In this study we investigated the effect of YHK supplementation on the activities of antioxidant, phase I and phase II metabolizing enzymes involved in detoxification as well on liver antioxidant defense system in rats. YHK was administered for four weeks to Wister rats. At the end of the treatment period, different cytochrome P450 (CYP) isoform and phase II enzyme activities were determined by incubation of the liver microsomes or cytosols with appropriate substrates Dietary supplementation of YHK (2%, w/v) to male rats for four weeks significantly increased the activities of glutathione peroxidase and catalase to 118% and 87% in liver as compared with corresponding normal diet fed control (P<0.05-0.001). CYP 1A2 activity was markedly increased in all the YHK treatment groups (P!0.05). CYP 1A1 activity was increased significantly in all the groups.Parallel to these changes, YHK feeding to mice also resulted in a considerable enhancement in the activity of phase I and II metabolizing enzymes such as glutathione S-transferase to 1.6 fold and 1.8 fold in liver as compared with corresponding normal diet fed control (P<0.05-0.01). The induction of such detoxifying enzymes by YHK suggest the potential value of this compound as protective agent against chemical carcinogensis and other forms of electrophilic toxicity. The significance of these results can be implicated in relation to cancer chemopreventive effects of YHK against the induction of tumors in various target organs.

OBJECTIVE: Acute radiation of the small intestine causes an immediate and potentially reversible effect on the sensitive regenerative epithelium of the intestinal mucosa while markedly altering the overall intestinal ecosystem. The aim of the present study was test a novel probiotic mixture formulation (Microflorana-F) in an experimental model of acute radiation enteritis with particular interest in endotoxinemia and bacterial translocation. MATERIALS ANDMETHODS: Male Wistar rats allocated to three groups were fed for 7 days with: (A) a standard balanced diet; (B) a standard diet with the addition of 1 mL t.i.d. of Microflorana-F and (C) the same probiotic but after heat inactivation. Under ketamine anesthesia, abdominal irradiation was performed at a single dose rate of 20 Gy. Sham-radiated healthy rats served as a control (D). Standard food and active/inactive probiotic supplementation schedule was maintained throughout the study period. When they were killed 14 days later a midline laparotomy and a medium sternotomy was carried out. The mesenteric lymph nodes, whole spleen and liver samples as well as blood, the portal vein and bile samples were cultured. Endotoxinemia was also measured.RESULTS: Early deaths (1 week) occurred mostly in rats fed standard food or inactivated probiotic. The endotoxin level significantly increased in irradiated rats fed standard food and inactivated probiotic while supplementation with the active form of the probiotic mixture significantly improved such parameters (P < 0.05). After radiation injury, mesenteric lymph nodes and portal blood were the samples most frequently yielding bacterial growth. Treatment with only the active form of probiotic significantly reduced the incidence of bacterial contamination in all samples.CONCLUSIONS: These data suggest that the manipulation of gut ecosystem by biologically effective probiotic preparations might be a worthwhile therapeutic and preventive tool in radiation-induced enteritis.

Hepatocytes isolated from 20- and 4-month Wistar rats and cultured with or without alpha-linolenic acid (LNA) were then added with nutraceutical YHK or sylibin before the test with iron or copper. Overall, YHK proved to be more effective than sylibin in Fe/Cu-induced peroxidative damage on normal and LNA-loaded hepatocytes (p < 0.05). YHK exerted a significant protection against DPPH radical-scavenging activity in the "old" group (p versus sylibin) and against lipophilic generators in both age groups (p < 0.05 versus sylibin). Both compounds were ineffective on age-related increase of surface-charge density. These preliminary data suggest that age per se enhances the vulnerability of hepatocytes to xenobiotics, whereas some safe nutraceuticals seem to exert significant protective effects.

