The amounts of reactive oxygen species generated in aqueous samples by irradiation with X-ray or clinical carbon-ion beams were quantified. Hydroxyl radical (•OH), hydrogen peroxide (H2O2), and the total amount of oxidation reactions, which occurred mainly because of •OH and/or hydroperoxy radicals (HO2•), were measured by electron paramagnetic resonance-based methods. •OH generation was expected to be localized on the track/range of the carbon-ion beam/X-ray, and mM and M levels of •OH generation were observed. Total •OH generation levels were identical at the same dose irrespective of whether X-ray or carbon-ion beam irradiation was used, and were around 0.28–0.35 µmol/L/Gy. However, sparse •OH generation levels decreased with increasing linear energy transfer, and were 0.17, 0.15, and 0.09 µmol/L/Gy for X-ray, 20 keV/µm carbon-ion beam, and >100 keV/µm carbon-ion beam sources, respectively. H2O2 generation was estimated as 0.26, 0.20, and 0.17 µmol/L/Gy, for X-ray, 20 keV/µm carbon-ion beam, and >100 keV/µm carbon-ion beam sources, respectively, whereas the ratios of H2O2 generation per oxygen consumption were 0.63, 0.51, and 3.40, respectively. The amounts of total oxidation reactions were 2.74, 1.17, and 0.66 µmol/L/Gy, respectively. The generation of reactive oxygen species was not uniform at the molecular level.
Oral cancer accounts for ~2% of all cancers worldwide, and therapeutic intervention is closely associated with quality of life. Here, we evaluated the effects of non-thermal plasma on oral squamous cell carcinoma cells with special reference to catalytic Fe(II). Non-thermal plasma exerted a specific killing effect on oral squamous cell carcinoma cells in comparison to fibroblasts. Furthermore, the effect was dependent on the amounts of catalytic Fe(II), present especially in lysosomes. After non-thermal plasma application, lipid peroxidation occurred and peroxides and mitochondrial superoxide were generated. Cancer cell death by non-thermal plasma was promoted dose-dependently by prior application of ferric ammonium citrate and prevented by desferrioxamine, suggesting the association of ferroptosis. Potential involvement of apoptosis was also observed with positive terminal deoxynucleaotidyl transferase-mediated dUTP nick end labeling and annexin V results. Non-thermal plasma exposure significantly suppressed the migratory, invasive and colony-forming abilities of squamous cell carcinoma cells. The oral cavity is easily observable; therefore, non-thermal plasma can be directly applied to the oral cavity to kill oral squamous cell carcinoma without damaging fibroblasts. In conclusion, non-thermal plasma treatment is a potential therapeutic option for oral cancer.
Plasma-activated medium (PAM) is a solution produced by exposing a liquid medium to non-thermal atmospheric pressure plasma (NTAPP). A number of reactive molecules, such as reactive oxygen species and reactive nitrogen species, are contained in PAM. Therefore, exposure to high doses of PAM results in cell death. We previously demonstrated that intracellular zinc (Zn2+) serves as an important mediator in PAM-induced cell death; however, the effects of sublethal treatment with PAM on cell functions are not fully understood. In the present study, we found that sublethal PAM treatment suppressed cell proliferation and induced senescence-like changes in lung adenocarcinoma A549 cells. Cell cycle analysis revealed that PAM induced cell cycle arrest at the G2/M phase. PAM increased the level of intracellular free Zn2+ and the Zn2+ chelator TPEN counteracted PAM-induced growth suppression, suggesting that Zn2+ functions in PAM-induced growth suppression. In addition, sublethal treatment with PAM induced phosphorylation of ATM kinase, accumulation of p53 protein, and expression of p21 and GADD45A, which are known p53 target genes, in a Zn2+-dependent manner. These results suggest that the induction of growth arrest and cellular senescence by sublethal PAM treatment is mediated by Zn2+-dependent activation of the ATM/p53 pathway.
