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The stilbene neutron detector which has been used for neutron emission profile monitoring in JT-60U has been improved, to respond to the requirement to observe the high-frequency phenomena in megahertz region such as toroidicity-induced Alfvén Eigen mode in burning plasma as well as the spatial profile and the energy spectrum. This high-frequency phenomenon is of great interest and one of the key issues in plasma physics in recent years. To achieve a fast response in the stilbene detector, a Flash-ADC is applied and the wave form of the anode signal stored directly, and neutron/gamma discrimination was carried out via software with a new scheme for data acquisition mode to extend the count rate limit to MHz region from 1.3 x 10(5) neutron/s in the past, and confirmed the adequacy of the method.

Background: Patients on chronic hemodialysis are likely to develop secondary hyperoxalemia. It is, however, difficult to measure plasma oxalate levels. To measure plasma oxalate levels, rapid plasma separation, deproteinization, and acidification are essential in preventing the formation of oxalate and the deposition of calcium oxalate within the test tube. The present study was undertaken to examine whether the oxalate level in dialyzer ultrafiltrate is potentially useful for estimating plasma oxalate levels., Methods: In nine patients on chronic hemodialysis, the plasma, after deproteinization with a filter, and the ultrafiltrate from the dialyzer before hemodialysis were acidified to a pH level of less than 3, followed by the measurement of oxalate levels by ion chromatography. Also, oxalate levels were compared between acidified and non-acidified ultrafiltrates from the dialyzer. In the second part of the study, seven patients on chronic hemodialysis receiving erythropoietin therapy, in whom the ferritin level was more than 300 ng/ml and transferrin saturation was less than 25%, were intravenously administered ascorbic acid, 100 mg, three times a week, after each dialysis session to facilitate the utilization of stored iron. This treatment was continued until the serum ferritin level decreased to a level below 300 ng/ml (for 3 months, at a maximum). The oxalate level in the dialyzer ultrafiltrate after this treatment was compared with that before treatment., Results: The mean +/- SE oxalate level in the dialyzer ultrafiltrate was 45 +/- 6 micromol/l, essentially equal to the plasma oxalate level (46 +/- 7 micromol/l). The plasma oxalate level had a significant positive correlation with the dialyzer ultrafiltrate oxalate level (plasma oxalate level = 0.99 x dialyzer ultrafiltrate oxalate level + 1.5; r = 0.95; P < 0.0001). The oxalate level in the acidified ultrafiltrate (45 +/- 6 micromol/l) did not differ significantly from that in the non-acidified ultrafiltrate (45 +/- 6 micromol/l). The mean +/- SE duration of ascorbic acid administration was 64 +/- 13 days. The hemoglobin level remained unchanged at 9.6 +/- 0.4 g/dl, whereas the serum iron level increased significantly, from 34 +/- 2 microg/dl to 43 +/- 4 microg/dl (P < 0.05), and serum ferritin levels decreased significantly, from 645 +/- 219 ng/ml to 231 +/- 30 ng/ml after the treatment (P < 0.05). The oxalate level in the acidified ultrafiltrate showed no significant change after ascorbic acid administration (31 +/- 8 micromol/l vs 47 +/- 7 micromol/l)., Conclusions: In patients on chronic hemodialysis, the oxalate level in acidified ultrafiltrate from the dialyzer was found to be useful for estimating the plasma level of non-protein-bound oxalate. When administering ascorbic acid to hemodialysis patients, the plasma oxalate level can be monitored using this method.