In the present study we tested a natural compound, L e. K-17.22, which is endowed by antioxidant properties and transaminases-lowering effect in HCV patients, on free radicals-related liver damage by ischemia/reperfusion injury. Wistar rats were put on a 2-week supplementation with either A) standard diet or B) standard diet added with 30mg of K-17.22 (Yojyo-Henshiko: K-17.22, Kyotsu Inc., Tokyo, Japan). A classical ischemia/reperfusion liver model was prepared and after 60min of reperfusion, hepatic tissue blood flow was measured and the rats were sacrificed. Sham-operated rats were used as controls. A separate set of forty rats undergoing liver ischemia. with or without dietary treatment were used for survival study. The following parameters were checked: liver tissue peroxides, SOD, Catalase, glutathione metabolism, hepatic tissue blood flow by H2 clearance method and radicals-trapping ability of K-17.22 by ESR. As compared to controls, after 60 min of reperfusion, B group rats showed a significantly lower MDA level (P<0.05) with an overall impairment of the liver antioxidant defense system (SOD, Catalase, GSH, GSSG and GSH-Px) (P<0.001). All these parameters were less affected in the B group (P<0.05). In particular, GSH and GSH-Px reverted to normal level with a significantly lower GPT level (P<0.05). However, K-17.22 didn't show any direct free radicals-trapping ability either on superoxide nor on hydroxyl radical generation systems. Ischemia-reperfusion phenomenon brought about a nearly 40% drop of the liver blood flow in the standard diet-fed rats (P<0.001 vs. sham-operated). Pre-treatment with K-17.22 enabled a recovery of such haemodynamic parameter (P<0.05 vs. untreated group). One-week survival study showed that, in the absence of any treatment, only 20% of rats survived after liver ischemia. while B group rats yielded a 45% survival rate (P<0.05). Even using a severe liver damage model as the present one, the present nutritional approach seems to offer some noteworthy boosting ability on the endogenous free radicals scavengers array which is unrelated to its direct in vitro action.

A bstract : The aim of this study was to test the effect of antioxidant supplementation on enzymatic abnormalities and free radical‐modified DNA adducts associated with premalignant changes in the gastric mucosa of elderly patients with HP‐negative atrophic gastritis (CAG). Sixty patients with atrophic gastritis and intestinal metaplasia underwent a nutritional interview and a gastroscopy with multiple biopsy samples in the antrum that were processed for histology and for assaying: alpha‐tocopherol, MDA, xanthine oxidase (XO), ornithine decarboxylase (ODC), and 8‐OHdG. Patients were randomly allocated into three matched groups and supplemented for 6 months with (1) vitamin E, 300 mg/day; (2) multivitamin, two tablets t.i.d.; and (3) Immun‐Age 6 g/day nocte (ORI, Gifu, Japan), a certified fermented papaya preparation with basic science‐validated antioxidant/immunomodulant properties. Ten dyspeptic patients served as controls. Histology and biochemistry were blindly repeated at 3 and 6 months. CAG patients showed a significantly ( P < .05 ) increased level of mucosal MDA and XO concentration that were reverted to normal by each supplementation ( P < .05 ). All supplements caused a significant decrease of ODC ( P < .01 ), but Immun‐Age yielded the most effective ( P < 0.05 ) and was the only one significantly decreasing 8‐OhdG ( P < 0.05 ). These data suggest that antioxidant supplementation, and, namely, Immun‐Age, might be potential chemopreventive agents in HP‐eradicated CAG patients and especially in the elderly population.

Healthy mice were allocated into three groups: A) control; B) infected (1) group and Q dietary supplement (MMT)-treated I-group. Mice were infected intranasally with 30 mul of 75HA units of virus. I-MMT groups received 5mg of MMT t.i.d. Signs of infection, total inflammatory cell counts in nasal washings, virus titres in lungs homogenates and plasma MDA were serially determined for 8 days. At sacrifice bronchoalveolar lavage fluid (BALF) collection and lung tissue were examined for superoxide radical production and for MDA, ascorbic acid and TNFalpha activity. MMT markedly blunted the nasal signs of virus infection and reduced the febrile response. Formazan-positive cells and MDA in the lung extract and in the plasma as well as of TNF-alpha in lung tissue significantly increased during viral infection but a significant improvement was observed in C group. SOD, catalase activities and ascorbic acid significantly decreased in the infected groups but not in C group which showed also a significant decrease of viral titre in the lung. No toxicity was detected up to dosages over 50-fold higher. It is suggested that a safe natural compound, i.e. MMT, has the potential to be applied in clinical practice while further studies are ongoing to elucidate its mechanism of action in more detail.

