Hydrogen peroxide is an endothelium-derived hyperpolarizing factor in mice.

The endothelium plays an important role in maintaining vascular homeostasis by synthesizing and releasing several endothelium-derived relaxing factors, such as prostacyclin, nitric oxide (NO), and the previously unidentified endothelium-derived hyperpolarizing factor (EDHF). In this study, we examined our hypothesis that hydrogen peroxide (H(2)O(2)) derived from endothelial NO synthase (eNOS) is an EDHF. EDHF-mediated relaxation and hyperpolarization in response to acetylcholine (ACh) were markedly attenuated in small mesenteric arteries from eNOS knockout (eNOS-KO) mice. In the eNOS-KO mice, vasodilating and hyperpolarizing responses of vascular smooth muscle per se were fairly well preserved, as was the increase in intracellular calcium in endothelial cells in response to ACh. Antihypertensive treatment with hydralazine failed to improve the EDHF-mediated relaxation. Catalase, which dismutates H(2)O(2) to form water and oxygen, inhibited EDHF-mediated relaxation and hyperpolarization, but it did not affect endothelium-independent relaxation following treatment with the K(+) channel opener levcromakalim. Exogenous H(2)O(2) elicited similar relaxation and hyperpolarization in endothelium-stripped arteries. Finally, laser confocal microscopic examination with peroxide-sensitive fluorescence dye demonstrated that the endothelium produced H(2)O(2) upon stimulation by ACh and that the H(2)O(2) production was markedly reduced in eNOS-KO mice. These results indicate that H(2)O(2) is an EDHF in mouse small mesenteric arteries and that eNOS is a major source of the reactive oxygen species.

Rat renin: purification and characterization.

In order to clarify the molecular basis of the unique features of rat renin (EC 3.4.99.19) and to provide materials and basic information for high blood pressure studies in rats, renin was purified from rat kidney. The final step of purification on CM-cellulose separated renin into three major isoenzyme peaks, R-I, R-II, R-III, and an additional minor peak. These preparations were judged homogeneous by multiple criteria, and the isoenzymes were found to have similar amino acid compositions. The amino acid composition is also closely analogous to hog renin, except that rat renin has a higher cysteine content. In contrast to hog renin, the rat enzymes do not contain amino sugars, yet are apparently glycoproteins as judged by their affinity for concanavalin A. The molecular weights of R-I, R-II, and R-III were estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 37 000, 36 000 and 35 000, respectively. The isoelectric points were 5.05, 5.15 and 5.22, respectively. The specific activities of the purified enzymes (determined using rat plasma as substrate) were 615, 626 and 452 Goldblatt units/mg, respectively. Comparison of activities with the hog- and rat-derived substrates indicated a preference for that from the rat. The reaction of the rat enzymes with a synthetic peptide substrate had a similar catalytic rate constant to the hog enzyme, indicating close similarity in the active site region of the two enzymes.

Obesity is associated with increased risk of allergy in Vietnamese adolescents.

BACKGROUND: The prevalence of allergic diseases has remarkably increased in the last decades in tandem with the number of obese individuals. Results of studies on obesity and allergic diseases are controversial, and most of them are related to asthma and asthma-like symptoms. OBJECTIVE: In our study, we evaluated the association of several obesity indices and the prevalence of allergic diseases, including bronchial asthma, allergic rhino-conjunctivitis, atopic dermatitis and food allergy. DESIGN: Cross-sectional study. RESULTS: From a total of 1185 adolescents (49.3% boys) aged 12 to 17 y old, 19.3% reported a physician-diagnosed allergic disease, and 35.3% reported undiagnosed allergic symptoms. Logistic regression analysis revealed a higher risk of allergy in children of middle (OR = 2.02, CI: 1.12-3.64) and high (OR = 2.40, CI: 1.25-4.61) compared to low socioeconomic status, in subjects reporting industrial emissions in the neighborhood (OR = 2.19, CI: 1.40-3.41), and in adolescents with parental history of allergy (OR = 1.92, CI: 1.26-2.92). Body mass index (BMI) and percentage of body fat (%BF) were significantly related to allergy (OR = 1.16, CI: 1.01-1.34 for BMI; OR = 1.03, CI: 1.01-1.06 for %BF). After controlling for socioeconomic status, industrial emissions and parental history of allergy, only %BF remained statistically significant (OR = 1.03, CI: 1.01-1.06). Food allergy was not related to obesity in our study population, and when adolescents with food allergy were excluded from the analysis, the odds of having allergy increased 28% with each increase in SD of BMI (OR = 1.28, CI: 1.05-1.57) and 5% for each unit increase in the percentage of body fat (OR = 1.05, CI: 1.01-1.08). CONCLUSION: In conclusion, being overweight was associated with an increased risk of allergy in our study population. Our results point towards an association between being overweight and rhino-conjunctivitis, but not food allergy. No association was observed with other allergic diseases.