We investigated the effect of the sodium glucose cotransporter-2 inhibitor (SGLT-2i) luseogliflozin on skeletal muscle. Eight-week-old mice were fed a standard diet or the standard diet with added luseogliflozin for 8 weeks. The mice were divided into the following four genotype/dietary groups: Db/m mice without SGLT-2i, Db/m mice with SGLT-2i inhibitor, Db/Db without SGLT-2i, and Db/Db with SGLT-2i. Among the mice with and without SGLT-2i, the ratio of soleus and plantaris muscle to body weight in the Db/Db mice was significantly lower than that in the Db/m mice. The cross-sectional area of soleus muscle in the Db/Db mice without SGLT-2i was significantly higher than that in the Db/Db mice with SGLT-2i. The expression of foxo1 in soleus muscle of the Db/Db mice was significantly higher than that of the Db/m mice, and the foxo1 expression of the Db/Db mice with SGLT-2i was significantly lower than that of the mice without SGLT-2i. The fluorescence intensity of foxo1 in the Db/Db mice fed SGLT-2i was significantly lower than that in the Db/Db mice without SGLT-2i. The administration of luseogliflozin resulted in the suppression of both the increased foxo1 expression and the reduced muscle cross-sectional area in the soleus muscle of Db/Db mice.
Cinnamtannin A2, an (−)-epicatechin tetramer, was reported to have potent physiological activity. Cinnamtannin A2 is rarely absorbed from the gastrointestinal tract into the blood and the mechanisms of its beneficial activities are unknown. Cinnamtannin A2 reported to increase sympathetic nervous activity, which was induced by various stressors. In present study, we examined the stress response in the mouse paraventricular nucleus following a single oral dose of cinnamtannin A2 by monitoring mRNA expression of corticotropin-releasing hormone (CRH) and c-fos using in situ hybridization. Corticotropin-releasing hormone mRNA showed a tendency to increase at 15 min and significantly increased at 60 min following a single oral administration of 100 µg/kg cinnamtannin A2. After a single dose of 10 µg/kg cinnamtannin A2, there was significant upregulation of CRH mRNA at 60 min. These results suggested that cinnamtannin A2 was recognized as a stressor in central nervous system and this may lead to its beneficial effects on circulation and metabolism.
Gut microbiota have profound effects on bile acid metabolism by promoting deconjugation, dehydrogenation, and dehydroxylation of primary bile acids in the distal small intestine and colon. High-fat diet-induced dysbiosis of gut microbiota and bile acid dysregulation may be involved in the pathology of steatosis in patients with non-alcoholic fatty liver disease. Epigallocatechin-3-gallate (EGCG), the most abundant polyphenolic catechin in green tea, has been widely investigated for its inhibitory or preventive effects against fatty liver. The aim of the present study was to investigate the effects of EGCG on the abundance of gut microbiota and the composition of serum bile acids in high-fat diet-fed mice and determine the specific bacterial genera that can improve the serum bile acid dysregulation associated with EGCG anti-hepatic steatosis action. Male C57BL/6N mice were fed with the control diet, high-fat diet, or high-fat diet + EGCG at a concentration of 0.32% for 8 weeks. EGCG significantly inhibited the increases in weight, the area of fatty lesions, and the triglyceride content in the liver induced by the high-fat diet. Principal coordinate analysis revealed significant differences in microbial structure among the groups. At the genus level, EGCG induced changes in the microbiota composition in high-fat diet-fed mice, showing a significantly higher abundance of Adlercreutzia, Akkermansia, Allobaculum and a significantly lower abundance of Desulfovibrionaceae. EGCG significantly reversed the decreased population of serum primary cholic acid and β-muricholic acid as well as the increased population of taurine-conjugated cholic acid, β-muricholic acid and deoxycholic acid in high-fat diet-fed mice. Finally, the correlation analysis between bile acid profiles and gut microbiota demonstrated the contribution of Akkermansia and Desulfovibrionaceae in the improvement of bile acid dysregulation in high-fat diet-fed mice by treatment with EGCG. In conclusion, the present study suggests that EGCG could alter bile acid metabolism, especially taurine deconjugation, and suppress fatty liver disease by improving the intestinal luminal environment.