The aim of this study was to test the effect of antioxidants on enzymatic abnormalities and free radicals-modified DNA adducts associated with gastric pre-malignant changes in patients with HP-negative chronic atrophic gastritis (CAG). Sixty patients with known atrophic gastritis and intestinal metaplasia underwent a gastroscopy with biopsy samples to be processed for histology and for assaying: alpha-tocopherol, MDA, xanthine oxidase (XO), ornithine decarboxylase (ODC) activity and 8-OHdG. Patients were randomly allocated into 3 matched groups which were supplemented for 6 months with: A) vitamin E 3001U/day; B) Multivitamin 2tabs/day and Q Immun-Agc FPP 6g/nocte, which is a certified fermented papaya preparation endowed of basic-science validated antioxidant/immunomodulant properties. Plasma oxidant/antioxidant status was within normal limits in all groups. CAG patients showed a significantly (p<0.05 vs control) increased level of mucosal MDA and XO concentration which were reverted to normal by each of the 3 supplementations (p<0.05). All 3 supplements caused a significant decrease of ODC activity (p<0.01) but at 6-month observation Immun-Age yielded the most significant effect (p<0.05 vs groups A and B) and was the only one determining a significant decrease of 8-OhdG (p<0.05 vs baseline and other groups). These data suggest that antioxidant supplementation, and namely Immun-Agc, might be potential chemopreventive agents in HP-eradicated CAG patients.

The aim of this study was to test the effect of antioxidants on enzymatic abnormalities and free radicals-modified DNA adducts associated with pre-malignant changes in HP-negative chronic atrophic gastritis (CAG) patients. 60 patients with CAG and intestinal metaplasia underwent a GI endoscopy with biopsy samples for histology and for: alpha-tocopherol, malonyldialdehyde, xanthine oxidase (XO), ornithine decarboxylase (ODC) and 8-hydroxydeoxyguanosine (8-OhdG). Patients were randomly allocated into three groups supplemented for 6 months with: (A) vitamin E, 300 mg/day; (B) Multivitamin, 2 tablets/day and (C) a certified fermented papaya preparation 6 g/nocte (Immune-Age FPP, Osato Research Institute, Gifu, Japan). Ten dyspeptic patients without histological abnormalities served as control. Histological and biochemical parameters were blindly repeated at 3 and 6 months. Plasma oxidant/antioxidant status was normal in all groups. CAG patients had a significantly (p<0.05 vs. control) increased mucosal level of MDA and XO concentrations which were returned to normal by each of the three supplementations (p<0.05). All three supplements decreased ODC activity (p<0.01) but at 6-month observation FPP yielded the most significant effect (p<0.05 vs. groups A and B) and was the only one achieving a significant drop of 8-OhdG (p<0.05 vs. baseline and other groups). The present data suggest that antioxidant supplementation, and namely Immune-Age FPP, might be a potential chemopreventive agent in HP-eradicated CAG patients.

