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Biomedical subjects

M Kambe

Publications and source records attributed to M Kambe.

At least 37 records · Page 2Linked to original sources

Expression and localization of angiotensin subtype receptor proteins in the hypertensive rat heart.

The cellular localization of the AT(2) receptor and the regulation of its expression in hypertrophied left ventricle are not well known. We compared the expression of the cardiac AT(1) and AT(2) receptor in spontaneously hypertensive rats/Izumo strain (SHR/Izm) and Wistar Kyoto rats/Izumo strain (WKY/Izm), ages 4, 12, and 20 wk, by means of immunohistochemistry and Western blot analysis. In SHR/Izm, compared with WKY/Izm, blood pressure (161 +/- 2 vs. 120 +/- 2 mmHg at 12 wk, P </= 0.01, and 199 +/- 3 vs. 123 +/- 3 mmHg at 20 wk, P </= 0.01) and heart-to-body weight ratio (3.76 +/- 0.07 vs. 3.06 +/- 0.06 mg/g at 12 wk, P </= 0.01, and 3.90 +/- 0.08 vs. 3.01 +/- 0.12 mg/g at 20 wk, P </= 0.01) were significantly elevated. There was no difference in these values between the two strains at 4 wk of age. Histologically, 20-wk-old SHR/Izm demonstrated myocardial hypertrophy, a thickening of the smooth muscle layer of the intracardiac arteries, and perivascular fibrosis. By immunohistochemistry, the AT(2) receptor was localized to cardiomyocytes and vascular endothelial cells, but not in the vascular smooth muscle cells. No major AT(2) receptor signal was observed in perivascular fibrosis at any age in either strain of rats. No difference was detected in this localization between the two strains. By Western blotting, a single 44-kDa band for the AT(2) receptor and a single 60-kDa band for the AT(1) receptor were detected in ventricles from both strains of rats at all ages. Densitometric analysis demonstrated that the AT(2) receptor 44-kDa band was decreased by 20% at 12 wk and 32% at 20 wk (P < 0.01) in SHR/Izm compared with WKY/Izm. The intensity of the AT(1) receptor 60-kDa band was increased by 57% in 20-wk-old SHR/Izm compared with WKY/Izm (P < 0.05). There was no significant difference in the intensity of the 44- or 60-kDa bands in 4-wk-old animals of either strain. We demonstrated a decrease in the AT(2) receptor and an increase in the AT(1) receptor protein with no change in their localizations in hypertrophied left ventricular myocytes of SHR/Izm.

Angiotensins↗

Abnormal Ca2+ handling and increased Mg2+ permeability in platelets of hypertensive rats.

In order to test the hypothesis that intracellular Na+ accumulation and cellular Mg2+ deficiency may be involved in the abnormalities in Ca2+ handling and reactivity in spontaneously hypertensive rats (SHR) platelets, the metabolism of Na+, Ca2+ and Mg2+ was determined in fluorescent dye loaded platelets from 15 SHR and 15 Wistar-Kyoto rats (WKY) at 12 weeks of age. Mg2+ leak was estimated as the Mg2+ influx induced by an increase in extracellular [Mg2+] (from 1 to 5 mmol/l) and Mg2+/Na+ exchange activity was estimated as the Mg2+ influx induced by a decrease in extracellular [Na+] (from 140 to 50 mmol/l). Cellular metabolism of the fluorescent dye was similar in the two groups. Mean platelet [Ca2+]i was significantly increased under basal and thrombin (0.1 U/ml)-stimulated conditions in SHR compared to WKY, both in the presence and absence of extracellular Ca2+. Mean Ca2+ discharge capacity was similar between the two groups. There was no difference in mean [Na+]i between the two groups. Basal [Mg2+]i was also increased in SHR platelets. Mg2+ leak was higher in SHR than in WKY, while Mg2+/Na+ exchange activity was similar in the two groups. There was no difference in serum Mg2+ concentration between SHR and WKY. These data suggest that abnormal Ca2+ handling is accompanied by elevation in [Mg2+]i via increased permeability of platelet cell membranes to Mg2+ in SHR without any alteration in [Na+]i, and do not support the Mg2+ deficiency hypothesis in genetically hypertensive rats.

