Effect of vitamin D supplementation on pegylated interferon/ribavirin therapy for chronic hepatitis C genotype 1b: a randomized controlled trial.

Chronic HCV-infected patients tend to have vitamin D deficiency, suggesting that vitamin D supplementation may enhance the efficacy of treatment with pegylated interferon (PEG-IFN) and ribavirin (RBV). We therefore assessed the effects of vitamin D supplementation on viral response to PEG-IFN/RBV. Eighty-four patients with HCV genotype 1b were randomized, 42 to oral vitamin D supplementation (1000 IU/day) and 42 to nonsupplementation (control), from week 8 to the end of PEG-IFN/RBV therapy. The primary end point was undetectable HCV RNA at week 24 (viral response [VR]). VR rate at week 24 was significantly higher in the vitamin D than in the control group (78.6% vs 54.8% P = 0.037). Adverse events were similar in both groups. When patients were subdivided by IL28B SNP rs8099917 genotype, those with the TT genotype group showed a significantly higher VR rate at week 24 with than without vitamin D supplementation (86.2% vs 63.3% vs P = 0.044). Although patients with the TG/GG genotype, who were relatively resistant to PEG-IFN treatment, had similar VR rates at week 24 with and without vitamin D supplementation, the decline in viral load from week 8 to week 24 was significantly greater with than without vitamin D supplementation. Multivariate analysis showed that rs8099917 genotype and vitamin D supplementation contributed significantly to VR at week 24. SVR rates were similar in the vitamin D and control groups [64.3% (27/42) vs 50% (21/42), P = 0.19]. Vitamin D supplementation may enhance the effects of PEG-IFN/RBV in HCV genotype 1b-infected patients.

Dietary gamma-linolenic acid dose-dependently modifies fatty acid composition and immune parameters in rats.

gamma-linolenic acid (GLA) has been reported to improve several inflammatory disorders through regulation of eicosanoid production. However, since GLA is a precursor of arachidonic acid, it may bring about increasing tissue arachidonic acid levels with subsequent pro-inflammatory events. To explore this possibility, we examined the effect of high-dose GLA acid on the fatty acid profile of immune cells, leukotriene B4 production by peritoneal exudate cells and immunoglobulin productivity of mesenteric lymph node lymphocytes of Sprague-Dawley rats. Male rats were fed 10% fat diets containing graded levels, 0, 20, 40 and 60% of GLA for 3 weeks. The results showed the distinction in activity of metabolizing GLA between immune cells and liver. Thus, in immune cells such as mesenteric lymph node and spleen lymphocytes and peritoneal exudate cells, more dihomo-gamma-linolenic acid was found than in the liver. Leukotriene B4 production by peritoneal exudate cells was significantly suppressed when fed the highest level of GLA suggesting a lower risk of allergic reaction. Moreover, immunoglobulin productivity in mesenteric lymph node lymphocytes was promoted by dietary GLA. The present study indicates that a high dose of GLA may exert anti-inflammatory effects through suppression of leukotriene B4 release and strengthening of gut immune system, thus ameliorating allergic reaction.

Mechanisms of phytosterolemia in stroke-prone spontaneously hypertensive and WKY rats.

Analysis of sterol composition in serum, liver, adipose tissue, adrenals, and abdominal aorta demonstrated that the contents of plant sterols, campesterol and sitosterol, were evidently higher in WKY and stroke-prone spontaneously hypertensive (SHRSP) rats than in Wistar and WKA rats fed a diet containing a 0.5% plant sterol mixture. Lymphatic 24-hour recovery of 3H-sitosterol was about 2-fold higher in the WKY and SHRSP rats than in the WKA rats. Lymphatic absorption of 14C-cholesterol was also higher in WKY and SHRSP rats compared with WKA rats, but the difference was smaller than in the case of sitosterol. The remarkable increase of sitosterol absorption in WKY and SHRSP rats was observed between 9 and 24 hours after the administration. In SHRSP rats, lymphatic absorption of sitosterol between 0 and 3 hours was also higher than those in the other rat strains. Markedly less esterified 3H-sitosterol was detected in lymph than 14C-cholesterol in all strains, and in WKY and SHRSP rats, only a small increase in the esterified forms of sitosterol and cholesterol was observed. Although the incorporation of micellar 3H-sitosterol and 14C-cholesterol into intestinal brush border membranes was higher in SHRSP rats than in WKA rats, no difference was observed between WKY and WKA rats. These observations suggest that the incorporation into the brush border membranes and the esterification of sterols are not the major determinants for the hyperabsorption of sitosterol and cholesterol in SHRSP and WKY rats. Secretion of sitosterol and cholesterol in the bile of rats fed a plant sterol mixture was lower in SHRSP than in WKA rats. These results suggest that WKY and SHRSP strains deposit plant sterols in the body by enhancing the absorption and lowering the excretion of plant sterols. These strains of rats may be suitable models for studying mechanisms of differential absorption of various sterols.

