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Study Objectives: We sought to assess the incidence of transient U-wave inversion during vasospasm of the left anterior descending coronary artery (LAD) with ST-segment depression as opposed to that with ST-segment elevation., Design: Retrospective study., Setting: Cardiology division of acute-care hospitals., Patients: We studied 49 patients with vasospastic angina whose vasospasm was induced in the LAD, not in the left circumflex coronary artery, by intracoronary injection of acetylcholine., Measurements and Results: The ECG traces obtained during acetylcholine-induced vasospasm of the LAD were examined. Based on the direction of ST-segment shift, the patients were categorized into two groups: the ST-segment elevation group (n = 27) and the depression group (n = 22). There were no differences in age, gender, or cardiovascular risk factors between the two groups. The distribution of the spastic site in the LAD was also similar. A total reduction in luminal diameter during a provoked attack was more often observed in the ST-segment elevation group than in the ST-segment depression group (37% vs 9%, p = 0.02). Collateral circulation to the LAD was found in only one patient in each group. There were no differences between the two groups in heart rate, systolic BP, and double product of heart rate and systolic BP during the attack. The incidence of acetylcholine-induced anginal attack with U-wave inversion in the ST-segment depression group was nearly as high as that in the ST-segment elevation group (77% vs 78%, p > 0.99)., Conclusions: The development of transient U-wave inversion during vasospasm of the LAD induced by intracoronary injection of acetylcholine does not depend on the magnitude of myocardial ischemia as judged by the direction of ST-segment shift.

Oxidative stress plays a critical role in normal functioning of cardiac and vascular cells as well as in the pathogenesis of cardiovascular disease. Growth arrest and DNA damage-inducible gene 153 (GADD153), which is upregulated by oxidative stress, regulates the cell cycle and apoptosis. Previously an AP-1 was reported to contribute significantly to GADD153 gene transcriptional activation by oxidative stress. Recently, we have reported that GADD153 gene promoter activity is negatively regulated by nuclear factor 1 (NF1), in vascular smooth muscle cells (VSMCs). The aim of this study was to elucidate the roles of AP-1 and NF1 in GADD153 gene induction by oxidative stress in VSMCs. H(2)O(2) induced GADD153 mRNA and reduced NF1 mRNA expression. In the electromobility shift assay, H(2)O(2) induced AP-1-binding activity and reduced NF1-binding activity. Overexpression of NF1 significantly suppressed the induction of the GADD153 gene after treatment with H(2)O(2). These results revealed that induction of the GADD153 gene by oxidative stress is regulated mainly by two nuclear factors, NF1 and AP-1., (©2002 Elsevier Science (USA).)

Background: There has been no report of ECG changes during anginal attacks in patients with coexistent hypertrophic cardiomyopathy (HCM) and vasospastic angina., Study Objectives: To elucidate the change in ST-segment during anginal attacks in patients with coexistent HCM and vasospastic angina (the HCM group) in comparison with that of patients with vasospastic angina and no left ventricular hypertrophy (the non-HCM group)., Design: Retrospective study., Patients: Twelve patients in the HCM group, and 28 patients in the non-HCM group., Measurements: The direction of ST segment shift, either ST-segment elevation or depression, on the ECGs recorded during vasospastic anginal attacks with severe vasoconstriction in the epicardial coronary artery after intracoronary injection of acetylcholine., Results: Age, male gender, and distribution of coronary arteries in which the vasospasm occurred were similar between the two groups. Collateral circulation to the affected arteries was absent in all the study patients. The prevalence of anginal attacks associated with ST-segment elevation was 2.7 times higher in the non-HCM group than in the HCM group (51. 5% [17 of 33 attacks] vs 18.8% [3 of 16 attacks], respectively; p = 0.03)., Conclusions: In the HCM group, myocardial ischemia associated with a transmural injury pattern seen on the ECG, which is represented as ST-segment elevation, seldom develops during vasospastic anginal attacks because of marked left ventricular hypertrophy.

