Distinct role of endothelial nitric oxide synthase gene polymorphisms from menopausal status in the patients with sporadic breast cancer in Taiwan.

Breast cancer has a high incidence in Taiwanese women and worldwide. Previous studies have indicated that NO has multiple independent roles in carcinogenesis; genetic polymorphisms in the endothelial nitric oxide synthase (eNOS) gene could modify its transcription and endogenous NO production. Previous studies have reported conflicting results for the relationship between polymorphisms in the eNOS gene and breast cancer risk. Estrogen levels are associated with eNOS expression; accordingly, variation in estrogen levels may contribute to the discordant results. Therefore, in this study, the effects of eNOS polymorphisms on breast cancer susceptibility were examined in terms of menopausal status in Taiwanese women. Three eNOS polymorphisms (-786T > C, VNTR, and 894G > T) were genotyped in 283 patients with breast cancer (139 premenopausal and 144 postmenopausal) and 200 cancer-free controls (100 premenopausal and 100 postmenopausal) by PCR-RFLP. There was a significantly higher breast cancer risk in premenopausal women carrying 894G > T T than in those with the 894G > T GG genotype; however, postmenopausal women carrying 894G > T T had a lower risk of developing breast cancer. In addition, based on a binary logistic regression analysis, interaction effects of these polymorphisms differed according to menopausal status. The relationship between eNOS polymorphisms and breast cancer hazard depended on menopause status, especially for the 894G > T polymorphism, which may provide an explanation for previous conflicting results.

Increased Igf-I/Igfbp-3 Ratios in Postmenopausal Taiwanese with Breast Cancer, Irrespective of Er and Pr Statuses and Her2 Expression in a Case-Control Study.

BACKGROUND: In most research, there were positive associations between the insulin-like growth factor I (IGF-I) status, including IGF-I, insulin-like growth factor binding protein-3 (IGFBP-3), and ratio of IGF-I/IGFBP-3, and risks of breast cancer (BC), which was influenced by many factors, including hormone statuses and ethnicity. Therefore, the alterations of the IGF-I status in Taiwanese women with BC by menopausal statuses and hormone receptors were investigated. METHODS: The levels of IGF-I and IGFBP-3 were determined by the enzyme-labeled chemiluminescent immunometric assay, and the protein expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor 2 (HER2) on paraffin-embedded sections of tissues with BC were analyzed by the immunohistochemical method. RESULTS: The ratios of IGF-I/IGFBP-3 were significantly higher in the women with BC than those in the controls, but not of the levels of IGF-I and IGFBP-3; furthermore, the significantly higher ratios were found only in the postmenopausal status. In addition, there was no significant difference between the IGF-I status and ER and PR statuses, and HER2 expression, respectively, in the women with BC. CONCLUSIONS: The ratios of IGF-I/IGFBP-3 were increased in postmenopausal Taiwanese women with BC, irrespective of their ages, ER and PR statuses, and HER2 expression.

The Immune Regulator VTCN1 Gene Polymorphisms and Its Impact on Susceptibility to Breast Cancer.

BACKGROUND: VTCN1, a T-cell regulator, belongs to the immunoglobulin superfamily. It is more highly expressed in tumor tissues than in normal tissues, which suggests that it could serve as a tumor-related agent. We hypothesize the gene variants for this coinhibitory molecule may be associated with the risk of breast cancer, given such gene polymorphisms could affect its related gene expression. METHODS: Genotypes of the VTCN1 gene variants (rs10754339, rs10801935, and rs3738414) were analyzed in 566 patients with breast cancer and 400 age-frequency-matched controls. RESULTS: Compared with the major allele, the minor alleles of rs10754339, rs10801935, and rs3738414 did modulate the risk of breast cancer with ORs (95% CI) of 1.42 (1.07-1.89), 1.39 (1.10-1.77), and 0.81 (0.67-0.99), respectively. Those with the rs10754339 genotype AG and rs10801935 AC genotype had significantly increased risks when compared with their major genotypes. However, in rs3738414, the AA genotype had a marginally significant decreased risk compared with its wild genotype. In the haplotype-based analysis, the GCG allele was associated with significantly increased risk (OR: 1.56, 95% CI: 1.09-2.22) based on the AAG reference. Further analyses of the haplotype pairs showed GCG carriers had a significantly increased risk. CONCLUSIONS: In this study, the VTCN1 genetic variants (rs10754339, rs10801935, and rs3738414) indicate they could be connected with the risk of breast cancer, which in turn provides indirect evidence that T-cell immunity could be involved in the development of breast cancer.

