WE1200 group in both light and dark periods. Moreover, rats in the WE1200 group had lower levels of serum free fatty acids and leptin levels. Rats in the WE600 had lower serum free fatty acids and leptin levels without showing significant effects on other parameters compared to controls. These results demonstrated that consumption of LWDH water extract suppressed weight gain, visceral fat mass and improved several phenotypes of metabolic syndrome in obese rats through increasing energy expenditure, decreasing energy intake and improving leptin sensitivity. ‐‐‐‐Research was supported by CIHR grant CCI‐92219.
Anti‐atherosclerotic property of emu oil in diet induced obese rats
Authors: Pichiah P.B. Tirupathi , Cha Youn-Soo
Published: June 2021

The present study was aimed to test the oil obtained from Emu bird Dromaius novaehollandiae for its anti‐atherosclerotic activity in diet induced obese (DIO) rat. Animals were divided into five groups randomly, normal diet group (ND), high fat diet group (CD), high fat diet plus emu oil 2.0 ml/kg BW (ED1), high fat diet plus emu oil 4.0 ml/kg BW (ED2), high fat diet plus emu oil 8.0 ml/kg BW (ED3). Oral treatment for 12 weeks was carried out. Body weight, liver and adipose tissue weight were measured, lipid profiles, C‐reactive proteins (CRP) followed by histopathology of thoracic aorta were carried out to confirm the anti‐atherosclerotic effect of emu oil. Sex hormones such as testosterone and luteinizing hormone were also measured. Results obtained revealed that emu oil supplementation reduced the lipid profiles and modulated the hormonal profiles, followed by reduced formation of atheromatous plaque or thickening of thoracic aorta, also there was a significant decrease in the body weight along with liver and adipose tissue weight indicating the significant anti‐atherosclerotic activity of emu oil. To conclude, we believe that high content of oleic acid could have orchestrated either solely or in combinations with other fatty acids such as linoleic and linolenic acids present in the emu oil towards up regulating testosterone biosynthesis and averting the atherosclerosis formation in diet induced obese animals.
Gene expression profiles of the colonic mucosa associated with phenotypic changes in mice fed high‐fat diet
Authors: Bae Yun Jung , Bak Youn-Kyung , Park Taesun , Choi Myung-Sook , Kim Jeongseon , Sung Mi-Kyung
Published: June 2021

Obesity is closely associated with pathological changes of the intestine including colon cancer, however little is known for possible molecular mechanisms. The objective of this study was to select candidate target genes which may be associated with abnormal cell growth and obesity‐related phynotipic biomarkers in the colon of mice fed high fat diet. C57BL/6J male mice were fed normal (ND) or high‐fat diet (HFD) over 8 weeks and sacrificed. Gene expressions of colon tissue were determined and those showing over 2‐fold changes in their signal intensity were analyzed for their correlations with obesity‐associated phenotypes including circulating adipokines. Mice fed with HFD exhibited significantly increased plasma leptin level compared with mice fed with ND. Expressions of Ces1 (carboxylesterase 1), Aldh1a1 (aldehyde dehydrogenase family 1) were down‐regulated while that of Gm5222 was up‐regulated in the colon tissue of HFD fed mice. The circulating leptin was negatively correlated with the expression of Aldh1a1 and Ces1. Aldh1a1 has been reported as a gene coding for a cytosolic enzyme involved in cell differentiation process, and therefore the down‐regulation of this gene may play a role in abnormal cell growth. [This research was supported by the SRC program (Center for Food & Nutritional Genomics: 2011‐0000912) of the NRF of Korea funded by the Ministry of Education, Science and Technology.]
Differential Effects of Added Dietary Fructose or/and Fat on Adipose Tissue Gene Expression in a Juvenile Pig Obesity Model
Authors: Solano-Aguilar Gloria Isabel , Jang Saebyeol , Laskman Sukla , Molokin Aleksey , Bruna Gonzalo , Mitchell Alva , Urban Joseph
Published: June 2021