A 54-year-old woman had been treated for hypertrophic cardiomyopathy and paroxysmal atrial fibrillation since 1992. She was admitted with paroxysmal atrial fibrillation which was resolved by medical treatment. However, on the next day, left lateral chest pain appeared. Computed tomography disclosed a low density area in the spleen. She received anticoagulant therapy under a diagnosis of splenic infarction, and the pain disappeared. Echocardiography showed hypertrophic cardiomyopathy with mid-ventricular obstruction. She was treated with cibenzoline to prevent paroxysmal atrial fibrillation attack and attenuate the hemodynamic load. After treatment, the pressure gradient decreased from 41 to 7 mmHg. This patient with hypertrophic cardiomyopathy suffered a rare isolated splenic infarction associated with paroxysmal atrial fibrillation.

The aim of this study is to investigate whether the cardiomyocyte apoptosis is induced after experimental right-sided pressure overload and whether the expression of proapoptotic factor is altered or not. Ten-week-old male Sprague-Dawley rats were subjected to right ventricular overload by experimental coarctation of the main pulmonary artery. In pulmonary artery-banded rats, TUNEL method revealed that positive nuclei were observed in cardiomyocytes exclusively in the right ventricle, and Northern blot analysis showed that p53 mRNA level in the right ventricle was 6.2-fold higher at the day 1 than that in sham-operated rats and its level decreased gradually. Bax mRNA in the right ventricle was also increased 3.3-fold at the day 1 in pulmonary artery-banded rats and also gradually decreased. The immunohistochemical study revealed that the immunoreactivity of P53 and Bax was observed exclusively in the right ventricle of the pulmonary artery-banded group. These results demonstrated that the occurrence of TUNEL-positive cardiomyocytes in the acute pressure overload was accompanied by the enhanced expression of apoptosis inducers. It is suggested that acute pressure overload is a potent apoptotic stimulus for cardiomyocytes.

A 38-year-old man underwent pericardiectomy because of effusive-constrictive pericarditis. The gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA) enhanced magnetic resonance imaging showed pericardial thickening separate from the effusion, which could not be shown by unenhanced computed tomography. Gd-DTPA enhanced magnetic resonance imaging could be useful for the early detection of effusive-constrictive pericarditis.

A 69-year-old woman was admitted because of increase of chest pain and dyspnea. Systolic murmur of Levine III/VI was heard. Electrocardiography showed ST depression caused by ischemia. Echocardiography revealed severe mitral regurgitation (MR) and inferoposterior hypokinetic wall motion. Left ventriculography revealed the presence of MR (II/IV). Coronary angiography showed severe organic stenosis of the right coronary artery. Based on these findings, the diagnosis was severe papillary muscle dysfunction caused by unstable angina. The lesion of the right coronary artery was successfully stented with a Palmaz-Schatz stent. During balloon inflation, the v wave of the pulmonary capillary pressure curve was greatly elevated. After the stent implantation, ST depression was normalized and MR improved dramatically. Therefore, we suppose that acute MR was induced by temporary papillary muscle dysfunction, and could be relieved with coronary angioplasty.

KL-6, a MUC1 mucin preferentially expressed in regenerating type 2 pneumocytes, has been reported to be a sensitive serum marker for evaluating the disease activity of interstitial pneumonitis (IP). Type III procollagen N-terminal peptide (PIIIP) and type IV collagen 7S (7S collagen) have also been reported to be useful in the serological evaluation of the activity. Their levels were measured and their serodiagnostic values were compared simultaneously in patients with IP and alveolar pneumonia. The study population was 45 patients with IP and 12 patients with alveolar pneumonia. Serum KL-6 levels were measured by a specific enzyme immunoassay, and both serum PIIIP and 7S collagen concentrations by their correspondent radioimmunoassay kits. There were no significant difference of serum C-reactive protein level, which was evaluated as an indicator of inflammatory process, between IP and alveolar pneumonia patients. In IP, the abnormally elevated rate of KL-6 [80% (36/45)] was significantly higher than those of PIIIP [40% (18/45)] and 7S collagen [40% (18/45)]. In alveolar pneumonia, the rate of KL-6 [0% (0/12)] was significantly lower than those of PIIIP [33% (4/12)] and 7S collagen [25% (3/12)]. There were no significant correlations among serum levels of the markers. These observations indicate that the serodiagnostic value of KL-6 for IP is superior to that of PIIIP and 7S collagen, and that KL-6 has a characteristic to discriminate IP from alveolar pneumonia.

