Japanese conference on cyclosporine blood level monitoring.
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Biomedical subjects
Publications and source records attributed to K Uchida.
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A 44-year-old female was admitted because of gross hematuria six years after beginning hemodialysis for chronic renal failure. There was a past history of hematuria one year before admission. Retrograde pyelography, computed tomographic scan and angiography had been performed, and a probable malignant tumor of left renal pelvis had been found in 1986. We recommended surgery at the time, but she refused treatment and was followed up. On June 22, 1987, the patient had a second attack of gross hematuria resulting in bladder tamponade, and hematuria from the left ureteric orifice was confirmed at cystoscopy. Ultrasound showed left obstructive nephropathy. Left nephrectomy was performed because of the presumptive diagnosis of malignant tumor. The histological diagnosis was transitional cell carcinoma of the left renal pelvis. Two courses of M-VAC (methotiexate, vinblastine, adriamycin and cisplatin) chemotherapy were accordingly given postoperatively. The incidence of renal pelvic tumor and its chemotherapy in patients with chronic renal failure are discussed.
Protein C is an anticoagulant protein that circulates in blood as a zymogen of a serine protease. Recently the exogenous Protein C activator has been obtained from the venom of Agkistrodon Contortrix Contortrix. We also reported a functional assay method which activates Protein C with PROTAC and then measure the amidolytic activity with chromogenic substrate (S-2366) by COBAS FARA. At present, S-2366 (Glu-Pro-Arg-pNA), CBS 65-25 (Lys-Pro-Arg-pNA) and SPECTROZYME PCa (Lys-Pro-Arg-pNA) are used as the chromogenic substrate for activated Protein C. We studied which substrate is more suitable for amidolytic activity assay for Protein C by PROTAC in 99 patients under long-term Warfarin therapy and 29 as normal subjects. Our results indicate that SPECTROZYME PCa and CBS 65-25 are unsuitable for Protein C activity assay with PROTAC. Because change of absorbance was detected in Protein C deficient plasma and we couldn't gain good correlation between amidolytic activity of activated Protein C measured with these chromogenic substrates and antigenicity of Protein C by EIA. On the other hand, S-2366 is the most specific for activated Protein C and we also obtained good correlation against Protein C antigen.
An outbreak of congenital hydranencephaly and cerebellar hypoplasia occurred between November 1985 and May 1986 in Miyazaki, South Kyushu, Japan. Seventy-three calves had nervous signs of varying severity such as inability to stand, locomotor difficulties, defective vision and difficulty in sucking. At necropsy, 62 calves had macroscopic lesions in the central nervous system: hydranencephaly accompanied by cerebellar hypoplasia in 47; hydranencephaly alone in eight; and dilatation of the lateral ventricle in seven; none had arthrogryposis. Microscopically, all 62 cases involved various degrees of hypoplasia of neural components, such as total or partial thinning of the cerebral or cerebellar laminae. Heterotopia, such as abnormal islands of granule cells or Purkinje cells was also observed. Fourteen of these animals had other lesions such as non-purulent encephalitis, focal gliosis, neuronal degeneration, calcification or pseudocalcification, and cholesterol deposits, activation of vascular endothelial cells and haemorrhage. From the findings, these cases were considered to represent mainly hypoplasia of nerve tissue due to infection with a virus different from Akabane virus.
Reported is a rare superficial papillary adenocarcinoma arising from a congenital cyst, along with a survey of pertinent cases in the world literature. A 43-year-old male with a huge hepatic cyst was admitted to hospital for further examination. Since the cystic drainage done for alcoholic fixation was seromucinous initially but later contaminated by bile, an operation was initiated for dome resection and the closure of the biliary communication. At the time of the deroofing, however, small granulomas were found. And those were confirmed as being malignant by frozen section. Thus, an extended right lobectomy was performed for therapy. From its history and nature, this was considered to be a cancer arising in a congenital hepatic cyst.
