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Biomedical subjects

M Sano

Publications and source records attributed to M Sano.

At least 487 records · Page 27Linked to original sources

A convenient bioassay for NGF using a new subline of PC12 pheochromocytoma cells (PC12D).

A useful bioassay for nerve growth factor (NGF) has been developed, based on the rapid outgrowth of neurites (within 24 h) from cells of a new subclone of PC12 cells (PC12D) in response to NGF. The sensitivity is similar to that of other bioassay systems that the the sensory ganglia of chick embryos of primed PC12 cells. The assay is readily adaptable for the purification of NGF and for the determination of levels of NGF in tissue, as shown by a comparison of results from this assay to the data obtained by an immunological assay.

Animals↗

Solubilization, purification, and characterization of a membrane-bound phospholipase A2 from the P388D1 macrophage-like cell line.

The release of free arachidonic acid from membrane phospholipids is believed to be the rate-controlling step in the production of the prostaglandins, leukotrienes, and related metabolites in inflammatory cells such as the macrophage. We have previously identified several different phospholipases in the macrophage-like cell line P388D1 potentially capable of controlling arachidonic acid release. Among them, a membrane-bound, alkaline pH optimum, Ca2+-dependent phospholipase A2 is of particular interest because of the likelihood that the regulatory enzyme has these properties. This phospholipase A2 has now been solubilized from the membrane fraction with octyl glucoside and partially purified. The first two steps in this purification are butanol extractions that yield a lyophilized, stable preparation of phospholipase A2 lacking other phospholipase activities. This phospholipase A2 shows considerably more activity when assayed in the presence of glycerol, regardless of whether the substrate, dipalmitoylphosphatidylcholine, is in the form of sonicated vesicles or mixed micelles with the nonionic surfactant Triton X-100. Glycerol (70%) increases both the Vmax and the Km with both substrate forms, giving a Vmax of about 15 nmol min-1 mg-1 and an apparent Km of about 60 microM for vesicles and a Vmax of about 100 nmol min-1 mg-1 and an apparent Km of about 1 mM for mixed micelles. Vmax/Km is slightly greater for vesicles than for mixed micelles. The lyophilized preparation of the enzyme is routinely purified about 60-fold and is suitable for evaluating phospholipase A2 inhibitors such as manoalide analogues. Subsequent steps in the purification are acetonitrile extraction followed by high performance liquid chromatography on an Aquapore BU-300 column and a Superose 12 column. This yields a 2500-fold purification of the membrane-bound phospholipase A2 with a 25% recovery and a specific activity of about 800 nmol min-1 mg-1 toward 100 microM dipalmitoylphosphatidylcholine in mixed micelles. When this material was subjected to analysis on a Superose 12 sizing column, the molecular mass of the active fraction was approximately 18,000 daltons.

Acetonitriles↗

The relationship of serotonin to depression in Parkinson's disease.

We have previously reported a correlation between depression in patients with idiopathic Parkinson's disease and decreased concentrations of the cerebrospinal fluid content of the serotonin metabolite, 5-HIAA. To further examine this relationship, we repeated the study in a new cohort of patients while they remained on dopaminergic medications, conducted follow-up interviews and examinations in our original cohort, and conducted an open trial of the serotonin precursor, 5-hydroxytryptophan in a group of new patients with depression. We were again able to demonstrate a significant reduction in cerebrospinal 5-HIAA in depressed patients in comparison to controls and patients without depression. Demented patients with Parkinson's disease, particularly those with concurrent depression, had the lowest values of 5-HIAA. No new cases of depression occurred in our original cohort after 2 1/2 years of follow-up, and depression remitted following conventional or experimental treatment in four patients. Depression improved in six of the seven new patients following oral 5-hydroxytryptophan. Three of these patients allowed a repeat lumbar puncture, and the concentration of 5-HIAA increased following 5-hydroxytryptophan. These three studies support our hypothesis that depression in idiopathic Parkinson's disease is associated with a reduction in brain serotonin. However, it also suggests that other factors, biological or environmental, may be causal factors.

5-Hydroxytryptophan↗

Generation of Listeria monocytogenes-specific T cells mediating delayed footpad reaction and protection in neonatally thymectomized mice but not in nude mice.

Neonatally thymectomized (NTx) mice, sham-operated control mice and congenitally athymic nude mice were immunized with viable Listeria monocytogenes and their spleen cells examined for the capacity to transfer both delayed footpad reaction and protection against challenge at the site of local transfer. Cells from immune NTx mice conferred significant degrees of delayed footpad reaction and protection comparable to sham mice, while cells from immune nude (nu/nu) mice did not. This ability was completely eliminated by the treatment of cells with anti-Thy1, anti-Lyt1 or anti-L3T4 antibody plus complement but not with anti-Lyt2 antibody plus complement. These results indicated that NTx mice can normally mount the immunity to L. monocytogenes by generating Lyt1+2-, L3T4+ T cells. Immune competence of NTx mice and thymus dependency of various immune responses are discussed.

Animals↗

Bilateral anterior opercular syndrome: localizing value of SPECT and MRI.

Bilateral anterior opercular syndrome is clinically characterized by facio-pharyngo-glossomasticatory diplegia due to bilateral opercular lesions. Reported cases of the syndrome have been relatively rare probably because of difficulty in determining opercular damage on clinical symptoms alone. We report a case of this syndrome in whom bilateral opercular damage could be detected not by CT, but suggested by SPECT, and confirmed by MR imaging.

Aged↗

Effects of enoxacin, ofloxacin and norfloxacin on theophylline disposition in humans.

