Shin-ichi Kubo

Changes in the Specificity of Blood Groups Induced by Enzymes from Soil Fungi

Three strains of Cunninghamella elegance (C. elegance), Penicillium simplicissimum (P. simpl.), and Aspergillus niger (A. niger) were isolated from soil that produced the enzymes acting on blood groups A and B red cells. Culture filtrates from these fungi contained alpha-N-acetyl-D-galactosaminidase as an A-decomposing enzyme, which resulted in an almost complete loss of A specificity and an enhancement of blood group H activity as measured by elution tests using monoclonal antibodies. They also contained an alpha-D-galactosidase and an alpha-L-fucosidase, which partially destroyed the blood group B specific activity, but did not influence the blood group H specific activity.

A demonstration of spermatozoa on vaginal swabs after complete destruction of the vaginal cell deposits.

The proteolytic enzyme, proteinase K, has been found to destroy all vaginal cells though it does not have the same effect on spermatozoa. In cases of sexual offenses, in which a swab has been used to wipe out the vagina, the female cells and their nuclei on that swab may also contain the heads of spermatozoa. After as short a time as 30 min of proteinase K treatment, the spermatozoa that had separated from the enzymatically destroyed vaginal cells were recovered. This proteinase destruction furnishes some spermatozoa with deformed heads and a somewhat greater number of isolated tails though a sufficient number of spermatozoan heads still remain for a reliable diagnosis. For detection of spermatozoa from a vaginal swab after proteinase K pretreatment, the heads of the spermatozoa are distinctly stained by Oppitzs method. Further, on prior treatment with proteinase K, the ABO blood grouping of the spermatozoa could also be determined on the vaginal swab by using the absorption-elution technique. The resistance of the spermatozoa to proteinase K is the basis for this method.

Hereditary recombined three-allele variant of the Gc system

A three-allele variant with Gc 2, Gc 1F and Gc 1A2 alleles was detected in both a baby and his mother during paternity testing by isoelectric focusing. His father had a normal Gc phenotype, Gc 2-1F. Further examination of his mothers relatives revealed that his grandfather also had the same three-allele variant, while his grandmother and his aunt had normal Gc 2-1F and Gc 2-2. From these results, it was considered that the Gc 1F and Gc 1A2 alleles were on the same single chromosome. It was suggested that recombination had occurred between two chromosomes that had the Gc 1F and Gc 1A2 allele, respectively, forming the variant allele Gc 1F1A2 on a single chromosome.

Immunohistological investigations of autopsied carotid bodies and their application to diagnosing strangulation

SummaryUsing immunohistochemical staining, the histological changes and the presence of neuropeptides (enkephalin and VIP) in the carotid body have been investigated in medico-legal autopsy cases, especially asphyxia cases. Only in cases of manual and/or ligature strangulation cases that sustained a force near the carotid body, were the chief cells mainly lightly stained, indicating that they had been “active” cells. Furthermore, these cells and their nuclei were enlarged in comparison to the chief carotid body cells in other autopsy groups. It was thus felt that these changes had resulted from the force that had directly affected the carotid body. Based on these findings, it was concluded that immunohistochemical investigation of the carotid body offers a useful possibility for diagnosing manual asphyxia, especially in autopsy cases involving strangulation.ZusammenfassungMit Hilfe immunhistochemischer Färbemethoden wurden die histologischen Veränderungen und das Vorhandensein von Neuropeptiden (Enkephalin und VIP) im Glomus caroticum bei rechtsmedizinischen Autopsie-Fällen, speziell bei Asphyxie-Fällen, untersucht. Lediglich in Fällen von manueller und werkzeugbedingter Strangulation, in denen die Gewalt in der Nähe des Glomus caroticum erlitten wurde, waren die Hauptzellen hauptsächlich leicht angefärbt, als Hinweis, daß sie „aktive” Zellen darstellten. Weiterhin waren diese Zellen und ihre Kerne vergrößert im Vergleich zu den Hauptzellen des Glomus caroticum in anderen Autopsie-Fällen. Es entstand daher der Eindruck, daß diese Veränderungen resultierten aus der Gewalt, die direkt das Glomus caroticum traf. Aufgrund dieser Befunde wurde geschlossen, daß die immunhistochemische Untersuchung des Glomus caroticum eine nützliche Möglichkeit darstellt, um die manuelle Asphyxie zu diagnostizieren, speziell in Autopsie-Fällen unter Einbeziehung der Strangulation.

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