Clinical effect of LM-001, a prostaglandin synthetic inhibitor developed from a drug delivery system, was evaluated in 54 patients with pain from urinary tract stones (stone pain) and 32 with vesical urgency after an operation on bladder or prostate. LM-001, felbinac ethyl incorporated in lipid microsphere, wes intravenously administered at the onset of stone pain or vesical urgency. Of 54 with stones and 32 with urgency, 53 and 29 were eligible for response, respectively. The symptoms improved or disappeared in some cases just after the administration and in the majority of patients within 15 minutes, in 49 of 53 patients with stone pain. Further, the effectiveness lasted over 24 hours in 26 of the 49 responding to this agent. On one hand, improvement or disappearance of vesical urgency was recognized in 25 of 29 patients, and the effectiveness was observed shortly after injection in 16 and lasted over 24 hours in 13 cases. Toxicities of this drug were investigated in 54 patients with stone pain and 32 with urinary urgency. Side effects consisted of pain at the injection site in 4, a slight fall of blood pressure in 1, slight visual disturbance in 1, body heat sensation in 1, leukocytosis in 3 and elevation of alkaline phosphatase in 1. These symptoms were transient and disappeared without use of any agent. LM-001 is concluded to be a useful drug for controlling stone pain and vesical urgency since an immediate effect, long durability and high response rates were obtained without severe side
The complex bis(phenanthrenequinone diimine)(bipyridyl)rhodium(III), Rh(phi)2(bpy)3+, cleaves DNA efficiently in a sequence-neutral fashion upon photoactivation so as to provide a novel, high resolution, chemical photofootpring reagent. Photofootprinting of two crystallographically characterized DNA-binding agents, distamycin, a small natural product which binds to DNA in the minor groove, and the endonuclease EcoRI, which binds in the major groove, gave respectively a 5-7 base pair footprint for the drug at its A6 binding site and a 10-12 base pair footprint for the enzyme centered at its recognition site (5'-GAATTC-3'). Both footprints agree closely with the crystallographic results. The photocleavage reaction can be performed using either a high intensity lamp or, conveniently, a simple transilluminator box, and the photoreaction is not inhibited by moderate concentrations of reagents which are sometimes required for examining interactions of molecules with DNA. When compared with other popular footprinting agents, the rhodium complex shows a number of distinct advantages: sequence-neutrality, high resolution, ability to footprint major as well as minor groove-binding ligands, applicability in the presence of additives such as Mg2+ or glycerol, ease of handling, and a sharply footprinted pattern. Light activated footprinting reactions furthermore offer the possibility of examining DNA-binding interactions with time resolution and within the cell.
Atrial-natriuretic-peptide (ANP) receptor, previously identified as a 140 kDa protein with a disulphide-linked homodimeric structure, was purified from bovine lung by (NH4)2SO4 fractionation and affinity chromatography on ANP-Affi-Gel 10. The purified receptor had a binding capacity of 4.2 nmol of ANP/mg of protein and an affinity constant of 6.5 pM. The isoelectric point of the receptor was 5.8, consistent with the acidic nature of the protein (amino acid analysis revealed a predominance of glutamic acid and aspartic acid residues). Treatment with endoglycosidase H and glycopeptidase F revealed that the receptor has three complex types of oligosaccharide chains per 70 kDa subunit. Deglycosylation of the receptor did not affect its binding activity. Reduction with dithiothreitol and reoxidation by dialysis revealed a strong tendency of the receptor subunits to dimerize via disulphide cross-linking; however, carboxymethylation of the reduced receptor indicated that the intersubunit disulphide bond is not necessary for the ligand-binding activity.
We previously have shown that urine components capable of stimulating ornithine decarboxylase activity of urothelium can enhance rat urinary bladder carcinogenesis, and that alpha-difluoromethylornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase, suppresses carcinogen-initiated rat urinary bladder carcinogenesis. The present investigation was conducted to determine whether DFMO's suppressive effect is stage specific during carcinogenesis and whether the suppressive effect lasts with its continued use. Following initiation with 0.05% N-butyl-N-(4-hydroxybutyl)-nitrosamine in drinking water for 6 wk, male Fischer 344 rats initially weighing 125 to 150 g were randomly divided into two groups, the first receiving 0.2% DFMO in drinking water ad libitum and the second receiving tap water only. Groups of animals were killed at regular intervals until the completion of the experiment at 75 wk. The effect of DFMO was evaluated by monitoring the incidence of tumors, the mean number of tumors per rat, the mean volume of individual tumors, and the mean total tumor volume per rat. The results showed that continuous treatment with DFMO significantly reduced tumor formation until 60 wk (P less than 0.017). The effect was only of borderline significance (0.017 less than P less than 0.035) at 75 wk. Discontinuation of DFMO treatment at 40 wk resulted in the loss of protective effect in all comparisons except for the borderline effect on the tumor number and total tumor volume per rat. DFMO had no significant effect on the incidence or development of preneoplastic early lesions. Mucosal polyamine (spermidine and spermine) levels were reduced and correlated well with the reduction in tumor growth, suggesting that the reduction in tumor growth rate by DFMO may be due to its ability to reduce polyamine levels in urothelium. There were no side effects attributable to DFMO treatment. DFMO may be a useful chemopreventive agent to retard the recurrence of human superficial bladder cancer.
In the presence of molecular oxygen and a catalytic amount of copper(II) ion, ascorbate almost completely degraded histamine (approx. 72%). The reaction was shown to occur at the imidazole group but not at the primary amino group in histamine. 4-[2-(Acetylamino)ethyl]-2,3-dihydroimidazol-2-one, a monooxygenated form of N-acetylhistamine, was first isolated as the primary product.
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A woman was operated on for a nonepithelial malignant tumor of the left leg and subsequently, for an epithelial carcinoma of the right breast and a borderline malignant tumor of the right ovary. She also developed a giant cavernous hemangioma that caused disseminated intravascular coagulation syndrome, which necessitated a left trisegmentectomy of the liver. Her family history suggested a hereditary predisposition to diverse malignant neoplasms, and also to giant cavernous hemangioma of the liver. Immunological evaluation disclosed selective inhibition of natural killer cell activity. Hormonal and hereditary factors are discussed in relation to the development of multiple primary tumors and giant cavernous hemangioma of the liver.
