Tatsuro Takino

Light and electron microscopic immunohistochemical studies of serotonin nerve fibers in the substantia nigra of the rat, cat and monkey

SummaryThe distribution of serotonin-containing nerve fibers in the substantia nigra of the rat, cat and monkey was studied with a highly sensitive peroxidase-antiperoxidase immunohistochemical method. Serotonin fibers in the substantia nigra of all species consisted of fine varicose fibers and formed a fine network. In the zona compacta of all species, serotonin fibers were sparsely distributed. In the zone reticularis of the rat and cat, these fibers were densely distributed and their distributional pattern was almost uniform, while in the monkey such fibers were unevenly distributed and high and low dense areas were intermingled. In the pars lateralis of all species, serotonin fibers were diffusely distributed, and the distributional density was much higher in the cat and monkey than in the rat. Immunoelectron-microscopic studies further revealed that a majority of the labeled varicosities in the rat substantia nigra were in close apposition to peridendritic axon terminals and were also free in the neuropil; occasionally they exhibited symmetrical synapses of “en passant” type with non-immunoreactive dendrites or somata. Our results support a functional significance of serotonergic regulation of the substantia nigra in mammals.

Prefixation treatment with ethidium bromide for high resolution banding analysis of chromosomes from cultured human bone marrow cells

Prefixation treatment of cultured human bone marrow cells with a DNA intercalating agent, ethidium bromide (EBr), induced a dose- and time-related elongation of chromosomes. When compared with EBr-free cultures, a 2.9-fold increase in the yield of early mitotic cells with more than 400 bands per haploid set of chromosomes was achieved by simply adding 10 micrograms/ml of EBr during the last 2 hours of culture. The proportion of early mitotic cells was equal to that obtained in methotrexate synchronized cultures. Fluorescence banding methods using base composition specific agents actinomycin D/DAPI for AT base pairs and chromomycin A3/distamycin A for GC suggested that EBr does not have base specificity, because EBr did not alter the banding patterns of chromosomes obtained with these staining procedures.

Chronic active hepatitis with histological features of primary biliary cirrhosis

SummaryA 51-year-old woman was evaluated because of Raynauds phenomenon, Sjögrens syndrome, and general malaise. There was neither skin itching nor jaundice. Endoscopic retrograde cholangiopancreatography showed a normal extrahepatic as well as intrahepatic biliary tree. Serum GOT and GPT fluctuated with episodes of marked increases. The alkaline phosphatase was slightly increased and total cholesterol was normal. There were marked increases of IgG and IgM. AMA was positive at a titer of 1:320, which was measured by an indirect immunofluorescence method. PBC-specific AMA (anti-M2) was positive, but mixed-form AMA (anti-M4) negative. An LE-cell test, ASMA, ANA, and anti-DNA antibody were all positive on several repeated tests. A wedge liver biopsy tissue showed mixed features of PBC and CAH. A diagnosis of CAH was made on the basis of the clinical, serological, and morphologic findings. The patient responded well to prednisolone treatments with prominent improvement of her symptoms and liver function tests. Subsequently, AMA fell to undetectable levels by indirect immunofluorescence method.

Immunohistochemical demonstration of serotonin nerve fibers in the subthalamic nucleus of the rat, cat and monkey

Distribution of serotonin-immunoreactive nerve fibers (5-HT fibers) in the subthalamic nucleus (STN) of the rat, cat and monkey was examined with a modified peroxidase-anti-peroxidase method using 5-HT antiserum without pretreatment. Fine varicose 5-HT fibers formed a fine network in the STN. Overall density of 5-HT fibers in the STN was highest in the monkey, and lowest in the rat. In the rat and cat, these fibers were distributed almost diffusely in the STN, whereas in the monkey they were particularly abundant in the ventral and medial parts of the STN. Many thick 5-HT fibers ran through the monkey STN.