Metabolic pathways related to oxidative stress in patients with hemoglobin h disease and iron overload.

BACKGROUND: Iron overload is a major complication in patients with hemoglobin H (Hb H) disease and causes damage of tissues. METHODS: We investigated 26 Hb H patients and 75 controls to evaluate their oxidative stress and antioxidant statuses. RESULTS: There were significantly increased levels of superoxide anion in leucocytes, nitrite (NO2-), and malondialdehyde (MDA) in plasma, and activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GRx) and oxidized glutathione (GSSG) in erythrocytes, decreased levels of nitrate (NO3-) and vitamin C in plasma, and reduced glutathione (GSH) in erythrocytes, in addition to the abnormal iron status in the patients when compared with those in the controls. Meanwhile, levels of serum ferritin were positively correlated with serum iron, plasma MDA, and erythrocyte SOD in the patients. In addition, the activities of SOD were positively correlated with those of GPx and GRx, and the levels of GSSG and MDA, but negatively correlated with those of GSH. Furthermore, the levels of MDA were negatively correlated those of vitamin C. CONCLUSIONS: These results demonstrate the presence of oxidative stress and decreased levels of antioxidants; moreover, the related metabolic antioxidant pathway is active in Hb H patients with iron overload.

Oxidative stress-related enzyme polymorphisms associated with the immunological biomarkers levels in heavy drinkers in Taiwan.

BACKGROUND: Excessive alcohol intake can result in the oxidative stress in cells and the genetic variations of alcohol-metabolizing enzymes are responsible for the different degrees of toxicity of alcohol in several organs, such as the liver and immunological systems. We hypothesized that the alteration of oxidative stress due to some genetic variations of oxidative stress-related enzymes could result in changes of specific biomarkers, and heavy drinkers could be cautioned about the predictive likelihood to induce drinking-induced diseases. METHODS: A total of 108 heavy drinkers and 106 nonheavy drinkers were enrolled and the hematological, biochemical, and immunological tests were measured; the genotypes of oxidative stress-related enzymes, including manganese superoxide dismutase (MnSOD1183T>C), glutathione peroxidase 1 (GPX1Pro198Leu), catalase (CAT-262C>T), and myeloperoxidase (MPO-463G>A), were assayed by real-time polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). RESULTS: For the males, the levels of carbohydrate-deficient transferrin (CDT), malondialdehyde (MDA), CD4(+), immunoglobulin G (IgG), immunoglobulin M (IgM), and IL-6 were significantly different between the two groups. Furthermore, there were higher proportions of CD19(+) cells and lower TNF-α levels in heavy drinkers with the MnSOD C carriers, and there were higher percentages of CD19(+) cells and IL-6 levels in heavy drinkers with the combined genotypes of MnSOD C carriers and MPO A carriers. CONCLUSIONS: Our findings indicate that heavy drinkers may be cautioned predictive likelihood for them to induce drinking-induced diseases by analyzing their MnSOD genotypes and immunological biomarkers.

Change of glycaemic control and predictors in diabetes patients: longitudinal observational study during the one year after hospital discharge.

Glycated haemoglobin (HbA1c) might reflect glycaemic control in persons with diabetes. Study aims were to identify changes in glycated haemoglobin values and predictors (baseline coping behaviour, fasting plasma glucose, disease-related and demographic factors) in patients during 1 year after hospital discharge. A longitudinal prospective design with convenience sampling was used. Subjects were recruited from a community hospital in Taiwan. Measures included Jalowiec Coping Scale, fasting plasma glucose, HbA1c values, and demographics. Generalized estimating equation was used to determine factors of change in glycated haemoglobin. A total of 57 patients completed 1 year of follow-up. Half did not receive diabetes mellitus education and regular exercise. Patients' glycated haemoglobin levels follow controls at 6 months after discharge. Patients with higher levels of blood glucose, less problem-focused coping and greater emotion-focused coping were associated with poor glycaemic control. Education programmes should involve individual-centred care and health behaviours for prevention of diabetes complications.