Changes in energy intake affects metabolic function in adipose tissue that plays a central role in lipid and glucose metabolism and the production of hormones and inflammatory cytokines. To determine the effects of excess calories from fructose and/or fat on adipose tissue and liver gene expression, 32 conventional crossbred juvenile pigs were randomized and fed 1) a basal calorie diet alone or 2) with added fructose (20% of total calories); or 3) a high calorie diet with added fat alone or 4) with added fat and fructose for 12 wk. Total body fat deposition detected by DXA‐scan and carcass digestion, and gene expression in liver and visceral and peripheral adipose tissue was measured. The results indicated that both high fat diets induced a significant body weight gain and fat deposition with expression of inflammatory genes in liver and visceral adipose‐derived stromal cells. Pigs on the basal diet with added fructose showed a marginal gain in bodyweight with no difference in fat deposition or induction of inflammation when compared to pigs on the basal diet. However, the combination of added fructose and fat induced expression of lipogenic genes (fatty acid binding protein (FABP4), stearoyl‐CoA desaturase (SCD), and leptin in adipocytes isolated from peripheral fat. Thus, excess dietary fructose combined with a high fat diet can exacerbate fat deposition and body weight gain by promoting lipidogenesis.
Whey based formula reduces intestinal atrophy and enhances antioxidant defenses against sepsis
Authors: Tsutsumi Rie , Sasaga Sayaka , Takegawa Mariko , Sakai Tohru , Nakaya Yutaka , Tsutsumi Yasuo M.
Published: June 2021

High quality whey protein has been shown that improve anabolism, GI tolerance, and antioxidant defenses. In this study, we assess the effect of whey based formula during the acute phase in the cecal ligation and puncture (CLP) mousemodel of sepsis. Male C57Bl/6 mice began whey based or casein based formula following induction of sepsis by CLP. Between 48 h to 7 days of sepsis, survival rate, intestinal histopathology, intestinal atrophy, antioxidant defense and serum levels of cytokines were assessed. Analyses were performed with StatView 5.0 (SAS Institute Inc., Cary, NC). Data are presented as mean±standard deviation (SD). P<0.05was considered statistically significant.

Serum cytokines were higher in mice that underwent whey based formula vs control (TNFα;34.3±6.54 vs.67.6±8.93pg/ml, IL6;230+37 vs.443+49 pg/ml after 48 h induction, respectively). Whey‐based group showed re‐established total hepatic glutathione and glutathione disulfide (GSH/GSSG)levels. Whey‐based formula significantly improved intestinal atrophy which was measured by diamine oxidase (DAO) activity. Our data indicate that administering whey‐based formula after sepsis has several benefits by decreasing cytokines, increasing antioxidants and reducing intestinal atrophy.
MicroRNA‐124 and its target gene are altered in the substantia nigra (SNc) of the brain of MPTP‐mouse model of Parkinson's disease
Authors: Kanagaraj Nandhini , Dheen S Thameem , Peng Zhao Feng , Srinivasan Dinesh Kumar , Tay Samuel S W
Published: June 2021

MicroRNAs (miRNAs) are small, non‐protein coding RNAs involved in post‐transcriptional gene expression regulation. Several recent studies have demonstrated the importance of miRNAs in diseases of the nervous system. Parkinson's disease (PD) is the second most common neurodegenerative disease characterized by the progressive loss of dopaminergic neurons in the substantia nigra (SNc) of the brain. The involvement of a few miRNAs in PD has been proven to be a promising potential for therapeutic targeting. The brain‐enriched miR‐124 has been shown to play a role in plasticity, inflammation and stress response in the brain. We used the 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridinium (MPTP)‐ induced mouse model to study if miR‐124 has any important role in PD pathogenesis and whether it can offer a new therapeutic target. We evaluated the expression of miR‐124 in the SNc of the PD model at different time points by Real‐time PCR. A significant decrease in expression of miR‐124 over time was observed. We also found an increase in the levels of NeuroD1 which has been shown to be a target of miR‐124. Preliminary data from the dopaminergic neuronal (MN9D) cell line treated with MPP iodide also showed a similar pattern. In summary, miR‐124 depletion plays an important role in the pathogenesis of PD. The exact mechanism involved remains to be elucidated. Supported by Grant MOE 2009‐T2‐1‐061 from the Ministry of Education.
Comparison of sterculic oil induced changes in the gut microbiota of lean and obese mice
Authors: Ghosh Shreya , Perfield James W. , Saikaly Pascal E. , Oerther Daniel B.
Published: June 2021

Recent obesity research has reported differences in intestinal microbiota between obese ob/ob mice and lean wild type mice suggesting a potential link between bacterial populations and metabolic health. Sterculic oil (SO) is a natural inhibitor of stearoyl‐CoA desaturase‐1, a potential therapeutic target for the treatment/attenuation of obesity and insulin resistance.

