[Probability of the standardization of control survey in local medical association].

This report discusses about the probability of the standardization of external quality control (control survey) in local medical area such as prefecture size. For example, our control survey by Hiroshima medical association in Hiroshima prefecture is selected and shown on their effort to the standardization of the control survey. This Hiroshima control survey is continuing for 27 years and its purpose is the improvement of the differences between laboratory facilities. For the all standardization of control survey system, tests, devices and reagents, the recent reports of Hiroshima control survey shows that the reference methods by Japan Society of Clinical Chemistry (JSCC) and the enzyme reference materials by Japan Committee of Clinical Laboratory Standardization (JCCLS) are very useful for the improvement of the differences between laboratory facilities in Hiroshima Prefecture.

[Relationship between flow volume curve and CT findings in non-smoking patients with long histories of bronchial asthma].

AIM: This study was conducted to verify whether bronchial asthma (BA) alone causes pulmonary emphysema (PE), and to examine the computed tomography (CT) findings in non-smokers with BA demonstrating the flow volume curve (FV curve) characteristic of PE. SUBJECTS AND METHODS: Non-smoking patients with a history of BA for more than 20 years were divided into 2 groups: the dogleg pattern group (n = 5), with an FV curve characteristic of PE, and the concave pattern group (n = 16) with an FV curve characteristic of BA. CT scans was performed using CT values (level, 900 H.U.; width, 400 H.U.) that facilitate detection of a low attenuation area (LAA), and using conventional CT values (level, 700 H.U.; width, 1,300 H.U.). LAA (including air trapping), thickness of the bronchial wall, and partial atelectasis were compared between the 2 groups. RESULTS: PE was not detected, although air trapping was found in all subjects. The thickness of the airway was greater in the dogleg pattern than in the concave pattern. The incidences of air trapping and partial atelectasis were higher in the former than in the latter. DISCUSSION: BA alone may not cause PE. Some BA patients without PE show the FV curve characteristic of PE, reflecting an increase in the thickness of the airway wall and a decrease in the pulmonary ventilation probably due to the air trapping and the partial atelectasis.

Intramolecular 1,3-dipolar cycloaddition strategy for enantioselective synthesis of FR-900482 analogues.

[reaction: see text] Enantioselective synthesis of FR-900482 analogues is described. The key reaction of the synthesis is intramolecular 1,3-dipolar cycloaddition of a highly functionalized nitrile oxide with complete stereo- and regioselectivities to construct the eight-membered benzazocine ring.

[Topics of clinical physiological tests--pulmonary function--proposal of physio-chemical tests].

Pulmonary functions are classified as respiratory and non-respiratory. Although the mechanisms of non-respiratory pulmonary function have already been established, no new tests of respiratory pulmonary function have been developed for a long time. Thus, we report here a new analytic test of respiratory pulmonary functions. The new analytic method is physio-chemical analysis, in which the chemical factors of lungs are estimated from the physiological characteristics of lung during respiration. As examples, this report shows the method of exchanging mechanical energy to chemical energy in a comparison of lung ventilatory work between smokers and non-smokers. This report discusses the relationship between chemical energy and ATP in the blood of pneumoconiosis patients.

IL-6 attenuates apoptosis, while neither IL-6 nor IL-10 affect the numbers or protease phenotype of fetal liver-derived human mast cells.

BACKGROUND: The combination of recombinant human stem cell factor (rhSCF), rh interleukin (IL)-6 and rhIL-10 was reported to be optimal for mast cell development from cord blood progenitors and to induce chymase expression in all such mast cells earlier in their development than tryptase. OBJECTIVE: The effects of rhIL-6 and rhIL-10 in various combinations on the rhSCF-dependent development of human mast cells from fetal liver progenitors were examined in serum-free media. METHODS: Dispersed fetal liver cells were cultured in serum-free AIM-V medium with rhSCF alone, or with combinations of rhIL-6 and rhIL-10. Tryptase and chymase expression, surface Kit expression, metachromasia with toluidine blue and apoptosis were measured. RESULTS: Neither rhIL-6 nor rhIL-10 nor the two interleukins together, when included from day 0 of culture, affected the number or protease phenotype of mast cells at 1 or 3 weeks. Expression of tryptase paralleled the appearance of metachromasia and surface Kit, both of which preceded chymase expression, regardless whether a rabbit polyclonal or mouse monoclonal anti-chymase antibody preparation was used. On the other hand, rhIL-6 markedly attenuated baseline levels of apoptosis in the presence of rhSCF as well as apoptosis occurring after withdrawal of rhSCF, whereas rhIL-10 had no effect. CONCLUSION: RhIL-6 protected fetal liver-derived mast cells from apoptosis, particularly after withdrawal of rhSCF, but neither rhIL-6 nor rhIL-10 nor the combination of these interleukins affected the numbers or protease phenotype of these mast cells.