Glutathione metabolism in red cell aging

Normal human red cells were centrifugally separated according to age by discontinuous density gradient of Percoll. Reduced glutathione (GSH), GSH stability and glucose-6-phosphate dehydrogenase (G6PD) activity in fractionated red cells decreased with age, while oxidized glutathione (GSSG) and methemoglobin (MetHb) increased with age.

Immunohistochemical demonstration of serotonin nerve fibers in the corpus striatum of the rat, cat and monkey.

SummaryThe distribution of serotonin-containing nerve fibers in the corpus striatum of the rat, cat and monkey was studied with modified peroxidase-antiperoxidase method using serotonin antiserum without any pretreatment. In the neostriatum (caudate nucleus and putamen) of all mammalian species investigated, the immunoreactive fibers were distinct varicose fibers in a fine network. The concentration of these fibers was high in the ventral, medial and caudal neostriatum. Especially in the area bounded by the globus pallidus, serotonin fibers were abundant and compactly arranged along the nucleus. In this area of the monkey, a few thick fibers (tract fibers) were intermingled; they ran along the lateral medullary lamina. Such tract fibers were also observed outside the medial medullary lamina and in the central portion of the medial pallidal segment. The paleostriatum (globus pallidus and entopeduncular nucleus) of the rat and cat as well as the medial pallidal segment of the monkey was diffusely innervated with serotonin fibers composed of numerous varicosities and fine intervaricose segments, while in the lateral pallidal segment of the monkey, the distribution of fibers was scantly and partial. Our results hint at the morphological basis of the serotonergic regulation of the extrapyramidal system in mammals.

Alteration of amino acid content of cerebrospinal fluid from patients with epilepsy

ABSTRACT‐ The alteration of amino acids content in cerebrospinal fluids (CSF) from 31 cases of epilepsy and 10 cases of headache (as control) was studied using high performance liquid chromatography (HPLC). In patients with epilepsy, it was found that the CSF levels of GABA and aspartic acid had a tendency to decrease, but these changes were not statistically significant. In simple partial seizures, the CSF levels of glutamic acid and glycine also showed a tendency to decrease. The decrease of CSF GABA found in epileptics had a tendency to normalize following treatment with valproic acid. At the same time, administration of valproic acid induced a decrease of aspartic acid in the CSF of epileptics. These results suggest that administration of valproic acid may induce an increase in GABA and a decrease in aspartic acid in the CSF of epileptics.

Alteration of amino acids in cerebrospinal fluid from patients with parkinson's disease and spinocerebellar degeneration

The alteration of amino acids in cerebrospinal fluids (CSF) from 14 cases of Parkinsons disease, five cases of cerebellar degeneration and five cases of headache (control) was studied using high performance liquid chromatography. In patients with Parkinsons disease, it was found that the CSF level of GABA showed a significant decrease, while that of taurine had an increase. The degree of disability in Parkinsons disease and the decreased GABA levels had a positive correlation, especially at its advanced stages. In patients with cerebellar degeneration, it was also found that there was a significant decrease in CSF GABA.

Detection of karyotypic abnormalities in most patients with acute nonlymphocytic leukemia by adding ethidium bromide to short-term cultures

A modified short-term culture method, in which cultured bone marrow cells were treated with ethidium bromide to prevent chromosome condensation was used to study the chromosomes of 70 patients with acute nonlymphocytic leukemia. Clonal karyotypic abnormalities were detected in 60 patients. Among these, 35 patients showed one of recurrent type specific alterations. A close relationship between karyotypes and clinical outcome was shown: thus, t(8;21) or a single miscellaneous chromosomal defect associated with a favourable prognosis whereas t(9;11) or a complex karyotype related to a poor prognosis. The ten cytogenetically normal patients did not appear to have a favourable prognosis.

Glutathione-linked enzyme activities in red cell aging

The enzyme activities of glucose-6-phosphate dehydrogenase (G6PD), 6-phospho-gluconate dehydrogenase (6PGD), glutathione peroxidase (GSH-Px) and glutathione reductase (GR) were measured in normal human red cells separated centrifugally in a discontinuous density gradient of Percoll. Activities of G6PD, GSH-Px and GR decreased with red cell aging.