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1.  Addition of strawberries to the usual diet increases postprandial but not fasting non-urate plasma antioxidant activity in healthy subjects 
Strawberries can augment plasma antioxidant activity, but this may be confounded by selection of methods, time of blood sampling and concomitant dietary restrictions. We examined the effect of strawberry consumption on ferric reducing ability (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (DPPH-test) of native and non-urate plasma in healthy subjects on their usual diet. Eleven subjects consumed strawberries (500 g daily) for 9 days. Fasting and 3-h postprandial plasma and 24-h urine collection were obtained before, during and after strawberry course for FRAP, DPPH-test and polyphenols determination. Fifteen subjects served as a control in respect to plasma antioxidant activity changes and effect of 300 mg of oral ascorbate. First, 5th and 9th strawberry dose increased 3-h postprandial DPPH-test by 17.4, 17.6 and 12.6%, and FRAP by 15.5, 25.6 and 21.4% in comparison to fasting values in non-urate plasma (p<0.05). In native plasma only a trend was observed to higher postprandial values for both tests. Strawberries increased urinary urolithin A and 4-hydroxyhippuric acid whereas plasma polyphenols were stable. No changes of FRAP and DPPH-test were noted in controls and after ascorbate intake. Strawberries transiently increased non-urate plasma antioxidant activity but this cannot be attributed to direct antioxidant effect of polyphenols and ascorbate.
doi:10.3164/jcbn.15-113
PMCID: PMC5110932  PMID: 27895386
strawberry; plasma antioxidant activity; phenolics; dietary antioxidants; bioactive compounds; supplements and functional foods
2.  Retention of acetylcarnitine in chronic kidney disease causes insulin resistance in skeletal muscle 
Insulin resistance occurs frequently in patients with chronic kidney disease. However, the mechanisms of insulin resistance associated with chronic kidney disease are unclear. It is known that an increase in the mitochondrial acetyl-CoA (AcCoA)/CoA ratio causes insulin resistance in skeletal muscle, and this ratio is regulated by carnitine acetyltransferase that exchanges acetyl moiety between CoA and carnitine. Because excess acetyl moiety of AcCoA is excreted in urine as acetylcarnitine, we hypothesized that retention of acetylcarnitine might be a cause of insulin resistance in chronic kidney disease patients. Serum acetylcarnitine concentrations were measured in chronic kidney disease patients, and were significantly increased with reduction of renal function. The effects of excess extracellular acetylcarnitine on insulin resistance were studied in cultured skeletal muscle cells (C2C12 and human myotubes), and insulin-dependent glucose uptake was significantly and dose-dependently inhibited by addition of acetylcarnitine. The added acetylcarnitine was converted to carnitine via reverse carnitine acetyltransferase reaction, and thus the AcCoA concentration and AcCoA/CoA ratio in mitochondria were significantly elevated. The results suggest that increased serum acetylcarnitine in CKD patients causes AcCoA accumulation in mitochondria by stimulating reverse carnitine acetyltransferase reaction, which leads to insulin resistance in skeletal muscle.
doi:10.3164/jcbn.15-146
PMCID: PMC5110933  PMID: 27895387
chronic kidney disease; insulin resistance; acetylcarnitine; skeletal muscle; mitochondria
3.  Non-invasive measurement of melanin-derived radicals in living mouse tail using X-band EPR 
The aim of this experiment is to measure in vivo generation of melanin-derived radicals non-invasively, as a quantifiable index of radio-biological effect. Melanin-derived radicals in a living intact mouse tail tip were non-invasively measured in very simple way using an X-band electron paramagnetic resonance spectrometer. Colored mouse strains, C57BL/6NCr, BDF1, and C3H/He, have clear EPR signal corresponding to melanin-derived radicals in the tail tip; however, albino mouse strains, BALB/cCr, ddY, ICR, have no EPR signals. An X-ray fraction of 2 Gy/day (1 Gy/min) was repeatedly irradiated to a C3H/He mouse tail skin every Monday to Friday for 4 weeks. In comparison to before starting irradiation, the C3H/He mouse tail skin became darker, like a suntan. The melanin-derived radicals in C3H/He mouse tail skin were increased in association with X-ray fractions. Melanin-derived radicals in mouse tail skin can be readily and chronologically measurable by using X-band EPR spectrometer, and can be a marker for a radiobiological effect in the skin.