Non-dipper phenomenon in essential hypertension is related to blunted nocturnal rise and fall of sympatho-vagal nervous activity and progress in retinopathy.

Although the relation between the blunted nocturnal decline in blood pressure and target organ damages is well established, the mechanism underlying these results has not been clarified. We investigated the relationship among heart rate variability, nocturnal change in blood pressure and the severity of cardiac and extracardiac target organ damages caused by essential hypertension. We studied 52 Japanese inpatients with essential hypertension (24 men and 28 women; mean age, 49+/-3 years). After a stabilization period of 1 week, ambulatory blood pressure monitoring (ABPM) and 24-h ECG monitoring were performed and analyzed. The non-dipper subjects were defined as those whose nocturnal decrease of mean BP was < 10% of daytime blood pressure (BP). The sex, age, body mass index. duration of hypertension, and 24-h BP were similar in dipper (n = 34) and non-dipper (n = 18) patients. The left ventricular mass index (LVMI) was significantly higher and the degree of hypertensive retinopathy was significantly worse in the non-dipper patients than that of the dipper patients. In the non-dipper patients, indexes of time-domain analysis such as the sum of differences between adjacent RR intervals (NNDrms), the number of pairs of adjacent RR intervals differing by more than 50 ms in the entire recording (RR 50) were significantly lower than that of the dipper patients. Additionally, as for spectral analysis, daytime low frequency/high frequency (LF/HF) was higher and nighttime high frequency (HF) was lower than that of the dipper patients. Independent predictors were the 24-h mean blood pressure (MBP) for left ventricular hypertrophy (LVH), nighttime systric BP (SBP) for progress in retinopathy and duration of hypertension for proteinuria. In conclusion, decrease in parasympathetic nervous function and increase in sympathetic nervous function may contribute to occurrence of non-dipper phenomenon, as well as progress in retinopathy.

Apoptosis is not increased in myocardium overexpressing type 2 angiotensin II receptor in transgenic mice.

To determine whether angiotensin type 2 (AT(2)) receptor stimulation induces apoptosis in cardiomyocytes in vivo, we developed transgenic mice overexpressing the AT(2) receptor in a cardiac-specific manner, using the alpha-myosin heavy-chain promoter. Ten- to 12-week-old male homozygous transgenic mice (n=44) and wild-type mice (n=44) were used. Both transgenic and wild-type mice were given either saline (control), a subpressor dose of angiotensin II (100 ng. kg(-1). min(-1)), a pressor dose of angiotensin II (1000 ng. kg(-1). min(-1)) for 14 days, a pressor dose of angiotensin II for 28 days to investigate the effects of stimulation on both angiotensin type 1 (AT(1)) and AT(2) receptors, the AT(1) antagonist L158809 alone, or a combination of angiotensin II (1000 ng. kg(-1). min(-1)) and L158809 for 14 days to investigate the effects of selective AT(2) receptor stimulation. Apoptosis was analyzed in paraffin-embedded ventricular sections by the terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling (TUNEL) technique. In both transgenic and wild-type mice, administration of a subpressor dose of angiotensin II, L158809, or a combination of angiotensin II and L158809 did not significantly affect the tail-cuff blood pressure or heart-to-body weight ratio, whereas administration of a pressor dose of angiotensin II for 14 or 28 days significantly increased blood pressure and the heart-to-body weight ratio. However, there was no statistical difference between the effects of angiotensin II in transgenic and wild-type mice. The number of TUNEL-positive nuclei was approximately 0 to 10 per 100 000 cardiomyocytes, with no difference between transgenic and wild-type mice, regardless of saline infusion or any stimulation. In infarcted canine myocardial tissue sections for positive control, the number of TUNEL-positive nuclei was increased by 13.8 to 19.1 times compared with those in the noninfarcted myocardium. In conclusion, angiotensin II infusion for a period of 28 days failed to induce cardiomyocyte apoptosis regardless of the presence or absence of cardiac AT(2) receptor overexpression. It is unlikely that in mice the AT(2) receptor is a strong signal to induce cardiomyocyte apoptosis in vivo.