Elevated lactate dehydrogenase activity and increased cardiovascular mortality in the arsenic-endemic areas of southwestern Taiwan.

Arsenic ingestion has been linked to increasing global prevalence of and mortality from cardiovascular disease (CVD); arsenic can be removed from drinking water to reduce related health effects. Lactate dehydrogenase (LDH) is used for the evaluation of acute arsenic toxicity in vivo and in vitro, but it is not validated for the evaluation of long-term, chronic arsenic exposure. The present study examined the long-term effect of chronic arsenic exposure on CVD and serum LDH levels, after consideration of arsenic metabolism capacity. A total of 380 subjects from an arseniasis-endemic area and 303 from a non-endemic area of southwestern Taiwan were recruited in 2002. Various urinary arsenic species were analyzed using high-performance liquid chromatography (HPLC) and hydride generation systems. Fasting serum was used for quantitative determination of the total LDH activity. A significant dose-response relationship was observed between arsenic exposure and LDH elevation, independent of urinary arsenic profiles (P<0.001). Furthermore, abnormal LDH elevation was associated with CVD mortality after adjustment for Framingham risk scores for 10-year CVD and arsenic exposure (hazard ratio, 3.98; 95% confidence interval, 1.07-14.81). LDH was elevated in subjects with arsenic exposure in a dose-dependent manner. LDH is a marker of arsenic toxicity associated with CVD mortality. Results of this study have important implications for use in ascertaining long-term arsenic exposure risk of CVD.

Toona sinensis modulates autophagy and cytokines in lipopolysaccharide-induced RAW 264.7 macrophages.

Toona sinensis (TS) is a medicinal herb possessing anti-apoptotic, anti-oxidant, and anti-inflammatory properties and is used to treat diabetes, cancer, and inflammatory diseases. In traditional Chinese medicine theory, TS clears dampness and heat, strengthens the stomach function, and regulates vital energy flow. TS is also used as an astringent and a pesticide. In this study, we aimed to evaluate how TS influences autophagy and cytokines during the inflammatory process in RAW 264.7 macrophages. The treatment groups were pre-supplemented with TS leaf extract; rapamycin was used to enhance autophagy and lipopolysaccharide (LPS) was used to induce inflammation. The expression of autophagy-related proteins was analyzed by western blotting. The survival rate of, and chemokine expression and oxidative stress in the cells were also assessed. TS leaf extract inhibited mammalian target of rapamycin (mTOR) phosphorylation at site S2448 in the macrophages. At relatively higher concentrations (50 and 75 µg/mL), TS elevated the expression of light chain 3 II (LC3-II), which further modulated autophagy. Pre-supplementation with TS leaf extract elevated the total glutathione (GSH) level and GSH/oxidized GSH (GSSG) ratio, but it decreased the GSSG, total nitric oxide, nitrate, nitrite, malondialdehyde, and superoxide anion levels. TS reversed the effects of LPS-induced cytokines, including interleukin (IL)-6 and IL-10. TS did not induce significant toxicity at the studied concentrations. In conclusion, TS leaf extract may modulate autophagy during inflammation. Furthermore, it may prevent cell damage via anti-inflammation and anti-oxidation. Thus, this study supports the ethnomedical use of TS in the prevention of inflammation-related diseases.

All-trans retinoic acid remodels extracellular matrix and suppresses laminin-enhanced contractility of cultured human retinal pigment epithelial cells.

