Masaichi Yamamura

Detection of Herpes Simplex DNA in Semen and Menstrual Blood of Individuals Attending an Infertility Clinic

Objective: To determine a possible link between herpes simplex virus 1 (HSV) and infertility.

Impaired superoxide production in peripheral blood neutrophils of germ-free rats

A method of separating neutrophils from the peripheral blood of rats with 95% purity is described. To determine the role of antigenic stimulation in neutrophil function, neutrophils front germ‐free (GF) rats were compared with those of conventional (CV) rats. Neutrophil counts were lower in GF rats but the total number of monocytic cells was the same.

Presence of HSV-1 DNA in semen and menstrual blood.

Using a specifically designed diagnostic PCR assay with nested primers the following could be achieved: (1) a group of 22 clinically indistinguishable women attending an infertility clinic, 18 with repeated embryo transfer failure, and asymptomatic for HSV-1 could be divided into two subgroups after testing their menstrual blood. An HSV-DNA positive (50%) and HSV-DNA negative group (50%) could be distinguished. None of the four controls were positive; (2) semen from 154 infertile and 24 fertile men was tested in relation to infertility. In the group of infertile men 37 (24%) were HSV-DNA positive but none of the fertile control (0%) was positive; (3) treatment of both partners with an antiviral drug resulted in two pregnancies; (4) HLA data on four of the couples in which the wifes menstrual blood was HSV positive was compared to seven HSV negative couples and 22 infertile, as well as 22 fertile couples. Clustering of class I HLA (B61 and Cw3) was found with a significant increase in Cw3 in both partners.

Effect of cyclic polylactates on tumor cells and tumor bearing mice

We studied the effect of cyclic polylactates ranging in size from a degree of polymerization number of 3 to 13 on pyruvate kinase, lactic dehydrogenase, anaerobic glycolysis, growth of tumor cells and survival of tumor bearing mice. Pyruvate kinase and lactic dehydrogenase activities were both inhibited by cyclic polylactates, and the inhibition mechanism of cyclic polylactates on lactic dehydrogenase was noncompetitive. About half the anaerobic glycolytic activity of FM3A ascites tumor cells was inhibited and tumor cell growth was also effectively inhibited by cyclic polylactates. Mice, which were treated with cyclic polylactates after inoculation of FM3A ascites tumor cells lived significantly longer than mice, which were treated with vehicle or non mice.

Congenitally abnormal plasminogen in juvenile ischemic cerebrovascular disease.

Background and Purpose Congenitally abnormal plasminogen is characterized by markedly decreased fibrinolytic activity and has been reported mainly in association with venous occlusive disease. Case Description We found three young adult patients (34, 45, and 27 years old at onset) with ischemic cerebrovascular disease, all of whom had congenital plasminogen abnormalities but no other known risk factors. Hemostatic tests of all three patients revealed plasma plasminogen activities at almost one half of the normal level despite normal plasma plasminogen antigen levels. They were found to be heterozygotes with abnormal plasminogen (normal Ala-601[GCT] to abnormal Thr-601[ACT]) by DNA sequence analysis after polymerase chain reaction. Conclusions Congenital plasminogen abnormalities could be one of the risk factors of juvenile ischemic cerebrovascular disease of the arterial as well as venous type.

Peripheral Blood Neutrophils of Germ‐Free Rats Modified by In Vivo Granulocyte–Colony‐Stimulating Factor and Exposure to Natural Environment

Peripheral blood neutrophils from germ‐free (GF), specific‐pathogen‐free (SPF) and conventional (CV) rats were compared. Besides neutropenia and impaired superoxide anion generation as previously reported, it was found that GF rats had lower phagocytic function (70%) and generated less nitric oxide than the other rats. GF and SPF rats were injected with recombinant human granulocyte–colony‐stimulating factor (rhG‐CSF), or were transferred to natural environment. It was found that total number, phagocytosis, intracellular killing, ratio of phagocytosis versus killing (killing efficiency) and nitric oxide production induced by recombinant rat interferon‐γ (rrIFN‐γ) were normalized upon injection of rhG‐CSF. These results indicate that rhG‐CSF may stimulate neutrophil production and induce the expression of neutrophil receptors for phagocytosis and nitric oxide production in GF rats. Although lower than in CV rats, the level of superoxide produced was sufficient for normal neutrophil‐killing efficiency in SPF and GF rats. In SPF rats, this could be amended by exposure to natural environment. However, neither rhG‐CSF injection nor transfer to natural environment could increase the generation of superoxide anion in GF rats.

