Hirofumi Suyama

Histochemical demonstration of methamphetamine by immunocytochemistry.

A method for the demonstration of methamphetamine (MA) by immunocytochemistry was established. The tissues of intoxicated mice, administered various amounts of MA in single doses of from 0.01 to 1 mg of MA-HCl, were fixed in glutaraldehyde-containing fixatives. Cryostat and paraffin slices gave a positive reaction of MA localization by staining the brain, liver, kidney, lung, stomach, spleen, and so forth, with the aid of the indirect immunoperoxidase technique. Those of animals administered a single dose of 0.1 mg or more (over 3 to 4 mg/kg--the usual dose of MA in acute intoxication death in forensic medicine), in particular, gave a strong strong reaction, so that the diagnosis of MA intoxication can be performed by macroscopic observation of stained slices. The histochemical diagnosis of MA intoxication in clinical toxicology and pathology might be regarded as a useful tool, especially in forensic pathology. The following cells gave a strong positive reaction: nerve cells and myelin sheaths, hepatocytes, epithelial cells of the distal part of the renal tubule and of the collecting tubule, alveolar and bronchial epithelial cells of the lung, chief and parietal cells of the gastric gland, capillaries of the renal glomerulus, macrophages in the blood and tissues, and striated muscle cells including cardiocytes. The morphological evidence of the pharmacodynamics and pharmacokinetics of MA can be determined at the cellular level by immunocytochemistry.

A new single band variant of the Gc subtypes determined by isoelectric focusing

SummaryA new single band variant (Gc Ta) of the Gc subtypes which does not correspond with the known Gc variants has been detected in a healthy blood donor by isoelectric focusing. Using this technique the variant is represented by a single band, which is more cathodically located than the 1A8(fast) band and more anodically located than the 1A3(fast) band.It is well known that the single band Gc phenotypes remain unaltered after neuraminidase treatment. Nevertheless, the new single band variant (Gc Ta) is altered after neuraminidase treatment. In this case the new single band Gc variant also contained n-acetylneuraminic acid (NANA).

Comparative immunological and electrophoretic analysis of Gc protein in sera from various animals

1. On immunodiffusion, using an anti-human Gc antibody, serum Gc in all mammals tested revealed a partial identity with human Gc. 2. The relative cross-reactivities of serum Gc in monkeys, dogs, cats and rats with human Gc antiserum were found to be more than 70% while the serum Gc in other mammals (pigs, cattle, goats and a guinea pig) was less than 50%. 3. Testing, using the isoelectrofocusing method, showed specific patterns of Gc in the mammals. In the sera of cats and cattle, Gc polymorphisms were detected. 4. Neuraminidase treatment affected the isoelectrofocusing Gc patterns of pigs, goats and cattle, whereas those in other mammals remained unchanged.

Detection of LeaSubstance in Saliva Stains by Enzyme-Linked Immunosorbent Assay (ELISA) Using Anti-Gum Arabic Serum

It is known that rabbit anti-gum arabic (GA) serum has cross-reactivity with Lea antigen, and that, by using this cross-reactive anti-Lea antibody, the presence of Lea antigen in red blood cells and saliva can be demonstrated with accuracy. We have devised a rapid and highly sensitive method for detecting Lea substance in human saliva by the enzyme-linked immunosorbent assay (ELISA) method using an anti-Lea antibody isolated from anti-GA serum by affinity chromatography on Synsorb Lea. The ELISA plate, coated with the specific anti-Lea antibody, adsorbed the Lea substance in saliva which was subsequently identified by adding enzyme labeled anti-Lea IgG in that order. The method could detect the Lea substance in Le(a+) saliva stains as small as 0.1 by 0.1 cm in size that had been stored at room temperature for three weeks and in Le(a+) saliva stains 0.7 by 0.7 cm in size that had been stored for ten years. This method seems to be useful for quantitative analyses of the Lea substance in various body fluids.

A new hereditary single band variant of the Gc system

SummaryA new single band variant (Gc Ar) or the Gc subtypes not identical with the known Gc variants has been detected in the plasma of a healthy blood donor by isoelectric focusing. Using this technique the variant is represented by a single band which has a similar isoelectric point to the Gc 1C2 anodal band. It is well known that the single band Gc phenotypes remain unaltered after neuraminidase treatment. Nevertheless, the new single band variant (Gc Ar) is altered after neuraminidase treatment as is Gc 2A3. After neuraminidase treatment, the Gc Ar band is affected and moved to the nearby position of the Gc 2 band. Investigation of the probands family shows that the variant occurs combined with the common alleles Gc 1F, Gc 1S and that it has an autosomal dominant inheritance.

Serum Gc, Hp and α2HS Phenotypes in Human T-Lymphotropic Leukemia Virus Type I Infection

The serum Gc, Hp and α2HS phenotypes were examined in 64 subjects known to have the human T-lymphotropic leukemia virus type I (HTLV-I) infection and in 60 uninfected subjects. There were n