doi:10.3164/jcbn.15-99
PMCID: PMC5110934  PMID: 27895382
melanin-derived radicals; electron paramagnetic resonance; non-invasive measurement; radio-biological effect; X-ray irradiation
4.  Involvement of reactive oxygen species derived from mitochondria in neuronal injury elicited by methylmercury 
Methylmercury induces oxidative stress and subsequent neuronal injury. However, the mechanism by which methylmercury elicits reactive oxygen species (ROS) production remains under debate. In this study, we investigated the involvement of mitochondrial ROS in methylmercury-induced neuronal cell injury using human neuroblastoma SH-SY5Y-derived ρ0 cells, which have a deletion of mitochondrial DNA and thus decreased respiratory activity. SH-SY5Y cells were cultured for 60 days in the presence of ethidium bromide to produce ρ0 cells. Our ρ0 cells showed decreases in the cytochrome c oxidase expression and activity as well as oxygen consumption compared with original SH-SY5Y cells. Methylmercury at a concentration of 1 µM induced cell death with oxidative stress in original SH-SY5Y cells, but not ρ0 cells, indicating that ρ0 cells are resistant to methylmercury-induced oxidative stress. ρ0 cells also showed tolerance against hydrogen peroxide and superoxide anion, suggesting that ρ0 cells are resistant to total ROS. These data indicate that mitochondrial ROS are clearly involved in oxidative stress and subsequent cell death induced by methylmercury. Considering that the dominant mechanism of ROS generation elicited by methylmercury is due to direct antioxidant enzyme inhibition, mitochondria might play a role in amplifying ROS in methylmercury-induced neurotoxicity.
doi:10.3164/jcbn.16-19
PMCID: PMC5110935  PMID: 27895385
methylmercury; reactive oxygen species; mitochondria; neurotoxicity; ρ0 cells
5.  Acetate alters expression of genes involved in beige adipogenesis in 3T3-L1 cells and obese KK-Ay mice 
The induction of beige adipogenesis within white adipose tissue, known as “browning”, has received attention as a novel potential anti-obesity strategy. The expression of some characteristic genes including PR domain containing 16 is induced during the browning process. Although acetate has been reported to suppress weight gain in both rodents and humans, its potential effects on beige adipogenesis in white adipose tissue have not been fully characterized. We examined the effects of acetate treatment on 3T3-L1 cells and in obese diabetic KK-Ay mice. The mRNA expression levels of genes involved in beige adipocyte differentiation and genes selectively expressed in beige adipocytes were significantly elevated in both 3T3-L1 cells incubated with 1.0 mM acetate and the visceral white adipose tissue from mice treated with 0.6% acetate for 16 weeks. In KK-Ay mice, acetate reduced the food efficiency ratio and increased the whole-body oxygen consumption rate. Additionally, reduction of adipocyte size and uncoupling protein 1-positive adipocytes and interstitial areas with multilocular adipocytes appeared in the visceral white adipose tissue of acetate-treated mice, suggesting that acetate induced initial changes of “browning”. In conclusion, acetate alters the expression of genes involved in beige adipogenesis and might represent a potential therapeutic agent to combat obesity.
doi:10.3164/jcbn.16-23
PMCID: PMC5110936  PMID: 27895388
acetate; beige adipogenesis; obesity; visceral fat; short chain fatty acid
6.  Preparation and antioxidant/pro-oxidant activities of 3-monosubstituted 5-hydroxyoxindole derivatives 
Antioxidant treatments have been expected to be a novel therapeutics for various oxidative stress-mediated disorders. Our previous study revealed that 5-hydroxyoxindole and its 3-phenacyl-3-hydroxy derivatives showed excellent antioxidant activities such as 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and lipid-peroxidation inhibitory activity. However, the DPPH radical scavenging activity of the 3,3-disubstituted derivatives was lower than that of the original 5-hydroxyoxindole. In the present study, we synthesized novel 3-monosubstituted 5-hydroxyoxindole derivatives that exhibited stronger DPPH radical scavenging activities and lipid peroxidation-inhibitory activities than the 3,3-disubstituted 5-hydroxyoxindoles. Moreover, the 3-monosubstituted 5-hydroxyoxindole derivatives showed neither an iron-mediated pro-oxidant effect nor a remarkable cytotoxicity against HL-60 cell lines except some of the highly lipophilic compounds. These results indicate that 3-monosubstituted 5-hydroxyoxindoles can be used as a promising antioxidant scaffold for drug discovery.