All-trans retinoic acid (atRA) has been reported to inhibit the proliferation of retinal pigment epithelial (RPE) cells and used in treatment of proliferative vitreoretinopathy (PVR) in animal model. This study aimed at examining the effectiveness of atRA in inhibiting the extracellular matrix (ECM) biosynthesis by RPE cells and the RPE cell-mediated collagen gel contraction. Cultured RPE cells were treated with atRA and the expression of four ECM proteins (collagen types I, III, IV and laminin beta1) was profiled. The results indicated that atRA treatment up-regulated de novo synthesis of collagen type I, but decreased that of laminin beta1 in a dose-dependent manner. Moreover, the effect of atRA on RPE cell contraction was evaluated by measuring the area of collagen gel where RPE cells populated. Treatment with atRA significantly inhibited RPE cell-mediated collagen gel contraction. Addition of exogenous laminin nonapeptide into gels promoted RPE cell contraction, while atRA reversed the laminin-enhanced contractility. atRA treatment significantly suppressed the gene expression of integrin beta3 but not alphaV subunit, and effectively inhibited the tyrosine phosphorylation of integrin beta3 at residue 747 in RPE cells grown on laminin-coated dish, suggesting that atRA may suppress the RPE contractility through either inhibiting integrin beta3 expression or abrogating the integrin beta3-mediated signaling. In conclusion, atRA pharmacologically possesses a tissue-remodeling capacity and inhibits contractility of RPE cells. Therefore, atRA might be potentially a therapeutic agent for certain ocular disorders such as PVR.

A case-control study of the HER2 Ile655Val polymorphism and risk of breast cancer in Taiwan.

OBJECTIVES: To investigate the HER2 Ile655Val polymorphism in relation to risk of breast cancer in a case-control study in Taiwan. DESIGN AND METHODS: The HER2 polymorphism at codon 655 was analyzed in 424 patients with breast cancer and 318 controls by using the polymerase chain reaction methodology, followed by the restriction fragment-length polymorphism (PCR-RFLP) analysis. RESULTS: There was a 1.48-fold (95% CI=1.00-2.24) increase in the risk of patients with breast cancer who are Val carrier (Ile/Val and Val/Val genotypes). Furthermore, for the early onset (less than 45 years old) breast cancers with Val carrier, there was a 2.24-fold (95% CI=1.17-4.34) increase in the risk of breast cancer. CONCLUSIONS: Our results indicate that the Val carrier was associated with increased risks in patients with breast cancer in Taiwan. The association was more apparent in patients who were younger than or equal to 45 years of age.

Roles of the genetic variation of alcohol-metabolizing enzymes on biomarkers in trauma patients with excessive alcohol intake at emergency department.

BACKGROUND: Alcohol abuse has been implicated as an important factor for accidents. We evaluated the roles of different genetic combinations of the ADH2 and ALDH2 genotypes on biomarkers in trauma patients with excessive alcohol intake at our emergency department. METHODS: Blood samples were obtained from 80 patients and 88 age-matched controls. The biomarkers, including AST, ALT, GGT, and MDA, were assayed. The polymerase chain reaction-restriction fragment length polymorphism method was used to determine the genetic polymorphisms of ADH2 and ALDH2. RESULTS: There were significant differences in the levels of AST, ALT, GGT, MDA, and AST/ALT ratios between the 2 groups. In addition, MDA values and AST/ALT ratios were significantly higher in the patients with normal activity of ADH2 than the patients with low activity of ADH2. Meanwhile, regarding ALDH2 genotypes, there were significantly higher ratios of AST/ALT in the patients with low activity of ALDH2. The highest AST/ALT ratios and MDA values were in the patients with ADH2 (*2/*2) and ALDH2 (*1/*2 and *2/*2). CONCLUSIONS: In conclusion, our results indicated that alcohol-induced liver damage or oxidative stress might be influenced by the genetic variation of ADH2 or ALDH2. Therefore, the combinations of different ADH2 and ALDH2 genotypes may be influential markers for susceptibility to alcohol-induced liver damage.

Distinct differences in the induction of stimulus-mediated superoxide generation by polymorphonuclear neutrophils isolated from patients with different types of leukemia.