Background: Upregulation of local monocyte chemoattractant protein-1 (MCP-1) production is involved in glomerular damage through macrophage recruitment and activation in diabetic nephropathy. Treatment of db/db mice with soluble receptor for advanced glycation end-products (RAGE) prevented recruitment of macrophages to the glomeruli and reduced albuminuria, suggesting that binding of ligands and RAGE may be involved in MCP-1 expression. Therefore, we investigated the role of advanced glycation end-products (AGEs) in MCP-1 production by podocytes and signalling events after RAGE activation., Methods: MCP-1 gene and protein expression were examined by using reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay in differentiated mouse podocytes. Dichlorofluorescein-sensitive intracellular reactive oxygen species (ROS) generation was measured by confocal microscopy. RAGE, phosphorylation of mitogen-activated protein kinases, nuclear factor (NF)-kappaB, c-Jun and Sp1 were studied using western blotting and immunocytochemistry., Results: Both differentiated and undifferentiated podocytes expressed RAGE. MCP-1 was induced by AGEs and carboxymethyllysine (CML) in a time-dependent and dose-dependent manner in differentiated podocytes. Neutralizing antibody for RAGE suppressed AGE- and CML-induced MCP-1 production. AGEs and CML rapidly generated intracellular ROS in podocytes. Blocking of ROS by using N-acetyl-l-cysteine abolished CML and H(2)O(2)-induced MCP-1 expression. Phosphorylated extracellular signal-regulated kinase (ERK) was found in podocytes incubated with CML and was prevented by N-acetyl-l-cysteine or 7'-amino 4 [trifluoromethyl]. PD98059, an inhibitor of ERK, partially prevented CML-induced MCP-1 gene expression. NF-kappaB and Sp1 were translocated into the nucleus after podocytes were incubated with CML for 60 min. Parthenolide and mithramycin A, inhibitors of NF-kappaB and Sp1, respectively, abolished CML-induced MCP-1 gene expression in a dose-dependent manner., Conclusions: These results suggest that AGEs and CML induce MCP-1 expression in podocytes through activation of RAGE and generation of intracellular ROS. NF-kappaB and Sp1 regulate MCP-1 gene transcription.

We propose and demonstrate a novel all-optical digital-to-analog (D/A) conversion using pulse pattern recognition based on optical correlation processing. It is composed of pulse pattern recognition based on correlation processing and intensity adjustment using an optical attenuator. We obtain a single pulse as a result of pulse pattern recognition by using correlation processing between a target digital signal and a prepared correlation filter function. The obtained single pulse can be promptly fed to an output port as a corresponding analog signal through adequate intensity adjustment. Experimental results show that four-bit digital signals with 1.65ps interval can be successfully converted to analog signals corresponding to input digital signals.

A 45-year-old woman was admitted to our hospital because of high fever and malaise. She also presented abnormal urine findings including macrohematuria and proteinuria with transient renal insufficiency. Salmonella Typhi was isolated from her blood and stool culture, and then she was diagnosed as having typhoid fever. Salmonella enterica serotype typhi infection can be complicated by renal involvement, although rare. In Japan, few cases have been reported about acute nephritic syndrome in typhoid fever. Here, we report a case of endemic acquired typhoid fever associated with acute renal failure probably due to acute nephritic syndrome successfully treated with levofloxacin.

Liddle's syndrome is an autosomal dominant form of salt-sensitive hypertension and has been shown to be caused by missense or frameshift mutations in the amiloride-sensitive epithelial sodium channel (ENaC), which is composed of three subunits: alpha, beta, and gamma. All disease mutations either remove or alter amino acids of the target proline-rich PPPxY sequence (PY motif) of beta- or gamma-ENaC and result in increased channel activity. In this report, we present a family with Liddle's syndrome whose abnormality is caused by a novel missense mutation, P616R, in the PY motif of the betaENaC. Functional studies using the P616R mutant expressed in Xenopus oocytes showed an approximately 6-fold increase in the amiloride-sensitive sodium channel activity compared with that of the wild type. These findings provide additional clinical evidence that a conserved PY motif is critically important for the regulation of ENaC activity.

There are several vacant channels for diagnosis, RF heating and so on through the shielding structure in fusion reactors. Some of them consist of dogleg ducts, through which neutrons stream in a complex manner. An experiment was conducted with the Fusion Neutronics Source (FNS) facility at JAERI to study the behaviour of neutrons in the duct and assess the reliability of calculation methods for the design of fusion reactors such as ITER. The assembly was an iron slab 180 cm in thickness with a doubly bent duct 30 x 30 cm2 in cross section. The experiment was analysed using a simple design code for radiation streaming, DUCT-III, and the Monte Carlo code MCNP. The results indicate that the simple design code is reliable enough to be used for shielding design analyses as well as the Monte Carlo method, which showed excellent agreement between calculated and measured values.

Three-dimensional Monte Carlo shielding analyses are conducted on the ITER Neutral Beam Injection (NBI) duct for the nuclear and bremsstrahlung radiation. The detailed distribution is evaluated about the nuclear heating rate and surface heat load of the NBI duct wall by the neutron and photon transport calculation. The analytical representations of these nuclear responses are established as a function of the distance from the blanket surface. It is clarified that these representations are different between the duct wall facing the plasma and that hidden from the plasma, and also between the duct wall in the blanket region and that in the vacuum vessel region. These results are very useful for the shielding design of the NBI duct wall in the nuclear fusion reactor.

