TAKAHASHI, T., ASANUMA, Y., KATO, T., HEBIGUCHI, T., KOYAMA, K. and KUDO, K. On-Line Plasma Exchange for Sepsis in Neonates and Infants. Tohoku J. Exp. Med., 1989, 159 (4), 249-256 - The present study way designed to assess the effectiveness of on-line plasma exchange (on-line P.E.) between septic infants and healthy adults using septic puppies. The plasma separation system consisted of a membrane plasma separator and a plasma treatment system, KM-9027 (Kurarey Co.) equipped with small 2 channel blood and plasma pumps. 5×109 CFU/ml/kg of E. coli with endotoxin was injected intravenously into 22 puppies and they were divided into 4 groups, namely untreated group (n=7), on-line P.E. group (n=5), P.E. group (n=5), sham group (n=5). In the on-line P.E. group, about 80ml/kg of plasma in septic puppy was replaced during 2hr with fresh plasma simultaneously obtained from healthy adult dog. As the results, 4 of 5 survived in the on-line P.E. group and 1 of 5 survived in the P.E. group, while all other puppies died within 24hr. In the P.E. group and on-line P.E. group, mean blood pressure and urinary output were significantly improved (p<0.05) by the treatment. The numbers of E. coli and endotoxin concentration in the blood were reduced significantly (p<0.05) only in the treatment with on-line P.E. opsonic activity recovered significantly (p<0.05) in P.E. group. This system appears to be effective and applicable for sepsis in infants and neonates.
SHIOI, A. and FUJIMOTO, T. Disorganization Process in the Development ofDiabetic Nodular Glomerulosclerosis. Tohoku J. Exp. Med., 1989, 159 (4), 257-275 -In order to clarify the mode of development of the diabetic nodular glomerulosclerosis, 38 kidney specimens of autopsied and biopsied diabetic cases were studied light and electron microscopically including serial sections. Disorganization process occurred chiefly at the capillary region of glomeruli. Early change of this process was proliferation of intramembranous cells following histolysis in the glomerular loop and it was characterized by transformation of the glomerular loop into enmeshed or reticular structure, that is, disorganization of the glomerular architecture. In the subsequent stage intercellular matrix production and hyalinization accompanied by axial crowding of proliferated intramembranous cells and peripheral recanalization of blood spaces were clarified. And in the later stage axially crowded intramembranous cells decreased in number and the matrix was increased and hyalinized, resulting in the formation of the axial hyaline nodule. The distribution of this process was focal and segmental, lesions of various stages coexisted, and it was suggested that glomerular lesions may spread all over the kidney by recurrence of this process. This process was quite similar to those seen in disorganization process of glomerulonephritis. But another characteristic changes were the presence of foam cells, intra- and intercellular deposition of lipid droplets, and increased matrix formation.
IIJIMA, T. and TAIRA, N. Mechanisms of β-Adrenoceptor Mediated Increase inDelayed Rectifier Potassium Current. Tohoku J. Exp. Med., 1989, 159 (4), 277-281 -Experiments were carried out in single ventricular cells of the guinea-pig heart. Isoproterenol, forskolin, intracellularly applied cyclic AMP and 3-isobutyl-1-methylxanthine increased the delayed rectifier potassium current (IK). The effect of isoproterenol was abolished by intracellularly applied guanosine 5'-O-(3-thio-triphosphate). These results indicate that isoproterenol stimulates β-adrenoceptors to activate adenylate cyclase by mediation through the stimulatory GTP-binding protein, and causes an increase in intracellular cyclic AMP levels. Then IK is probably increased by phosphorylation of the IK-channel protein by cyclic AMP-dependent protein kinase.
INABA, T., YAMASHITA, M., KAWASE, Y., NAKAHASHI, H. and WATANABE, H. Quantitative Measurement of Renal Plasma Flow by Positron Emission Tomographywith Oxygen-15 Water. Tohoku J. Exp. Med., 1989, 159 (4), 283-289-We successfully determined renal plasma flow in the human kidney by means of positron emission tomography (PET) with oxygen-15 water using a one compartment model.
OHNEDA, A., KOIZUML, F. and OHNEDA, M. Effect of Porcine Pancreastatin onEndocrine Function of Canine Pancreas. Tohoku J. Exp. Med., 1989, 159 (4), 291-298 - Controversial results concerning the modulation of insulin release with pancreastatin prompted us to investigate the effect of this peptide upon the endocrine function of the pancreas using an in situ perfusion method in dogs. Administration of porcine pancreastatin into the pancreaticoduodenal artery in a dosage of 0.34μg/kg of body weight during arginine infusion increased slightly plasma insulin and lowered minimally plasma glucagon in the pancreaticoduodenal vein. Pancreastatin infusion in a dosage of 500ng/min did not modify glucose-induced insulin release and glucagon suppression by glucose. Theophylline infusion induced a slight increase in plasma insulin and glucagon in the pancreaticoduodenal vein. Pancreastatin infusion accelerated slightly these changes in plasma insulin and glucagon following theophylline infusion. From the present experiment it is concluded that porcine pancreastatin did not inhibit glucoseinduced insulin release from the canine pancreas, inconsistent with the first report on insulin release from the rat pancreas. Alternatively, porcine pancreastatin rather stimulated insulin release during arginine infusion or following theophylline administration.
SAIJO, Y., KUMANO, N., TOKUE, Y., SATOH, K., OIZUMI, K. and MOTOMIYA, M. Characterization of Resistance to VP-16 in Human Leukemic Cell Line. Tohoku J. Exp. Med., 1989, 159 (4), 299-306-Resistance mechanism was studied in the VP-16-resistant human leukemic cell line (THP-1/E) which was developed by continuous drug exposure. The drug uptake and efflux studies revealed no decrease in net cellular drug accumulation. VP-16-induced DNA single- and double-strand breaks in the whole THP-1/E cells decreased significantly compared to the sensitive counterpart as assessed by alkaline elution methods. Decrease in DNA SSBs was also observable in the isolated nuclei from the THP-1/E cells. The resistance to VP-16 in THP-1/E appeared to be independent of altered membrane permeability, and more likely to be associated with decreased VP-16-mediated DNA cleavage.