Animals↗

Abnormal magnesium status in patients with cardiovascular diseases.

To investigate magnesium status in patients with cardiovascular diseases and in those presenting high factors for these diseases, we measured the concentrations of serum total Mg, serum ionized Mg and intra-erythrocyte Mg. Mg is an important cofactor for many enzymes, especially those involved in phosphate transfer reactions. Mg deficiency has been shown to be associated with fatal cardiovascular diseases, as well as with risk factors for these diseases. Only measurement of the serum concentration of total Mg is routinely available, but ionized Mg is the physiologically active component. Furthermore, most of the body's Mg is present in the intracellular space. Subjects included patients with ischaemic heart disease (n=80), cardiac arrhythmia (n=60), diabetes mellitus (n=36), essential hypertension (n=194) and hypercholesterolaemia (n=60). The same measurements were made in healthy controls (30 men and 26 women; mean age 58+/-11 years). The serum ionized Mg concentration was measured with a selective ion electrode. The intra-erythrocyte Mg concentration was measured by atomic absorption. No gender difference was found for any Mg parameter, nor was age related to any Mg parameter. The serum albumin concentration was positively correlated only with the serum total Mg concentration. Although the serum total Mg concentration was similar in all groups, patients with diabetes mellitus and arrhythmia had lower serum levels of ionized Mg. Patients with essential hypertension exhibited higher intra-erythrocyte Mg concentrations than the healthy controls. Thus the measurement of serum total Mg concentration may obscure the presence of extracellular Mg deficiency in patients with arrhythmia and diabetes mellitus. Furthermore, the intracellular accumulation of Mg does not support the hypothesis of Mg deficiency in patients with essential hypertension.

Biomarkers↗

[Evaluation of vascular endothelial function].

Endothelial function was evaluated in renal and forearm vessels from patients with essential hypertension. First, the increase in renal blood flow evaluated by the clearance of para-aminohippurate, serum c-GMP and urinary NOx during L-arginine infusion was significantly attenuated in essential hypertension. This attenuated endothelium-dependent vasodilation was improved by angiotensin converting enzyme inhibitor, but unchanged by calcium antagonist. Second, the increase in forearm blood flow evaluated by plethysmography during acetylcholine infusion or reactive hyperemia was attenuated in essential hypertension. This attenuation was abolished by NO synthesis inhibitor. Forearm endothelial function was improved by angiotensin converting enzyme inhibitor, daily aerobic exercise and body weight reduction by low calorie diet. In conclusion, endothelium-dependent vasodilation was attenuated in renal and forearm vasculature of essential hypertensives via reduction of NO synthesis. This attenuation can be improved by several treatments.

Acetylcholine↗

[Development of a case of Mycobacterium avium complex disease from right pleural effusion].

Moist pleurisy in patients with Mycobacterium avium Complex (MAC) is rarer than tuberculosis. We encountered an extremely rare case of MAC disease in a 75-year-old man who initially had only right pleural effusion. Gaffky VII was detected in the pleural effusion, and Mycobacterium avium was identified by culture and PCR. Although administration of antitubercular agents (RFP, INH, EB, and SM) + CAM and thoracic lavage were repeated, the Gaffky persisted strongly. Accordingly, pulmonary decortication and filling of the cavity with an omental flap were performed as surgical treatments. However, fistulas were formed between the remaining empyema cavity and the surgical wounds. Fenestration was also carried out. Postoperatively, centriacinar abnormalities appeared on computed tomography (CT). It has been reported that MAC disease begins with centriacinar abnormalities and the incidence of the lymphatic developmental pattern was low. Tuberculosis (the idiopathic pleuritis type) is considered to be caused this pattern from the primary infection focus. Therefore, the onset of unilateral effusion is extremely rare in patient with MAC disease, suggesting that the lymphatic developmental pattern occurs less frequently in patients with MAC disease. Furthermore, in this case, we speculated that centriacinar abnormalities were the MAC infection foci and could be detected by CT due to surgical invasion.