Angiotensin II type 2 receptor inhibits cell proliferation and activates tyrosine phosphatase.

The angiotensin II type 2 (AT2) receptor inhibits basic fibroblast growth factor-induced proliferation of R3T3 fibroblast cells and transiently stimulates a vanadate-sensitive phosphotyrosine phosphatase, strongly suggesting that AT2 is a mitogen inhibitor. We generated AT2 gene-null mice that showed increased blood pressure, indicating the hypotensive action of AT2. However, inhibition of renomedullary AT2 by selective antagonists, as reported by Sassard and associates, show that AT2 suppresses pressure natriuresis. Thus, both AT1 and AT2 work in the direction of sodium retention, suggesting a unique role for angiotensin II in the kidney in terms of blood pressure regulation and sodium metabolism.

Increased left ventricular function as an adaptive response in vibration disease.

Vibration disease results from the long-term use of vibrating tools. Vibration, noise, and cold are stressors that impair the human body, inducing vibration disease. From echocardiographic methods, the left ventricular ejection fraction in vibration disease was 79 +/- 4%, a significantly higher value than that in control subjects (75 +/- 6%) (p less than 0.01). The increase in ejection fraction appeared to be due mainly to an increase in left ventricular end-diastolic dimension. The value of the ejection fraction was proportional to the activity of the autonomic nerves. The stroke volume index in patients with vibration disease was also significantly larger than that in the control subjects (p less than 0.001). Electrocardiograms revealed a significantly lower heart rate at rest and an increase in the ratio of T waves to R waves in precordial lead V6. These data suggest that the cardiovascular system in patients with vibration disease provides an adaptive response to the stressors.

Substrate specificity of carboxypeptidase from Watermelon.

The substrate specificity of carboxypeptidase (F-II) purified from watermelon for various synthetic peptides and esters was examined kinetically. The enzyme showed a broad substrate specificity against various carbobenzoxy- and benzyl-dipeptides. Peptides containing glycine or proline were hydrolyzed slowly by the enzyme. Peptides containing hydrophobic amino acids were hydrolyzed rapidly. The presence of hydrophobic amino acid residues, not only at the C-terminal position but also at the second position and probably the third position from the C-terminal resulted in an increase in the rate of hydrolysis. Inhibition studies with diisopropyl flurophosphate and diastereomers of carbobenzoxy-Phe-Ala demonstrated that the peptidase and esterase activities of the enzyme are both catalyzed by the same site of the enzyme molecule, but the binding sites for peptides and esters seem not to be the same. The enzyme also had amidase activity, which was optimal at pH 7.0.

Subunit structure of pig kidney cathepsin A.

The two forms of pig kidney cathepsin A (A,L and A,S) were found to be composed of three kinds of polypeptides as common structural subunits with molecular weights of 20,000, 25,000, and 55,000. [32P]-DFP was incorporated into the subunit with a molecular weight of 25,000. This subunit seems to be a catalytic one, on which active serine is present. A subunit composition ratio of 1 : 1 : 1 in native cathepsin A,S was estimated by its molecular weight and those of the subunits. Native cathepsin, A,L also seems to have a subunit composition ratio of 1 : 1 : 1. These results suggest a monomer-polymer relationship between both enzymes.

Purification and some properties of cathepsin A of small molecular size from pig kidney.

Cathepsin A [EC 3.4.2.-] of small molecular size (cathepsin A, S) has been purified about 800-fold from pig kidney by procedures including chromatographies on DEAE-Sephadex, SP-Sephadex, and Sephadex G-150. 1. The homogeneity of the purified enzyme was proved by ultracentrifugation and polyacrylamide gel electrophoresis. The molecular weight (100,000) and isoelectric point (pI=5.0) were estimated. 2. The enzyme was remarkably stabilized by sucrose and KCl, and was most stable at pH 5-5.5 in the presence of both stabilizers. The enzyme had not only peptidase activity but also esterase and amidase activity; it was optimally active at pH 5.2 for peptide hydrolysis and at pH 8 for the hydrolysis of esters and amides. 3. Diisopropyl fluorophosphate and iodoacetamide completely inhibited these three activities. 4. The enzyme hydrolyzed various benzoyl- and benzyloxycarbonyl-dipeptides with neutral, acidic, and basic amino acids, and proline in the C-terminal position. The carboxypeptidase nature of the enzyme was proved by its action on an oligopeptide. 5. Several enzymatic properties of cathepsin A, S were almost the same as thoas of cathepsin A of large molecular size (cathepsin A, L) and the crude homogenate.