CCAAT-enhancer binding protein-delta (C/EBP delta) is a transcriptional nuclear factor, and belongs to basic region-leucine zipper class DNA binding proteins. One genomic clone containing a 12-kb sequence of the C/EBP delta gene was isolated from a rat genomic library, and a 2,056-bp fragment containing the 5'-flanking region was characterized. Sequence analysis of this fragment revealed that there were a TATA-like sequence (TAGAAAA) and many transcriptional regulatory elements. The transcription start site of the gene was determined by both primer extension analysis and riboprobe mapping. Both analyses indicated that the transcription start site was located at 31-bp downstream of the TATA-like sequence. Transient transfection experiments showed that the fragment cloned in this study was able to act as a functional promoter in rat vascular smooth muscle cells. The 5'-deletion analysis of this fragment revealed that the sequence spanning -235 through -82, which was designated as an upstream control element (UCE), remarkably increased a basal promoter activity of the C/EBP delta gene, and was also able to act as a promoter by itself. In addition, we also studied effects of the UCE on the heterologous gene promoter including rat alpha-actin gene promoter or SV40 virus promoter. Interestingly, the UCE specifically increased the promoter activity of the rat alpha-actin gene suggesting that the C/EBP delta gene may be positively controlled by the UCE via a cell-type or promoter-type specific manner.

Carcinoembryonic antigen (CEA) has been clinically used as a serum marker for lung cancer, especially for pulmonary adenocarcinoma. Since we had developed a tumor marker against pulmonary adenocarcinoma, CAM 123-6 belonging to KL-6 MUC 1 mucin, we compared its clinical value with that of CEA, in combination with squamous cell carcinoma-related antigen (SCC) and neuron-specific enolase (NSE). The subjects were 30 patients with adenocarcinoma, 23 with squamous cell carcinoma, 16 with small cell carcinoma, and 20 with benign lung disease. For discriminating lung cancer from benign lung disease, the combination of CAM 123-6, SCC, and NSE showed 64% diagnostic accuracy, and the combination of CEA, SCC, and NSE 65%. In differentiating pulmonary adenocarcinoma from other lung cancer, the diagnostic accuracy of CAM 123-6 was 81% and that of CEA was 74%, although there was no significant difference between them. These observations indicate that the diagnostic accuracy of CAM 123-6 is similar to that of CEA. Neither marker is sufficiently specific to differentiate pulmonary adenocarcinoma from other forms of lung cancer.

A 38-year-old man was admitted to our hospital for detailed examination of fever, cough and yellow sputum. At the age of 32, be had mitral prosthesis for the first time, because of mitral regurgitation due to mitral valve prolapse. Four years previously, he had again undergone mitral prosthetic valve replacement due to prosthetic valve endocarditis due to staphylococcus epidemidis. This occasion, staphylococcus aureus was isolated by arterial blood culture. Transesophageal echocardiography detected vegetation attached to the mitral prosthetic valve and paravalvular leakage. The diagnosis was prosthetic valve endocarditis. He underwent a third mitral prosthetic valve replacement. Detection of the source of infection was difficult only by transthoracic echocardiography, and immediate transesophageal echocardiography seemed mandatory to diagnose bacterial endocarditis.

Generation of soluble cytokine receptors is a general phenomenon, and the roles of several such receptors have been investigated in clinical settings. Unlike other soluble cytokine receptors, soluble interleukin-6 receptor (sIL-6R) can act as an agonist and thus is implicated as an important modulator in the acute-phase reaction of prolonged inflammation. The purpose of the present study was to determine the roles of pleural sIL-6R in both differential diagnosis of pleural diseases and in the induction of acute-phase protein. Specific sandwich enzyme-linked immunosorbent assays were used to determine sIL-6R and IL-6 in 19 tuberculous, 48 malignant and 10 transudative effusions. Although IL-6 levels in pleural effusions were strikingly different, no significant differences in pleural sIL-6R levels were found between the groups. Pleural levels of IL-6 were invariably much higher, whereas those of SIL-6R were invariably lower than serum levels. Furthermore, IL-6, but not sIL-6R, levels in effusions correlated significantly with serum C-reactive protein levels. These results suggest that: (1) pleural levels of sIL-6R are not increased even in strong inflammation such as tuberculous pleurisy, nor significantly different among pleural diseases; and (2) the local levels of sIL-6R are not as important as expected for the induction of acute-phase proteins in patients with pleural diseases.

