SummaryThe influence of the chronic intake of a newly developed sweetener named "Neosugar" (fructooligosaccharide) on body weight gain, organ weight, serum lipids, fecal excretion and intestinal function was investigated in rats. The following results were obtained. 1) Body weight gain was diminished more severely in rats fed on 20% Neosugar diet than in rats fed on 10% Neosugar diet. 2) The wet weights of cecum and colon were greatly increased by Neosugar feeding. 3) Fecal wet weight was significantly increased and gastrointestinal transit time was shortened by Neosugar feeding compared with those of the control group. 4) Serum triacylglycerol levels were significantly lower in rats fed Neosugar, whereas serum cholesterol levels were similar to those of the control group. 5) Fecal excretions of neutral sterol and volatile fatty acids were significantly increased by Neosugar feeding. These results were quite similar, with the exception of diarrhea to those obtained using a dietary fi ber such as glucomannan. Therefore, Neosugar with a pleasant-tasting sweetness appears to be an unavailable oligosaccharide with a dietary fi ber-like action. Key Words Neosugar, fructooligosaccharide, dietary fiber, chronic in take, sweetener, undigestibility, gastrointestinal A dietary preference for sweet foods accompanied by an excessive intake of energy leads inevitably to obesity in people who continue to consume more energy than they burn. In this sense, pleasant-tasting, non-energy sweetener that is entirely safe would make it possible to enjoy sweetened food and at the same time, to avoid both obesity and its complications such as diabetes mellitus. The newly developed 111

Mitochondrial DNA (mtDNA) of peripheral blood mononuclear cells (PBMCs) collected from Human immunodeficiency virus 1 (HIV-1)-infected patients and healthy controls were measured longitudinally using real-time polymerase chain reaction to evaluate the effects of antiretroviral agents on mtDNA synthesis in vivo and to assess the value of monitoring mtDNA in PBMCs to predict adverse events amongst these patients. MtDNA levels in PBMCs were significantly decreased in treatment-naive HIV-1-infected patients compared with healthy people. MtDNA levels were not only significantly correlated with CD4(+) T-cell count, but also inversely correlated with HIV-1 viral load. MtDNA levels in untreated patients and healthy controls were stable during the period of observation. On the other hand, amongst patients treated with regimens containing AZT/3TC or d4T/3TC, mtDNA increased during treatment and recovered to levels comparable to healthy controls. In contrast, mtDNA decreased immediately after the initiation of an AZT/ddC-containing regimen. We did not find a correlation between mtDNA levels and changes in clinical parameters. There was no significant difference in mtDNA levels between patients with and those without lipoatrophy. Furthermore, there was no obvious difference in mtDNA levels amongst those patients exhibiting signs and symptoms of peripheral neuropathy. In conclusion, the decrease in mtDNA levels in PBMCs amongst HIV-1-infected patients and its amelioration by antiretroviral therapy may suggest the influence of direct effects on mitochondria or mtDNA by HIV-1 infection. Further investigations are needed to elucidate the mechanisms contributing to decreased mtDNA and the value of mtDNA measurement in the care of HIV-1-infected individuals.

Although Japan is classified as a country with a low prevalence of human immunodeficiency virus type 1 (HIV-1), domestic sexual transmission has been increasing steadily. Because 70% of the Japanese population expresses HLA-A24 (genotype HLA-A*2402), we wished to assess the effect of the dominant HLA type on the evolution and transmission of HIV-1 among the Japanese population. Twenty-three out of 25 A24-positive Japanese patients had a Y-to-F substitution at the second position [Nef138-10(2F)] in an immunodominant A24-restricted CTL epitope in their HIV-1 nef gene (Nef138-10). None of 12 A24-negative Japanese hemophiliacs but 9 out of 16 patients infected through unprotected sexual intercourse had Nef138-10(2F) (P < 0.01). Two of two A24-positive but none of six A24-negative Australians had Nef138-10(2F). Nef138-10(2F) peptides bound well to the HLA-A*2402 heavy chain; however, Nef138-10(2F) was expressed poorly on the cell surface from the native protein. Thus, HIV-1 with Nef138-10(2F) appears to be a cytotoxic-T-lymphocyte escape mutant and has been transmitted frequently by sexual contact among the highly A24-positive Japanese population

The digestion of Neosugar, a mixture of 1F-(1-beta-fructofuranosyl)n-1 sucrose [n = 2, 1-kestose (GF2); n = 3, nystose (GF3); n = 4, 1F-beta-fructofuranosyl nystose (GF4)] was investigated in vitro and in vivo by using the rat. The results obtained were as follows. GF2 and GF3 were not hydrolyzed by a pancreatic homogenate. The GF2- and GF3-hydrolyzing activities of the enzymes in the intestinal mucosa homogenate were negligible compared with the activities of maltase and sucrase. GF2 and GF3 added to the incubation mixture did not affect the activities of sucrase and maltase in the intestinal mucosa. Long-term ingestion of Neosugar did not cause induction or suppression of GF2- and GF3-hydrolyzing enzymes in the small intestine. [U-14C]Neosugar injected intravenously was rapidly excreted in the urine without having undergone any degradation. These results indicate that Neosugar, which consists of GF2, GF3 and GF4, is scarcely hydrolyzed by the digestive enzymes of the gastrointestinal tract and internal organs, and that suggests to us that Neosugar is not utilized as an energy source in the body.

Immune response enhanced by therapeutic HIV-1 vaccine may control viral proliferation after discontinuation of highly active antiretroviral therapy (HAART). Although which strategies for therapeutic vaccination are feasible remains controversial, application of dendritic cells (DCs) as a vaccine adjuvant represents a promising approach to improving deteriorated immune function in HIV-1-infected individuals. The safety and efficacy of DC-based vaccine loaded with HIV-1-derived cytotoxic T lymphocytes (CTL) peptides were thus investigated in this study. Autologous DCs loaded with seven CTL peptides with HLA-A*2402 restriction were immunized to four HIV-1-infected individuals under HAART. In terms of safety, peptide-loaded DCs were well tolerated, and only mild local and general symptoms were observed during vaccine administration. ELISPOT assays to detect IFN-gamma production in CD8(+) lymphocytes revealed a limited breadth of responses to immunized peptides in two of four participants, but no response in the remaining two participants. Differences in immunological response might be attributable to the fact that responders displayed higher nadir CD4 counts before starting HAART and were immunized with a larger number of DCs per reactive peptide than non-responders. Discontinuation of HAART after vaccination failed to lower viral set points compared to those before starting HAART. This early outcome warrants further exploration to elucidate the therapeutic value of vaccination with DCs in HIV-1 infection.