BACKGROUND/AIMS:It has been shown that alcohol impairs erythrocyte (red blood cell) membrane fluidity and lipid composition. The aim of this study was to test the effect of a novel acid-resistant antioxidant on the hemorrheology in alcoholics.METHODOLOGY:Thirty alcoholics (25 males, 5 females; mean age: 42 years; range: 31-54; 150 g ethanol/day for 3-5 years) were enrolled into the study. Patients were randomly and double-blindly allocated into 2 groups which were given, for a 2 week period, 18 g/day of Bionormalizer (obtained from biofermentation of carica papaya, pennisetum purpureum, sechium edule, Osato Res. Foundation, Gifu, Japan) dissolved in 5 mL of water at bedtime and 3 hours prior to examination. Placebo consisted of flavored sugar. Healthy teetotalers served as control. On the examination day, blood samples were taken for testing: routine tests, plasma glutathione, ascorbic acid, selenium, plasma lipid hydroperoxides and alpha-tocopherol. Erythrocytes were separated and tested for red blood cell malonyldialdehyde and glutathione content. The hemorheological studies were as follows: blood and plasma viscosity, whole blood filterability, red blood cell membrane fluidity by electron spin resonance, red blood cell aggregation index by photometric rheoscopy and red blood cell deformability by ektacytometry.RESULTS:As compared to healthy controls, alcoholics on placebo treatment showed no change of plasma viscosity but a significantly higher red blood cell malonyldialdehyde, blood viscosity (P < 0.05) and lower plasma glutathione, whole blood filterability and red blood cell fluidity (P < 0.01). No relationship appeared between biochemical tests and red blood cell membrane fluidity. Bionormalizer group showed a significant recovery to control values of either blood viscosity and whole blood filterability (P < 0.01) and a partial, although significant, improvement of red blood cell membrane fluidity, red blood cell malonyldialdehyde and plasma glutathione (P < 0.05). As compared to healthy control, red blood cell aggregation decreased in alcoholics (P < 0.05) and was not affected by Bionormalizer. However, Bionormalizer significantly improved the reduced red blood cell deformability (P < 0.05 vs. alcoholics) and this parameter correlated with red blood cell malonyldialdehyde (r: 0.62. P < 0.05).CONCLUSIONS:These preliminary data suggest that an effective antioxidant supplementation is able to improve the hemorrheology in alcoholics either by directly affecting the ethanol-related lipoperoxidation and xanthine oxidase system activation and/or by modifying red blood cell membrane characteristics.

BACKGROUND/AIMS:Thirty alcoholic patients and 24 teetotaler dyspeptic patients were considered and underwent baseline blood chemical evaluation and the Schilling test.METHODOLOGY:During gastroscopy, biopsy samples were taken to assay: routine histology, malonyldialdehyde, vitamin E and glutathione concentration and for testing vitamin B12-Intrinsic Factor binding. Examinations were repeated after 1-week supplementation with Bionormalizer.RESULTS:Plasma malonyldialdehyde level and lipid hydroperoxides concentration as well as either malonyldialdehyde and xanthine oxidase concentration in the gastric mucosa in alcoholics were significantly higher than in controls and despite unchanged alcohol consumption, significantly decreased after Bionormalizer supplementation. Gastric mucosal glutathione was markedly depressed in alcoholics and partly recovered after Bionormalizer supplementation. Although the alcoholics showed a normal intrinsic factor secretion in the gastric juice, they exhibited a markedly depressed intrinsic factor-cobalamin binding on the "ex vivo" study. Moreover, nearly 23% of them had an abnormal Schilling test. Both these impairments reverted to normal after Bio-normalizer supplementation.CONCLUSIONS:It can be postulated that the antioxidative action played by Bionormalizer, possibly due to its availability substrates for glutathione synthesis as well as to its effects on local oxidative burst from neutrophil, is able to recover a normal cobalamin absorption.

therapy 175 Bile lipids 515 -secretion 515 Binary complexes 305 Bionormalizer 538 Bisacodyl 69 Bosentan 484 Bullfrog duodenum 324 Caco-2 cells 238 Caerulein 56 Calcitonin gene-related peptide 338 cAMP 324 Cancer 216 -, gastrointestinal tract