BACKGROUND/PURPOSE: Accumulating literature has documented that there exists a distinct difference in nitro blue tetrazolium reduction capacity by the polymorphonuclear neutrophils (PMNs) from patients with different types of leukemia. The underlying mechanism associated with this observed phenomenon remains to be clarified. METHODS: The production of O2-, monitored by a validated probe (lucigenin)-based ultraweak chemiluminescence, in resting and/or phorbol-1,2-myristate-1,3-acetate (PMA)- and zymosan-stimulated systems of various leukemic PMNs was measured. In parallel with these studies, we also quantified superoxide dismutase isozymes (Cu, Zn-SOD, Mn-SOD) from these isolated PMNs by established methods. RESULTS: A marked increase was observed in O2- generation by the PMNs from patients with acute myeloid leukemia (AML), and chronic myeloid leukemia (CML), but not from patients with acute lymphocytic leukemia (ALL) when compared with controls either in the resting condition or after being stimulated by either PMA or zymosan. In parallel, we also quantified SOD isozyme activities and found that the total and CuZn-SOD of PMNs from AML were indeed significantly lower than either controls or ALL, implying that higher levels of O2- generation might result from a deficiency in this O2- metabolizing enzyme. CONCLUSION: Our data suggest that a distinct difference in the capability of O2- generation under stimulated conditions between PMNs from ALL and AML (or CML) may be of potential taxonomic or even therapeutic usefulness.

Antioxidant status and superoxide anion radical generation in acute myeloid leukemia.

OBJECTIVES: This study was undertaken to investigate if there is a disparity in the antioxidant status and the ability of superoxide anion (O(2)(-)) generation in the patients with acute myeloid leukemia (AML). DESIGN AND METHODS: The peripheral blood samples from thirty AML patients and thirty-six healthy subjects were collected and leukocytes, erythrocytes and plasma were separated for use in various parameter measurements. RESULTS: The generation of O(2)(-), as reflected by lucigenin-based CL (LBCL), by the leukocytes of patients with AML was found to be significantly elevated either in resting or stimuli-elicited condition as compared with that of healthy controls (p<0.05). Coincidentally, these data were matched up with the suppressed SOD activities, notably in Cu/Zn SOD isoform found in AML patients (p<0.05). Conversely, SOD and GPx activities in erythrocytes of patients with AML were shown to be significantly higher than their normal counterparts (p<0.05). CONCLUSIONS: These data suggest that altered expression of antioxidant enzymes and higher capability of O(2)(-) generation by leukocytes seem to be a distinct feature of AML.

Glutathione status in the blood and tissues of patients with virus-originated hepatocellular carcinoma.

OBJECTIVES: Glutathione status can be regarded as the redox status for many diseases. This study was performed to investigate the glutathione status in virus-originated hepatocellular carcinoma (HCC). DESIGN AND METHODS: The blood and tissue samples were obtained from 24 patients. Blood samples were also obtained from 137 controls for comparison. RESULTS: The GSH level and the ratios of GSH/GSSG and GSH/total glutathione of the blood samples from the patients were significantly lower than those of the controls, while the GSSG values were significantly higher. Meanwhile, levels of GSH and total glutathione, as well as the ratios of GSH/GSSH and GSH/total glutathione, were significantly decreased, whereas GSSG levels were significantly higher, in the HCC tissues than those of the adjacent cancer-free tissues. CONCLUSIONS: Glutathione status in the HCC suggested that the antioxidant system is severely impaired, supporting a consistent role of the free radical cytotoxicity in the pathophysiology of the disease.

Distribution of alcohol-metabolizing enzyme genotypes in trauma patients with excessive alcohol consumption in the emergency department.