doi:10.3164/jcbn.16-24
PMCID: PMC5110937  PMID: 27895383
antioxidant; radical scavenging activity; lipid peroxidation; pro-oxidant effect; 5-hydroxyoxindole
7.  Exendin-4 induces extracellular-superoxide dismutase through histone H3 acetylation in human retinal endothelial cells 
Extracellular-superoxide dismutase (genetic name SOD3) is a secreted anti-oxidative enzyme, and its presence in vascular walls may play an important role in protecting the vascular system against oxidative stress. Oxidative stress has been implicated in the pathogenesis of diabetic retinopathy; therefore, increases in extracellular-superoxide dismutase have been suggested to inhibit the progression of diabetic retinopathy. Incretin-based drugs such as glucagon-like peptide-1 receptor agonists are used in the treatment of type 2 diabetes. Glucagon-like peptide-1 receptor agonists are expected to function as extrapancreatic agents because the glucagon-like peptide-1 receptor is expressed not only in pancreatic tissues, but also in many other tissue types. We herein demonstrated that exendin-4, a glucagon-like peptide-1 receptor agonist, induced the expression of extracellular-superoxide dismutase in human retinal microvascular endothelial cells through epigenetic regulation. The results of the present study demonstrated that exendin-4 induced the expression of extracellular-superoxide dismutase through histone H3 acetylation at the SOD3 proximal promoter region. Moreover, plasma extracellular-superoxide dismutase concentrations in diabetic patients were elevated by incretin-based therapies. Therefore, incretin-based therapies may exert direct extrapancreatic effects in order to protect blood vessels by enhancing anti-oxidative activity.
doi:10.3164/jcbn.16-26
PMCID: PMC5110938  PMID: 27895384
extracellular-superoxide dismutase; incretin-based therapy; exendin-4; epigenetics; diabetic retinopathy
8.  Reactive oxygen species induce neurite degeneration before induction of cell death 
Reactive oxygen species (ROS) induce neuronal cell death in a time- and concentration-dependent manner. Treatment of cultured cells with a low concentration of hydrogen peroxide induces neurite degeneration, but not cell death. Neurites (axons and dendrites) are vulnerable to ROS. Neurite degeneration (shrinkage, accumulation, and fragmentation) has been found in neurodegenerative disorders, such as Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease. However, the mechanism of ROS-related neurite degeneration is not fully understood. Many studies have demonstrated the relationship between mitochondrial dysfunction and microtubule destabilization. These dysfunctions are deeply related to changes in calcium homeostasis and ROS production in neurites. Treatment with antioxidant substances, such as vitamin E, prevents neurite degeneration in cultured cells. This review describes the possibility that ROS induces neurite degeneration before the induction of cell death.
doi:10.3164/jcbn.16-34
PMCID: PMC5110939  PMID: 27895381
ROS; neurite degeneration; neuron; mitochondria; calcium
9.  Maslinic acid in olive fruit alleviates mild knee joint pain and improves quality of life by promoting weight loss in the elderly 
Consumption of olives (Olea europaea L.) is associated with a low incidence of inflammation-related diseases. Olive fruit is rich in bioactive pentacyclic triterpenoids, mainly maslinic acid. This study, a randomized, double-blind, and placebo-controlled trial, examined the effects of an orally administered maslinic acid supplement, olive fruit extract, on 20 middle-aged and elderly volunteers with mild knee joint pain. Each subject (58 ± 7 years) received either olive fruit extract, containing 50 mg maslinic acid (n = 12), or placebo (n = 8) daily for 12 weeks and evaluated for pain and physical functions as primary outcome measures. Secondary outcome measures included body composition and inflammatory biomarkers in serum. Although both groups exhibited improved pain visual analogue scale score and quality of life after supplementation, symptoms were better in the maslinic acid group than in the placebo group. After 12 weeks, maslinic acid group exhibited significant decrease in body weight and body mass index suggesting that maslinic acid affected the weight of volunteers with mild knee joint pain. Therefore, olive products containing maslinic acid may be useful as a new preventive and therapeutic food ingredient for arthritic diseases. Since this clinical study is a preliminary study, it was not registered in a publicly accessible database.
doi:10.3164/jcbn.16-40
PMCID: PMC5110940  PMID: 27895390
olive fruit extract; maslinic acid; knee joint pain; weight loss; anti-inflammation
10.  Characteristics of gastric cancer detected within 1 year after successful eradication of Helicobacter pylori 
Gastric cancers are sometimes diagnosed in patients who have successfully undergone Helicobacter pylori (H. pylori) eradication. We analyzed the clinicopathological features of gastric cancers detected after eradication to clarify their characteristics. We reviewed 31 patients with 34 cases of gastric cancer detected after successful H. pylori eradication. Clinicopathological characteristics analyzed included interval since eradication, interval since last endoscopy, tumor size, and depth of invasion. Patients were classified into two groups: early detection (<1 year since eradication) and delayed detection (≥1 year since eradication). The interval since last endoscopy was significantly shorter in the early detection group than in the delayed detection group. However, gastric cancers were significantly larger and more invasive in the early detection group than in the delayed detection group. In conclusion, diligent endoscopy is necessary during the first year after successful H. pylori eradication. (The University Hospital Medical Information Network clinical trial registration number is UMIN000018541.)
