[Cerebral infarction due to bacterial emboli associated with methamphetamine abuse].

The number of stimulant-drug addicts has recently been on the rise again, and they are being increasingly encountered in the emergency room. There are also frequent reports of cerebrovascular disorders complicating drug toxicity. These cerebrovascular disorders have included subarachnoid hemorrhage, intracranial hematoma, and a few cases of cerebral infarction. Here, we report the case of a 37-year-old male with drug toxicity, consciousness disorder, and hyperthermia. He was in a coma with a temperature of 43.1 degrees C and blood pressure of 58/35 mmHg when brought to our hospital. His condition worse rapidly deteriorated, and he died the same day. Cerebral infarction caused by gram-positive bacillus embolism, not necrotizing angiitis, was found at autopsy. Because drug addicts, especially stimulant-drug addicts, tend to inject themselves drug under unsanitary conditions, the possibility of this type of complication is always present. This is the first such case ever reported, and is therefore regarded as a rare complication of stimulant-drug intoxication.

Estimation and comparison of the contents of blood group B antigens in selected human tissues by microphotometric quantification of Griffonia simplicifolia agglutinin I-B4 staining with or without prior alpha-galactosidase digestion.

Griffonia simplicifolia agglutinin I-B4 (GSAI-B4) has broader specificity for B antigen variants and can recognize the antigens in a wide variety of human tissues. Thus, the concentration range of GSAI-B4 required for staining and the susceptibility of staining to alpha-galactosidase digestion is presumed to correlate well with the density of B antigens in tissue sections. By microphotometric quantification of staining intensity at different concentrations of GSAI-B4 with or without alpha-galactosidase digestion, concentration of B antigens in selected tissues was evaluated and compared. Based on the present results and the previous ones of direct measurement of galactose of B antigens in sublingual glands and red blood cells (Ito et al., 1993), the order of concentration of B antigens in tissues examined was estimated as follows; mucous cells of sublingual glands from German nonsecretors < red blood cells and vascular endothelial cells (= 2.7 x 10(-3) nmole/cm2), thyroid papillary carcinomas and Hassall's corpuscles from nonsecretors < mucous cells of sublingual gland from Japanese nonsecretors < pancreatic acinar cells from both secretor and nonsecretors, Hassall's corpuscles and kidney collecting tubules form secretors < mucous cells of sublingual gland from secretors (> 8.5-11.7 nmole/cm2) and mucous cells of Brunner's gland from nonsecretors < mucous cells of Brunner's gland from secretors. From the above estimation, it is apparent that the expression of B antigen in Brunner's gland is partly dependent on the secretor status of individuals and that Japanese nonsecretors secrete substantial amounts of B antigens from sublingual gland while German nonsecretors do not. The present results also revealed an unexpected staining behavior of GSAI-B4 in some tissues, i.e. in mucous cells of sublingual glands and collecting tubules of kidney from secretors, staining intensity was markedly depressed at higher concentration of the lectin and this depression was recovered by prior alpha-galactosidase digestion. In addition, the present method was successfully applied for the estimation of the content of B antigens neo-expressed in thyroid papillary carcinomas, showing that the content of B antigen had a similar level to that of red blood cells and vascular endothelial cells.

Localization of binding sites of Ulex europaeus I, Helix pomatia and Griffonia simplicifolia I-B4 lectins and analysis of their backbone structures by several glycosidases and poly-N-acetyllactosamine-specific lectins in human breast carcinomas.