The aim of this study was to investigate the response of the obese and lean gut microbiota to a SO supplemented diet.

Five‐week old male lean (WT) and obese (ob/ob) mice were fed either a control AIN‐93G diet or an AIN‐93G diet containing 0.5% SO for 9 weeks Barcoded pyrosequencing of the V4‐V5 region of the 16S rRNA gene was employed to characterize cecal microbiota.

Intestinal microbiota from WT mice fed a control diet were more diverse, although there were no significant differences in the proportions of major phyla from WT and ob/ob mice. Irrespective of genotype, SO was negatively correlated with the relative abundances of Actinobacteria, Bacilli and Erysipelotrichi. This shift was not associated with a change in body weight, however a statistically significant improvement in glucose tolerance and insulin sensitivity was observed in the ob/ob mice fed SO. In our study, diet had a greater impact on gut microbiota as compared to genotype and these changes were associated with an improved metabolic health.

Grant Funding Source : Missouri University of Science and Technology
Initial lesions of the elastic fibers and extracellular matrix in varicose veins: an inmunohistochemical and confocal microscopy study
Authors: Regadera Javier , Prachaney Parichat , España Gabriel , Condezo-Hoyos Luis , Rubio Maria , Gonzalez Maria C , de Pablo Angel Luis Lopez , Arribas Silvia M
Published: June 2021

Chronic venous insufficiency (CVI) has high morbidity in middle and advanced age population. We studied varicose vein alterations in early stages to identify possible therapeutic targets. Saphenous veins from 10 patients were classified according to Color Doppler in proximal segment with lesions (Varicose vein, VV) and distal (normal vein, NV). We analyzed extracellular matrix (ECM), elastic fibers, collagen I and IV and SMC in sections stained with Masson trichrome, orcein, sirius red and collagen I and IV antibodies. Elastin was studied with confocal microscopy in intact veins (autofluorescence). Compared to NV, VV intima shows multifocal areas of endothelial denudation, deposition of powdery collagen and elastic material and short and thin elastic fibers. VV media has SMC atrophy, increased immature ECM deposition with few and small collagen I fibers and incomplete elastogenesis. Adventitia is thick with abundant mature collagen I and deposition and increased thick and irregularly distributed elastic fibers. Some areas of the distal NV segment also exhibited some of these abnormalities. We conclude that initial multifocal degeneration of adventitial elastic fibers, intimal elastogenesis and progressive atrophy of SMC are processes perpetuating CVI and irreversible lesions. Funding: Universidad Autónoma de Madrid (UAM/32).
Potentially protective dual oxidase enzymes (Duox1 and Duox2) in the normal murine and bovine reproductive tracts
Authors: Adu-Addai Benjamin , Mackenzie Charles D , Langerveld Anna J , Agnew Dalen W
Published: June 2021

Vaginal and uterine mucosae are portals for sexually transmitted diseases (STD) in both humans and animals. Understanding innate immunity at mucosal surfaces is essential to enhancing protection and controlling STD infections. The recent discovery of the dual oxidases (Duox1/Duox2) capable of H2O2 associated killing of microbes (Harper et al 2008) in different mucosal membranes suggests that these enzymes may be important in the innate defenses of the reproductive tract through the associated action of ROS and cytokines. Dual Oxidase 1 and 2 (Duox1 and Duox2) have been well studied in the respiratory and the digestive tract in multiple species, but no report concerning the reproductive tract has been made so far. This current study addresses the role that Duox 1 and Duox 2 may play in the reproductive tract of mouse and the cow through immunohistochemistry (IHC) and rtPCR, and by examining the production of Duox1 and 2 in vitro during acute infection of cultured bovine endometrial cells with T. foetus using IHC and PCR. Ovaries, oviduct, uterine mucosa, vagina, and intestine were collected in RNAlater from cattle and mice for analysis of Duox1 and 2 gene expression. For IHC, samples were labeled with Duox1 and 2 monoclonal antibodies and examined for specific binding. qPCR results indicated expression in all reproductive tissues of both mouse and cow with highest concentration in the vagina. Expression was also confirmed in both bovine trophoblast and endometrial cell lines. When bovine endometrial cells were infected with T. foetus for 6 hours, however, there were no significant changes in Duox 1 and 2 levels, suggesting that if these molecules play a role in innate immunity, it may either be earlier or later than 6 hrs post infection studies here. This work was supported financially by the CVM MSU, Endowment Sterner Fund.
Reduction in the Intestinal Epithelial Glutathione Redox Potential May Regulate Proliferation in the Neonatal Murine Gut
Authors: Richardson Arena , Myers Loren S , Song Shuh-Chyung , Ray Laurie , Berardinelli Andrew , Hansen Jason , Denning Patricia Wei
Published: June 2021