doi:10.3164/jcbn.16-43
PMCID: PMC5110941  PMID: 27895391
gastric cancer; H. pylori; eradication
11.  Determination of the adequate dosage of rebamipide, a gastric mucoprotective drug, to prevent low-dose aspirin-induced gastrointestinal mucosal injury 
Small intestinal mucosal injury caused by low-dose aspirin is a common cause of obscure gastrointestinal bleeding. We aimed to investigate the protective effects and optimal dose of rebamipide for low-dose aspirin-induced gastrointestinal mucosal injury. In this prospective randomized trial, 45 healthy volunteers (aged 20–65 years) were included and divided into three groups. The groups received enteric-coated aspirin 100 mg (low-dose aspirin) plus omeprazole 10 mg (Group A: proton pump inhibitor group), low-dose aspirin plus rebamipide 300 mg (Group B: standard-dose group), or low-dose aspirin plus rebamipide 900 mg (Group C: high-dose group). Esophagogastroduodenoscopy and video capsule endoscopy were performed, and the fecal occult blood reaction and fecal calprotectin levels were measured before and two weeks after drug administration. Although the fecal calprotectin levels increased significantly in Group A, they did not increase in Groups B and C. The esophagogastroduodenoscopic and video capsule endoscopic findings and the fecal occult blood test findings did not differ significantly among the three groups. In conclusion, standard-dose rebamipide is sufficient for preventing mucosal injury of the small intestine induced by low-dose aspirin, indicating that high-dose rebamipide is not necessary.
doi:10.3164/jcbn.16-49
PMCID: PMC5110942  PMID: 27895392
rebamipide; low-dose aspirin; gastrointestinal mucosal injury; fecal calprotectin; capsule endoscopy
12.  Comparisons of dietary intake in Japanese with non-alcoholic fatty liver disease and type 2 diabetes mellitus 
Non-alcoholic fatty liver disease (NAFLD) is a multifactorial disease that involves a complex interaction between genetics, diet, and lifestyle, all of which combine to form the NAFLD phenotype. In Japan, medical nutrition therapy for NAFLD has not yet been established, so NAFLD patients are instructed in the dietary modifications used for type 2 diabetes mellitus (T2DM). Because points of difference may exist in the effects of dietary choices on NAFLD and T2DM, the present study aimed to compare and assess the dietary intake of Japanese individuals with NAFLD and T2DM. This cross-sectional study involved 219 patients (77 NAFLD subjects; 33 males, 44 females; 142 T2DM subjects: 76 males, 66 females) aged 40–79 years. Dietary intake was assessed using a validated self-administered diet history questionnaire. Among the results, the most notable in NAFLD patients relative to T2DM patients were: 1) the low intake of vegetables that can reduce the overall energy density; 2) the high consumption of fruits and confectionery containing simple carbohydrates such as fructose; and 3) BMI may be higher. We demonstrated differences in dietary selection between the two groups. NAFLD patients were more likely to have dietary habits that promote fat accumulation in the body.
doi:10.3164/jcbn.16-7
PMCID: PMC5110943  PMID: 27895389
non-alcoholic fatty liver disease; diabetes mellitus; dietary intake; dietary habits; vegetables; fructose
13.  Relationship between homeostasis model assessment of insulin resistance and beta cell function and serum 25-hydroxyvitamin D in non-diabetic Korean adults 
The purpose of this study is to look at these relationships in non-diabetic Korean adults. This study was based on data from the KNHANES V-1, which is representative of the population of Korea. A total of 5,492 participants (≥20 years in age) without type 1 or type 2 diabetes, assessed for serum 25-hydroxyvitamin D [25(OH)D], fasting blood glucose and insulin, as well as anthropometric variables, were included in the analyses. The key study results were as follows: First, vitamin D status [vitamin D deficient, 25(OH)D <25 nM; vitamin D insufficient, 25(OH)D ≥25, <50 nM; vitamin D sufficient, 25(OH)D ≥50 nM] was inversely associated with homeostasis model assessment of insulin resistance (HOMA-IR) and beta cell function (HOMA-B) in model 2 (adjusted for age and gender) and 3 (further adjusted for smoking, alcohol drinking, regular exercise, systolic and diastolic blood pressure, waist circumference, and body mass index). Second, in model 4, when further adjusted for total cholesterol, triglycerides, and HDL-C, vitamin D status was inversely associated with HOMA-B. However, association of vitamin D status and HOMA-IR was no longer significant. In conclusion, vitamin D was inversely associated with beta cell function in non-diabetic Korean adults but was not associated with insulin resistance.