The nucleotide and deduced amino acid sequences of human basic and acidic calponins were determined. The basic calponin cDNA from human aorta (1496 bp) contained a single open reading frame (ORF) which encodes 297 amino acids (33,169 Da). The acidic calponin cDNA from human kidney (1607 bp) contained a single ORF which encodes 329 amino acids (36,412 Da). Basic calponin mRNA was expressed in only smooth muscle tissues, but acidic calponin mRNA was expressed in non-smooth muscle tissues as well as smooth muscle tissues. Fluorescent in situ hybridization revealed that basic and acidic calponin genes localize in 19p13.1-13.2 and 1p21-22 of human chromosomes, respectively.

We present the first case (to our knowledge) of acetaminophen-induced eosinophilic pneumonia. Although the patient had taken an acetaminophen-containing combination drug, Kinuya-chinnetsu, for only three days, the abnormal shadow on his chest radiograph continued for about a month. Lymphocyte stimulation tests showed that both Kinuya-chinnetsu and acetaminophen induced the proliferation of peripheral blood lymphocytes.

A simplified and sensitive method for measuring expression levels of type-1 angiotensin II (AT1) receptor subtypes, AT1A and AT1B, was established. The two receptor cDNAs were co-amplified and measured by polymerase chain reaction using primers based on the corresponding receptor subtype genes. Both AT1A and AT1B mRNAs were widely expressed in the rat tissues including adrenal gland, kidney, heart, aorta, lung, liver, testis, pituitary gland, cerebrum and cerebellum. AT1A mRNA was predominantly expressed in the rat tissues examined except adrenal gland and pituitary gland where AT1B mRNA was predominantly expressed. Sodium depletion did not change mRNA levels of AT1A and AT1B in the all tissues. However, both AT1A and AT1B mRNA levels in the heart and aorta were down-regulated by treatment with AT1 specific antagonist, TCV 116. In contrast, AT1B mRNA in the adrenal gland was mainly reduced by the treatment. These results suggest that the expression level of AT1B mRNA in the adrenal gland depends on the activity of the renin-angiotensin-aldosterone system (RAAS) and both receptor subtypes mediate contraction and hypertrophy of the smooth and cardiac muscles via the RAAS.

We measured interleukin 6 (IL-6) concentrations in the pleural fluid of various patients to determine its role in pathophysiology and diagnosis by using specific functional bioassay. IL-6 levels were significantly higher in exudate than in transudate (79.3 +/- 176.2 U/ml [n = 55] vs 1.7 +/- 1.8 U/ml [n = 12]; p < 0.01). Tuberculous effusion contained a significantly higher amount of IL-6 than malignant effusion (181.3 +/- 176.2 U/ml [n = 13] vs 29.4 +/- 71.5 U/ml [n = 29]; p < 0.005). Pleural IL-6 levels were invariably higher than serum IL-6 levels, and both were significantly correlated (n = 21, r = 0.632; p < 0.02). Pleural IL-6 levels were significantly correlated with lactate dehydrogenase (LDH) in pleural fluid (r = 0.392; p < 0.01), ratio of pleural/serum LDH (r = 0.571; p < 0.01), pleural adenosine deaminase activity (r = 0.599; p < 0.01), and serum C-reactive protein (r = 0.494; p < 0.01). Furthermore, IL-6 levels were significantly correlated with peripheral blood platelet counts (r = 0.447; p < 0.001). These results suggest that (1) IL-6 is produced locally in pleural space, (2) pleural IL-6 level is helpful for differential diagnosis, and (3) locally produced IL-6 could leak to circulation and cause systemic effects such as the induction of C-reactive protein and thrombocytosis.

To determine the sensitivity of serum KL-6 and serum lactate dehydrogenase for detecting the contraction of radiation pneumonitis, 15 patients with lung cancer who had radiation therapy were monitored. Six of the patients contracted radiation pneumonitis (pneumonitis group) and the other patients did not (control group). Serum levels of KL-6 were significantly (p less than 0.05) elevated according to the complication of radiation pneumonitis in all patients of the pneumonitis group. In the control group, however, one-sided changes of KL-6 level were not observed. In the pneumonitis group, serum LDH levels were not significantly changed. However, there was a strong correlation between the altered levels of KL-6 and those of LDH (r = 0.992). These observations indicate that the same cytopathologic changes may cause the elevation of serum KL-6 level and the elevated activity of serum LDH in the patients with radiation pneumonitis, and that KL-6 is much more sensitive than LDH for detecting radiation pneumonitis.