Objective: Adiponectin, an adipocyte-derived protein, has been suggested to enhance insulin sensitivity and prevent atherosclerosis. Circulating adiponecin levels are reduced in states of insulin resistance such as type 2 diabetes. We examined transcardiac utilization of adiponectin in patients with and without type 2 diabetes., Research Design and Methods: A total of 17 male type 2 diabetic patients and 17 male nondiabetic patients were investigated. Venous blood samples were taken to measure glucose and lipid variables. Blood samples for the measurement of adiponectin were collected simultaneously from the aortic root and coronary sinus. Angiographic semiquantitative stenosis score of coronary artery was also evaluated., Results: The adiponectin levels in both the aortic root and coronary sinus in the diabetic patients were significantly lower than those in the nondiabetic patients. The adiponectin level was significantly lower in the coronary sinus than in the aortic root in the nondiabetic patients, but there was no significant difference between adiponectin levels in the aortic root and coronary sinus in the diabetic patients. The total stenosis score, as an index of severity of coronary artery stenosis, was significantly higher in the diabetic patients than in the nondiabetic patients. The stenosis score was correlated with the degree of transcardiac utilization of adiponectin from the aortic root to coronary sinus in the nondiabetic patients but not in the diabetic patients., Conclusions: Diabetic patients not only have a decreased adiponectin level in the basal state compared with nondiabetic patients but also have impaired utilization of adiponectin in the coronary artery and/or the heart, which may promote the development of atherosclerosis.

In this study spatial and time distribution of neutrons leaking from Hokkaido University 45 MeV Electron Linac facility have measured and compared with the Monte Carlo simulations. The neutron transport processes inside and outside the facility building has been simulated using MCNP. The neutrons have measured by BF3 counters and 3He counters with polyethylene moderators up to the distance of 330 m from the facility. The spatial distribution of ambient dose equivalent converted from the counts has been compared with the simulation. The distribution estimated from the counts by the BF3 counter has been shown fairly good agreement with the calculation. The spatial distribution of counts obtained at the 45 MeV Electron Linac facility has been compared with that obtained at the Fusion Neutronics Source (FNS) facility of JAERI. The difference between the propagation characteristics of neutrons leaking from those facilities has been discussed.

Superheated emulsions being inexpensive, easy to fabricate, and having tissue equivalent composition make them as one of the popular neutron dosemeters. One more advantage is that they can be made insensitive to gamma rays by the choice of the sensitive liquid. It is observed that the response of commercially available bubble detector to neutron decreases above 20 MeV while its response is roughly flat in the 0.1-15 MeV region. This restricts its application as a dosemeter to high-energy neutrons. The response of bubble detector from Bubble Technology Industries, has been observed by using Pb-breeder for high-energy neutrons from different facilities in Japan. It is observed that 2-3 cm Pb-breeder is effective in increasing the response of the detector to the nominal value. Theoretical calculation using MCNPX code indicates an increase in neutrons in the energy range of 0.1-10 MeV with Pb-breeder. The present work indicates the possibility of using the bubble detector as a dosemeter to high-energy neutron using a Pb-breeder of proper thickness.

Microtubule-associated protein 1 light chain 3 (LC3) is a unique modifier protein. LC3-I, the cytosolic form, is modified to LC3-II, the membrane-bound form, by a mechanism similar to ubiquitylation by E1- and E2-like enzymes, Apg7p and Apg3p, respectively. In the present study, we found that LC3-I is processed to LC3-II during the differentiation and recovery from puromycin aminonucleoside-induced nephrosis of podocytes. LC3 is especially expressed in the podocytes of rat kidney as the membrane-bound form LC3-II. Biochemical analysis using a conditionally immortalized mouse podocyte clone (MPC) revealed that LC3-I is processed to LC3-II during the differentiation of cells into mature podocytes and accumulates in the membrane-rich fraction of the cell lysate. LC3-II-localized vesicles, which differ from lysosomes and endosomes, in differentiated MPC cells are morphologically similar to autophagic vacuoles during starvation-induced autophagy. During starvation-induced autophagy, autophagosomes fuses with lysosome and LC3-II on autophagosomes is finally degraded by lysosomal proteases. However, in differentiated MPC cells, little LC3-II on the vesicles is degraded by lysosomal proteases, suggesting that little LC3-II-localized vesicles in differentiated MPC cells fuse with lysosome. Furthermore, the LC3-II level in differentiated MPC cells increases with recovery from damage caused by experimental puromycin aminonucleoside-induced nephrosis. These results suggest that LC3-II-localized vesicles play an important role in the physiological function of podocytes.