The intracellular redox potential of the glutathione (GSH)/glutathione disulfide (GSSG) couple regulates many cellular processes. In vitro studies indicate that a reduced GSH/GSSG redox potential favors proliferation while an oxidized redox potential favors differentiation. Developmental regulation of intestinal growth and adaptation depends upon an appropriate balance between the two. However, how the ontogeny of intestinal epithelial cellular (IEC) GSH/GSSG redox regulates these processes in the developing intestine has not been characterized in vivo. IEC GSH/GSSG redox potential becomes increasingly reduced (primarily driven by a more than 4‐fold increase in IEC GSH concentration) over the first 3 weeks of life in all regions of the murine intestine with the biggest change occurring between 1 and 2 weeks. These changes are accompanied by an increase in intestinal growth and IEC proliferation as assessed by intestinal length, villus height/crypt depth, and Ki67 staining. Understanding how IEC GSH/GSSG redox potential is developmentally regulated may provide insight into how premature human intestinal redox states can be manipulated to optimize intestinal growth and adaptation.
Effects of claudins‐6, ‐7 or ‐9 overexpression on migration and invasiveness of breast and hepatocellular cancer cells
Authors: Salas Berenice , Rendón-Huerta Erika P. , Montaño-Estrada Luis F.
Published: June 2021

Transmembrane proteins claudins preserve cell polarity and determine the size and charge of the molecules that pass through the paracellular space in epithelial tissues. Altered claudin expression in early and late stages of several cancers has been related to increased invasiveness and metastatic potential. We have reported an expression of claudin‐6, ‐7 and ‐9 in human gastric adenocarcinomas and their role in the human adenocarcinoma cells invasiveness. To determine if claudin overexpression confere advantages to breast and hepatocellular cancer cell lines, we evaluated the invasive capability of claudins transfected cells in Matrigel invasion chambers and by wound‐healing scratch assay. Proliferation rate by Brdu incorporation, and apoptosis of TNF treated cells by Anexin V detection were also measured. Overexpression of claudin‐6 or ‐9 resulted in increased migration and invasive potential; however claudin‐9 transfected cells migrated faster than the others.

This work was supported by PAPIIT IN208910‐2.
Leptin Increases HER2 Stability through HSP90 in Breast Cancer Cells
Authors: Giordano Cinzia , Vizza Donatella , Rovito Daniela , Barone Ines , Bonofiglio Daniela , Panza Salvatore , Lanzino Marilena , Fuqua Suzanne , Catalano Stefania , Andò Sebastiano
Published: June 2021

High serum levels of the adipocyte‐derived hormone leptin in obesity are a risk for breast cancer in postmenopausal women. The HER2/Neu receptor, overexpressed in 25% of human breast cancers, has been associated with endocrine resistance.

Here, we evaluated the link between leptin and HER2 in human MCF‐7 breast cancer cells and in MCF‐7/HER2‐18 cells stably overexpressing HER2.

We found that leptin treatment increased HER2 protein levels, without affecting HER2 mRNA levels. Leptin enhanced the expression of HSP90, a chaperone protein that regulates HER2 degradation, and increased HER2/HSP90 interaction. These events were dependent on JAK2/STAT3 activation, since inhibition of this cascade by AG490 or a STAT3 dominant negative abrogated leptin‐induced HER2 expression. Moreover, leptin treatment increased EGF‐mediated HER2, AKT and MAPK phosphorylation. Anchorage‐independent growth assays revealed that long term leptin treatment reduced sensitivity to the antiestrogen tamoxifen, while increased responsiveness to herceptin, a monoclonal antibody against HER2.

