Immunohistochemical identification of proteoglycan form of macrophage colony-stimulating factor on bone surface.

Several studies using op/op mice have shown that macrophage colony-stimulating factor (M-CSF) was necessary for osteoclast formation in vivo. Previously we reported that osteoblastic cells produced two molecular forms of M-CSF; one is an 85-kDa M-CSF, and the other is a proteoglycan form of M-CSF (PG-M-CSF) which has a binding affinity for bone-derived collagens and is extractable from human bone. In this study, we performed immunostaining of human bone using a newly established anti-PG-M-CSF antibody, and showed positive staining PG-M-CSF, probably produced by bone lining cells, on the bone surface.

Reproducibility of subjectively graded voluntary isometric muscle strength in unilateral and simultaneous bilateral exertion.

Reproduction error of voluntary isometric muscle strength that was graded subjectively by the subject was investigated under unilateral and bilateral conditions. Six kinds of tasks, i.e.

Monocyte chemoattractant protein-1 stimulates tumor necrosis and recruitment of macrophages into tumors in tumor-bearing nude mice: increased granulocyte and macrophage progenitors in murine bone marrow.

Monocyte chemoattractant protein-1 (MCP-1) belongs to the newly recognized "chemokine" superfamily of activation-inducible cytokines. We report here that MCP-1 gene-transferred mouse myeloma cells modulate tumor necrosis in myeloma-bearing nude mice. We established an MCP-1-producing myeloma cell line (X63-MCP-1) by transfection with human MCP-1 cDNA as well as interleukin-8-producing X63 cells (X63 IL-8).

Flow cytometric analysis of nuclear DNA content in the endoscopic biopsy tissues of gastric cancer.

The nuclear DNA content was measured by flow cytometry in gastric cancer patients using endoscopically biopsied tissue specimens. When the specimens were classified into diploid and aneuploid according to the DNA histogram, 56% (65/117) of the specimens were aneuploid, and advanced cancer was clearly more often aneuploid than early cancer. The frequency of aneuploidy appeared to be higher as the histologic depth of cancer was greater.

Flow cytometric analysis of nuclear DNA content in tissues of colon cancer using endoscopic biopsy specimens.

Flow cytometric assay of nuclear DNA in endoscopic biopsy specimens was evaluated in colon cancer patients. When the cells were divided into diploid cells and aneuploid cells, aneuploidy was observed in 63% (58 of 92) of the colon cancer patients. However, no clear relation was observed between the frequency of aneuploidy and the invasive depth, size, or histological type of colon cancer.

Cry j 2, a major allergen of Japanese cedar pollen, shows polymethylgalacturonase activity.

We examined Cry j 2, a major allergen of Japanese cedar (Cryptomeria japonica) pollen, for polygalacturonase enzyme activity, since a nucleotide sequence of cDNA of Cry j 2 showed a significant homology with that of tomato polygalacturonase. Polygalacturonase is well known to depolymerize preferentially polygalacturonic acid (PGA) by hydrolysis. However, Cry j 2 did not act on PGA, but was found to depolymerize pectin and methylesterified PGA in a dose-dependent manner.

Mitochondrial lipid peroxidation and superoxide dismutase in rat hypertensive target organs.

Mitochondrial respiratory chains leak a large amount of superoxide anion radicals, which chain react with membrane phospholipid to develop lipid peroxidation. Manganese superoxide dismutase (MnSOD) is then inducible and catalyzes superoxide detoxification within mitochondria. We examined mitochondrial thiobarbituric acid-reactive substance, an end product of lipid peroxidation, and MnSOD concentration in hypertensive target organs of spontaneously hypertensive and deoxycorticosterone acetate salts-induced hypertensive rats.

Reperfusion induces 5-lipoxygenase translocation and leukotriene C4 production in ischemic brain.

5-Lipoxygenase (5-LO) converts arachidonic acid, released from membrane phospholipids upon external stimulation, to leukotriene C4 (LTC4), which induces various kinds of cellular and molecular responses. We examined the effects of 5 min of ischemia on brain 5-LO and LTC4 during reperfusion using the gerbil model of transient forebrain ischemia that develops neuronal necrosis selectively in the hippocampus. Neurons exhibited dense 5-LO immunoreactivity; 5-LO was partially redistributed from cytosolic to particulate fractions 3 min during reperfusion.

Hypoxia/reoxygenation-mediated induction of astrocyte interleukin 6: a paracrine mechanism potentially enhancing neuron survival.

To elucidate mechanisms underlying neuroprotective properties of astrocytes in brain ischemia, production of neurotrophic mediators was studied in astrocytes exposed to hypoxia/reoxygenation (H/R). Rat astrocytes subjected to H/R released increased amounts of interleukin (IL) 6 in a time-dependent manner, whereas levels of tumor necrosis factor and IL-1 remained undetectable. IL-6 transcripts were induced in hypoxia and the early phase of reoxygenation, whereas synthesis and release of IL-6 antigen/activity occurred during reoxygenation.

Bioactive glass-ceramic containing crystalline apatite and wollastonite initiates biomineralization in bone cell cultures.

Rat bone cells were cultured in the presence of bioactive glass-ceramic containing crystalline apatite and wollastonite. Scanning electron microscopy observations of the surface of the seeded ceramic disks revealed that cells attached, spread, and proliferated on the material surface. Soaking in cell-free culture medium showed that no change occurred in the surface structure.

Proteoglycan form of macrophage colony-stimulating factor binds low density lipoprotein.