A dynamic membrane rearrangement occurs in cells during autophagy to form autophagosomes. In this dynamic process, two ubiquitin-like modifications, Apg12p-conjugation and LC3-modification, are essential for the formation of autophagosomes. Apg7p and Apg10p catalyze the conjugation of Apg12p to Apg5p. The same Apg7p and Apg3p catalyze the processing of LC3 to a membrane-bound form, LC3-II. In this paper, we investigated whether Apg12p has an influence on the second LC3-modification system. A cross-linking experiment revealed that Apg3p interacts with the endogenous Apg12p.Apg5p conjugate. However, Apg3p itself interacts with free Apg12p more preferentially than the Apg12p.Apg5p conjugate, when free Apg12p exists. When Apg12p was overexpressed, LC3 processing was significantly enhanced in the presence of Apg7p. In contrast, when the Apg12p.Apg5p conjugate itself was accumulated by the overexpression of Apg12p and Apg5p, LC3 processing was dominantly inhibited, even in the presence of Apg7p. These results indicate that both Apg12p and the Apg12p.Apg5p conjugate are regulatory factors for LC3 processing.

Apg7p is a unique E1 enzyme which is essential for both the Apg12p- and Apg8p-modification systems, and plays indispensable roles in yeast autophagy. A cDNA encoding murine Apg7p homolog (mApg7p) was isolated from a mouse brain cDNA library. The predicted amino acid sequence of the clone shows a significant homology to human Apg7p and yeast Apg7p. Murine Apg12p as well as the three mammalian Apg8p homologs co-immunoprecipitate with mApg7p. Site-directed mutagenesis revealed that an active-site cysteine within mApg7p is Cys(567), indicating that mApg7p is an authentic E1 enzyme for murine Apg12p and mammalian Apg8p homologs. The mutagenesis study also revealed that Apg12p has a substrate preference for mApg7p over the three Apg8p homologs, suggesting that the Apg12p conjugation by Apg7p occurs preferentially in mammalian cells compared with the modification of the three Apg8p homologs. We also report here on the ubiquitous expression of human APG7 mRNA in human adult and fetal tissues and of rat Apg7p in adult tissues., ((C)2002 Elsevier Science (USA).)

Pyrroles were synthesized by three-component coupling reaction of aldehydes, amines, and nitroalkanes in the presence of a catalytic amount of a samarium species under mild conditions. The reaction is considered to involve the coupling of alpha,beta-unsaturated imines, which are provided by the samarium-catalyzed aldol-type condensation of imines generated from amines and aldehydes, with nitroalkanes. In the case of the three-component coupling of alpha,beta-unsaturated aldehydes (or ketones) with amines and nitroalkanes, alkylpyrroles were obtained by only heating in the absence of any catalyst. For instance, a mixture of butylamine, 2-butylidenecyclohexanone, and nitroethane, allowed to react at 60 degrees C for 15 h, produced isoindole, 4r, which is difficult to prepare by conventional methods, in 39% yield.

The sea urchin (Hemicentrotus pulcherrimus) arylsulfatase (HpArs) gene, expressed specifically in aboral ectoderm, contains a 229-bp enhancer in its first intron that is required for the activation of HpArs gene expression. Deletion analysis shows that a tandem repeat of orthodenticle-related protein (Otx) binding sites are responsible for the activity of this enhancer. Gel mobility shift analysis reveals that three types of Otx-proteins, which show different mobilities in gel shift assays, form complexes with the enhancer. Band I appears before hatching and gradually decreases by the gastrula stage. Band III appears at the blastula stage and Band II appears at the mesenchyme blastula stage; the levels of Band II and III remain constant until the gastrula stage. Two distinct types of HpOtx cDNA clones have been isolated from cDNA libraries of unfertilized eggs and gastrulae. Nucleotide sequences of the homeobox and downstream regions are well conserved in the two types of HpOtx cDNAs, while the region upstream from the homeobox has different nucleotide sequences. By genomic Southern blot analysis, only a single copy of HpOtx gene is detectable in the Hp genome, making it likely that two HpOtx isoforms are generated from the same gene. Results from Northern blot analysis confirm the presence of two types of HpOtx transcripts. Transcriptional regulation of the HpArs gene may, in part, be carried out through switching of Otx isoforms.