Retrospective study on the effects of lipiodolization before a potentially curative hepatectomy for colorectal liver metastases: long-term results of a pilot study.

BACKGROUND/AIMS: Lipiodolization, a selective regional cancer chemotherapeutic modality using lipiodol plus anticancer drugs, can prolong the survival time of patients with unresectable liver cancer. A preliminary study was conducted with adjuvant lipiodolization before a potentially curative hepatectomy for patients with metachronous colorectal liver metastases. The ultimate aim of this study was to improve the long-term survival after hepatectomy. METHODOLOGY: Twenty-one consecutive patients with colorectal hepatic metastases were included in this study. Seven patients underwent preoperative lipiodolization, while the remaining 14 patients did not receive any preoperative adjuvant therapy. The clinicopathological features and prognoses of these patients were investigated. The median follow-up period after a curative hepatectomy was 56 months. RESULTS: The clinicopathological factors did not differ markedly between the 2 groups. However, the cumulative survival rate of the 7 patients receiving preoperative lipiodolization was significantly (P < 0.05) better than that in those not receiving any preoperative treatment. CONCLUSIONS: Based on the above encouraging findings, we therefore propose that a prospective randomized trial should be carried out to confirm the beneficial effects of our adjuvant chemotherapeutic modality on patient survival following a curative hepatectomy for the patients with colorectal liver metastases.

Interactions of dietary fats and proteins on fatty acid composition of immune cells and LTB4 production by peritoneal exudate cells of rats.

The interaction of dietary fats and proteins on lipid parameters of rats was studied using safflower oil (linoleic acid-rich), borage oil (gamma-linolenic acid-rich) or perilla oil (alpha-linolenic acid-rich) in combination with casein or soybean protein. The experiment was focused on the fatty acid composition of immune cells and the leukotriene B4 production by peritoneal exudate cells. Serum total cholesterol, triglyceride, and phospholipid levels were low in perilla oil-fed or soybean protein-fed rats. Fatty acid compositions of serum and liver phospholipids reflected those of dietary fats. However, feeding borage oil resulted in a marked increase in the proportion of dihomo-gamma-linolenic acid in phospholipids of peritoneal exudate cells, spleen lymphocytes, and mesenteric lymph node lymphocytes in relation to those of liver and serum. It is suggested that activities of metabolic n-6 polyunsaturated fatty acids are different between immune and other tissues. In addition, the magnitude of the reduction of the proportion of linoleic acid of perilla oil in immune cells was considerably more moderate than serum and liver, indicating a different degree of interference of alpha-linolenic acid with linoleic acid metabolism. Leukotriene release from peritoneal exudate cells was in the order of safflower oil > borage oil > perilla oil groups as reflecting the proportion of arachidonic acid, and tended to be lower in soybean protein-fed groups. These suggest an anti-inflammatory property of gamma-linolenic acid as well as alpha-linolenic acid tended to be strengthened when they were combined with soybean protein than with casein.

Cloning and characterization of the Caenorhabditis elegans CeCRMP/DHP-1 and -2; common ancestors of CRMP and dihydropyrimidinase?

The vertebrate CRMP (collapsin-response-mediator protein) gene family comprises at least four members. These CRMPs exhibit about 60% amino acid identity with vertebrate dihydropyrimidinase (DHP), an amidohydrolase involved in the pyrimidine degradation pathway. CRMP is also referred to as DRP (DHP-related protein), TOAD-64 (turned on after division, 64 kDa) and Ulip (Unc-33-like phosphoprotein). These vertebrate CRMPs are expressed mainly in early neuronal differentiation, which suggests that they play a role in neuronal development. In this study we isolated two cDNA clones from nematode C. elegans based on their sequence homology to vertebrate CRMPs and DHP. These two molecules, termed CeCRMP/DHP-1 and -2, turned out to be Ulip-B and -A, respectively, which were previously identified in the C. elegans genomic database by Byk et al. (1998). These newly isolated molecules were believed to represent a common ancestral state before the gene duplication between CRMPs and DHP. CeCRMP/DHP-1 and -2 protein retained all putative zinc-binding residues thought to be essential for the amidohydrolase activity of DHP and exhibited a weak amidohydrolase activity when 5-bromo-dihydrouracil was used as a substrate. Whole-mount in situ hybridization and expression analysis using GFP fusions revealed that CeCRMP/DHP-1 was transiently expressed in the hypodermis of C. elegans during the early larva stage. CeCRMP/DHP-1 was also expressed in a single nerve cell between the pharynx and ring neuropil. On the other hand, expression of CeCRMP/DHP-2 was observed in the body wall muscle throughout the lifespan of C. elegans. These results indicate that a major site of CeCRMP/DHP-1 and -2 expression is non-neuronal. Targeted gene disruption of CeCRMP/DHP-2 caused no particular difference in appearance or movement phenotype.