In our previous report, we showed the effect of astaxanthin intake on VEGF level in the aqueous humor and the relationship between VEGF level and reactive oxygen species-related parameters and other relevant factors. VEGF level is associated with total hydroperoxide level, and a multivariate analysis identified sex as a secondary factor affecting these relationships. Here, we analyzed the effects of astaxanthin on the relationship between VEGF level and reactive oxygen species-related parameters by sex. Patients (16 males and 19 females, aged 71.3 and 70.6, respectively) underwent bilateral cataract surgery on one side before and the other side after astaxanthin treatment (6 mg/day for 2 weeks). Levels of VEGF, hydrogen peroxide, and total hydroperoxide, and O2•− scavenging activity, were measured in the aqueous humor. In females only, VEGF level was negatively correlated with O2•− scavenging activity before the astaxanthin intake (r = −0.6, p<0.01) and positively correlated with total hydroperoxide level before and after the astaxanthin intake (r = 0.7 and 0.8, respectively, p<0.01). In conclusion, astaxanthin appears to affect O2•− scavenging activity in the aqueous humor in females, and is likely to be involved in the control of VEGF levels in the anterior eye.
The objective of this study is to elucidate the effect of a supplement enriched with l-leucine, l-arginine, and l-glutamine on body compositions/skin conditions. Healthy young women (n = 29) were allocated to a group (n = 14) receiving an amino-acid supplement (600 mg l-leucine, 250 mg l-arginine, and 300 mg l-glutamine) and a placebo group (n = 15) receiving a supplement not-containing the amino acids. The amino-acid supplement and placebo were given twice/day for 6 weeks. After a wash-out (2 months) from the 1st test, the amino-acid group received the placebo and the placebo group the amino-acid supplement. The body compositions/skin conditions were measured 4 times (day 1 and weeks 2, 4, and 6) in each test. Percentage-change of muscle mass in the amino-acid group increased up to 4 weeks (p = 0.05) and was higher than that in the placebo group (p = 0.09). Skin texture estimated by the image processing of neck skin replica tended to increase in the amino-acid group at 6 weeks compared with that at 0 week, though there was no significant intergroup difference. In conclusion, the young adult women having no fitness habit showed the significant increase of the muscle amount and improvement tendency of the skin texture by the continuous intake of the amino-acid supplement.
We have previously reported that the risk of type 2 diabetes, early impaired glucose tolerance, and insulin resistance can be predicted using fasting levels of adiponectin, leptin, and insulin. Here, we aimed to evaluate the utility of hemoglobin A1c in detecting the risk of type 2 diabetes compared with other well-known biomarkers. We randomly enrolled 207 volunteers with no history of diseases, who underwent 75-g oral glucose tolerance tests and were stratified into normal, borderline, abnormal, or diabetic groups. Eighty-one participants with normal baseline levels of hemoglobin A1c (<6.0%) were included in the normal groups of both glucose tolerance and insulin resistance. Hemoglobin A1c was significantly correlated with the plasma glucose and insulin resistance index. Leptin, adiponectin, glycoalbumin, and body mass index also were correlated well with plasma glucose levels and insulin resistance index. Normal hemoglobin A1c levels with abnormal glucose tolerance and insulin resistance were noted in 85 and 67 participants, respectively. Hemoglobin A1c did not strengthen the prediction algorithm of diabetes, determined by our proposed biomarkers, leptin, adiponectin, and insulin. In conclusion, hemoglobin A1c is a surrogate biomarker for risk of diabetes, with inadequate predictive value, and should be used in combination with other biomarkers.