The case of a 16-year-old female patient with the giant cell type of malignant fibrous histiocytoma (MFH) in the left posterior mediastinum is reported. This is a very unusual site for MFH. Immunotherapy induced a prolonged partial remission lasting 15 months with a marked reduction in tumor size. The usefulness of immunotherapy for MFH needs to be evaluated.

Serum marker KL-6 antigen has been reported to be a valuable indicator of the disease activity of interstitial pneumonia. It is not clear how sensitive the serum KL-6 antigen level is in reflecting histologic changes in lung tissues. We report here the results of serial measurements of serum KL-6 antigen in a 76-year-old male patient with radiation pneumonia. Serum KL-6 antigen levels were more sensitive than lactate dehydrogenase and procollagen type III N-terminal peptide. The level of serum KL-6 antigen appears to reflect the histologic changes of the lung more sensitively than does C-reactive protein.

We established a simplified method of the quantitative measurement of extra-renal renin messenger RNA using polymerase chain reaction system. We could detect the renin messenger RNA in the kidney, heart, aorta and adrenal gland from a single RNA sample obtained from a Wistar-Kyoto rat. In the kidney, heart, aorta and adrenal gland, the contents of renin messenger RNA were found to be 55.8 +/- 17.8, 0.15 +/- 0.05, 0.11 +/- 0.03 and 0.16 +/- 0.07 pg/micrograms of total RNA (n = 5, mean +/- s.d.), respectively. The present method is very useful to study the extra-renal renin-angiotensin system.

The activity of angiotensin-converting enzyme in human prostate was measured by spectrophotometric method using Hippuryl-L-histidyl-L-leucine (Hip-His-Leu) as substrate. Benign hypertrophic prostate had significantly higher concentration of angiotensin-converting enzyme (1.99 +/- 0.36 (S.E.M.) units/g of tissue, n = 13) than normal prostate (0.44 +/- 0.1 units/g of tissue, n = 5) (p less than 0.01). More than 80% of the enzyme activity occurred in soluble fraction of normal and hypertrophied prostates. Human prostatic angiotensin-converting enzyme, partially purified from benign hypertrophic prostates, had an apparent molecular weight of 290,000 and pI of 4.1. Optimum pH of the enzyme with KM value of 1.0 mmol/l was 7.8 in 0.1 mmol/l borate/sodium/carbonate buffer. The enzyme was competitively inhibited by captopril (Ki, 1.8 nmol/l).

Human membrane-bound neutral arylamidases were solubilized from small intestinal mucosa, lung, kidney, liver and placenta with trypsin. These five membrane-bound neutral arylamidases were identified by polyacrylamide gel-disc electrophoresis. The heat sensitivity of each enzyme was in the order, liver and placenta greater than kidney greater than lung greater than small intestine. This order correlates with that of electrophoretic mobility, except for the placental membrane-bound neutral arylamidase. Five membrane-bound neutral arylamidases have the same molecular weight, 240 000, as estimated by Sephadex G-200 gel filtration. The five membrane-bound neutral arylamidase have very similar KM values (8.7 x 10(-5) M towards L-alanyl-beta-naphthylamide), optimal pH values, hydrolysis ratios towards L-alanyl-beta-naphthylamide and L-leucyl-beta-naphthylamide, and sensitivities of inhibition by chelators or amino acids. These results suggest that the multiple forms of membrane-bound neutral arylamidase found in five different human organs are organ-specific isoenzymes.

