Isolation and characterization of hemorrhagic factors a and b from the venom of the Chinese habu snake (Trimeresurus mucrosquamatus).

Hemorrhagic factors a and b were isolated from Trimeresurus mucrosquamatus venom by Sephadex G-100, CM-Sephadex C-50 and DEAE-Sephacel column chromatographies. The hemorrhagic factors were homogeneous, as established by a single band on acrylamide gel electrophoresis, isoelectric focusing and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Molecular weights of 15 000 and 27 000 were found for hemorrhagic factors a and b, respectively. Factor a possesses proteolytic activity hydrolyzing the His(10)-Leu(11), Tyr(16)-Leu(17) and Arg(22)-Gly(23) bonds of oxidized insulin B chain, whereas, factor b hydrolyzed only the Ala(14)-Leu(15) bond. Hemorrhagic activity of these hemorrhagic factors was inhibited by ethylenediaminetetraacetic acid, 1,10-phenanthroline or p-chloromercuribenzoate, but not by soybean trypsin inhibitor or diisopropyl fluorophosphate. The hemorrhagic factors were injected into the skin of the back of albino rabbits, and the minimum hemorrhagic dose of factors a and b was 1.7 and 2.3 micrograms, respectively. These purified hemorrhagic factors were not lethal at 15 micrograms/g in mice. Factor a hydrolyzed the B beta chain of fibrinogen, while factor b hydrolyzed the A alpha chain. Hemorrhagic factor a was shown to differ immunologically from factor b. Factors a and b produced systemic hemorrhage in internal organs such as the heart and stomach of mice. Moreover, factor b produced hemorrhage in the liver.

Establishment of a new scirrhous gastric cancer cell line with loss of heterozygosity at E-cadherin locus.

Scirrhous gastric carcinoma, characterized by carcinoma cell proliferation and infiltration with extensive fibrosis in the stroma, frequently causes peritoneal metastasis. We describe here a newly established cell line, OCUM-6, derived from ascites effusion of a scirrhous gastric cancer patient. The cells are floating and round shape, similar to other scirrhous gastric carcinoma cell lines previously reported. Histologic findings of xenografted tumor obtained from OCUM-6 cells showed medullary growth with a poorly differentiated adenocarcinoma containing signet ring cells. LOH at E-cadherin locus 16q22 was observed in the OCUM-6 cells. LOH at E-cadherin locus might be closely associated with histologic findings and metastatic process of scirrhous gastric cancer. The scirrhous gastric cancer cell line, OCUM-6, may be useful for investigation of the mechanisms of peritoneal dissemination and carcinogenesis.

sigma receptor ligands attenuate N-methyl-D-aspartate cytotoxicity in dopaminergic neurons of mesencephalic slice cultures.

We investigated the potential neuroprotective effects of several sigma receptor ligands in organotypic midbrain slice cultures as an excitotoxicity model system. When challenged with 100-microM N-methyl-D-aspartate (NMDA) for 24 h, dopaminergic neurons in midbrain slice cultures degenerated, and this was prevented by (5R, 10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,b]-cyclohepten-5, 10-imine (MK-801; 1-10 microM). Concomitant application of ifenprodil (1-10 microM) or haloperidol (1-10 microM), both of which are high-affinity sigma receptor ligands, significantly attenuated the neurotoxicity of 100 microM NMDA. The sigma(1) receptor-selective ligand (+)-N-allylnormetazocine ((+)-SKF 10047; 1-10 microM) was also effective in attenuating the toxicity of NMDA. The effect of R(-)-N-(3-phenyl-1-propyl)-1-phenyl-2-aminopropane hydrochloride ((-)-PPAP), a sigma receptor ligand with negligible affinity for the phencyclidine site of NMDA receptors, was also examined. (-)-PPAP (3-100 microM) caused a concentration-dependent reduction of NMDA cytotoxicity, with significant protection at concentrations of 30 and 100 microM. In contrast, (+)-SKF 10047 (10 microM) and (-)-PPAP (100 microM) showed no protective effects against cell death induced by the Ca(2+) ionophore ionomycin (1-3 microM). These results indicate that sigma receptor ligands attenuate the cytotoxic effects of NMDA on midbrain dopaminergic neurons, possibly via inhibition of NMDA receptor functions.