We demonstrate that leptin, through HSP90, enhances HER2 stability, providing further insights into the crucial role of leptin in modulating tumor growth and aggressiveness. Clinically, our results suggest that leptin and HSP90 might be novel targets for therapeutic intervention in breast cancer, especially in obese patients.
Chronic Heart Failure is Associated with Elevated Skeletal Muscle Inflammation and Toll‐Like Receptor 4 Signaling
Authors: Timmerman Kyle L , West Jeremiah N , Markofski Melissa M , Dhanani Shaheen , Rasmussen Blake B , Ckoksi Kashyap B , Barbagelata N Alejandro , Volpi Elena
Published: June 2021

Chronic heart failure (CHF) is associated with increased inflammation. As inflammation impairs skeletal muscle protein maintenance, elevated skeletal muscle inflammation may increase the risk of developing CHF‐associated cachexia. Elevated expression and activation of the innate immune receptor, toll‐like receptor 4 (TLR4) may contribute to skeletal muscle inflammation. The purpose of the present study was to compare skeletal muscle expression of TLR4, phosphorylated‐IκBα (a marker of TLR4 activity), and the inflammatory cytokines IL‐6 and TNF‐α between CHF patients and healthy, age‐matched controls. Biopsies were taken from the vastus lateralis of CHF patients (CHF, N=8) and healthy controls (CON, N=8), and Western blotting was used to analyze skeletal muscle TLR4, phosphorylated‐IκBα, IL‐6, and TNF‐α. Additionally, serum levels of IL‐6 and TNF‐α were measured using enzyme linked immunosorbent assays. Compared to CON, CHF exhibited elevated skeletal muscle expression of phosphorylated‐IκBα and TNF‐α. There were trends for skeletal muscle TLR4 (P=0.15), serum IL‐6 (P=0.10) and serum TNF‐α (P=0.09) to be elevated in CHF compared to CON. These preliminary data show that CHF patients exhibit greater skeletal muscle inflammation than healthy controls, and that this difference may be partially explained by elevated TLR4 signaling. Funding: P30 AG024832.
Calcium channel blocking drugs ameliorates the experimental septic cardiomyopathy induced by cecal ligation and puncture in mice
Authors: Celes Mara Rubia N , Prado Cibele M , Campos Erica C , Malvestio Lygia M , Ferezin Patricia , Freitas Ana C.S. , Dias Priscila P. , Tanowitz Herbert B. , Rossi Marcos A
Published: June 2021

Objective

This study tested the hypothesis that verapamil (VP), an L‐type voltage‐dependent calcium channel inhibitor, and dantrolene (DT), a sarcoplasmic reticulum calcium inhibitor, could attenuate calpain overexpression, dystrophin loss/reduction and cardiac dysfunction in experimental sepsis by CLP.

Methods and Results

Male C57Bl/6 mice were subjected to sham and severe septic injury (SSI) induced by CLP; half of animals from each group were treated with either verapamil (5mg/kg, SSI+VP) or dantrolene (10mg/kg, SSI+DT). Reduced myocardial levels of NF‐kB and calpain‐1 were detected in SSI+DT and SSI+VP mice associated with increased actin and myosin expression as compared to SSI mice. Consonantly, DT and VP significantly prevented the loss of dystrophin in comparison with SSI mice. Concurrently, SSI+DT and SSI+VP mice presented ejection fraction and fractional shortening improvement associated with increased survival rate.

Conclusions

These observations give support to the opinion that increased intracellular calcium concentration induces calpain activation that causes dystrophin loss/reduction and consequent myocardial dysfunction in CLP experimental sepsis. Studies are needed to further elucidate these new possible interventional pathways to prevent septic cardiomyopathy/heart failure.
Escherichia coli (E. coli) antihistone activity (AHA) can modify the chromatin organization in rat testicular germ cells
Authors: Loginova Anastasia K. , Plotnikov Andrey O. , Nemtseva Natalia V. , Klimushkin Alexey V. , Stadnikov Alexander A.
Published: June 2021