Background and Aims: To determine whether serum acylated ghrelin levels were associated with anxiety, clinical symptoms, depressive status, quality of life, gastric motility and endoscopic findings based on Kyoto classification in functional dyspepsia (FD) patients. Methods: We enrolled three groups, FD patients (n = 15) with high levels of acylated ghrelin, FD patients (n = 33) with normal levels of acylated ghrelin and FD patients (n = 35) with low levels of acylated ghrelin. There was no significant differences in the positivity of Helicobacter pylori infection among the three groups. Clinical symptoms were evaluated by Gastrointestinal Symptom Rating Scale (GSRS) and FD symptoms based on Rome III classification. Acylated ghrelin levels were measured by ELISA methods. Depressive status, anxiety, sleep disturbance were respectively asscessed by Self-rating questionnaire for depression (SRQ-D) score, STAI-state/-trait, Pittsburgh sleep quality index (PSQI) scores. Endoscopic findings were evaluated based on Kyoto classification. Results: Body Mass Index (BMI) in FD patients with low levels of acylated ghrelin was significantly higher (p<0.001 and p = 0.008, respectively) compared to those in FD patients with high and normal levels of acylated ghrelin. SRQ-D scores in FD patients with low levels of acylated ghrelin was significantly lower (p = 0.008 and p<0.001, respectively) compared to those in FD patients with high and normal levels of acylated ghrelin. Scoring of gastric atrophy, intestinal metaplasia, xanthoma and mucus based on Kyoto classification in FD patients with low levels of acylated ghrelin were significantly higher (p<0.001, p = 0.0077, p = 0.036 and p = 0.0063, respectively) compared to those in FD patients with more than low levels of acylated ghrelin. Conclusion: Acylated ghrelin levels were associated with BMI, depressive status, and endoscopic findings based on Kyoto classification in FD patients.
We evaluated serological Helicobacter pylori and cytotoxin-associated gene A (CagA) antibodies and endoscopic atrophy after eradication to identify factors predicting post-eradication gastric cancer development. Thirty-five patients with successful eradiation were divided into the post-eradication gastric cancer (13 cases) and non-gastric cancer (22 cases) groups. Serum Helicobacter pylori and CagA antibody titers and endoscopic atrophy before and six years after eradication were examined. Median Helicobacter pylori antibody titers had decreased significantly from baseline at 0.5–2 years after eradication in both groups (gastric cancer group, from 39.0 to 11.0 U/ml, p = 0.011; non-gastric cancer group, from 29.6 to 4.97 U/ml, p<0.001), but were significantly higher in the gastric cancer than in the non-gastric cancer group (p = 0.029). Median serum CagA antibody titers had also decreased significantly at 0.5–2 years after eradication (gastric cancer group, from 6.35 to 3.23 U/ml, p = 0.028; non-gastric cancer group, from 9.88 to 1.21 U/ml, p = 0.0045). Serum CagA in each group showed no significance. Endoscopic atrophy improved significantly after eradication in the non-gastric cancer, but not the gastric cancer, group (p = 0.0007). In conclusion, changes in Helicobacter pylori and CagA antibody titers and endoscopic atrophy after eradication might be useful as predictive factors for post-eradication gastric cancer.
Although magnesium oxide is widely used as a laxative, alterations in serum magnesium concentrations among patients taking daily magnesium oxide have not been clarified. The present retrospective, cross-sectional study investigated the risk factors for hypermagnesemia in patients taking daily oral magnesium oxide. Of 2,176 patients administered daily magnesium oxide, 193 (8.9%) underwent assays of serum magnesium concentrations and were evaluated. High serum magnesium concentration and hypermagnesemia were defined as serum magnesium concentrations ≥2.5 mg/dl and ≥3.0 mg/dl, respectively. Of the 193 patients taking daily magnesium oxide, 32 (16.6%) had high serum magnesium concentration and 10 (5.2%) had hypermagnesemia. Factors associated with hypermagnesemia included chronic kidney disease (CKD) grade 4 (p = 0.014) and magnesium oxide dosage (p = 0.009). Factors associated with high serum magnesium concentration included magnesium oxide dosage >1,000 mg/day (p = 0.004), CKD grades 4 (p = 0.000) and concomitant use of stimulant laxatives (p = 0.035). Age, however, was not associated with hypermagnesemia or high serum magnesium concentration. In conclusion, renal function and magnesium oxide dosage, but not age, were associated with hypermagnesemia and high serum magnesium concentration in patients with functional constipation taking daily magnesium oxide.