Requirement of neural activity for the maintenance of dopaminergic neurons in rat midbrain slice cultures.

Chronic treatment of organotypic midbrain slice cultures with L-type Ca(2+) channel blocker nicardipine (3-10 microM) or verapamil (10 microM) for 18 days resulted in a drastic decrease in the number of dopaminergic neurons. A voltage-dependent Na(+) channel blocker tetrodotoxin (1 microM) was also effective in decreasing the number of dopaminergic neurons. Concurrent application of forskolin (20 microM) or dibutyryl cyclic AMP (1 mM) counteracted the effects of nicardipine and tetrodotoxin. These results suggest that spontaneous neuronal activity within midbrain slice cultures, causing Ca(2+) influx through L-type Ca(2+) channels that maintains intracellular cyclic AMP levels, is required for the maintenance of dopaminergic neurons.

Distribution of cesium-137 in Japanese forest soils: correlation with the contents of organic carbon.

The spatial and vertical distributions of 137Cs in surface soils were surveyed and analyzed then correlated with the contents of organic carbon in the hinoki (Chamaecyparis obtusa Sieb. et Zucc.) plantation forest and secondary forest dominated by red pine (Pinus densiflora Sieb. et Zucc.) in Japan. The spatial variation of 137Cs activity was observed in the surface soil around the red pine. The average activity of 16 samples around the tree is 42.4 Bq/kg and the standard deviation is 25.9 Bq/kg. This finding indicates the importance in the selection of a sampling site and the number of samples from the surface soils especially around a tree. For the vertical distribution of 137Cs activity, it was found that the concentration in the surface soil is highest, 149 Bq/kg in the hinoki stand and 101 Bq/kg in the red pine stand, and decreases with depth. The relationship between 137Cs activity and carbon content in the forest soil was investigated in two undisturbed forest stands. The relations were more precisely expressed using an exponential equation than by a linear equation. From the same forest, similar regression equations were obtained. This indicates that the distribution of 137Cs could be characterized by the organic carbon content in an undisturbed forest. It is also suggested that the coefficient values in the regression equation help to define the movement of 137Cs accompanying the decomposition of organic matter.

[Analysis of factors influencing the self-regulation of medication and construction of an educational program for patients with chronic hepatic diseases].

The purpose of this study is to construct an educational program for patients with chronic hepatic diseases to avoid self-regulation of medication. We have statistically analyzed the influencing factors of self-regulation from the following 3 viewpoints. First, in the investigation of the number of and the kind of prescribed medicines, it was found that the number of prescribed medicines increased sharply at the stage of Liver Cirrhosis b. Secondly, in the survey of patients' awareness of medication, it was found that self-regulation was influenced by lack of understanding the effects of medicines and the necessity of medication, and by lack of understanding the clinical conditions. Especially, at the stage of Liver Cirrhosis b, self-regulation was strongly influenced by lack of understanding the clinical conditions. Lastly, in the evaluation of patients' characteristics by pharmacists, self-regulation was also strongly influenced by the level of understanding the necessity of medication, understanding the clinical conditions, a patient-doctor relationship, and anxiety about side-effects of medicines. Therefore we have constructed an educational program for patients from the viewpoint of medication. The information consists of two articles; one is the action mechanism of the medicine connected with the clinical conditions to help the patients understand the necessity of medication; the other is how to check the effect and adverse reaction of the medication by themselves to decrease the anxiety about side-effects. Consequently, the medication management and the consultation for inpatients with chronic hepatic diseases were standardized by incorporating the education with the information described above.

[Statistical study of aqueous flare in rhegmatogenous retinal detachment].