doi:10.3164/jcbn.15-143
PMCID: PMC5018567  PMID: 27698542
vitamin D; HOMA-IR; HOMA-B; non-diabetic adult
14.  Raf kinase inhibitor protein regulates oxygen-glucose deprivation-induced PC12 cells apoptosis through the NF-κB and ERK pathways 
Raf-1 kinase inhibitory protein (RKIP) is a critical molecule for cellular responses to stimuli. In this study, we investigated whether RKIP is responsible for neural cell apoptosis induced by oxygen-glucose deprivation (OGD) and explored the role of NF-κB and ERK pathways regulated by RKIP under OGD stimuli. RKIP was overexpressed or knocked down using lentivirus in PC12 cells, which were then challenged by OGD. RKIP overexpression significantly increased the cell viability of OGD cells, and attenuated apoptosis, cell cycle arrest, and reactive oxygen species generation. RKIP knockdown induced reverse effects. Moreover, we found that RKIP interacted with TAK1, NIK, IKK, and Raf-1 and negatively regulated the NF-κB and ERK pathways. RKIP overexpression significantly inhibited IKK, IκBα, and P65 phosphorylation in NF-κB pathway and MEK, ERK, and CREB phosphorylation in ERK pathway, respectively. RKIP knockdown induced reverse effects. Furthermore, a NF-κB inhibitor BAY 11-7082 and a MEK inhibitor U0126 blocked the changes caused by RKIP down-regulation after OGD. In conclusion, these results demonstrate that RKIP plays a key role in neural cell apoptosis caused by OGD partly via regulating NF-κB and ERK pathways. The present study may provide new insights into the role of RKIP in ischemic stroke.
doi:10.3164/jcbn.15-128
PMCID: PMC5018566  PMID: 27698534
RKIP; ERK pathway; NF-κB pathway; PC12 cells; OGD
15.  Effects of isoflavones on lipid and apolipoprotein levels in patients with type 2 diabetes in Heilongjiang Province in China 
In this study, we examined the association between soy isoflavones and lipid profiles, apolipoprotein levels in patients with type 2 diabetes in China. The study population was composed of 120 cases (80 women with type 2 diabetes and 40 healthy women). Objects in treatment group received isoflavones 435 mg/day for 2 months, then lipid profiles were analyzed by the colorimetry method and apolipoprotein levels were determined by immune turbidimetric method. And all the indexes were determined after oral glucose tolerance test. The levels of total cholesterol, triglyceride and LDL-C significantly reduced and the levels of HDL-C and apolipoprotein A1 significantly raised in the treatment group after intervention (p<0.05). After oral glucose tolerance test, the level of total cholesterol was lower at postprandial 6 h than at empty stomach in treatment group, it had significantly difference (p<0.05). LDL-C levels in the treatment group not only decreased after intervention, but also was significantly lower at postprandial 4, 6 h than in non-intervention group. The ratio of apolipoprotein A1/apolipoprotein B at postprandial 2 h was the highest after treatment in isoflavone group. Supplementation with 435 mg/day of isoflavones exerted favorable effect on the blood total cholesterol, LDL-C levels and the ratio of apolipoprotein A1/apolipoprotein B in Chinese type 2 diabetes women.
doi:10.3164/jcbn.15-147
PMCID: PMC5018568  PMID: 27698541
isoflavones; lipid profiles; apolipoprotein; type 2 diabetes; China
16.  Vitamin A deficiency impacts the structural segregation of gut microbiota in children with persistent diarrhea 
To investigate whether gut microbiota is associated with vitamin A nutritional levels in children with persistent diarrhea, a total of 59 pediatric patients with persistent diarrhea aged 1–12 months were selected from the Department of Gastroenterology at the Children’s Hospital of Chongqing Medical University, China. Subjects were hospitalized and divided into VA-deficient (n = 30) and VA-normal (n = 29) groups according to their venous serum retinol levels. Fecal samples from all 59 subjects were collected immediately after admission and analyzed by Illumina MiSeq for 16S rRNA genes to characterize the overall microbiota of the samples. The gut microbiota of the VA-deficient and VA-normal groups were compared using a bioinformatic statistical approach. The Shannon index (p = 0.02), Simpson index (p = 0.01) and component diagram data indicated significantly lower diversity in the VA-deficient than the VA-normal group. A metagenome analysis (LEfSe) and a differentially abundant features approach using Metastats revealed that Escherichia coli and Clostridium butyricum were the key phylotypes of the VA-normal group, while Enterococcus predominated the VA-deficient group. In conclusion, the diversity of gut microbiota and the key phylotypes are significantly different in children with persistent diarrhea at different VA nutritional levels.