Substrate specificities of cathepsin A,L and A,S from pig kidney.

The substrate specificities of two different molecular sizes of cathepsin A, A,L (large form) and A,S (small form), for synthetic substrates were examined kinetically. Both enzymes showed a similar broad substrate specificity against various acyl dipeptides, amino acid esters, and amino acid amides. Z-Phe-Ala and Ac-Phe-OEt were good substrates. Peptides containing hydrophobic amino acids were hydrolyzed rapidly. The presence of hydrophobic amino acid residues, not only at the C-terminal position but also at the second position and probably the third position from the C-terminal, resulted in an increase in the rate of hydrolysis. Peptides containing glycine and proline were hydrolyzed slowly. Inhibition studies with Z-D-Phe-D-Ala and Z-Phe suggested that the peptidase and esterase activities of the enzymes are both catalyzed by the same site of the enzyme molecule, but it remains to be elucidated whether or not the binding sites for peptides and esters are the same.

Physiological methods used in Japan for the diagnosis of suspected hand-arm vibration syndrome.

In the pathogenesis of hand-arm vibration syndrome, vibration affects the peripheral system, especially the circulatory, nervous, and musculoskeletal systems. Medical questionnaires, physical examinations, and laboratory tests are used to diagnose the hand-arm vibration syndrome. The laboratory tests are satisfactory diagnostic aids for detecting slight disorders of the peripheral circulatory, nervous, and musculoskeletal systems. Peripheral circulatory function tests include skin temperature measured by a thermistor under specific temperature conditions, the nail compression test, and the cold provocation test (10-min immersion in cold water at 5 or 10 degrees C). To assess peripheral nervous function, tests for the pain and vibration senses are included. Peripheral musculoskeletal function tests consist of grip strength, pinch strength and tapping counts. These physiological tests are applied in worker screening twice a year. These tests have been authorized by the Ministry of Labour since 1973. Several criteria for the tests, although not authorized, are widely used. The diagnostic implication of each separate test may be low. Consequently, some laboratory tests should be evaluated, along with medical questionnaires and physical examinations, in the diagnosis of suspected hand-arm vibration syndrome.

Pathophysiology and clinical picture of hand-arm vibration syndrome in Japanese workers.

Hand-arm vibration syndrome is an occupational disease induced by long-term use of vibratory tools such as rock drills and chain saws. The three major stressors of vibration, noise and cold may produce various symptoms and signs not only with peripheral circulatory, nervous and muscle-joint disorders but also with general disorders. It is a point of controversy whether the symptoms and signs should be limited in the peripheral disorders. The question may involve differences in definition: vibration alone or work mode with vibratory tools. There are two viewpoints in the staging: peripheral and general viewpoints. The key concept in the peripheral viewpoint staging is a checkup to find disorders at the early stage and classify the peripheral disorders of the digits in detail. The general viewpoint staging seeks to grasp the general pictures of patients, and to classify from light to severe cases in the treatment. Clinical manifestations may include the general disorders in proportion to the severity of the syndrome according to our clinical experience. A differential diagnosis should be carefully made in the light of legal, medical and economic compensation. The treatments that we have used for approximately 20 years have beneficial effects on the whole-body, which include 1) physiobalneotherapy (therapeutic exercise, exercise in a pool and physiotherapy), 2) drug therapy (vasodilating drugs, autonomic stabilizers, etc.), 3) nerve blocking therapy, 4) surgical therapy for ulnar nerve paralysis or paresis, and 5) education for patients. Even with these therapies, a beneficial effect may not be observed in a short period. The recovery may be slow.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of diltiazem on occupational Raynaud's syndrome (vibration disease).

An open trial on the therapeutic effect of diltiazem, a calcium antagonist, was conducted on occupational Raynaud's syndrome (vibration disease). Seventeen men with vibration disease were studied. The mean age was 54 years. No patient had cardiovascular disease. Primary and secondary cases of Raynaud's phenomenon were excluded. Diltiazem, 30mg orally 3 times daily, was given to the patients for 6 weeks. Before and after this treatment period, we evaluated the subjective symptoms (11 items including Raynaud's phenomenon, numbness of hands and arms, stiffness of shoulder and neck, and others) and peripheral function (by cold water immersion test, vibration and pain sensations for fingers, tapping test, and momentary grasp strength test), and did laboratory blood tests (12 variables). The "effective" rates of subjective symptoms evaluating by the method of 5 steps and the peripheral function was 69.6% and 52.9%, respectively. The overall effectiveness of diltiazem therapy was assessed by collectively evaluating the changes in subjective symptoms, peripheral functions, and the occurrence of side effects. No side effects occurred during the treatment period. Thus, the collective effectiveness of diltiazem was 64.7%. In conclusion, diltiazem, a calcium antagonist, can be effective in long-term treatment of patients with vibration disease.