TAKASU, N., MASUKO, T., HOJO, H. and HASHIMOTO, Y. A Microtestplate-Immunofluorescence Assay for Anti-Trypanosoma cruzi. Antibodies. Tohoku J. Exp. Med., 1989, 159 (4), 307-312-A new microassay method for the detection of anti-Trypanosoma (T.) antibodies was developed by using Terasaki's microtestplates, and T. cruzi or other parasites were immunofluorescently labeled with polyclonal or monoclonal anti-T. cruzi antibodies. The fluorescence intensity of immunofluorescently stained parasites was assessed by either visual observation or quantitative analysis using a microscopic spectrophotometer. Although the sensitivity of the microtestplate assay was nearly equal to that of the conventional glass slide assay or ELISA, this method is easier in the assay procedures and spares the amount of antibodies and the number of target parasites requisite to the assay.
TAKASU, N., MASUKO, T., SUGAHARA, K. and HASHIMOTO, Y. Production andCharacterization of Monoclonal Antibodies against Trypanosoma cruzi-AssociatedAntigens. Tohoku J. Exp. Med., 1989, 159 (4), 313-321 - Nine monoclonal antibodies (mAb) recognizing characteristic antigens in Trypanosoma (T.) cruzi were obtained from hybridomas which had been established by a fusion between mouse myeloma cells and the spleen cells of a mouse immune to the epimastigote form of T. cruzi (Tulahuen strain). Antigen specificities of these mAb were assessed by an indirect immunofluorescence (IIF) method using T. cruzi in different life cycles (amastigote and trypomastigote) and other members of the Trypanosomatidae. The mAb were classified into 3 groups from their reaction patterns to different parasites: 1) The strain specific mAb that reacted only with T. cruzi Tulahuen epimastigote, 2) the species specific mAb that reacted with all T. cruzi strains but not with other species of parasites, and 3) the mAb that were cross-reactive with other species of Trypanosomatidae. Most mAb were specific to epimastigote form of T. cruzi, but some reacted weakly with trypomastigote and amastigote form of the parasites. Immunoblotting and glycolipid analyses of the membrane fraction of homogenized parasites using the mAb identified at least 3 distinct antigenic molecules; those of protein nature having Mr. 43, 000 and 58, 000 and Mr. 43, 000 and 62, 000 and molecule(s) of glycolipid nature.
YOSHIDA, K., SAKURADA, T., KAISE, K., KAISE, N., NOMURA, T., ITAGAKI, Y., YAMAMOTO, M., SAITO, S. and YOSHINAGA, K. Relationship between Serum FreeThyroid Hormone Concentrations and Target Organ Responsiveness in ThyroidDisease Patients before and after Treatment. Tohoku J. Exp. Med., 1989, 159 (4), 323-331 - Relationship between serum free thyroid hormone concentrations and several markers of peripheral tissue response to thyroid status was studied in 24 patients with hyperthyroidism, 17 with euthyroidism with goiter and 10 with primary hypothyroidism before and after the initiation of standard forms of treatment. Before treatment, serum free T4 correlated significantly, either positively or negatively, with the basal metabolic rate (BMR), duration of the achilles tendon reflex (ATR), resting pulse rate (RPR), serum total cholesterol (T-cholesterol), ratio of the pre-ejection period to the left ventricular ejection time (PEP/LVET), serum high density lipoprotein (HDL) and creatine phosphokinase (CPK) in the descending order. Serum free T3 also correlated significantly all the these parameters. After treatment, changes in both serum free T4 and free T3 correlated well with those of the BMR and ATR in the patients with hyperthyroidism, and with those of the ATR, serum CPK and T-cholesterol in the patients with hypothyroidism. These results indicate that these peripheral parameters correlated well with serum free thyroid hormone concentrations in thyroid disease patients. Therefore, these parameters can be used to assess thyroid functions in patients with thyroid hormone resistance.
TAKEDA, A., MATSUNAGA, J., TOMITA, Y., TAGAMI, H. and SHIBAHARA, S. Molecular Analysis of the DNA Segments Cross-Hybridizable to the TyrosinaseGene in Patients Affected with Oculocutaneous Albinism. Tohoku J. Exp. Med., 1989, 159 (4), 333-340 - The human tyrosinase gene is greater than 35kb and is organized in four introns and five exons [Tomita et al. (1989) Biochem. Biophys. Res. Commun., 164, 990-996]. Using the full-length cDNA encoding human tyrosinase and its exon-specific fragments as hybridization probes, we show that overall structural organization of the tyrosinase gene is unchanged in three patients affected with tyrosinase-negative oculocutaneous albinism (OCA). Moreover, we are able to show the presence of additional DNA segments cross-hybridizable to exon 4 or exon 5 of the tyrosinase gene in the genome of three OCA patients and a healthy individual. Namely, the exon 4-specific probe detected two bands in the DNA digested with EcoRI or HindIII and the exon 5-specific probe detected two bands in the Bgl II-digested DNA, in spite of the facts that no recognition sites for these enzymes are present in each exon. We have then isolated two phage clones harboring the distinct DNA segments, hybridizable to the exon 5 probe. Southern blotting analysis of each cloned DNA digested with Bgl II showed two different hybridization patterns as those detected in genomic DNA digested with Bgl II, confirming that there are at least two DNA segments hybridizing to the exon 5 sequence in human genome.