The effect of three new fluoroquinolones on theophylline kinetics and the urinary excretion of metabolites was studied in 5 healthy subjects (3 male, 2 female). All subjects received serial, single i.v. infusions of theophylline (aminophylline, 250 mg) over 60 min after 200 mg doses of a quinolone (enoxacin, ofloxacin, norfloxacin) every 8 h for 3 consecutive days, the quinolone being administered up to the day following theophylline administration. Pretreatment with ofloxacin and norfloxacin did not influence theophylline disposition, but theophylline clearance fell from 0.054 to 0.027 l.h-1.kg-1 in the presence of enoxacin, without a change in the apparent volume of distribution. Enoxacin, too, was the sole compound to increase the urinary excretion of theophylline (33.2 vs 43.9 mg, before vs after treatment), and significantly to decrease the excretion of 3-methylxanthine (3-MX), 1-methyluric acid (1-MU) and 1,3-dimethyluric acid (1,3-DMU) in 24-h urine samples (from 19.8 to 7.16 mg, from 28.3 to 10.3 mg and from 68.8 to 49.5 mg, respectively). The effect of the quinolones on hepatic drug metabolizing enzyme activity was investigated in each subject using the ratios of 6-hydroxycortisol to total 17-hydroxycorticosteroids and to free cortisol in 24-h urines as an index of the hepatic P-450-dependent enzyme system. No significant difference in ratio was observed between control and other treatments. It is concluded that the theophylline-enoxacin interaction was largely due to inhibition of a metabolic system other than the common hepatic P-450 system.

17-Hydroxycorticosteroids↗

Immunolocalization of cathepsins B, H and L in skeletal muscle of X-linked muscular dystrophy (mdx) mouse.

The amounts of non-collagen proteins (muscle structural proteins) and the activity of creatine kinase were significantly decreased in muscles of 28-day-old mdx mice. The activities of lysosomal thiol proteases such as cathepsins B and L were increased in muscles of mdx mice at as early as 10 days of age. Endogenous thiol proteinase inhibitor and various lysosomal hydrolases also showed increased activities. The localization of cathepsins B, H and L, and endogenous thiol proteinase inhibitor was investigated using the respective specific antibodies. While only invading macrophages were stained strongly with anticathepsin B and H, and anti-thiol proteinase inhibitor antibodies, cathepsin L was localized in muscle cells as well as in invading macrophages. Cathepsin L in muscle cells itself may initially degrade muscle structural proteins, before lysosomal thiol proteases, mainly derived from macrophages, degrade them in skeletal muscles of mdx mice.

Age Factors↗

Homocitrullinuria and homoargininuria in hyperargininaemia.

A four-year-old boy with hyperargininaemia had increased urinary excretion of homocitrulline and homoarginine. A single oral lysine load created a marked increase in these amino acids in plasma. A daily oral lysine supplementation resulted in a remarkable urinary leakage of homocitrulline and homoarginine. These findings suggest that the patient had an enhanced synthesis of these amino acids.

Amino Acid Metabolism, Inborn Errors↗

Efficacy of praziquantel against Schistosoma nasale infection in cattle.

Cattle naturally infected with nasal schistosomiasis were treated with a single oral dose of praziquantel at 20 mg/kg body weight. The drug was highly effective causing a considerable reduction in egg counts, cessation of clinical signs and progressive regression of the nasal granulomatous growths.

Animals↗

Long-term administration of oral physostigmine in Alzheimer's disease.

Minimal short-term benefits with oral physostigmine have been reported in Alzheimer's disease. We examined long-term oral physostigmine therapy in 14 patients with probable Alzheimer's disease in an extended double-blind crossover trial in which they received physostigmine for five intervals of 4 to 6 weeks in length and placebo for one interval determined at random. At each interval a Selective Reminding Test was administered and three indexes were examined: total recall, long-term recall, and intrusions. As a group, patients' memory test scores were significantly better during the drug period. Two patients who previously had performed worse on drug than on placebo in a short-term trial continued to do so. Nine of the remaining 12 patients performed better on two or more measures on physostigmine compared with placebo. In some cases scores improved up to 50% over placebo values. This improvement was not predicted by response to medication in the earlier short-term trial. Seven patients completed an additional six-interval crossover trial. All but one continued to have improved performance with oral physostigmine and, in grouped data, memory measures remained significantly improved. These results suggest that long-term administration of physostigmine in Alzheimer's disease may be more effective than short-term. This benefit can be sustained for up to a year in some patients.

Administration, Oral↗

[Six-month chronic intravenous toxicity study of cefodizime sodium in dogs].

The chronic intravenous toxicity of cefodizime sodium (THR-221) was studied in beagle dogs. Groups of 6 males and 6 females were treated with THR-221 at doses of 0 (saline), 200, 400, 800 and 1600 mg/kg/day for 6 months. The THR-221 related symptoms were vomiting, excessive drinking behavior and salivation. The paleness of the visible mucosa and discoloration of vascular color by funduscopy due to systemic anemia were observed in one animal each of 800 and 1600 mg/kg/day groups. Body weight was depressed transiently or continuously in a few animals of 400-1600 mg/kg/day groups. The hematological, serum chemical and urinalysis findings in a few animals of 400-1600 mg/kg/day groups revealed decreases in RBC count, PCV and hemoglobin, an increase in reticulocyte count, a decrease in WBC count, a decrease in platelet count, slight increase in TP, and albumin, a decrease in AlP, and an increase in urinary Na. Light microscopically, deposition of hemosiderin and increased extramedullary hematopoiesis in the liver and spleen, and deposition of fibroid substance in the white pulp of the spleen and diffuse fibrosis in the bone marrows were detected in a few animals of 800 and 1600 mg/kg/day groups. Electron microscopically, no significant toxic changes were observed. The maximum nontoxic doses of THR-221 are estimated as 200 mg/kg/day in male and less than 200 mg/kg/day in female.

Anemia↗