Responses of myocardial blood flows to whole-body vibration in the dog.

Experimentally acute exposure to whole-body vibration resulted in changes in myocardial blood flows in the in vivo dog. Regional myocardial blood flows measured by a hydrogen gas clearance method were increased at a vibration frequency of 120 Hz and decreased at 50 Hz. No appreciable changes were found in presence of a beta-adrenergic blocker. The values of plasma cyclic nucleotides were increased at both frequencies of vibration, more at 50 Hz than at 120 Hz. No significant changes were observed in heart rates and arterial pressure during and after vibration load. These findings suggest that increased myocardial blood flows in response to vibration may be related more to the frequency of vibration than to beta-adrenergic actions.

Noninvasive assessment of the autonomic nervous tone in angina pectoris: an application of digital plethysmography with auditory stimuli.

To assess the relationship between autonomic nerve activity and the anginal attack, we examined the daily variation of the autonomic tones employing digital plethysmography with auditory stimuli, and also tested the daily variation of exercise capacity. Thirteen of 45 cases (28.9%) of spontaneous angina, excluding Prinzmetal's angina, complained of typical chest discomfort or pain especially in the morning. They manifested an augmented level of the autonomic nerve activity and positive stress testing. The remainder did not demonstrate these abnormal findings in the morning. For the 13 cases, calcium antagonists were given orally immediately after awakening. This resulted in the complete relief from chest discomfort or pain. These findings suggest that an autonomic imbalance plays an important role in the pathogenesis of this type of anginal attack. Thus, the estimation of the daily variation of autonomic nervous tone would be valuable for the diagnosis and treatment of angina pectoris.

Further observations of the digital plethysmography in response to auditory stimuli and its clinical applications.

Further observations of the digital plethysmography with auditory stimuli and its clinical applications were performed in patients with vibration disease and heart diseases. The responsive pattern to the auditory stimuli in the digital plethysmogram could be faithfully reproducible if it elapses more than 5 minutes apart between the first stimulus and the second one. The responsive patterns were divided into four types: normal (N), hyperreactive (I and D) types and hyporeactive (P) type. The values of urinary catecholamine increased in parallel to the activity level of the autonomic nerve. The hyperreactive type (D) had the highest value of urinary catecholamine, and vice versa. With regard to the age, the elderly was prone to show hyporeactive (P) type, and the young tended to be hyperreactive (D) type. Psychologic factors were examined by Cornell medical index. Neurosis was not necessarily related to the hyperreactive type. In clinical applications, it was observed by this method that the autonomic imbalance in patients with vibration disease, angina pectoris, or hypertension was recovered by the treatment, and the abnormal types of the response recovered to N type. In conclusion, the digital plethysmography with auditory stimuli as one of the autonomic nerve function tests would be much useful and non-invasive method for observation of clinical course and decision of the severity.

Whole-body vibration suppresses gastric motility in healthy men.

The influence of whole-body vibration on gastric motility was investigated by using an electrogastrography (EGG) in seven healthy men. The EGG is usually referred to as a noninvasive technique of recording gastric myoelectrical activity by means of placing electrodes on the abdominal surface. Sinusoidal vertical vibration at each of 3 different frequencies (10 Hz, 20 Hz, 40 Hz) were randomly given to the subject seated on a platform of vibrator for 5 min. The vibration magnitude was kept at a constant of 2.0 msec-2 (r.m.s.) during operation. The mean dominant frequency of EGG at control period was prior to operation 3.3 cycles per min (cpm). During vibration exposure at 10 Hz, the peak of dominant frequency increased to 3.9 cpm, and the relative power of slow wave showed the statistically significant decrease (45.8%, p < 0.05). The mean relative power of slow wave which is composed of frequencies ranged from 2.0 to 5.0 cpm was 56.6% at control period. On the contrary the mean relative power of frequencies ranging from 5.0 to 9.0 cpm, tachygastria increased from 29.5% to 39.1%. These results suggest that the short-term exposure to whole-body vibration effects on the gastric myoelectrical activity.

Cardiovascular reflexes during vibration stress.