This study examines the hypothesis that a testicular infection with the B120 strain of E. coli positive for AHA (+) can alter germ cell‐specific chromatin organization. In two experimental groups, the adult male albino rats received a single 0.1 mL intratesticular injection of bacterial suspension containing 5·108 E. coli/mL in sterile saline as follow: 1) AHA+ and 2) AHA− clones. Control rats were injected with saline only. The paraffin‐embedded histological sections of testes were examined on days 3 and 7 with H&E staining, the Feulgen reaction for DNA, and the Alfert‐Geschwind technique for histones. The clones of E. coli that survived in the testes were assessed for their AHA level. We found that after 3 days in the testes AHA of the AHA+ clones was reduced from 6.8% to 2.0%, whereas in the AHA− clones it was increased from 0.0% to 1.8%. At the same time, both experimental groups have demonstrated similar reactive changes in spermatogenic cells 7 days post‐injection, which included chromatin decondensation and its translocation to the nuclear periphery, as well as the presence of numerous multinucleated cells. Moreover, the staining for histones revealed their degradation in germ cells (based on the intensity of intranuclear staining on a scale from 0 to 3 per 100 cells): 0 (27%), 1 (46%), 2 (27%) and 3 (0%). Therefore, we concluded that the AHA+ clones of E. coli are able to change the genetic apparatus of testicular germ cells.
Immune brinksmanship: a conceptual model of endogenous stressors of the acute‐phase response
Authors: LeGrand Edmund Kenwood , Alcock Joe
Published: June 2021

Many features of the acute‐phase response, including fever, nutrient restriction, and anemia, are stressors that cause self‐harm, affecting both the host and the pathogens contained within. However, pathogens and infected/neoplastic cells are additionally subjected to the stressors of local inflammation and the vulnerability associated with proliferation/growth and invasion. Therefore, additional non‐specific stress will typically harm the pathogens relatively more than the host. The immune brinksmanship model of the acute‐phase response proposes that hosts evolved multiple endogenous stressors as a systemic defense that supplements local defenses, gambling that the host can handle the coordinated stressors better than can the pathogens. This research was not dependent on external funding.
Expression of peroxisome proliferator activated receptor gamma coactivator 1 (PGC‐1) on the immune response to bacteria
Authors: Del Nero Luis Guilherme , Marta Guilherme , Lima-Salgado Thais , Kubo Sueli , Llimona Flávia , Velasco Irineu Tadeu , Souza Heraldo
Published: June 2021

Introduction/Objective

Inflammatory response during sepsis requires energy expenditure from immune cells. Since the transcription coactivator PGC‐1 controls cell energy homeostasis, we aimed to determine whether PGC‐1 may modulate the immune response in an experimental model of sepsis.

Methods

Live bacteria were injected intraperitoneally in Balb/C mice and PGC‐1β expression was evaluated by quantitative PCR in peritoneal macrophages. Further, mice (20 animals in each group) were submitted to cecal ligation and puncture (CLP). Group 1 received intraperitoneal injections of antisense oligonucleotide (ASO) against PGC‐1β (3,0 nmol) three days before and three days after CLP. Group 2 was given no specific treatment. The animals were followed for 72 hours and mortality was evaluated every 12 hours.

Results

Increased PGC‐1β expression was observed in macrophages from animals exposed to bacteria compared to controls (2.7±0.5 vs. 1.0±0.2, respectively, p<0.05). We further evaluated whether PGC‐1β absence would interfere with the response to acute bacterial aggression in vivo. Mice from Group 1, where PGC‐1β expression was blocked, showed an 80% mortality after 72 hours, while animals from Group 2 had a mortality rate of 63% (p=0,7714).

Conclusions

PGC‐1β is up‐regulated during bacterial infection, however, abolishing PGC‐1β expression did not affect mortality of septic mice.

Financial support: FAPESP
Interaction of the human Prostacyclin receptor with the PDZ adapter protein PDZK1: Role in Endothelial Cell Migration and Angiogenesis
Authors: Kinsella B Therese , Mulvaney Eamon P , Turner Elizebeth C , Reid Helen M
Published: June 2021

