Differential effects of body weight, hyperinsulinemia and oral glucose load on serum C-peptide/insulin molar ratio.

Serum C-peptide immunoreactivity (CPR)/immunoreactive insulin (IRI) molar ratio was determined in 136 subjects without renal, hepatic and thyroid disorders, at fasting, and during the initial period of 75 g-oral glucose tolerance test. The subjects were divided into 4 groups based on their body weight and age; Group A, young (< 55 years) and normal body weight (body mass index [BMI, kg/m2] < or = 25) subjects; Group B, young and overweight (BMI > 25) subjects; Group C, aged (> or = 55 years) and normal body weight (BMI < or = 25) subjects; Group D, aged and overweight subjects. Fasting CPR/IRI ratio and absolute CPR level negatively correlated in Groups B and D but not in A and C. After oral glucose load with elevation of insulin, CPR/IRI ratio invariably declined in all groups and significant negative correlation between CPR/IRI and CPR was found in Groups A, B and D but not in C. Slope of the regression lines obtained for correlation between CPR/IRI ratio and CPR were significantly steeper at fasting compared to the post-stimulation phase. CPR/IRI ratio is affected by hyperinsulinemia and oral glucose load but not by obesity alone. Assuming that CPR/IRI ratio reflects hepatic extraction of insulin, the insulin clearance at fasting is progressively reduced with increasing insulin secretion in overweight subjects: failure to detect such phenomenon in normal body weight subjects may be due to a narrower CPR range in this population. Insulin metabolism at fasting and during glucose stimulation is likely to be regulated by distinct factors.     

The augmentative effect of a protein-bound polysaccharide (PSK) on tumoricidal activity of the soluble factor produced by a streptococcal preparation (OK-432)-activated macrophages

The enhancement of antitumor activities of the tumoricidal soluble factor (SF) from a streptococcal preparation (OK-432)-activated macrophages by the pretreatment with a protein-bound polysaccharide (PSK) was investigated in tumor-bearing mice.Two-step stimulations with OK-432 atin vivo priming andin vitro eliciting were required for the production of the tumoricidal SF by macrophages, and the tumoricidal activity of the SF apparently correlated with the uptake of OK-432 by macrophages at priming phase.Tumoricidal activity of the SF from OK-432-activated macrophages in proteose-peptone (P-P)-pretreated mice significantly decreased with the development of the tumor, whereas in PSK-pretreated mice did not. Pretreatment of tumor-bearing mice with PSK saved a decrease in the macrophages carrying Ia^k or asialo GM1 antigens and an increase in wheat germ agglutinin (WGA) receptors. Furthermore, the uptake of OK-432 by macrophages at priming phase was enhanced. The tumoricidal activity of the SF from OK-432-activated macrophages was augmented.Thus, PSK may restore the depressed functions of macrophages, and the combination therapy with PSK and OK-432 may be effective to enhance the production of tumoricidal SF in tumor-bearing mice.     

Effects of chair restraint on plasma enzyme values in the rhesus monkey (Macaca mulatta)

The purpose of this paper was to examine the effect of chair restraint on plasma enzyme values in the rhesus monkey. Six monkeys were restrained to the monkey chair for eight hours. Creatine phosphokinase (CK) value increased significantly three hours after the onset of restraint and LDH value did eight hours after the onset of restraint. The increase in CK, GOT and GPT values continued for 1 or 2 days after the release from restraint. On the other hand, these plasma enzyme values in non-restraint monkeys showed almost no changes. These results indicate that it is necessary to establish a proper method for the adjustment to chair restraint in the rhesus monkey.     

Ascaris suum: suppression of reaginic and hemagglutinating antibody responses in the mouse by crude extract and maintenance fluid.

Intraperitoneal administration in mice of crude extract (CE) or maintenance fluid (MF) of Ascaris suum in Freund's incomplete adjuvant (FICA) in doses of 200 and 2 (CE) and 4 μg (MF) on Days ?4, 0, and +4 relative to the day of the immunization with 10 μg of hen egg white lysozyme (HL) resulted in the suppression of anti-HL reaginic antibody responses at varying degrees depending on the dose and their time of administration. Hemagglutinating antibody responses were also affected but in a different manner. Treatment with CE on Day ?4 resulted in complete suppression of reaginic antibody responses and some degree of suppression of hemagglutinating antibody responses depending on the size of the CE dose. In mice pretreated with MF, transient suppression was found only for reaginic antibody responses. In mice receiving the treatment of CE on Day 0, 200 μg of CE caused complete suppression of reaginic antibody responses, while 2 μg was less effective. Hemagglutinating antibody responses were also suppressed in proportion to the dose. Simultaneous treatment with MF did not cause any suppression of either reaginic or hemagglutinating antibody responses. In mice treated with CE on Day +4, reaginic antibody responses were not markedly suppressed and hemagglutinating antibody responses were also not altered. In contrast, treatment with MF on Day +4 resulted in suppression of reaginic antibody responses during the whole course of the primary response, but had no effect on hemagglutinating antibody responses. When MF was administered 7 days after the priming, no suppressive effect on the antibody responses was demonstrated. On the other hand, if a lower dose (1 μg of HL) was used for the priming, the effect of MF treatment with Day +4 was more pronounced in the primary reaginic antibody response and the secondary response was also affected. A comparable suppression of hemagglutinating antibody responses also was observed.     

Comparative studies on antibody formation against hen egg white lysozyme and its split fragment.

A peptide fragment Fr. 17 (Lys1-Cys-Asn27 Leu129-Cys-Ala122) of hen egg white lysozyme (HL) was previously found to retain at least one antigenic determinant region of the native protein. In this work a highly purified preparation of Fr. 17, contaminated with less than 0.01% HL and less than 1% of other fragments was found to be strongly immunogenic to rabbits. The kinetic patterns of antibody formation against Fr.17, assayed by passive hemagglutination (PHA), were quite different from those of antibody formation against HL. The specificity of the antibody elicited to Fr. 17 was mainly directed to the Fr. 9-10-a region (Ala11-Asn27) while that of the antibody elicited to the Fr. 17 region in native HL was directed to the Fr. 15-b region (Lys1-Cys-Ala10 Leu129-Cys-Trp123). It is concluded that in the process of antibody formation, the recognition of the Fr. 17 region in native HL is different from that of fragment Fr. 17.     

Synthesis and Bioactivity of Novel Acetylenic Compounds

The synthesis of novel acetylenic ketone compounds and anti-inflammatory and antimicrobial activities are herein described.     

Monitoring Autophagy Flux and Activity: Principles and Applications

Macroautophagy is a major degradation mechanism of cell components via the lysosome. Macroautophagy greatly contributes to not only cell homeostasis but also the prevention of various diseases. Because macroautophagy proceeds through multi-step reactions, researchers often face a persistent question of how macroautophagic activity can be measured correctly. To make a straightforward determination of macroautophagic activity, diverse monitoring assays have been developed. Direct measurement of lysosome-dependent degradation of radioisotopically labeled cell proteins has long been applied. Meanwhile, indirect monitoring procedures have been developed. In these assays, autophagosome marker proteins, microtubule-associated proteins 1A/1B light chain 3B-II (LC3B-II) and gamma-aminobutyric acid receptor-associated protein-II (GABARAP-II) have been analyzed and the validity of the assays strongly depends on appropriate assessment of the fluctuation of LC3-II and/or GABARAP-II levels in the presence or absence of lysosomal inhibitors. This article describes these monitoring methods, paying special attention to the principles and characteristics of each procedure.