doi:10.3164/jcbn.15-148
PMCID: PMC5018569  PMID: 27698538
vitamin A deficiency (VAD); gut microbiota; key phylotype; persistent diarrhea (PD); children
17.  Energy metabolism and nutritional status in hospitalized patients with lung cancer 
This study aimed to investigate the energy metabolism of patients with lung cancer and the relationship between energy metabolism and proinflammatory cytokines. Twenty-eight patients with lung cancer and 18 healthy controls were enrolled in this study. The nutritional status upon admission was analyzed using nutritional screening tools and laboratory tests. The resting energy expenditure and respiratory quotient were measured using indirect calorimetry, and the predicted resting energy expenditure was calculated using the Harris–Benedict equation. Energy expenditure was increased in patients with advanced stage disease, and there were positive correlations between measured resting energy expenditure/body weight and interleukin-6 levels and between measured resting energy expenditure/predicted resting energy expenditure and interleukin-6 levels. There were significant relationships between body mass index and plasma leptin or acylated ghrelin levels. However, the level of appetite controlling hormones did not affect dietary intake. There was a negative correlation between plasma interleukin-6 levels and dietary intake, suggesting that interleukin-6 plays a role in reducing dietary intake. These results indicate that energy expenditure changes significantly with lung cancer stage and that plasma interleukin-6 levels affect energy metabolism and dietary intake. Thus, nutritional management that considers the changes in energy metabolism is important in patients with lung cancer.
doi:10.3164/jcbn.16-1
PMCID: PMC5018572  PMID: 27698539
energy metabolism; nutritional status; indirect calorimetry; lung cancer
18.  Analysis of negative result in serum anti-H. pylori IgG antibody test in cases with gastric mucosal atrophy 
The purpose is to elucidate factors related to negative results of anti-H. pylori antibody test in cases with gastric mucosal atrophy. A total of 859 individuals without past history of eradication therapy for H. pylori (545 males, 314 females; mean age 52.4 years) who underwent an upper GI endoscopy examination and serological test were enrolled as subjects. Serological testing was performed using SphereLight H. pylori antibody J®, and endoscopic findings of gastric mucosal atrophy by the classification of Kimura and Takemoto and post-eradication findings were analyzed. The positive rates for the anti-H. pylori antibody test in subjects with and without gastric mucosal atrophy were 85.6% and 0.9%, respectively. In analysis of subjects with gastric mucosal atrophy, a low positive rate and serum titer was observed in subjects with C1, C2 and O3 atrophy. When the analysis was performed separately in male and female subjects, low positive rate was observed in males with O3 atrophy and females with C2 atrophy. Suspected post-eradication endoscopic findings were more frequently observed in cases with C2 atrophy. In conclusion, negative result of anti-H. pylori antibody test was frequently observed in middle-aged subjects with C1, C2 and O3 gastric mucosal atrophy.
doi:10.3164/jcbn.16-13
PMCID: PMC5018573  PMID: 27698543
Helicobacter pylori; diagnosis; serologic tests; endoscopy; atrophy
19.  l-Theanine protects against excess dopamine-induced neurotoxicity in the presence of astrocytes 
l-Theanine (γ-glutamylethylamide), a component of green tea, is considered to have regulatory and neuroprotective roles in the brain. The present study was designed to determine the effect of l-theanine on excess dopamine-induced neurotoxicity in both cell culture and animal experiments. The primary cultured mesencephalic neurons or co-cultures of mesencephalic neurons and striatal astrocytes were pretreated with l-theanine for 72 h, and then treated with excess dopamine for further 24 h. The cell viability of dopamine neurons and levels of glutathione were evaluated. Excess dopamine-induced neurotoxicity was significantly attenuated by 72 h preincubation with l-theanine in neuron-astrocyte co-cultures but not in neuron-rich cultures. Exposure to l-theanine increased the levels of glutathione in both astrocytes and glial conditioned medium. The glial conditioned medium from l-theanine-pretreated striatal astrocytes attenuated dopamine-induced neurotoxicity and quinoprotein formation in mesencephalic neurons. In addition, replacement of l-glutamate with l-theanine in an in vitro cell-free glutathione-synthesis system produced glutathione-like thiol compounds. Furthermore, l-theanine administration (4 mg/kg, p.o.) for 14 days significantly increased glutathione levels in the striatum of mice. The results suggest that l-theanine provides neuroprotection against oxidative stress-induced neuronal damage by humoral molecules released from astrocytes, probably including glutathione.