We have previously shown the existence of the major substrate protein of Mr 100,000 (substrate 100 K protein) for Ca2+/calmodulin (CaM)-dependent protein kinase in rat adrenal glomerulosa cells. In the present study, the identity of the substrate 100 K protein to elongation factor 2 (EF-2) was investigated. In a 105,000 g-supernatant fraction (cytosol), the protein of Mr 100,000 with the pI (isoelectric point) value of 6.7 was phosphorylated in the presence of calcium and CaM. The optical densities of this phosphorylated band were greatly enhanced in the presence of the EF-2 purified from pig liver (1 microgram) [20-23-fold, n = 5] when compared with those in the absence of the component. In the presence of the purified EF-2, the phosphorylation of Mr 100,000 was detected only in the presence of calcium alone or calcium plus CaM. This phosphorylation in the presence of calcium alone was completely inhibited in the presence of the CaM antagonist pimozide (500 microM), showing the existence of endogenous CaM in the cytosol. In the same fraction, the ADP-ribosylated protein of Mr 100,000 was detected in the presence of diphtheria toxin (fragment A) and (adenylate-32P) NAD, indicating the presence of EF-2 in the cytosol from rat adrenal glomerulosa cells. These results suggest that the substrate 100 K protein may be identical to EF-2 in rat adrenal glomerulosa cells.
Successful surgical treatment of a case of aneurysm of the vein of Galen is presented. An 11-month-old boy was admitted with episodes of convulsive attacks. Neuroradiological examination revealed an aneurysm of the vein of Galen filled by branches of posterior cerebral arteries and left thalamoperforating artery. Regional cerebral blood flow, which was measured by a method of xenon-enhanced computed tomography, showed a low flow area in the frontal and occipital regions. The aneurysm was approached through the bilateral parietooccipital interhemisphere, freed from the feeders, and ligated. During the operation, cortical blood flow was monitored for prevention of normal perfusion pressure breakthrough. After the feeders were successfully clipped, the blood flow increased and became normalized in the related cortex. There was no postoperative neurological deterioration.
Using heterotopically transplanted rat urinary bladder, experiments were conducted to develop a reproducible animal model of bacterial cystitis-associated urothelial hyperplasia without calculus formation, and to elucidate which bacterial component(s) might induce urothelial hyperplasia. Bladder instillation of live Escherichia coli (E. coli) resulted in persistent infection and inflammation and also diffuse urothelial hyperplasia. Instillation of killed E. coli also induced diffuse hyperplasia. Hyperplastic changes regressed following withdrawal of the killed E. coli treatment. Urothelial hyperplasia was also induced by repeated instillation of protein-rich lipopolysaccharide (LPS), the endotoxin derived from gram-negative bacterial wall component, but not by protein-free LPS. A finding common to bladders showing hyperplasia was the infiltration of neutrophils into intercellular spaces of the urothelium. We conclude that urothelial hyperplasia is induced by E. coli cystitis, that LPS plays a significant role in the hyperplastic response, and that neutrophils may mediate the response.
Using a monoclonal antibody to bromodeoxyuridine and immunohistochemistry, we measured the incorporation of this thymidine analogue into the deoxyribonucleic acid of human prostate adenocarcinoma cells exposed in situ. Fifteen patients with prostate cancer were given an intravenous infusion of 500 mg. bromodeoxyuridine at needle biopsy to label tumor cells in the deoxyribonucleic acid synthesis phase (S phase). The tumor specimens were fixed with 70 per cent ethanol, embedded in paraffin, sectioned and stained by an indirect immunoperoxidase method using anti-bromodeoxyuridine monoclonal antibody as the first antibody. The results showed that this method demonstrated bromodeoxyuridine-labeled nuclei satisfactorily in tissue section. The bromodeoxyuridine labeling index, S phase fraction, was determined by counting the number of bromodeoxyuridine-labeled cells in the tissue sections. Grade 3 tumors averaged 4.37 +/- 0.48 per cent labeling versus 2.41 +/- 0.49 per cent in grade 2 tumors, and grade 1 tumor in the series had an S phase fraction of 1.36 +/- 0.39 per cent. The average S phase fractions for single gland, cribriform, fused and medullary were 1.16, 2.30, 3.74 and 4.95 per cent, respectively. The results obtained with S phase fraction measured with bromodeoxyuridine labeling proved to be comparable to the results of histological grade and growth pattern. Thus, the higher S phase fraction may indicate biological malignancy. Moreover, the degree of heterogeneity concerning S phase fraction distribution within prostate cancer tissue could be compared to the morphological appearance. Our preliminary results suggest that the measurement of bromodeoxyuridine labeling index in prostate cancer may prove to be a new objective and quantitative assay of biological potential of individual tumor.