A series of arylnaphthalene lignans were synthesized and tested for hypolipidemic activity. The most potent compound (4b) (TA-7552) not only reduced serum cholesterol, but also increased high-density lipoproteins cholesterol in rats. The effective dose of 4b is 100 times less than that of cholestyramine. Structure-activity relationships are discussed.

2,2,-Disubstituted norbornenes (1, 2), 2,2-Disubstituted norbornane (3), 2,2,3-trisubstituted norbornenes (4, 5), oxanorbornenes (6) and azanorbornenes (7) were synthesized by the Diels-Alder reaction using alpha, beta-dehydroamino acids as a key step, and their antiulcer activities were examined. The oxazolidine derivative (1h) exhibited the most potent activities against several ulcer-models in rat.

A series of 2-aminoethanol derivatives was synthesized and their inhibitory activities against protein kinase C were investigated. Among these compounds, 2-endo-hexadecylamino-5-norbornene-2- exo-methanol (4h) and 2-endo-hexadecylamino-5-norbornene-2,3-exo-dimethanol (4i) inhibited protein kinase C at the IC50 values of 2 x 10(-5) and 1 x 10(-5) M, respectively, but not protein kinase A at a concentration of 1 x 10(-3) M. The structure-activity relationships are discussed.

2-endo-Hexadecylamino-2-exo-hydroxymethylnorbornene (1a) was synthesized from 2-acetamidonorbornene-2-carboxylic acid methyl ester (2) in a good overall yield. 2-endo-Hexadecylamino-2,3-exo-bis(hydroxymethyl)norbornene (1b) was synthesized starting from dimethyl acetamidofumarate based on Diels-Alder strategy. 1a and 1b inhibited protein kinase C at the IC51 values of 2 x 10(-5) and 1 x 10(-5) M, respectively, but not protein kinase A at a concentration of 1 x 10(-3) M. The structure-activity relationships are discussed.

In order to investigate the antidiuretic hormone (ADH) in essential hypertension and secondary hypertension, plasma ADH levels were measured in normal subjects, in patients with normal and low essential hypertension, and in other patients with various forms of secondary hypertension. Plasma ADH levels were significantly lower in low renin essential hypertension and higher in malignant hypertension than in normal subjects. The plasma ADH levels tended to be lower in renal hypertension and primary aldosteronism, and higher in renovascular hypertension, but these differences were not statistically significant. From these results, it appeared that ADH might play a role in malignant hypertension, but not in the other hypertensive diseases.

In order to investigate the relationship between the kallikrein-kinin (K-K) system and renin-angiotensin (R-A) system, plasma kinin (pKIN), plasma angiotensin II (pAII), plasma renin activity (PRA) and angiotensin converting enzyme activity (ACEA) were determined in 19 essential hypertensives (EHT). pKIN and pATII measurements were performed by highly sensitive radioimmunoassay (RIA), both of which were established in our laboratory. The assay sensitivity of pKIN and pAII were 0.5 pg/tube and 0.1 pg/tube, respectively. In pKIN RIA, pKIN was extracted by ethanol from 0.8 ml of plasma obtained with a syringe containing kinin generating and destroying enzyme inhibitors. In pAII RIA, pAII was measured directly in small amounts of 50-100 microliter, unextracted plasma samples. The level in pAII was significantly higher (p less than 0.05) in the normal renin group (NRH: n = 13) as compared with low renin group of EHT (LRH: n = 6). However, no significant difference was found in pKIN and ACEA between these two groups. Although a significantly positive correlation was observed between pAII and PRA (p less than 0.001) in EHT, the ratio of pAII/PRA tended to be higher in LHR than in NRH. ACEA correlated positively with pAII (p less than 0.01) or PRA (p less than 0.02), respectively. On the other hand, a significant negative correlation was also found between pKIN and pAII (p less than 0.05). From these findings, it was assumed that there was a closed relationship between R-A and K-K systems, and that angiotensin converting enzyme (kininase II) might play some role in the interrelation between both systems.