We recently isolated a proteoglycan form of macrophage colony-stimulating factor (PG-M-CSF) that carries a chondroitin sulfate glycosaminoglycan chain. Here, we examined the interaction of PG-M-CSF with low density lipoprotein (LDL). When LDL preincubated with PG-M-CSF was fractionated by molecular size sieving chromatography, it was eluted earlier than untreated LDL.

Characteristics of aspartate aminotransferase binding immunoglobulin determined by the isotope method.

A woman who had no known underlying diseases showed a persistent elevation (about 300 U/L) of serum aspartate aminotransferase (AST) without other abnormal laboratory findings. Cellolose gel electrophoresis showed that the AST activity in the patient had an atypical band with slower mobility than normal AST. When the sera from the patient and from a patient with acute hepatitis were mixed, the atypical band increased in density and the band of normal size AST disappeared.

Quantitative analysis of the two macrophage colony-stimulating factor mRNA expressed in a human stromal cell line by reverse transcription-polymerase chain reaction (RT-PCR).

We established a quantitative analysis system for 4.0 kb and 1.6 kb macrophage colony-stimulating factor (M-CSF) mRNA, using reverse transcription-polymerase chain reaction.

Alteration of the proteoglycan form of macrophage colony-stimulating factor produced by a human stromal line stimulated by tumor necrosis factor-alpha.

Immunoblot analysis of macrophage colony-stimulating factor (M-CSF) in KM 102 cell-conditioned medium showed the presence of two M-CSF molecular types, one being 85-kd M-CSF, the other a proteoglycan form (PG-M-CSF) carrying a chondroitin sulfate chain of variable length. When KM 102 cells were stimulated by TNF-alpha, they produced more M-CSF than that produced in unstimulated condition, in which PG-M-CSF had a shorter chondroitin sulfate chain. Although PG-M-CSF has binding affinity for type V collagen, the PG-M-CSF with the shorter chondroitin sulfate chain shows lower affinity.

Metabolic and biosynthetic alterations in cultured astrocytes exposed to hypoxia/reoxygenation.

To investigate the astrocyte response to hypoxia/reoxygenation, as a model relevant to the pathogenesis of ischemic injury, cultured rat astrocytes were exposed to hypoxia. On restoration of astrocytes to normoxia, there was a dramatic increase in protein synthesis within 3 h, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of metabolically labeled astrocyte lysates showed multiple induced bands on fluorograms. Levels of cellular ATP declined during the first 3 h of reoxygenation and the concentration of AMP increased to approximately 3.

Lack of task-related motor unit activity in human triceps brachii muscle during elbow movements.

Surface electromyographic (EMG) recordings have been associated with the acceleration and deceleration characteristics of single joint elbow movements [J. Neurophysiol., 63 (1990) 465-472].

A new gerbil model of hindbrain ischemia by extracranial occlusion of the bilateral vertebral arteries.

A new gerbil model of hindbrain ischemia was induced by extracranial occlusion of the bilateral vertebral arteries just before their entry into the transverse foramen of the cervical vertebra. Carbon black studies, performed at 5 min after occlusion, revealed that the pons-medulla oblongata, and the cerebellum were quite ischemic in all animals. Cardiovascular changes in mean arterial blood pressure (MABP) and heart rate were recorded until 30 min after occlusion, and revealed that the typical cerebral ischemic response (i.

An ischemic opening of the blood-brain barrier may deteriorate brain stem auditory evoked potentials following transient hindbrain ischemia in gerbils.

To clarify the effect of vasogenic brain edema on the brainstem, the relationships between waveform changes in brainstem auditory evoked potentials (BAEP) and blood-brain barrier (BBB) disturbance following transient hindbrain ischemia were investigated. Hindbrain ischemia was induced in gerbils by bilateral occlusion of the vertebral arteries. The animals were divided into three groups subjected to 0, 5, and 30 min of bilateral vertebral occlusion (BVO-0',-5', and -30' groups; n = 4 in each group).

Internucleosomal DNA cleavage involved in ischemia-induced neuronal death.

Pyramidal neurons of the hippocampal CA1 are known to be particularly vulnerable to transient ischemia resulting in "delayed neuronal death". Recent studies using aurintricarboxylic acid suggested that ischemia- or excitotoxin-induced neuronal death should share intracellular mechanisms in common with apoptosis. It is, however, unclear about involvement of endonucleases.

Differential vulnerability in the hindbrain neurons and local cerebral blood flow during bilateral vertebral occlusion in gerbils.

Differential vulnerability in the hindbrain neurons was examined immunohistochemically during hindbrain ischemia in the gerbil. Hindbrain ischemia was produced by extracranial occlusion of the bilateral vertebral arteries just before their entry into the transverse foramen of the cervical vertebra. Local cerebral blood flow was measured by quantitative autoradiographic technique after 5 min of ischemia and was reduced to less than 5 ml/100 g per min in the cerebellum, the pons, and the medulla, indicating that severe and reproducible hindbrain ischemia was induced immediately after occlusion.

Effect of transient forebrain ischemia on superoxide dismutases in gerbil hippocampus.

Substantial generation of oxygen-derived free radicals has been implicated in pathophysiology of ischemic brain damage. Immunoreactive mitochondrial manganese and cytosolic copper-zinc superoxide dismutases, initial and essential enzymes to scavenge superoxide radical anions, increased in the gerbil hippocampal neurons after transient forebrain ischemia. Neuronal cells responded to oxidative stress in ischemia and induced the protective mechanism to increase superoxide dismutases.