Vibration disease due to hand-held vibratory tools has various symptoms and signs which can be characterized by the severity. They include disorders of the central and autonomic nervous systems, as well as peripheral system disorders. The mechanism of Raynaud's phenomenon in vibration disease is proposed to be: Vibration and cold affect the local vessels and nerves directly, leading to enhanced release of chemical vasoconstrictors. Vibration, noise, cold, ergonomic and biodynamic conditions, and emotional stress during work result in disorders of the central and autonomic nervous systems. In the early stages, the autonomic nervous system may be stimulated, and in the later stages it is suppressed. When local vessel injuries and disorders of the central and local autonomic nervous mechanisms controlling the vessels occur, vasospasms in the fingers develop when the whole body is exposed to cold. The cardiovascular system, other than the peripheral circulatory system, may adapt to vibration stress. The adaptation subsides 7 to 8 years after discontinuation of the use of hand-held vibratory tools.

A revised cold water immersion test for assessing peripheral circulatory function.

To improve the conventional cold water immersion test for assessing peripheral circulatory function, the immersed area of the hand was changed in this study. Twenty-seven healthy medical students with a mean age of 24 years participated in the study. The cold water immersion tests were carried out using two methods (in 5 degree C water for 1 min), in which the immersed area was up to the metacarpophalangeal (MP) joints (hereafter, MP-method) which is a new method, or to the wrist (hereafter, W-method) which is a conventional method. The recovery of skin temperature, vibratory perception threshold and complaints of finger pain were determined during cold immersion tests with the two methods. The mean skin temperature for the MP-method after immersion recovered more quickly than that for the W-method. However, the time courses of recovering were parallel for the two methods. Significant differences were noted at 3 min, 5 min and 10 min after cessation of immersion. There was a significant positive correlation in the recovery rates at 5 min after immersion between the two methods (correlation coefficient = 0.65, p < 0.01). The mean vibratory thresholds for the W-method were higher than those for the MP-method, and the difference was significant (p < 0.01). All subjects preferred the MP-method to the W-method because there was less finger pain. These results indicate that the MP-method, immersion up to the MP joint of a hand, for assessing peripheral circulatory function is better than the W-method.

Frequency dependence of the suppressive effects of vibration on atherosclerosis in the rabbit.

Whole-body vibration suppresses the development of atherosclerosis in the rabbit (Oki and Matoba, 1987). The present study was designed to clarify whether the effect of vibration on atherosclerosis depends on the frequency of vibration. Longitudinal vibrations at a frequency of 30 or 60 Hz was applied to 12 New Zealand white rabbits for 12 weeks. The gradual decrease in body weight and blood hematocrit in the vibration groups with time were parallel to the changes in the controls. The rate of increase in serum lipid concentrations induced by a cholesterol-rich diet was significantly suppressed in the vibration groups, as compared to the controls. This may be due to the vibration and not the diet. The aortic wall was thinner at 60 Hz than at 30 Hz, whereas the ratios of trace metals (Ca/Mg and Zn/Cu) in the aortic tissues were smaller at 30 Hz. The area of plaque formation in the intima was smaller at 60 Hz than at 30 Hz (p less than 0.05). Thus, the suppressive effect of vibration on the development of atherosclerosis in the aorta may be greater at a frequency of 60 Hz than at 30 Hz. Vibration may play an important role in lipid metabolism.

Application of Stockholm criteria to patients with hand-arm vibration syndrome in a follow-up study.

We applied Stockholm criterion to patients with hand-arm vibration syndrome (HAVS) for assessing the therapeutic effects in comparison with the criterion of the Ministry of Labour in Japan (MLJ). Forty male patients recognized as an occupational disease were examined in two times (mean interval periods 3.0 years). The subjects were free from vibratory tasks during the mean period of 8.3 years. The clinical examinations and some laboratory tests were conducted with the same procedures by the two same occupational physicians. The criterion of MLJ was graded more severe than Stockholm one for the vascular component at the first examination. According to Stockholm criterion, 16 patients (40%) were classified into stage 0 in the vascular component, 11 (70%) of which complained of coldness in the fingers. At the second examination, 9 of 16 patients in stage 0 had Raynaud's phenomenon (RP). On the other hand, the criterion of MLJ could reflect an improvement of symptoms, for example, coldness. The findings may indicate that Stockholm criterion for vascular component is not enough to assess the clinical changes for light stages according to the severity of RP alone. For the sensorineural (SN) component, both criteria were almost compatible. From the viewpoint of preventive medicine, it was concluded that Stockholm criterion was not suitable for evaluating their clinical severity in the early stage of HAVS.