Acute-Phase Plasma Proteins in Gastric Cancer: Association with Metastatic Potential and Prognosis

We evaluated the possibility that acute-phase plasma proteins such as alpha 1-antichymotrypsin (ACT) and immunosuppressive acidic protein (IAP) might be useful predictors of lymph node metastasis and prognosis in patients with gastric cancer. Both ACT and IAP levels generally increased according to the pTNM stage. Patients with both abnormal IAP and ACT levels showed a high risk of lymphatic and hepatic metastasis as well as peritoneal dissemination, with a resultant poor prognosis. Patients who had abnormal IAP levels with or without abnormal ACT levels had a significantly higher risk of lymph node metastasis, as well as more invasive tumors and a worse prognosis than those who had normal IAP levels with or without abnormal ACT levels. In combination group 4 [IAP(+) ACT(-) vs. IAP(-) ACT(+)] lymphatic metastasis was seen more often with isolated IAP(+) (76.4%) than with ACT(+) (52.9%) (p < 0.0045), especially in the subgroup of poorly differentiated adenocarcinoma (POR; p < 0.0177). However, this does not demonstrate that ACT(+) is a protective factor against lymphatic invasion, because the results of combination group 6 [IAP(-) ACT(+) vs. IAP(-) ACT(-)] show that isolated ACT(+) is also significantly related to lymphatic metastasis (p < 0.001). The same is true for the subgroup of signet ring cell carcinomas (p = 0.038), but it has not been tested versus the POR subgroup.

Detection of a new mutant α‐1‐antichymotrypsin in patients with occlusive‐cerebrovascular disease

A new mutant α‐1‐antichymotrypsin (variant ACT) was found by direct sequencing and PCR‐single strand conformation polymorphism (PCR‐SSCP). This variant ACT was a point mutation of exon V of ACT, with the substitution of Met by Val. Four out of six individuals with this variant ACT had occlusive‐cerebrovascular disease, leading to one hypothesis that there might be an association between this mutation and occlusive‐cerebrovascular disease.

Heat production as a cell cycle monitoring parameter

A microcalorimetric method was applied to define the cell cycle by measuring heat production of mouse breast cancer cell line, FM3A. FM3A cells, were synchronised at the Go phase and produced 13.0 mu Watts (W) per 1 X 10(6) cells. Although the number of cells in fresh medium remained unchanged during the following 24 hour, a dramatic increase of heat production was observed and maximum heat (46.2 mu W) was produced by the cells at 24 hours when the cell cycle was presumably at the G2 phase. At 26 hours, although cell number increased, heat production decreased. Since the cells were not treated in any manner, this microcalorimetric method of measuring the cell cycle by monitoring heat production can be a very useful tool.

Glucose metabolism and protective biochemical mechanisms in a rat brain affected by kaolin-induced hydrocephalus.

Abstract To clarify glucose metabolism in a hydrocephalic rat brain, substances related to glycolytic metabolism were biochemically measured. Kaolin-induced hydrocephalic rats were sacrificed and lactate dehydrogenase (LDH), LDH isozyme, lactate, adenosine triphosphate (ATP), and isocitrate dehydrogenase (ICDH) were measured in the following regions: cortex, thalamus, midbrain, hippocampus, cerebellum, and pons with medulla. During the development of hydrocephalus, lactate and LDH increased in most regions, the LDH M-subunit increased in the cortex, and ICDH decreased in most regions. However, ATP levels did not change. The increases in lactate, LDH and M-subunit suggested an anaerobic environment in the cell leading to activation of the anaerobic glycolysis. The decrease in ICDH represented a diminution of the tricarboxylic acid cycle. Through these alterations, the ATP level can be kept constant during the course of hydrocephalus, allowing the brain to create a better biochemical milieu.