In vitro fitting accuracy of copy-milled alumina cores and all-ceramic crowns.

PURPOSE: The purpose of this study was to evaluate the marginal and internal fit of alumina cores and all-ceramic crowns in a new preparation design for abutment teeth devised for copy milling. MATERIALS AND METHODS: Using conventional dies and dies from the new type of preparation design, 20 alumina cores were produced by copy milling, 10 cores in each die type. Also using the new type of die, another 10 final crowns were fabricated. Each of the 30 specimens was returned to its original die and embedded in epoxy resin. Each specimen was then sectioned labiolingually and measured for gaps at the margin, in the central area, and at the incisal edge. RESULTS: The alumina cores fabricated using the new type of die had mean gaps of 30 to 40 microns at the margins of the labial and lingual sides, which was significantly smaller than the gaps (67 to 130 microns) found in the alumina cores produced with the conventional type of die. The final crowns had mean gaps of 44 to 63 microns at the margins and on the inner surfaces. There was no significant difference in the gaps between the alumina cores produced using the new type of die and those found in the final crowns. CONCLUSION: The new type of die was effective in allowing the fabrication of all-ceramic crowns that provide excellent marginal and internal fit by copy milling.

Effect of dietary fats and sesamin on the lipid metabolism and immune function of Sprague-Dawley rats.

We examined the effect of three dietary fats, safflower oil (SAF) rich in linoleic acid, borage oil (BOR) rich in gamma-linolenic acid, and perilla oil (PER) rich in alpha-linolenic acid, on the lipid metabolism, and chemical mediator and immunoglobulin levels in Sprague-Dawley rats, as well as the dietary effect of sesame-derived antioxidative sesamin. The serum cholesterol, phospholipid, triglyceride, prostaglandin E2 level and splenic leukotriene B4 level were lower in the rats fed on BOR or PER than in those fed on SAF. SES feeding suppressed the expression of the lipid-decreasing effect of BOR, but not in the rats fed on PER. In respect of the fatty acid composition of the liver and spleen, PER feeding gave a lower arachidonic acid level, and higher eicosapentaenoic and docosahexaenoic acid levels than SAF feeding did, while the effect of BOR feeding was marginal. The effect of SES feeding on fatty acid composition was much smaller than that of dietary fats. In respect of immunoglobulin production, PER + SES feeding gave the lowest IgE productivity in the mesenteric lymph node lymphocytes. These results suggest that PER feeding regulated lipid metabolism and exerted an anti-allergic effect by a different mechanism from that with BOR feeding.

Identification of novel mutations in the dihydropyrimidine dehydrogenase gene in a Japanese patient with 5-fluorouracil toxicity.

5-Fluorouracil (5-FU) is used widely in the treatment of several common neoplasms. Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme in the catabolism of 5-FU. Several recent studies have described a pharmacogenetic disorder in which cancer patients with decreased DPD activity develop life-threatening toxicity following exposure to 5-FU. We reported recently the first Japanese case of decreased DPD activity accompanied by severe 5-FU toxicity. The present study describes the results of molecular analysis of this patient and her family, in which three novel mutations (Arg21Gln, Val335Leu, and Glu386Ter) of the gene coding for DPD were identified. We also revealed that Arg21Gln and Glu386Ter are on the same allele and that Val335Leu is on the other allele, on the basis of analysis of the family genome. Expression analysis in Escherichia coli showed that Val335Leu and Glu386Ter led to mutant DPD protein with significant loss of enzymatic activity and no activity, respectively. The Arg21Gln mutation, however, resulted in no decrease in enzymatic activity compared with the wild type. The present data represent the first molecular genetic analysis of DPD deficiency accompanied by severe 5-FU toxicity in a Japanese patient.

Effects of dietary sesaminol and sesamin on eicosanoid production and immunoglobulin level in rats given ethanol.

The effects of sesaminol and sesamin on the ethanol-induced modulation of immune indices related to food allergy were examined in rats given a low (10%)-casein diet. Chronic ethanol drinking, at the dietary level of 23% (w/w), significantly increased the plasma IgA and IgM concentrations, irrespective of the presence of 0.1% and 0.2% sesaminol, but the effects disappeared with 0.2% sesamin. A significant IgG-elevating effect of these lignans was also found. In contrast, the concentration of plasma IgE was not influenced by the dietary manipulation. Although ethanol drinking did not influence splenic leukotriene B4 production, sesaminol tended to decrease it dose dependently, while sesamin increased the plasma prostaglandin E2 concentration. These results suggest that sesaminol and sesamin seems to have a diverse effect on the plasma levels of immunoglobulins and eicosanoids.