A highly sensitive and specific radioimmunoassay for kinin (minimal detectable amount, 0.5 pg/tube) was applied to measure the blood kinin level. A five ml blood sample was collected with a siliconized needle and plastic syringe which contained 2.5 ml of 0.8 N-HCl. The blood kinin was extracted with butanol, following reextraction with water. According to this procedure, the mean recovery (mean +/- SE) calculated from added 125I-bradykinin (500 CPM) and the known amounts of cold bradykinin were 50.4 +/- 0.8% and 51.1 +/- 2.2%, respectively. In comparison with other sampling methods in 6 normal subjects, the blood samples taken without HCl in syringes showed a higher level (24.4 +/- 10.1 pg/ml) than the samples with HCl (5.3 +/- 1.3 pg/ml). And very high levels were obtained in the plasma samples collected by the method of Talamo or Vinci (0.53 +/- 0.24 ng/ml and 3.5 +/- 1.3 ng/ml, respectively). The kinin content in blood samples taken with HCl was stable at -20 degrees C for at least one month, but increased significantly at room temperature or 4 degrees C for 48 hours. Blood samples were obtained from 17 normal subjects, and 3 patients with acute myocardial infarction. Blood kinin levels in the patient with acute myocardial infarction, 121 +/- 20.9 pg/ml, were significantly higher than those in normal subjects (3.8 +/- 0.5 pg/ml). From these results, it was concluded that high levels of blood kinin reported previously may have resulted from inadequate sampling procedures. Thus, in order to measure blood kinin accurately, inactivation of the kinin generating and destroying enzymes must be done immediately after the sampling. In addition, this radioimmunoassay method should be very useful in investigating the pathophysiological role of blood kinin in various diseases.

A very sensitive and simplified direct radioimmunoassay system for plasma angiotensin II was developed using the antiserum against synthetic angiotensin II (final dilution = 1: 1,500,000) in combination with 125I-labeled angiotensin II (specific activity = 1,600 microCi/micrograms). In this assay system, it was possible to carry out a direct assay using 100 microliter of plasma without any extraction procedure. This conclusion was supported by 100% recovery, parallelism of plasma samples against the standard curve, and no difference in hormone levels, there was also a high positive correlation between the plasma angiotensin II levels measured by this direct assay and the dowex column extraction method. The sensitivity of this assay system was 0.1 pg/tube, which is the highest sensitivity in studies reported to date. The cross-reactivities of angiotensin III and I against this antiserum were 100% and less than 0.1%, respectively, suggesting that the antiserum was very specific for the C-terminal of angiotensin II. Plasma angiotensin II levels in normal subjects after overnight fasting ranged from 3.0 to 21.3 pg/ml (12.0 +/- 2.1 pg/ml, mean +/- SE). By comparison, plasma angiotensin II levels of patients with essential hypertension were similar to those in the normal renin group, lower than those in low renin group and higher than those in high renin group. In patients with secondary hypertension, levels were lower in those with primary aldosteronism and higher in those with renovascular hypertension when compared to normal subjects.

In order to investigate the role of plasma angiotensin I (pAI) converting enzyme on the activity of the renin-angiotensin system, we measured plasma renin activity (PRA), plasma angiotensin II (pAII), and plasma angiotensin I converting enzyme (pACE) activity in fourteen patients with essential hypertension before and after two hours of ambulation combined with intravenous furosemide administration. Significant increases were observed in the values of PRA (p less than 0.005), pAII (p less than 0.005) and ACE activity (p less than 0.05) after ambulation. The ratios of log pAII/log PRA were increased significantly after ambulation (p less than 0.005), and a significantly positive correlation was observed between log pAII/log PRA and ACE activity (p less than 0.01). These results suggested that not only PRA but also ACE activity contributed to the activity of renin-angiotensin system through the generation of AII.