doi:10.3164/jcbn.16-15
PMCID: PMC5018574  PMID: 27698535
l-theanine; dopamine; astrocyte; glutathione; neuroprotection
20.  Esomeprazole- or rabeprazole-based triple therapy eradicated Helicobacter pylori comparably regardless of clarithromycin susceptibility and CYP2C19 genotypes 
The aim of this study was to assess the efficacy of esomeprazole-based triple therapy compared with rabeprazole-based triple therapy according to CYP2C19 genotype and clarithromycin susceptibility status for first-line eradication therapy of Helicobacter pylori (H. pylori) in Japan. We enrolled 219 H. pylori-infected patients, and randomly allocated patients to the EAC group (esomeprazole 20 mg, clarithromycin 200 mg, amoxicillin 750 mg for one week, with all drugs given twice daily) or RAC group (rabeprazole 10 mg, clarithromycin 200 mg, amoxicillin 750 mg for one week, with all drugs given twice daily). The H. pylori eradication rate according to the PP analyses was 75.0% (95% CI: 65.2–82.8%) in the EAC group and 71.4% (95% CI: 61.4–79.1%) in the RAC group. There were no statistically significant differences. The eradication rates of the clarithromycin-resistant/-sensitive strains were, respectively, 45.0% (95% CI: 30.7–60.2%)/98.0% (95% CI: 88.7–100%) in the EAC group and 39.5% (95% CI: 25.6–55.3%)/93.5% (95% CI: 81.9–98.4%) in the RAC group. The eradication rate of the clarithromycin-sensitive strains was significantly higher than that of the resistant strains in both groups. In conclusion, EAC and RAC therapies show a comparable efficacy regardless of the CYP2C19 genotype and clarithromycin susceptibility status in Japan.
doi:10.3164/jcbn.16-18
PMCID: PMC5018575  PMID: 27698544
Helicobacter pylori; eradication; esomeprazole; rabeprazole; CYP2C19
21.  Influence of regular exercise on gastric emptying in healthy men: a pilot study 
Functional gastrointestinal disorders (FGIDs), including functional dyspepsia (FD), are common chronic disorders even in the younger population. Physical activity is advocated for patients with FGIDs, although the evidence is insufficient. We investigated the association between the intensity of regular exercise and gastric emptying to determine the effect of physical activity on dyspeptic symptoms. Thirty healthy individuals were selected and divided into three groups (low, moderate, and high) using the index of total exercise intensity in a week. Gastric emptying was evaluated by the 13C-acetate breath test. Gastroesophageal reflux symptoms, dyspeptic symptoms, stool forms, scores of anxiety and depression, and scores of sleep quality were also compared. Baseline scores of gastroesophageal reflux symptoms, anxiety, depression, and sleep quality were not different among the three groups. Gastric emptying was significantly faster in low-intensity exercise group than the moderate-intensity exercise group. Although the presence of loose stool and alcohol consumption were also associated with the intensity of regular exercise, these variables were not confounders. In conclusion, the intensity of regular exercise was independently associated with gastric emptying in healthy individuals. These baseline data would be useful for consideration of an optimal exercise intervention for the treatment of FD.
doi:10.3164/jcbn.16-29
PMCID: PMC5018576  PMID: 27698540
exercise; gastric emptying; functional dyspepsia; irritable bowel syndrome; 13C-acetate breath test
22.  Resveratrol improves salivary dysfunction in a non-obese diabetic (NOD) mouse model of Sjögren’s syndrome 
Resveratrol is a natural polyphenol produced by plants in response to environmental stress. This compound has been shown to have pharmacological effects against a wide range of diseases including neurological, hepatic, cardiovascular and autoimmune conditions. The non-obese diabetic (NOD) mouse, in which loss of lacrimal and salivary gland function occurs, has been studied as an animal model for Sjögren’s syndrome. In this study, we confirmed that administration of resveratrol results in increased secretion of saliva in NOD mice. Although resveratrol enhanced Sirt1 activity, inflammatory cell infiltration was not affected. Moreover, expression of the anti-inflammatory cytokine IL-10 in salivary glands was enhanced in the resveratrol-administered group. Thus, we confirmed a novel therapeutic effect for resveratrol on salivary dysfunction in Sjögren’s syndrome.