OBJECTIVES: The objective of this study was to investigate the distribution of genetic polymorphisms of alcohol-metabolizing enzymes in trauma patients with excessive alcohol consumption in the emergency department (ED). DESIGN AND METHODS: A total of 100 trauma patients and age-matched control subjects composed of 98 participants were enrolled in this study. The activities of liver enzymes and genotypes of alcohol-metabolizing enzymes, including ADH2, ALDH2, and CYP2E1, were analyzed with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: There was a significant difference in the allele frequencies of ALDH2 between the two groups. For the genotypes, there were significant differences in the genotype frequencies of ADH2 and ALDH2. There was also a significantly lower frequency in patients with the ALDH2*2 phenotype than those of the controls. For the activities of liver enzymes, there were significant differences between the two groups. For ADH2 and ALDH2, there were significantly higher ORs (odds ratios) in trauma patients with normal activity than those with weak or intermediate activity but there were no significant difference in CYP2E1 genotype between two groups. To investigate the interaction of alcohol-metabolizing enzyme genotypes, we have estimated the odds ratios in two alcohol-metabolizing pathways. The ORs of the combined genotypes of ADH2 (*1/*1+*1/*2) and ALDH2 (*1/*1) and the combined genotypes of either CYP2E1 (*c1/*c1) or CYP2E1 (*c1/*c2+*c2/*c2) and ALDH2 (*1/*1) were significantly higher than that of the reference group in the major and the minor pathway, respectively. CONCLUSIONS: Genetic variation of alcohol-metabolizing enzymes, especially ALDH2, may play an important role on the occasions of alcohol problems in the emergency department.

Whey Protein Concentrate Renders MDA-MB-231 Cells Sensitive to Rapamycin by Altering Cellular Redox State and Activating GSK3ß/mTOR Signaling.

Whey protein concentrate (WPC) is an amino acid-rich supplement that has been shown to increase cellular antioxidant capacity. Mammalian target of rapamycin (mTOR) is a crucial regulator of signaling in mammalian cells, and serves as a therapeutic target for triple-negative breast cancer (TNBC). This study was designed to investigate the effect of combining WPC with rapamycin on MDA-MB-231 human breast cancer cells. These cells were found to be insensitive to rapamycin and exhibited higher glutathione (GSH) and reactive oxygen species levels than non-tumorigenic MCF-10A cells. However, for MDA-MB-231 cells, the half maximal inhibitory concentration of rapamycin was lower when this drug was administered in combination with WPC than when used alone. Furthermore, combining WPC with rapamycin depleted GSH levels and reduced Nrf2 nuclear accumulation. In addition, WPC activated GSK3ß/mTOR signaling, and GSK3ß appeared to be involved in the WPC-mediated Nrf2 reduction and mTOR activation. In conclusion, WPC induced rapamycin sensitivity in MDA-MB-231 cells by altering their redox state and activating GSK3ß/mTOR signaling. These results not only suggest a novel therapeutic approach for breast cancer treatment, but also provide insight into the critical pathways affecting the resistance to mTOR inhibition observed in a subgroup of TNBC patients.

Selective effects of whey protein concentrate on glutathione levels and apoptosis in rats with mammary tumors.

Glutathione (GSH) plays an important role in antioxidant defense and regulation of apoptosis. GSH deficiency is related to many diseases, including cancer, and increased GSH levels in cancer cells are associated with chemotherapy resistance because of resistance to apoptosis. In this study, we investigated the effects of whey protein concentrate (WPC), a precursor of GSH, in rats with mammary tumors induced by treatment with 7,12-dimethylbenz(a)anthracene (DMBA). DMBA treatment results in cellular changes that mimic the initiation and promotion of carcinogenesis of breast tissue. We aimed to examine the possible preventive effects of diets containing whey protein on DMBA-induced mammary tumors in rats. The results indicate that WPC (0.334 g/kg) supplementation significantly increased the liver GSH levels by 92%, and were accompanied by low Bax/Bcl-2 ratio (from 5 to 3) and cleaved caspase-3/procaspase-3 ratio (from 2.4 to 1.2) in DMBA-treated rats. Furthermore, tumor GSH levels were decreased by 47% in WPC-supplemented rats, which resulted in increased Bax/Bcl-2 ratio (from 0.9 to 2) and cleaved caspase-3/procaspase-3 ratio (from 1.1 to 2.7). In conclusion, supplementation with WPC could selectively deplete tumor GSH levels and, therefore, WPC supplementation might be a promising strategy to overcome treatment resistance in cancer therapy.