Aged↗

[Results of a questionnaire survey about "standardization" of connection methods in Laboratory Automation System or Laboratory Information System by the National University Hospital Clinical Laboratory Divisions].

"Standardization" is very important in the field of clinical laboratory medicine. Enzyme reference materials(ERM) and standard plasma proteins(CRM470) have already been developed. Reference methods for some clinical chemical tests have also been developed. We are studying "standardization" of electric communication methods between computers and automatic analyzers in Laboratory Automation System(LAS) or Laboratory Information System(LIS). We present the results of a questionnaire survey of 73 LAS or LIS making Companies in this paper. Although "standardization" of electric communications or local area network in LAS or LIS has been done in only 22 companies(34.9%), we are planning more functional standard electric communication methods such as Health Level 7(HL7) or American Society for Testing and Materials(ASTM).

Autoanalysis↗

Effect of L-arginine infusion on systemic and renal hemodynamics in hypertensive patients.

This study was designed to compare the renal endothelial function in patients with essential hypertension and normal renal function with that in hypertensive patients with renal insufficiency. We studied the effects of L-arginine (500 mg/kg intravenously over 30 min) on renal hemodynamics in 30 normotensive control subjects, 32 patients with mild to moderate essential hypertension who had normal renal function, and seven hypertensive patients with renal insufficiency who had a serum creatinine concentration >2.0 mg/mL and a glomerular filtration rate <50 mL/min/1.48 m2. L-Arginine infusion similarly reduced the mean blood pressure between the three groups (normotensive: -9.7% +/- 0.7%, hypertensives with normal renal function: -10.2% +/- 0.8%, and hypertensives with renal insufficiency: -8.2% +/- 1.3%). The L-arginine-induced decrease in renal vascular resistance was smaller in essential hypertensive patients than in normotensive subjects (-11.0% +/- 2.2 v -19.8% +/- 2.1%, P <.05). However, L-arginine had no effect on the renal vascular resistance in hypertensive patients with renal insufficiency (1.6% +/- 4.8%). Urine nitrite/nitrate levels in response to L-arginine significantly increased in the three groups in the following order: patients with renal insufficiency (47% +/- 15%), essential hypertensive patients (87% +/- 10%), and normotensive subjects (129% +/- 12%). The glomerular filtration rate was unaffected by L-arginine in normotensive and essential hypertensive patients (3.1% +/- 2.4% and 4.2% +/- 2.5%), but significantly decreased in hypertensive patients with renal insufficiency (-13.7% +/- 6.1%). These findings suggest that the ability of the L-arginine-nitric oxide-cGMP pathway to relax the renal vascular tone may be impaired in essential hypertensive patients and more markedly blunted in hypertensive patients with renal insufficiency, in parallel with increasing serum creatinine concentrations.

Arginine↗

[The present and future of the pulmonary function tests. The relationship between respiratory and non-respiratory function].

Pulmonary function is classified as respiratory and non-respiratory function. Respiratory pulmonary function involves exchange of O2 and CO2 gas at alveolar units in the lung and non-respiratory pulmonary function concerns metabolism of enzymes, hormones, surfactants vaso-active substances and immunological factors in the lung. The purpose of this study was to examine the relationship between respiratory and non-respiratory function using our newly developed expired NO gas continuous analytic method and the calculation method of ventilatory energy of the lung. Furthermore, we analyzed the relationship between arterial blood O2 pressure (PaO2) and mitochondrial function.

Humans↗

[Magnesium (Mg) status in patients with cardiovascular diseases].

Mg is the fourth most abundant total cation in the human body and the second most abundant intracellular cation. Moreover, Mg is an important cofactor for many enzymes especially those involved in phosphate transfer reactions. Mg is therefore essential in the regulation of the metabolism of other ions and cellular functions. Mg deficiency has been shown to be associated with fatal cardiovascular diseases such as cardiac arrhythmias and coronary heart disease, as well as with risk factors for these diseases, such as hypertension, and diabetes mellitus. Clinical evaluation of Mg status has been limited by the lack of suitable technology for measuring this cation. Although the measurement of serum total Mg is routinely available, ionized Mg is physiologically active. Furthermore, most of the body's Mg is present in the intracellular space. Our findings showed that serum total Mg was similar in all groups, but patients with arrhythmias and diabetes mellitus revealed lower levels of serum ionized Mg. On the other hand, patients with essential hypertension exhibited higher intraerythrocyte Mg concentrations than healthy controls. The measurement of serum total Mg may obscure the diagnosis of an abnormality in Mg metabolism in patients with arrhythmias and diabetes mellitus. Furthermore, the intracellular accumulation of Mg does not support the hypothesis of Mg deficiency in patients with essential hypertension.