doi:10.3164/jcbn.16-31
PMCID: PMC5018577  PMID: 27698537
salivary gland; resveratrol; NOD mouse; salivary secretion; Sjögren’s syndrome
23.  Astaxanthin affects oxidative stress and hyposalivation in aging mice 
Oral dryness, a serious problem for the aging Japanese society, is induced by aging-related hyposalivation and causes dysphagia, dysgeusia, inadaptation of dentures, and growth of oral Candida albicans. Oxidative stress clearly plays a role in decreasing saliva secretion and treatment with antioxidants such astaxanthin supplements may be beneficial. Therefore, we evaluated the effects of astaxanthin on the oral saliva secretory function of aging mice. The saliva flow increased in astaxanthin-treated mice 72 weeks after administration while that of the control decreased by half. The plasma d-ROMs values of the control but not astaxanthin-treated group measured before and 72 weeks after treatment increased. The diacron-reactive oxygen metabolites (d-ROMs) value of astaxanthin-treated mice 72 weeks after treatment was significantly lower than that of the control group was. The plasma biological antioxidative potential (BAP) values of the control but not astaxanthin-treated mice before and 72 weeks after treatment decreased. Moreover, the BAP value of the astaxanthin-treated group 72 weeks after treatment was significantly higher than that of the control was. Furthermore, the submandibular glands of astaxanthin-treated mice had fewer inflammatory cells than the control did. Specifically, immunofluorescence revealed a significantly large aquaporin-5 positive cells in astaxanthin-treated mice. Our results suggest that astaxanthin treatment may prevent age-related decreased saliva secretion.
doi:10.3164/jcbn.15-150
PMCID: PMC5018570  PMID: 27698533
astaxanthin; aquaporin-5; hyposalivation; oral dryness; inflammation
24.  Synergistic antioxidative effect of astaxanthin and tocotrienol by co-encapsulated in liposomes 
Astaxanthin and vitamin E are both effective antioxidants that are frequently used in cosmetics, as food additives, and in to prevent oxidative damage. A combination of astaxanthin and vitamin E would be expected to show an additive anntioxidative effect. In this study, liposomes co-encapsulating astaxanthin and the vitamin E derivatives α-tocopherol (α-T) or tocotrienols (T3) were prepared, and the antioxidative activity of these liposomes toward singlet oxygen and hydroxyl radical was evaluated in vitro. Liposomes co-encapsulating astaxanthin and α-T showed no additive anntioxidative effect, while the actual scavenging activity of liposomes co-encapsulating astaxanthin and T3 was higher than the calculated additive activity. To clarify why this synergistic effect occurs, the most stable structure of astaxanthin in the presence of α-T or α-T3 was calculated. Only α-T3 was predicted to form hydrogen bonding with astaxanthin, and the astaxanthin polyene chain would partially interact with the α-T3 triene chain, which could explain why there was a synergistic effect between astaxanthin and T3 but not α-T. In conclusion, co-encapsulation of astaxanthin and T3 induces synergistic scavenging activity by intermolecular interactions between the two antioxidants.
doi:10.3164/jcbn.15-153
PMCID: PMC5018571  PMID: 27698536
astaxanthin; vitamin E; tocotrienol; intermolecular interaction; synergistic activity
25.  Pre-germinated brown rice prevented high fat diet induced hyperlipidemia through ameliorating lipid synthesis and metabolism in C57BL/6J mice 
Pre-germinated brown rice (PGBR) can ameliorate hyperlipidemia, but the action mechanism is not clear. We focus the mechanisms of PGBR prevented hyperlipidemia. Six-week-old mice were divided into: standard-regular diet (SRD), high-fat diet (HFD) and HFD with PGBR (HFD + PGBR) groups for 16 weeks. The HFD group has higher concentrations of TG, TC, HDL and Non-HDL in the blood, and a higher atherosclerosis index (AI). The TG levels in the liver, and TG, bile acid levels in the feces were enhanced; and the total adipocytokines level in adipose tissue was reduced. The HFD group had higher protein expressions of SREBP-1, SCD-1, FAS, LDLR, and CYP7α1 in the liver. Moreover, the greater expressions of SREBP-1, SCD-1, FAS and the less expressions of PPAR-α and adiponectin were in adipose tissue. In the HFD + PGBR group, the PGBR regulated the levels of TG, TC, HDL, Non-HDL, AI and adipocytokines. PGBR increased more cholesterol and bile acid exhaust in feces. The SREBP-1, SCD-1, FAS, HMGCR, LDLR, CYP7α1 and PPAR-α proteins in the liver; and the SREBP-1, SCD-1, FAS, PPAR-α and adiponectin proteins in adipose tissue were reversed by PGBR. Taken together, PGBR can improve lipid synthesis and metabolism, and we suggest PGBR is a recommendable food for controlling hyperlipidemia.
doi:10.3164/jcbn.15-117
PMCID: PMC4933684  PMID: 27499577
pre-germinated brown rice; hyperlipidemia; lipid synthesis and metabolism

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