Blood-brain barrier dysfunction occurring in mice infected with Angiostrongylus cantonensis.

Several indices were used to assess whether blood-brain barrier (BBB) damage occurs in neurological disorders. Dysfunction of the BBB was surmised to be involved in the pathological changes of eosinophilic meningitis caused by the infection of Angiostrongylus cantonensis. The mean concentration of protein and albumin in the cerebrospinal fluid (CSF) of infected mice gradually increased from days 0 to 18 after infection and then rapidly increased 21 days after infection. The concentrations of protein and albumin in the CSF of infected mice 15 days after infection were all significantly higher than those in uninfected mice (all P-values at least <0.05). Parallel with the increase in protein and albumin in the CSF, infected mice showed a gradual increase in their CSF/serum protein and albumin ratios. The increase became significant at days 21 and 18 after infection, respectively (P<0.01 and P<0.05, respectively). The higher the worm counts in the brain, the higher the CSF/serum albumin ratio was observed in infected mice at day 21 after infection (P<0.001). In addition, the ratios of the CSF/serum albumin were positively correlated with the worm counts in the brain (P<0.001). The total leukocyte and eosinophil counts were also positively correlated with ratios of CSF/serum albumin (P<0.01). The amount of Evans blue in the brain of mice 21 days after infection from peripheral blood via BBB became significantly increased than those in uninfected mice (P<0.001). Thus, the evidence of high concentrations of protein and albumin, high leukocyte counts in CSF, high ratio of CSF/serum protein and albumin, and high permeability of BBB show that dysfunction of the BBB occurred in mice infected with A. cantonensis.

Development of probe-based ultraweak chemiluminescence technique for the detection of a panel of four oxygen-derived free radicals and their applications in the assessment of radical-scavenging abilities of extracts and purified compounds from food and herbal preparations.

We report here the development of a probe-based ultraweak chemiluminescence (uwCL) method capable of detecting a panel of four oxygen-derived free radicals (ODFRs) including superoxide (O2-), hydrogen peroxide (H2O2), hydroxyl radical (*OH), and peroxyl radical (ROO*) using different probes specific for these radicals performed by the same uwCL analyzer. The selected radical-generating systems and their corresponding uwCL-probing emitters were validated. These ODFR-detecting systems were subsequently utilized by us to assess the radical-scavenging ability (RSA) of a variety of extracts and purified constituents derived from foods and herbal preparations. Our approach for assessing RSA for these constituents is based on the suppression of uwCL generated by each ODFR, and the degrees of inhibition have been shown to be dose-dependent. For this reason, the estimation of IC50 for each testing compound can be obtained from the curve constructed based on the percent of inhibitions of uwCL versus the concentrations of the compound tested. To illustrate the practical applications of our devised methodology, data for comparative studies of RSA activities of fermented extracts of Cordeceps sinensis, purified methylgallate isolated from Toona sinesis, resveratrol purified from grape seeds, plus epimedin C from the aerial part of the Epimedium plant (yinyanghuo) are to be presented.

Whey protein concentrate promotes the production of glutathione (GSH) by GSH reductase in the PC12 cell line after acute ethanol exposure.

Excessive ethanol consumption may increase the production of reactive oxygen species (ROS), which results in the damage of tissues, especially the neurons and glial cells in the central nervous system (CNS). The purpose of this study is to evaluate the effects of whey protein concentrate (WPC) on the glutathione (GSH) status after acute ethanol exposure in the pheochromocytoma (PC12) cell line. In this study, we assayed the cell viability, the percentage of lactate dehydrogenase released (% LDH released), the level of GSH, and the activity of GSH reductase (GRx). The results showed that with the supplement of WPC, the cell viability displayed no significant difference after acute exposure of ethanol in groups with or without ethanol treatment. The ethanol-induced cytotoxicity showed a slight decrease ,and the level of GSH showed a significant increase. The activity of GRx significantly increased when 0.1, 10mg/ml of WPC was supplied. In conclusion, these results suggest that WPC in a moderate concentration should be a precursor agent to promote the production of GSH and will enhance the antioxidant capacity in the PC12 cell line.