Calcium↗

[Significance of area under the flow volume curve--useful index of bronchial asthma].

Some studies have reported that area under the flow volume curve (AUFVC) can be an index of pulmonary function. However, the significance of AUFVC remains to be clarified. We have clarified that AUFVC reflects the momentum of expired air. Size of flow volume curve (= AUFVC) has been commonly recognized to be changeable visually in patients with asthma. To clarify whether size of flow volume curve (= AUFVC) is a useful index of the course of bronchial asthma, we compared the rate of improvement forced expiratory volume in 1 second (FEV1.0), peak expiratory flow rate (PEFR) or forced vital capacity (FVC) to the rate of improvement in AUFVC after admission in 20 patients with bronchial asthma. The rate of improvement in AUFVC positively correlated with the rate of improvement in FEV1.0, PEFR or FVC. AUFVC demonstrated more marked improvement than other indices. Since AUFVC reflects the momentum of expired air, bronchial asthma's patients could understand visually that the momentum of expired air decreased due to airway stenosis by presenting flow volume curve. Therefore, visual size of flow volume curve helped patients with asthma to understand the condition of asthma. Size of flow volume curve (AUFVC) may be useful index of the course of bronchial asthma.

Adult↗

[The future of clinical laboratory database management system].

To assess the present status of the clinical laboratory database management system, the difference between the Clinical Laboratory Information System and Clinical Laboratory System was explained in this study. Although three kinds of database management systems (DBMS) were shown including the relational model, tree model and network model, the relational model was found to be the best DBMS for the clinical laboratory database based on our experience and developments of some clinical laboratory expert systems. As a future clinical laboratory database management system, the IC card system connected to an automatic chemical analyzer was proposed for personal health data management and a microscope/video system was proposed for dynamic data management of leukocytes or bacteria.

Clinical Laboratory Information Systems↗

[Expired nitric oxide concentration derived from nasal cavity and that derived from airway and lung in Japanese cedar pollinosis].

AIM: Japanese cedar pollinosis (Type I allergy to Japanese cedar pollen) shows a high incidence in spring in Japan, demonstrating symptoms such as pituita, rhinostenosis, lacrimation and sometimes pharyngeal pain. The following study investigated whether expired nitric oxide (NO) concentration is increased in Japanese cedar pollinosis, and whether expired NO concentration can be used to discriminate from cold syndrome which symptoms are similar to Japanese cedar pollinosis. SUBJECTS AND METHODS: In 20 healthy subjects, 20 patients with Japanese cedar pollinosis and 20 patients with cold syndrome, expired NO concentrations derived from nasal cavity and those derived from airway and lung were measured. Expired NO concentrations derived from nasal cavity and those derived from airway and lung were compared among the groups. RESULTS: In patients with Japanese cedar pollinosis expired NO concentrations derived from nasal cavity and those derived from airway and lung markedly increased. In patients with cold syndrome there were no significant increases in these concentrations. DISCUSSION: It is considered that expired NO concentration is useful for discriminating Japanese cedar pollinosis from cold syndrome. It seems interesting that not only expired NO concentrations derived from nasal cavity but also those derived from airway and lung were clevated. Generalized airway inflammation may be present, even without clinical asthma, in patients with Japanese cedar pollinosis.

Adult↗

Characterization of the integrin alpha(v) gene promoter in mice.