Prostacyclin is increasingly implicated in re‐endothelialization and angiogenesis but through largely unknown mechanisms. Herein the high‐density lipoprotein (HDL) scavenger receptor class B, type 1 (SR‐B1) adapter protein PDZ domain‐containing protein 1 (PDZK1) was identified as an interactant of the human prostacyclin receptor (hIP) involving a Class I PDZ ligand at its carboxyl terminus and PDZ domains 1, 3, and 4 of PDZK1. The interaction is constitutive, but may be dynamically regulated following cicaprost activation of the hIP through a mechanism involving cAMP‐dependent protein kinase (PK)A‐phosphorylation of PDZK1 at Ser505. Although PDZK1 did not increase overall levels of the hIP, it increased its functional expression at the cell surface, enhancing ligand binding and cAMP generation. Consistent with a role in re‐endothelialization and angiogenesis, cicaprost activation of the hIP increased endothelial cell migration and tube formation/in vitro angiogenesis, effects abrogated by the specific IP antagonist RO1138452. Furthermore, similar to HDL/SR‐B1, small interfering RNA (siRNA)‐targeted disruption of PDZK1 abolished cicaprost‐mediated endothelial responses but did not affect VEGF responses. The identification of PDZK1 as a functional interactant of the hIP sheds significant mechanistic insights into the protective roles of these key players within the vascular endothelium. Funded by SFI.
Modulation of CB1R expression, trafficking and signaling by HSP90 isoforms
Authors: Filipeanu Catalin , Bailey Evangeline M
Published: June 2021

We recently demonstrated that the levels of AHA1, a HSP90 cochaperone, were increased by chronic Δ9‐THC in female rat cerebellum (Filipeanu et al., J. Neurochem., 2011). In turn, AHA1 overexpression upregulated the plasma membrane levels and signaling of CB1 cannabinoid receptor (CB1R) in HEK293T and Neuro2A cell lines. To precisely define the roles of HSP90 chaperone machinery in the regulation of cannabinoid receptors expression levels and signaling, we generated stable HEK293T cells with specific reduction of HSP90α or β cellular levels. Downregulation of HSP90α was accompanied by an increase in the total CB1R cellular levels, whereas the receptor plasma membrane levels were increased in both HSP90α and β deficient cells. Similar results were obtained after pharmacological inhibition of HSP90 activity using 17‐DMAG (0.5 μM, 18h). Fluorescent microscopy analysis of CB1R subcellular localization indicated that in basal conditions this receptor is present not only at the plasma membrane, but also in the early endosomes. Reduction of the HSP90 levels decreased the CB1R pool present in the endosomes. These changes in the CB1R total cellular levels and subcellular localization determined alterations in the P‐ERK 1/2 activation by acute Δ9‐THC treatment. In conclusion, these results together with our previous publications (Filipeanu et al., BBA., 2011), indicate that HSP90 chaperone machinery is a novel modulator of G protein‐coupled receptors trafficking and signaling. Supported by NIH Grant1R03DA031596‐01.
Cannabinoid Receptor Interacting Protein 1a (CRIP1a) regulates CB1 signaling and transcriptional activity
Authors: Blume Lawrence Christopher , Bass Caroline E. , Dalton George D. , Selley Dana E. , Howlett Allyn C.
Published: June 2021

The CB1 cannabinoid receptor is the most abundantly expressed GPCR in the CNS and modulates many diverse neuronal functions related to synaptic plasticity and neuroprotection. The cellular actions of CB1 ligands on neuroprotection are poorly understood. A recent demonstration that the cannabinoid receptor interacting protein (CRIP1a) had the ability to switch CB1 neuroprotection from an agonist‐ to antagonist‐driven mechanism is intriguing (Stauffer et al., 2011 Neuroscience Letters). To better understand the role of CRIP1a in cytoprotection, we developed neuronal cell models to over‐express or knock‐down CRIP1a in order to identify changes in CB1‐mediated signal transduction and gene transcription. Data from On‐cell‐Western experiments indicate that over‐expression of CRIP1a significantly reduces the levels of phosphoERK, a downstream kinase and transcript factor activated by CB1. Additional studies focusing on phospholipase C and NFkappaB pathways demonstrated that modulation of CRIP1a produces alterations in the mRNA levels of c‐Fos, CCAAT/Enhancer binding protein (Cebpb), NF‐kappaB pathway and genes involved in cell survival and neuroprotection. These experiments suggest that CRIP1a may modulate CB1 signaling and transcription of genes that are important in stress and cell survival pathways.

Supported by NIH grants T32‐DA007246, R01‐DA003690, R21‐ DA025321, K01‐DA024763.