Dietary fibers modulate indices of intestinal immune function in rats.

To explore the immunological roles of dietary fiber, male 4-wk-old Sprague-Dawley rats were fed for 2 wk cellulose (water-insoluble), konjak mannan (water-soluble), pectin (water-soluble) or chitosan (acid-soluble) at 5 g/100 g diet. Serum IgE concentrations in rats fed konjak mannan, pectin and chitosan were significantly lower than in those fed cellulose (mean +/- SEM: 5.0 +/- 1.1, 3.6 +/- 1.3, 3.0 +/- 1.2 and 9.6 +/- 1.9 microg/L, respectively). Rats fed pectin had significantly higher serum IgA and IgG concentrations (358 +/- 38 and 424 +/- 36 mg/L for IgA and IgG, respectively) than those fed cellulose (240 +/- 31 and 337 +/- 25 mg/L) or chitosan (176 +/- 22 and 379 +/- 23 mg/L), while the IgM concentration did not differ among the groups. Concentrations of IgA, IgG and IgM in mesenteric lymph node (MLN) lymphocytes generally were greater, while IgE concentration was lower, in rats fed pectin and chitosan than in those fed cellulose. The proportion of CD4+ T-cells in MLN lymphocytes was also dietary fiber-dependent, and the CD4+/CD8+ ratio was significantly higher in the pectin fed group than in all other groups. Under certain experimental conditions, MLN lymphocytes from rats fed pectin had markedly greater interferon-gamma concentration than cells from other groups, while the effect on tumor necrosis factor-alpha concentration was less marked. Thus, dietary fiber may have an immunoregulatory effect on the intestinal immune system of rats.

Linoleic and alpha-linolenic acids differently modify the effects of elaidic acid on polyunsaturated fatty acid metabolism and some immune indices in rats.

To explore whether the metabolic responses to trans, compared with cis, fatty acids depend on the source of dietary polyunsaturated fatty acids (PUFA), male Sprague-Dawley rats, 5 weeks old, were fed on diets containing 30 g oleic (cis) or elaidic (trans) acids/kg in combination with either 70 g perilla oil (alpha-linolenic acid) or safflowerseed oil (linoleic acid)/kg for 3 weeks in separate experiments. The dietary fats were adjusted to have the same level of total PUFA. The dietary manipulation did not influence the growth indices, but spleen weight was greater when the dietary PUFA source was perilla oil. The incorporation of trans fatty acid into liver phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylserine and adipose tissue lipids, particularly phospholipids, was significantly higher when rats were fed on safflowerseed oil compared with perilla oil. However, only limited differences were observed in the effects of cis and trans fatty acids on the proportions of PUFA in liver phospholipids. Splenic production of prostaglandin E2 was reduced by trans fatty acid when safflowerseed oil was the PUFA source, but no trans effect was observed on leukotriene C4 production. Dietary PUFA significantly influenced the concentration of plasma immunoglobulins (Ig) but the effect of geometry was only seen in IgG which was increased by trans acid. Dietary trans fatty acid increased the CD4+:CD8+ T-lymphocyte ratio in the spleen, reflecting a decreasing trend of the proportion of CD8+, when combined with perilla oil. These observations indicate that the type of PUFA simultaneously ingested specifically influences the effect that trans acid exerts on PUFA metabolism, eicosanoid production and some immune indices.

Effect of dietary fats on cholesterol metabolism and eicosanoid production in hamsters fed undigested fraction of soybean protein.

The undigested fraction of soybean protein (UDF) exerts a markedly greater hypocholesterolemic effect than soybean protein itself in rats. The present study was undertaken to confirm the effect in hamsters, a more appropriate animal model for human cholesterol metabolism. Hamsters were given diets containing UDF at a nitrogen level equivalent to the 20% casein diet. Dietary fats, at the 10% level, were perilla oil and safflower oil. There was apparently no increase in the serum and liver cholesterol levels in both groups of animals cholesterol-enriched diets that had been fed for 38 days. Fecal excretion of neutral and acidic steroid tended to be higher in the perilla oil group than in the safflower oil group. The perilla oil group significantly increased 20:5n-3 in liver phosphatidylcholine and phosphatidylethanolamine accompanying a decrease in 20: 4n-6. Such changes were not so evident in liver phosphatidylinositol. The production of leukotriene B4 and the concentration of prostaglandin E2 in the spleen were higher in the safflower oil group than in the perilla oil group. Thus, the hypocholesterolemic effect of the undigested fraction of soybean protein was apparently reproduced even in hamsters. Dietary fat-induced changes in lipid parameters in hamsters resembled those observed in rats.