The neutral arylamidase activity in urine of healthy and nephrotic children was determined using L-leucyl-beta-napthylamide as substrate. The neutral arylamidase activity in urine samples from 13 nephrotic children (22.4+/-3.0 (S.E.) mumol/min/g of creatinine) was significantly higher (p less than 0.001) than that from 27 normal children (8.5+/-0.4). Two forms of neutral arylamidase were demonstrated by polyacrylamide gel electrophoresis and Sephadex G-200 gel filtration in urine samples from normal children. One was a high molecular weight arylamidase (more than 1 000 000), the other had a molecular weitht of 240 000 as estimated by Sephadex G-200 gel filtration. Two additional electrophoretically distinct froms of neurtral arylamidase were demonstrated on polyacrylamide gel in urine samples from nephrotic children. Two neutral arylamidases from normal children, one additional arylamidase from normal children, one additional arylamidase from nephrotic children and kidney membrane-bound neurtral arylamidase had identical KM values, effect of inhibition by L-methionine and heat stability. One of four urinary arylamidases from nephrotic children was suggested to be derived from plasma. It is concluded that an increase in urinary neutral arylamidase activity in nephrotic children is not only due to increased released from kidney membranes, but also due to leakage of plasma neutral arylamidase in the urine.

We isolated a 34,000-dalton protein from the heat-soluble fraction of avian smooth muscle using the procedures of ammonium sulfate fractionation, cation exchange chromatography and gel filtration. The amount of 34,000-dalton protein in the muscle homogenate was as much as tropomyosin. The 34,000-dalton protein bound to F-actin and F-actin-tropomyosin in a Ca2+-independent manner, but it Ca2+-dependently interacted with calmodulin. We tentatively named the 34,000-dalton protein gizzard p34K.

A placental type of membrane-bound arylamidase was detected in serum and ascites from a patient with renal cell carcinoma by isoelectric focusing. The enzyme had the same molecular weight, electrophoretic mobility on polyacrylamide gel, isoelectric point and KM value as placental membrane-bound arylamidase. The new type of membrane-bound arylamidase was named Shiba-isoenzyme after the patient's name.

Experiments were performed to determine whether the renal nerve mediates the renin release induced by intracerebroventricular administration of angiotensin II (ANG II), bradykinin (BK), leucine-enkephalin (Leu-ENK) and neurotensin (NT). In sham-operated rats, both ANG II and NT suppressed plasma renin activity (PRA), BK did not affect PRA, and Leu-ENK increased PRA. Renal denervation abolished the increase in PRA by Leu-ENK. Suppression of PRA by ANG II was attenuated in denervated rats. Renal denervation did not influence the renin release by BK or NT. These results suggest that the renal nerve plays an important role in elevating PRA after central stimulation by Leu-ENK. Although suppression of PRA is mainly mediated by mechanisms other than the renal nerve, the renal nerve partially participates in suppression of PRA by ANG II.

Serum cholesterol, triglyceride, high density lipoprotein cholesterol (HDL-C) and apolipoprotein (apo) A-I, A-II and B concentrations were measured in 109 first-degree relatives of patients with angiographically defined coronary artery disease (CAD). Age- and sex-matched healthy factory employees were chosen as a control group. Male relatives of the CAD patients had significantly higher serum triglyceride and apoB levels, and significantly lower serum HDL-C and apoA-I levels than the controls. Female relatives of the CAD patients also showed similar differences in serum HDL-C, apoA-I and apoB levels. Discriminant analysis indicated that apolipoproteins were better discriminators than lipids in both patients with CAD and their relatives. In univariate analysis, the best discriminator was apoB between male relatives and the controls, and apoA-I between female relatives and the controls. The percentage of exact classification achieved using three variables (serum cholesterol, triglyceride and HDL-C) was 74% in male relatives and 70% in female relatives. By adding variables of apoA-I and apoB, the percentage of correctly classified subjects was increased to 82% and 80%, respectively. These results indicate that serum apolipoprotein abnormalities are prevalent in relatives of the CAD patients. These abnormalities may explain the familial aggregation of CAD.

In our previous paper (Biochem. Biophys. Res. Commun. 141, 20-26 (1986) we reported the isolation of a 34000-dalton protein (p34K) which binds to calmodulin and F-actin from chicken gizzard smooth muscle. We examined the distribution of the immunoreactive component of gizzard p34K in bovine tissues by immunoblot analysis using a rabbit polyclonal antibody raised against gizzard p34K. The immunoreactive components with molecular weights of 33000-35000 were detected in all smooth muscles from aorta, esophagus, stomach, trachea and uterus. In non-smooth muscle tissues, a 36000-dalton cross-reactive protein was present in adrenal medulla and cortex. The immunoreactive form of gizzard p34K occurred in large amounts in smooth muscles from various bovine tissues.