Human plasma low molecular weight kininogen was purified with ammonium sulfate fractionation, DEAE-cellulose, CM-Sephadex C-50 and aprotinin-agarose affinity column chromatography, after which this was further purified with Sephadex G-150 and DEAE-Sephadex A-50 column chromatographies. Kallikrein activity was measured as the kininogenase activity reflecting the kinin-producing capacity from kininogen. The purification factor from crude plasma to purified substrate was 44-fold, and the recovery was 18%. The purified human substrate did not contain kinin-generating or destroying enzymes which would interfere with kininogenase activity, and showed a cross-reactivity of less than 0.1% against kinin antiserum. In the kininogenase assay, all kininogen was removed by adding ethanol to terminate the enzyme reaction. Because of the high sensitivity of kinin radioimmunoassay, the kinin levels in urine could be determined in very small amounts of samples (0.5 to 2.0 nl of the original urine). These findings indicated that kinin levels in incubation solution could be measured directly, and the control tubes are unnecessary in this assay procedure. In a comparison among human, dog and bovine low molecular weight kininogen as the substrate for human urinary kallikrein, the enzyme activity was 5 and 80 fold higher in the human low molecular weight kininogen, respectively, suggesting that a human substrate is the best for human enzymes. This simple, specific, sensitive and homologous kininogenase assay system seems to be very useful investigating the physiological or pathophysiological role of the renal kallikrein-kinin system in hypertensive and renal diseases.

In order to clarify the mechanism of the hypotensive action of captopril, the acute and chronic effects of this drug on the kallikrein-kinin and renin-angiotensin systems were investigated respectively in 14 and 19 patients with hypertension. To determine the acute effect, a dose of 50 mg of captopril was administered once orally. For the chronic effect, 75-300 mg of the drug was administered daily for 14 days. In observations of the acute effect, blood pressure decreased significantly at 30 min. and maximally at 60-180 min. after administration with no change in heart rate. Significant increases in blood kinin levels and plasma renin activity (PRA), and a decrease in plasma angiotensin II levels were also observed. A marked augmentation was also found in urinary kinin excretion, but not in urinary kallikrein excretion. Moreover, the changes in blood pressure significantly correlated negatively with basal PRA, basal plasma angiotensin II and the changes in blood kinin levels, and positively with the changes in plasma angiotensin II. In our study of the chronic effect of captopril, similar changes in blood kinin levels, PRA, plasma angiotensin II levels, blood pressure and heart rate to the acute effect study were observed. Significant correlations of the changes in blood pressure were found negatively with basal PRA, basal plasma angiotensin II levels and the changes in blood kinin levels and positively with the changes in plasma angiotensin II levels. In addition, significant increases in urine volume and urinary sodium excretion occurred following administration of captopril for 14 days, and both increases negatively correlated with the changes in blood pressure.(ABSTRACT TRUNCATED AT 250 WORDS)

The method of measurement for urinary total kallikrein (KAL) and preKAL in human was developed, and daily excretions of urinary total KAL, KAL and preKAL were investigated in patients with essential hypertension. Forty microliter of urine samples were incubated with or without 120 micrograms of chymotrypsin-free trypsin for total KAL or KAL, respectively. KAL was measured with direct radioimmunoassay and kininogenase assay. PreKAL was calculated by the subtraction of KAL from total KAL. The subjects of this study included 7 normotensives (NT) and 8 essential hypertensives (EHT). Daily excretions of total KAL, KAL and preKAL were significantly lower in EHT than those in NT. KAL/total KAL ratio, which reflects the conversion rate from preKAL to KAL in the kidney, was not significantly different between EHT and NT. From these results, it is suggested that decreased urinary KAL excretion in EHT is mainly caused by reduced preKAL production rather than the impaired conversion from preKAL to KAL in the kidney. It is emphasized that this method of measurement for urinary total KAL and preKAL may be a very useful tool for research of the renal kallikrein-kinin system.

Changes in plasma ADH levels were investigated in human male subjects whose blood volume was altered under isotonic circumstances. Blood volume was reduced by ambulation and increased by isotonic saline infusion in an overnight dehydrated state, and determinations were made on plasma ADH levels, plasma osmolality and hematocrit values. Plasma ADH levels were clearly affected by the small changes in blood volume, and significantly negative correlation was found between plasma ADH levels and the percent changes in blood volume under isotonic circumstances. From these findings, it was concluded that ADH release in human subjects is also controlled by the changes of the blood volume factor in addition to osmotic stimuli.