Inflammation of the anterior chamber in rhegmatogenous retinal detachment was studied in 338 eyes with a laser flare cell meter. The mean flare value, 32.84 +/- 81.39 pc/msec, was significantly higher (p < 0.0001) than the 5.01 +/- 2.05 pc/msec of the control group of 133 normal eyes. Clinical findings of retinal detachment were grouped into 11 factors and each factor was divided into 34 details. The following details were proven by t-test to cause significant elevation of the flare value: (1) duration of more than 3 months, (2) hypotony of less than 9 mmHg, (3) aphakia, (4) retinal breaks in the ora area or the posterior area, (5) extension of retinal detachment with more than 2 quadrants, and (6) choroidal detachment. The correlation coefficients of the 6 clinical factors showed significant elevation of flare value in the following order: (1) presence or absence of choroidal detachment (0.396), (2) extension of retinal detachment (0.375), (3) intraocular pressure (-0.28), (4) duration of retinal detachment (0.18), (5) location of the break (0.15), and (6) presence or absence of the lens (0.134).

Air pump for retinal detachment with macular hole.

From 1984 to 1992, 128 eyes with substantial retinal detachment due to macular hole underwent vitrectomy at the Ideta Eye Hospital. In the first 76 eyes treated, air was injected by syringe after removal of the vitreous (group A); in the subsequent 52 eyes, air was injected by a pump while subretinal fluid was aspirated through the macular hole (group B). The rates of retinal reattachment after the first surgery were 63.2% in group A and 82.7% in group B (P < .05). Thus, using the air pump was more effective than air injection.

Anterior chamber inflammation after transconjunctival cryosurgery.

BACKGROUND: Inflammation caused by transconjunctival cryotherapy for prophylactic retinal detachment surgery was measured in various conditions. METHODS: Thirty-four eyes of 28 patients with peripheral retinal lesions predisposing to retinal detachment were studied by laser flare cell meter before and after treatment. RESULTS: The mean flare value for 34 eyes was 4.06 +/- 1.45 photon counts/ms before surgery and 5.72 +/- 2.52 pc/ms after surgery (p < 0.05). Flare value was elevated at 1, 2, and 3 weeks after treatment, peaking at 2 weeks (p < 0.05), and normal again at 4 weeks. There were no significant differences in flare increase between eyes with and without retinal breaks, eyes with and without limited retinal detachment, eyes with myopia more and less than -8.0 D, and eyes with a treatment area limited to one quadrant and extending over more than one quadrant. CONCLUSION: Transconjunctival cryosurgery caused mild inflammation in the anterior chamber of the eye for 3 weeks. The inflammation was not affected by the presence of retinal break or limited retinal detachment, the degree of myopia, or the extent of the treatment area.

Superoxide dismutase activity in organotypic midbrain-striatum co-cultures is associated with resistance of dopaminergic neurons to excitotoxicity.

We have previously demonstrated that dopaminergic neurons in midbrain-striatum slice co-cultures are more resistant to NMDA cytotoxicity than the same neuronal population in single midbrain slice cultures. Here, we show that dopaminergic neurons in midbrain-striatum co-cultures also exhibit resistance to the cytotoxicity of nitric oxide donors, 2,2'-(hydroxynitrosohydrazono)bis-ethanamine (NOC-18) and 3-morpholinosydnonimine (SIN-1). The cytotoxicity of NMDA (30 microM) in single cultures was significantly attenuated by the nitric oxide synthase (NOS) inhibitor N(omega)-nitro-L-arginine (100 microM), whereas the toxicity in co-cultures was not. The levels of tyrosine residue nitration of tyrosine hydroxylase, a hallmark of the occurence of peroxynitrite anion in dopaminergic neurons, were lower in co-cultures than those in single cultures. Single cultures and co-cultures did not show appreciable differences in the number or distribution of NOS-containing neurons as assessed by NADPH diaphorase histochemistry. On the other hand, midbrain slices cultured with striatal slices showed higher levels of superoxide dismutase (SOD) activity as well as increased protein levels of Cu,Zn-SOD, than midbrain slices cultured alone. These results suggested that the generation of NO is involved in NMDA cytotoxicity on dopaminergic neurons, and that increased activity of SOD in co-cultures renders dopaminergic neurons resistant to NMDA cytotoxicity by preventing the formation of peroxynitrite.