Several studies have shown the deletion of blood group A or B antigens and the accumulation of H antigens in human breast carcinomas. Other studies have independently demonstrated that the binding sites of lectins such as Helix pomatia agglutinin (HPA) and Griffonia simplicifolia agglutinin I-B4 (GSAI-B4) are highly expressed in these cells. In order to clarify the molecular mechanisms of malignant transformation and metastasis of carcinoma cells, it is important to understand the relationship between such phenotypically distinct events. For this purpose, we examined whether the binding sites of these lectins and Ulex europaeus agglutinin I (UEA-I) are expressed concomitantly in the same carcinoma cells and analyzed their backbone structures. The expression of the binding sites of these lectins was observed independently of the blood group (ABO) of the patients and was not affected by the histological type of the carcinomas. Observation of serial sections stained with these lectins revealed that the distribution of HPA binding sites was almost identical to that of GSAI-B4 in most cases. Furthermore, in some cases, UEA-I binding patterns were similar to those of HPA and GSAI-B4 but in other cases, mosaic staining patterns with these lectins were also observed, i.e., some cell clusters were stained with both HPA and GSAI-B4 but not with UEA-I and adjacent cell clusters were stained only with UEA-I. Digestion with endo-beta-galactosidase or N-glycosidase F markedly reduced the staining intensity of these lectins. Together with the reduction of staining by these lectins, reactivity with Griffonia simplicifolia agglutinin II appeared in carcinoma cells following endo-beta-galactosidase digestion. Among the lectins specific to poly-N-acetyllactosamine, Lycopersicon esculentum agglutinin (LEA) most vividly and consistently stained the cancer cells. Next to LEA, pokeweed mitogen agglutinin was also effective in staining these cells. Carcinoma cells reactive with these lectins corresponded well to those stained with both HPA and GSAI-B4, and in some cases, with UEA-I. These results demonstrate that the binding sites of UEA-I, HPA, and GSAI-B4 are expressed concomitantly in the same carcinoma cells and all carry linear and branched poly-N-acetyllactosamine on N-glycans, suggesting that the synthesis of this complex carbohydrate is one of the most important and basic processes leading to the malignant transformation of cells, invasion, and metastasis of carcinoma cells.

Simultaneous expression of keratan sulphate epitope (a sulphated poly-N-acetyllactosamine) and blood group ABH antigens in papillary carcinomas of the human thyroid gland.

The monoclonal antibody 5-D-4 recognizes heavily sulphated forms of keratan sulphate epitope. It reacted strongly with the cell surfaces of most thyroid papillary carcinomas from all the individuals examined, independently of the blood group of the patients. Cells of follicular variants of papillary carcinomas were also labelled by 5-D-4. In contrast, no labelling with this antibody was observed in other types of thyroid neoplasms, or in normal tissues. The reactivity of 5-D-4 with papillary carcinomas was markedly reduced or abolished by prior digestion with endo-beta-galactosidase, keratanase II, or N-glycosidase F. Although keratanase digestion had no effect on 5-D-4 labelling, it revealed the binding sites of Griffonia simplicifolia agglutinin II (GSA-II), which recognizes terminal N-acetylglucosamine in a limited number of carcinoma cells from some individuals. Blood group ABH antigens, which are simultaneously expressed together with keratan sulphate epitope in cancer cells, were eliminated by digestion with endo-beta-galactosidase and N-glycosidase F, but were resistant to keratanase and keratanase II treatment. These results indicate that keratan sulphate oligosaccharides are cancer-associated and are probably oncofoetal antigens, as are the blood group antigens in human thyroid glands. The results suggests that poly-N-acetyllactosamine, which is ubiquitously and consistently produced in papillary carcinomas, is modified in two different ways: sulphation on the 6-position of at least some units of either galactose or N-acetylglucosamine or both, and decoration of non-reducing termini with the blood group antigens. Along with the endo-beta-galactosidase-GSA-II labelling procedure, labelling with 5-D-4 may be a useful diagnostic means for distinguishing papillary carcinoma from other types of thyroid neoplasms.

[Paternity probability in the cases of incest].

To calculate the paternity probability in the cases of incest where the alleged father was either the father or the brother of the plaintiff's mother, some algebraic expressions applicable to a simple codominant diallelic genetic marker system were derived by modifying the formulas of Essen-Möller and Komatsu (the both formulas gave the same result). The paternity probability in the incest case is generally lower than that in usual case, because in the former case an allele present in the mother is sometimes found in both the alleged father and the child (plaintiff), even if the alleged father is not true father. The paternity probability in the incest case, however, becomes higher than that in usual case when an allele is common to both the alleged father and the child but not to the mother. The mean value of paternity probability becomes lower, as the relationship becomes closer between the alleged father and the mother.

[A method to calculate the probability of paternity between relatives--a paternity case where the putative father was a deceased granduncle].

To test paternity in a case where the putative father was a deceased uncle of mother (plaintiff's granduncle), we designed a new method to calculate the probability of paternity likelihood. The putative father's genotypes of red cell antigens, HLA and short tandem repeat (STR) polymorphism were estimated from those of mother and sister of the plaintiff. When the probability was calculated from the frequencies in the unrelated individuals (the standard method), a significant bias might be introduced since the putative father and the plaintiff were likely to have the same alleles come from their common ancestry. Therefore, we designed a new method to calculate the likelihood ratio from the frequencies in the group of mother's uncles estimated from mother's genotypes. The probability (0.9299) calculated with our method was found to be lower than that (0.9992) done with the standard method indicating that the new method could remove the bias introduced from the incest.