Twenty-two healthy teetotal volunteers underwent gastroscopy during which biopsy samples from the antrum and body were taken for chemiluminescence assay, routine histology, and for malonyldialdehyde, xanthine oxidase and glutathione determination. Subjects were divided into 2 groups which, in a double-blind fashion, were randomly and orally given either (a) Bionormalizer 9 g at bedtime and 3 h prior examination, or (b) flavored sugar 9 g as placebo. During the second gastroscopy 40 ml of 80% ethanol were sprayed perendoscopically. Gastroscopy with biopsy was repeated 60 min later. As compared to the placebo group, subjects given Bionormalizer showed significantly reduced gastric mucosal damage at endoscopy and the histological level. When considering the placebo group, ethanol administration brought about a significant increase in the luminol-amplified chemiluminescence response in gastric mucosa as compared to the baseline value which was correlated with the histological score. The mean chemiluminescence value in the Bionormalizer group was significantly lower than in the placebo group. Ethanol ingestion brought about a significant increase in xanthine oxidase and malonyldialdehyde together with a decreased glutathione concentration. Bionormalizer significantly prevented such changes. The present data suggest that the natural antioxidant Bionormalizer when given orally promotes an effective protection against ethanol-induced gastric mucosal damage.

Aim. The aim of this study was to assess the possible effect of a probiotic preparation (Microflorana-F: L. acidophilus, L. helveticus, Bifidobacterium in a vitamin-ion-enriched medium, Named s.r.l., Lemo, Italy) on the age-associated decline of some immunomodulatory properties. Methods. 45 young (7-wk old) and 45 old (19-mo. Old) Swiss mice were split into 2 groups given perorally for up to 14 day: 1ml/day of Microflorana-F; as A, but with heat-killed bacteria. After sacrifice at 2, 7 and 14 day, splenocytes were stimulated with 5 μg/ml ConA and with 2 μg/ml LPS for IFN-gamma and IFN-alfa assay. Peritoneal macrophages aliquots (1x10 6 cells/ml) were used for enzymatic (LDH, β-glucuronidase, β-galactosidase), phagocytosis assays (vs. 1 × 10 9 cell/ml of S. typhi) and O 2 -generation. Results. After either stimulation, baseline production of IFN-gamma and IFN-alfa from splenocytes of aged mice was negligible as compared to young mice (p aged mice: p<0,005). Furthermore the IFN values in the aged mice under treatment were comparable to baseline values in the young group. Microflorana-F enhanced both the enzymatic activity (p<0,05 vs. control) and the opsonized phagocytosis (p<0,05) and O 2 -production (p<0,01; the response in the young mice was comparable to that in the aged mice). Conclusions. These data suggest that an effective probiotic preparation such as Microflorana-F is able to boost the immune system by increasing both IFN production and macrophage activity and, not least, by reverting to normal the age-related decline of IFN production.

BACKGROUND/AIMS The aim of this investigation was to study the oxidative phenomena which take place in the early recovery phase after alcohol withdrawal. Furthermore, the effects of a novel natural antioxidant, Bionormalizer (BN), in such a clinical setting was studied. METHODOLOGY Forty-six alcoholics with moderate drinking habits (daily ethanol intake: > 80g to < 120g) were enrolled in the study, divided into two groups and given either a placebo or 9g of BN by mouth every night for one week. The patients agreed to stop drinking alcohol, and daily blood sampling was obtained for routine tests and to check plasma and erythrocyte levels of MDA, SOD, GPX and the hydroperoxide level. The groups were comparable in terms of initial biochemical parameters. RESULTS BN prevented the early increase of plasma TBARS observed in the placebo group, enabling a near-to-normal level of plasma and erythrocyte MDA by the fourth day. BN also prevented the significant drop of erythrocyte GPX and the transient decrease of plasma SOD observed in the placebo group. Despite alcohol withdrawal, plasma lipid hydroperoxide remained significantly elevated in the placebo group, but this phenomenon was rapidly improved by BN. CONCLUSIONS To a significant extent, BN is able to prevent the free radical-mediated lipoperoxidative changes that occur soon after alcohol withdrawal, while fastening the recovery mechanisms.