[Detection of antigenic determinants of autoantibodies in idiopathic thrombocytopenic purpura using anti-platelet monoclonal antibodies and flow cytometry in whole blood].

To detect antigenic sites of platelet-bound autoantibodies in chronic ITP, whole blood or platelet-rich plasma from patients was stained with monoclonal antibodies (MoAbs) directed against platelet membrane glycoproteins (GPs) and with FITC-conjugated anti-mice IgG in an unwashed system followed by flow cytometric analysis. Platelets were identified by light-scattering profile and the amount of anti GP MoAb bound to platelets was measured and expressed as mean fluorescence intensity. This system enabled to quantitate the amount of intact GPs on platelet surface, regardless of activation of platelet.

[Analysis of cytolytic activity and cell surface phenotypes of lymphokine activated killer cells stimulated with R-IL 2 and an anti-CD 3 antibody].

We analyzed cytolytic activity and cell surface phenotypes of LAK (lymphokine activated killer) cells which were stimulated with recombinant IL-2 and an anti-CD3 antibody. This study involved 9 patients with astrocytomas, one patient with ganglioglioma, and one patient with medulloblastoma and their ages ranged between 5 and 80yr. Peripheral blood lymphocytes were separated from about 40 ml venous blood on a Ficoll-Paque gradient.

Biochemical pathogenesis of demyelination in globoid cell leukodystrophy (Krabbe's disease): the effects of psychosine upon oligodendroglial cell culture.

The effects of psychosine on the metabolism of myelin associated glycolipids such as galactocerebroside and sulfatide in mouse brain cell cultures were investigated in order to clarify the mechanism of demyelination in globoid cell leukodystrophy (Krabbe's disease). The incorporation of 3H-galactose into cerebroside and sulfatide was studied in the presence of psychosine (1-3 micrograms/ml medium). These data indicated that psychosine inhibited the incorporation of 3H-galactose into cerebroside and sulfatide not in astroglial cell culture but in oligodendroglial cell culture.

Galactosylceramide and galactosylsphingosine loading studies in cultured skin fibroblasts in human and murine globoid cell leukodystrophy.

Cell level studies of 3H-galactosylceramide(GalCer) and 3H-galactosyl sphingosine (GalSph) have been carried out in cultured skin fibroblasts from human and murine globoid cell leukodystrophy (GLD). GalCer loading studies disclosed that the hydrolysis rates of GalCer in human control and GLD were 72% and 45%, respectively, and those from the murine control and GLD cells were 77% and 21%, respectively, on the 5th day of culture. On the other hand, GalSph loading studies showed that the hydrolysis rate of GalSph in the human control and GLD were 40% and 10%, respectively, and those from murine control and GLD cells were 38% and 10% on the 12th day of culture.

Biochemical and morphological studies of dorsal root ganglion and its cultured cells from twitcher mouse (murine globoid cell leukodystrophy).

The biochemical pathogenesis of globoid cell leukodystrophy (GLD) (Krabbe disease) was investigated in vitro using the cultured neural cells obtained from dorsal root ganglion (DRG) of twitcher mouse (murine Krabbe disease). Electron microscopic examination of twitcher DRG of 30 days old showed the demyelination and abnormal inclusion bodies, whereas mitochondrial structure appeared to be intact. Cultured neural cells from control mice were well proliferated to form the network processes, while those from twitcher were decreased in cell numbers and showed the vacuolation of cell body, degeneration of processes, and finally died after three weeks.

[Surgical indication to deep-seated arteriovenous malformation].

The natural history of cerebral arteriovenous malformation (AVN) is still a subject of dispute. In the case of deep seated AVM ruptures, the problem is more serious because the hemorrhages often give rise to severe disability or death, depending on their location. Total extirpation of the nidus is fundamentally the best choice for treatment of AVM.

Immunocytochemical and ultrastructural studies of the histogenesis of Ewing's sarcoma and putatively related tumors.

The histogenesis of Ewing's sarcoma (EW) and extraskeletal Ewing's sarcoma (EEW) is still disputable. Their relationship to the so-called Askin's tumor, neuroectodermal tumor of bone, and peripheral neuroblastoma remains to be established. In an attempt to clarify these points, immunocytochemical and ultrastructural studies were done on tissues from 14 cases of EW, 4 cases of EEW, and 9 cases of primitive neuroectodermal tumor (PNET) and compared with neuroblastoma and olfactory neuroblastoma.

Fetal GM1-gangliosidosis: morphological and biochemical studies.

A 23-week fetus with GM1-gangliosidosis type 1 was studied morphologically and biochemically. The GM1-ganglioside content in the brain was approximately twice that of a control. A GM1-ganglioside comprised about 25% of the total ganglioside NANA (N-acetyl-neuraminic acid), whereas in control fetus brain the proportion was 14.

Accumulation of phosphorus compounds in tissues and cultured skin fibroblasts in patients with hypophosphatasia.

Patients with hypophosphatasia caused by a deficiency of alkaline phosphatase first showed marked accumulation of phosphoethanolamine and other phosphorus compounds in kidney and liver, while in placenta and intestine contents of these compounds were within a normal range. Furthermore, 32P-incorporation in cultured skin fibroblasts of patients with hypophosphatasia was increased about two to three times of control. FPLC chromatographic analysis also indicates that the accumulated phosphorus compounds in hypophosphatasia was smaller molecular phosphorus containing compounds.

Liposome targeting to mouse brain: mannose as a recognition marker.

Liposomes prepared from lecithin:cholesterol:p-aminophenyl-alpha-mannoside (7:2:1, v/v/v) were efficiently incorporated into the mouse brain across the blood brain barrier. Furthermore, liposomes injected intraperitoneally were exclusively distributed into lysosome rich fraction and also taken up by glial cells. These data suggest that blood brain barrier cells and glial cells recognize mannose molecule on the surface of the membrane and can be used for the treatment of brain damage by lysosomal storage disease.

Clinical and biochemical studies of Japanese neuronal ceroid-lipofuscinosis.

We studied clinical manifestations of Japanese patients with neuronal ceroid-lipofuscinosis (NCL). The onset of the disease and initial symptoms were almost identical to those reported previously in Caucasians. Japanese patients with NCL were not significantly clinically different from Caucasian cases.

Partial deficiency of beta-hexosaminidase activity in canine GM2-gangliosidosis.

4-Methylumbelliferyl-N-acetylglucosamine-6-sulfate (4MUGLc6S) which is known to be a specific substrate for human hexosaminidase A was used to determine enzymatic features of canine GM2-gangliosidosis. The enzyme activity using 4MUGlc6S in affected dog brain and liver was less than 20 to 30% of control tissues, whereas total 4-methylumbelliferyl beta-glucosaminidase activity in canine GM2-gangliosidosis was normal or elevated. However, when beta-hexosaminidase was fractionated by DEAE-Sepharose column chromatography, beta-hexosaminidase A like fraction in affected dog tissues was reduced to 20 to 30% of control.

Pathochemistry, pathogenesis and enzyme replacement in multiple-sulfatase deficiency.

Multiple-sulfatase deficiency (MSD) is now considered to be heterogeneous and could be classified into three or four clinical phenotypes according to the onset of the disease: neonatal, late infantile, juvenile and possibly adult type. Neonatal-type MSD shows severe clinical involvement and practically no arylsulfatase A, B and C activities in cultured skin fibroblasts. Furthermore, arylsulfatase A activity in neonatal-type MSD was not enhanced by the addition of thiosulfate.