It was demonstrated that angiotensin I-converting enzyme was excreted in human urine. The mean activity of the enzyme in normal urine was found to be 0.38 +/- 0.04 (S.E.M.) units/day (n = 18) and the enzymic activity correlated well with the concentration of the excreted sodium (r = 0.76, p less than 0.005). Urinary angiotensin I-converting enzyme was partially purified. Three different molecular weights of enzyme (greater than 400 000, 290 000 and 140 000) were demonstrated by Sephadex G-200 gel filtration. The enzymic properties of these three enzymes were identical with those of angiotensin I-converting enzyme from human lung with regard to inhibitory effects (bradykinin potentiator c and Arg-Pro-Pro), Cl- dependency, pH optimum and KM value.

A placental form of membrane-bound neutral arylamidase was found in the tissue of renal cell carcinoma. Membrane-bound neutral arylamidases from renal cell carcinoma, an intact part of the same kidney and placenta, had the same molecular weight (240 000) and were identical with respect to KM value and effect of inhibition by chelators or amino acids. Membrane-bound neutral arylamidase from renal carcinoma had the same electrophoretic mobility and heat stability as placental membrane-bound neutral arylamidase, but differed from kidney membrane-bound neutral arylamidase with respect to electrophoretic mobility, heat stability and susceptibility to urea inactivation. Results suggest the carcinoplacental alterations in membrane-bound neutral arylamidase isoenzyme in renal cell carcinoma.

Soluble and membrane-bound neutral arylamidases from renal cell carcinoma were partially purified and their properties were compared. Soluble neutral arylamidase was a heat labile and SH-dependent metalloenzyme. Membrane-bound neutral arylamidase was a rather heat stable metalloenzyme and was activated by Co2+ with changing KM and VMAX. KM values for soluble and membrane-bound neutral arylamidases were different. L-Methionine (5mM) did not affect the enzymic activity of soluble neutral arylamidase, but inhibited 82 percent of the enzymic activity of membrane-bound neutral arylamidase. Molecular weights of soluble and membrane-bound neutral arylamidases by Sephadex G-200 gel filtration were 140000 and 240000, respectively. Soluble and membrane-bound neutral arylamidases from renal cell carcinoma appear to be distinct enzymes.

We designed aldehyde derivatives of small peptides representing the C-terminal portion of angiotensin I sequence as an inhibitor of human renin. Among compounds that we synthesized, benzyloxycarbonyl (Z)-Phe-His-Leucinal (compound V), Z-Pro-Phe-His-Leucinal (Compound IV) and Z-[3-(1'-naphthyl)Ala]-His-Leucinal (compound VII) markedly inhibited human renin (IC50, 7.5 X 10(-7), 3.2 X 10(-7) and 8.0 X 10(-8) mol/l, respectively). Compound VII was shown to be noncompetitive (Ki = 2.4 X 10(-7) mol/l). It did not inhibit either cathepsin D or pepsin. Compound V had slight or no inhibitory effect at the concentration of 10(-5) mol/l on six animal renins except for monkey and rabbit renins. Results obtained show that these aldehyde compounds are highly selective and species specific inhibitors for human and monkey renins.

A high molecular weight angiotensinogen (Mr. 332,000 daltons) was prepared from plasma of pregnant women by gel filtration on Sephacryl S-300. The molecular weight was reduced to 81,000 by treatment with dithiothreitol (DTT), but not by treatment with SDS. DTT-treated high molecular weight (HMW) angiotensinogen was very similar to low molecular weight (LMW) angiotensinogen with respect to molecular weight, pH profile for angiotensin formation by human kidney renin, thermostability, Km value and isoelectric point. The antibody against LMW-angiotensinogen completely cross-reacted with HMW-angiotensinogen. These results suggest that HMW-angiotensinogen is probably a complex of LMW-angiotensinogen and other protein(s) which might be bound by disulfide bond.

Anti-placental and anti-kidney membrane-bound arylamidase antibodies were prepared in rabbits. Both antibodies showed similar titration curves against both the original antigens, and the membrane-bound arylamidases partially purified from small intestine, lung, liver and renal cell carcinoma. Double immunidiffusion tests showed that six membrane-bound arylamidases were immunochemically indistinguishable.