To investigate the relationship between plasma kinin and the sympathetic nervous system, we measured plasma kinin, norepinephrine, and angiotensin-I-converting enzyme (ACE:kininase II) activity in normal subjects. In six normal subjects, subpressor and pressor doses of norepinephrine (10 ng/kg/min and 50 ng/kg/min, respectively) were infused for 1 hour. In five normal subjects, 60-degree head-up tilting was performed for 20 minutes. After norepinephrine infusion, plasma kinin levels were decreased significantly (p less than 0.01) with the subpressor dose of norepinephrine and were further decreased (p less than 0.01) with the pressor dose. Plasma ACE activity, however, showed no significant changes following these infusions. During the tilting, plasma kinin levels decreased significantly (p less than 0.01), whereas plasma norepinephrine levels increased significantly (p less than 0.01). From these results, the assumption of elevated levels in plasma endogenous or exogenous norepinephrine leads to a decrease in plasma kinin levels; this suggests that these two systems may be closely interrelated.

Two autopsy cases of Cronkhite-Canada syndrome were reported. The caused of hypoproteinemia, electrolyte imbalance and ectodermal changes were discussed with reference to previously reported cases. The mechanism of protein loss was probably due to outflow into the intestinal lumen of the mucous substance in the cystically dilated glands, directly and/or indirectly followed by loss of mucosal surface. Electrolyte imbalance probably developed from gastrointestinal loss as well as poor substitution. The ectodermal changes were probably not a subsequent part of the emaciation or hypoproteinemis, but an inherent part of this disease. Therapy, whether substitution or surgical procedure, should be selected in order to control the general condition of the patient.

Twenty liver cancer patients, including 9 hepatocellular carcinoma and 11 with metastatic liver cancer, were treated by intra-arteral one-shot chemotherapy. Alterations in blood coagulation and fibrinolysis were observed serially after one-shot chemotherapy by testing the levels of PT, APTT, FDP, fibrinogen, AT III, alpha 2-macroglobulin, and plasminogen. APTT was prolonged, FDP increased, Fbg increased after a transicent decrease, and AT III, alpha 2-M, and plasminogen decreased. The peaks of these alterations occurred within 7 days after the one-shot treatment; recovery was observed after about weeks. The more advanced the cancer, the greater were the alterations.

In order to investigate the role of the renal kallikrein-kinin (K-K) system in normal (NRH) and low renin (LRH) subgroups of essential hypertension (EHT), daily urinary excretions of renal K-K system components including kallikrein (KAL), total KAL, pre-KAL, kinin (KIN) and kininase (total, I and II), were measured in 21 normotensives (NT) and 45 patients with EHT (NRH: 29, LRH: 16). Urinary KAL and KIN quantities, KAL activity, total and pre-KAL, and kininase (total, I and II) were measured by direct RIA, kininogenase assay, direct RIA of KAL after trypsin treatment, and KIN destroying capacity, respectively. The daily excretions of KAL quantity and activity, total and pre-KAL, and KIN were significantly lower in EHT than in NT. That of total kininase and kininase I were significantly higher in EHT than in NT while no significant difference was found in kininase I between EHT and NT. In comparing NRH and LRH, the urinary KAL activity and KIN were lower in LRH than in NRH, and kininase I was higher in LRH than in NRH. No significant difference, however, was found in total and pre-KAL, KAL quantity and kininase II between NRH and LRH. The ratio of KAL quantity/total KAL which reflects the conversion rate from pre-KAL in the kidney, did not show any significant difference among NT, NRH and LRH.(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma prolactin level and plasma renin activity were determined in normal subjects and patients with low and normal renin essential hypertension, renal hypertension, renovascular hypertension, primary aldosteronism, Cushing syndrome, pheochromocytoma and malignant hypertension. In both normal subjects and the normal renin essential hypertensives, plasma prolactin was significantly higher in females than in males. Plasma prolactin was also significantly higher in the normal renin essential hypertensives than in normal subjects of both sexes, while no significant difference was found between the low renin group and normal subjects of either sex. A significantly positive correlation was observed between plasma renin activity and the plasma prolactin level in male essential hypertensives, but not in females. Although no significant difference in plasma prolactin level could be detected between patients with secondary hypertension and normal subjects, this level was significantly higher in malignant hypertensives than in normotensives. From these results, it was shown that significant differences of plasma prolactin levels exist between normal renin essential hypertensives, and low renin essential hypertensives or normal subjects, and that these differences may partly depend on renin status which might be related to the central dopaminergic activity. In malignant hypertensives, the high level of plasma prolactin may be caused by diminished renal function, but the suppression of central dopaminergic activity cannot be excluded in the mechanism of plasma prolactin increment.