Histochemical demonstration and analysis of poly-N-acetyllactosamine structures in normal and malignant human tissues.

Poly-N-acetyllactosaminyl structures carry a variety of physiologically and pathologically important carbohydrate antigens and are presumed to have essential roles in the process of cellular recognition, differentiation, malignant transformation and cancer metastasis. Monoclonal antibodies, lectins and endo-beta-galactosidase are useful histochemical tools for detecting and analyzing poly-N-acetyllactosamines in tissue sections. I (branched structure) and i (linear structure) antigens recognized by monoclonal antibodies have been shown to be differentiation antigens in mouse embryo and mouse and human teratocarcinoma cells as well as in human erythrocytes. They are also oncofoetal antigens and are expressed in carcinoma cells in several tissues and organs. Immobilized lectins specific to poly-N-acetyllactosamine structures have been successfully applied for fractioning glycoproteins with poly-N-acetyllactosamine, but histochemical use of these lectins has been restricted to some animal tissues. Among them, pokeweed mitogen agglutinin was used to detect branched poly-N-acetyllactosamine in normal and malignant human colon, demonstrating that it has a highly selective affinity for colorectal carcinomas. Griffonia simplicifolia agglutinin-II staining following endo-beta-galactosidase digestion procedure revealed the presence of poly-N-acetyllactosamine structures with or without blood group-specificities in several normal human tissues. By using this procedure, it was demonstrated that the blood group-related antigens oncofoetally expressed in thyroid carcinoma cells are carried by poly-N-acetyllactosamines containing a domain susceptible to the enzyme digestion. Staining with lectins specific to poly-N-acetyllactosamine in combination with endo-beta-galactosidase digestion demonstrated that poly-N-acetyllactosaminyl structures ubiquitously and consistently produced in thyroid papillary carcinomas are highly heterogeneous in their chain length and branching status and quite different from those produced in other thyroid neoplasms. Staining with monoclonal antibodies or lectins combined with endo-beta-galactosidase digestion procedures have been proven to be powerful tools for localizing and analyzing different types of poly-N-acetyllactosamine structures in normal and malignant tissues.

Histochemical demonstration of different types of poly-N-acetyllactosamine structures in human thyroid neoplasms using lectins and endo-beta-galactosidase digestion.

Blood-group-related antigens expressed in papillary carcinomas and other types of neoplasm of the human thyroid glands have been shown to be carried by poly-N-acetyllactosamines containing a linear domain susceptible to endo-beta-galactosidase digestion. To make clear more precisely the backbone poly-N-acetyllactosamine structures, labelled lectins specific to different types of these structures and specific to core structures with beta 1-6GlcNAc branching of N- and O-linked glycoproteins were employed in conjunction with prior endo-beta-galactosidase digestion on formalin-fixed, paraffin-embedded neoplasms of the human thyroid glands. In papillary carcinomas, Datura stramonium agglutinin (DSA) and succinyl wheat germ agglutinin (Suc-WGA) reacted most consistently and frequently with papillary carcinomas from all the individuals examined. Pokeweed mitogen (PWM) likewise stained the cells of papillary carcinomas from all the individuals examined, but in some individuals the number of lectin-reactive cells were very small. Lycoperscion esculentum aggultinin (LEA), Solanum tuberosum agglutinin (STA), Phaseolus vulgaris agglutinin L (PHA-L) and Artocarpus integrifolia agglutinin (jacalin) similarly bound to the cancer cells from most of the individuals, and in these cases the number of reactive cells was usually much more restricted than was the case with DSA or PWM. In adenoma and other types of carcinoma, such as follicular carcinomas, these lectins specific to poly-N-acetyllactosamine exhibited slight or no reactivity with the cells, whereas PHA-L and jacalin similarly bound to the cells of adenomas and carcinomas from most of the individuals examined. Prior digestion with endo-beta-galactosidase completely eliminated or markedly reduced the reactivity with PWM and LEA in papillary carcinomas. Reactivity with DSA, Suc-WGA, STA, PHA-L and jacalin was slightly reduced or not at all affected by enzyme digestion. These results confirmed that poly-N-acetyllactosamine species found in papillary carcinomas are quite different from those in other types of thyroid neoplasm, suggesting that at least three different types of poly-N-acetyllactosamine, that is, linear unbranched short and long sequences and highly branched ones are produced in these cells.