To characterize a promoter for integrin alpha(v) in mice, we cloned a 5'-flanking region of the mouse integrin alpha(v) gene. The nucleotide sequence of the promoter region lacks both TATA or CCAAT boxes, and has the consensus sequence for Sp1, Ets, AP-2 and GATA-1. The transcription initiation site was mapped as 287 base pairs (bp) upstream from the translation initiation site by primer extension analysis. To analyze the promoter activity of the integrin alpha(v) gene, the successive 5'-deletions which were cloned upstream of the luciferase reporter gene were transfected into mouse melanoma B16F10 cells with high metastatic properties. The luciferase assay reveals that a region (positions -108 to +22) has important cis-acting elements for the promoter activity. Further, three distinct protein-DNA complexes in the promoter region were detected in a gel shift assay using nuclear extracts from B16F10 cells. To identify positions where the complexes are formed, the promoter region (positions -108 to +97) was subjected to DNaseI footprinting analysis. The analysis showed that a 17 bp-element (positions -31 to -15) interacts with putative transcription factors. Deletion of the 17 bp-element from the promoter resulted in a reduction of about 80% in the promoter activity when compared with the wild type promoter, and a gel shift assay using the deleted mutant of the promoter showed only one protein-DNA complex. These results reveal that the 17 bp-element contains cis-acting elements, and plays an important role in the transcription initiation of the integrin alpha(v) gene.

Animals↗

DD genotype of the angiotensin I-converting enzyme gene is a risk factor for early onset of essential hypertension in Japanese patients.

Although angiotensin-converting enzyme (ACE) plays an important role in blood pressure regulation, no relationship between the insertion/deletion (I/D) polymorphism of the ACE gene and essential hypertension has been observed. However, as the pathogenesis and genetic background of essential hypertension are heterogeneous, we investigated whether the ACE gene is a marker of subgroups of patients with essential hypertension. A group of 178 patients with essential hypertension (90 men/88 women; 53 +/- 13 years of age, mean age +/- SD ) and 101 normotensive control subjects (54 men/47 women; 51 +/- 14 years of age, mean age +/- SD ) were included in the study. The allele frequencies of the two groups were similar. There were no differences in age, blood pressure, retinopathy grade, presence of proteinuria, or resting plasma renin activity (PRA) among the hypertensive patients with the II, ID, and DD genotypes. However, the age of onset of hypertension of patients with the DD genotype was lower (p < 20.05) and their left ventricular mass index was higher (p < 2 0.05) than those in patients with the non-DD genotype. These data suggest that the I/D polymorphism of the ACE gene is associated with an early onset of hypertension and left ventricular hypertrophy in Japanese patients with essential hypertension.

Adult↗

Cloning and characterization of a novel zinc finger protein that associates with nuclear matrix.

We have recently reported that the nuclei of B16 melanoma cells are intensely stained with anti-rat vitronectin (Vn) antibody, which reacts with both mouse and rat Vn. In the present study, we characterized the protein immunoreactive with the antibody using NIH3T3 cells, whose nuclei were also stained with the antibody. Western blot analysis showed that a protein with an approximate molecular weight of 75 kDa (p75), which was distinct from Vn, existed in the nuclear fraction, and, more specifically, in the nuclear matrix fraction, of NIH3T3 cells. Screening of an NIH3T3 cDNA library resulted in the isolation of a nearly full-length cDNA clone encoding p75. A database search revealed that the cDNA represents a novel gene. The deduced amino acid sequence showed that the protein is 580 amino acids long and contains two C2H2-type zinc finger motifs and glutamic acid-rich domains in the C-terminal region. When a fusion protein of green fluorescence protein and p75 was expressed in NIH3T3 cells, fluorescence was preferentially observed in the nuclei, demonstrating that the protein has a nuclear localization signal. The p75 protein, termed ZAN75, exhibited DNA-binding activity in a zinc-dependent manner. Southern blot analysis demonstrated that the ZAN75 gene exists in a single copy in the mouse genome and that a closely related gene is also present in chicken, rat, and human. Northern blot analysis showed that the ZAN75 gene is ubiquitously expressed in adult mouse tissues. In the cell cycle of NIH3T3 cells, expression was low in the G0/G1 phase, increased during the G1 phase, and persisted during the S and G2/M phases, suggesting that ZAN75 plays a role in regulating cell growth.

3T3 Cells↗