A placental type of membrane-bound arylamidase (Shiba isoenzyme) was found in the tissue of adenocarcinoma of the lung. Cancerous lung membrane-bound arylamidase had the same electrophoretic mobility, isoelectric point (pH 4.2), and susceptibility to SDS inactivation as placental membrane-bound arylamidase. Cancerous lung membrane-bound arylamidase had molecular weight and kinetic properties similar to normal lung membrane-bound arylamidase, but differed from the normal lung membrane-bound arylamidase with respect to electrophoretic mobility, isoelectric point, heat stability and inactivation by SDS.

Human plasma renin substrate was purified and separated from renin by a concanavalin A-Sepharose affinity column. Human renin substrate as well as renin were bound to concanavalin A. Renin substrate was eluted with 0.1 M D-glucose in 20 mM Tris/HCl buffer, pH 8.0. The specific activity increased from 38.5 to 653 ng of angiotensin-I equivalents per mg of protein (17-fold) and the recovery was 85%. Renin was eluted completely with 0.2 M alpha-D-methylglucoside and 0.2 M alpha-D-methylmannoside.

To examine the relationship of hypertriglyceridemia to coronary artery disease (CAD), we measured serum cholesterol, triglyceride, high density lipoprotein cholesterol (HDL-C) and apolipoproteins (apo) A-I, A-II and B in 82 male patients with angiographically defined CAD and 140 age-matched healthy controls. The CAD patients had significantly lower apo A-I and A-II and HDL-C levels, but had higher apo B and triglyceride levels than the controls. After adjustments of apolipoproteins for serum triglyceride, CAD patients had significantly higher apo B and lower apo A-I and A-II levels than the controls. Discriminant analysis showed that apo B was the best discriminator and that apo A-I was next. In the normotriglyceridemic subgroup HDL-C also had a sufficient power for discrimination between CAD patients and the controls, but in the hypertriglyceridemic subgroup HDL-C had no discriminative power. Both apo A-I and B had significant discriminative power between CAD patients and the controls, independently of the serum triglyceride level. These results indicate that measurements of serum apo A-I and apo B are useful for the study of coronary risk factor in hypertriglyceridemic subjects. Finally, it is necessary to sub-classify dyslipoproteinemia by serum apolipoprotein levels for predicting the future occurrence of CAD in the general population.

Oral administration of clonidine lowered the high blood pressure in two-kidney, one-clip hypertensive rats in chronic stage (three to four months), but not in one-kidney, one-clip hypertensive rats. This reduction of blood pressure was well correlated with decrease in plasma renin concentration, but not with decrease in urinary noradrenaline excretion. These results suggest that the reduction of blood pressure in two-kidney, one-clip hypertensive rats may be due to reduced renin release through the action of clonidine, while in one-kidney, one-clip hypertensive rats the sympathetic nervous activity may play little or no role in the maintenance of high blood pressure.

Angiotensin-converting enzyme (ACE, EC 3.4.15.1) was demonstrated in the tissue of human renal cell carcinoma and its cultured cells by immunofluorescence and immunohistochemistry using a specific rabbit antibody to human kidney ACE. The biochemical properties of the partially purified enzyme from the tumor tissue were identical with the pure enzyme from normal kidney with respect to Km, optimal pH, chloride ion activation and heat sensitivity. Catalytic activities of tumor and normal kidney enzymes were similarly inhibited by an ACE inhibitor, captopril, and by the antibody to human kidney ACE. However, double immunodiffusion analysis showed spur-forming immunoprecipitin bands between the tumor and normal kidney enzyme. These data suggest that human renal cell carcinomas have a new isozyme of ACE differing in part immunologically from the enzyme of normal kidney.

A membrane-bound arylamidase from well-differentiated hepatocellular carcinoma having the same electrophoretic mobility as placental membrane-bound arylamidase was found. The enzyme was found to be a sialoprotein and was activated by Co2+. Hepatoma membrane-bound arylamidase had a similar molecular weight (240 000) and kinetic properties to normal liver membrane-bound arylamidase, but differed from the liver enzyme with respect to electrophoretic mobility, heat stability and urea inactivation.