Human skin-derived mast cells can proliferate while retaining their characteristic functional and protease phenotypes.

Human mast cells in adult tissues have been thought to have limited, if any, proliferative potential. The current study examined mast cells obtained from adult skin and cultured in serum-free medium with recombinant human stem cell factor. During the first 4 weeks of culture, the percentages of mast cells increased from 10 to almost 100. After 8 weeks, a 150-fold increase in the number of mast cells was observed. When freshly dispersed mast cells were individually sorted onto human fibroblast monolayers and cultured for 3 weeks, one or more mast cells were detected in about two thirds of the wells, and in about two thirds of these wells the surviving mast cells showed evidence of proliferation, indicating most mast cells in skin can proliferate. Such mast cells all expressed high surface levels of Kit and Fc epsilon RI, each of which were functional, indicating IgE was not required for Fc epsilon RI expression on mast cells. Such mast cells also retained the MC(TC) protease phenotype of mast cells that normally reside in the dermis. After 4 to 8 weeks of culture these mast cells degranulated in response to substance P and compound 48/80, characteristics of skin-derived mast cells that persist outside of the cutaneous microenvironment. (Blood. 2001;97:2045-2052)

Coronary atherosclerosis and oxidative stress as reflected by autoantibodies against oxidized low-density lipoprotein and oxysterols.

Clinical studies and animal experiments have demonstrated that oxidized low-density lipoprotein (oxLDL) and oxysterols play important roles in atherogenesis. OxLDL is immunogenic, and autoantibodies (Ab) against oxLDL are detectable in serum. We investigated the relevance of oxysterols and Ab against-oxLDL to coronary artery disease (CAD) in 183 patients undergoing coronary angiography. Patient groups included angiographically normal subjects (< 75% stenosis), others with spasm (> 75% narrowing in response to acetylcholine), and some others with fixed stenosis (> 75%). The group with stenosis was subdivided into patients with stable and unstable angina. Serum concentrations of autoantibodies and 25-, 27-, and 7-beta-hydroxycholesterols were significantly higher in the stenotic group than in the normal group (P < 0.01, P < 0.05, P < 0.05, and P < 0.05, respectively). Antibodies, but not oxysterol concentrations, were significantly greater in subjects with unstable than with stable angina (P < 0.01). We conclude that anti-oxLDL antibody and oxysterol concentrations are associated with coronary artery stenosis, and that oxidative stress may be greatly increased in unstable angina.

Disialoganglioside GD3 is selectively expressed by developing and mature human mast cells.

BACKGROUND: Disialoganglioside GD3 is expressed on the surface of selected cell types. Anti-GD3 mAb administered to human subjects with malignant melanoma produces signs and symptoms of immediate hypersensitivity reactions. OBJECTIVE: The expression of GD3 by human mast cells was assessed during mast cell development in vitro and in samples of lung and skin. METHODS: GD3 on tissue- and in vitro-derived mast cells was analyzed after double labeling of cells for tryptase (G3 mAb) or Kit (YB5.B8 mAb) and GD3 (R24 mAb). Glycolipids in extracts of fetal liver-derived mast cells were examined by using high-performance thin-layer chromatography. RESULTS: Flow cytometry showed that the percentage of GD3+ cells increased in parallel to Kit+ cells during the recombinant human stem cell factor-dependent development of fetal liver-derived mast cells. Double-labeling experiments showed that GD3+ cells were also surface Kit+ and granule tryptase positive, identifying them as mast cells in preparations of lung-, skin-, fetal liver-, and cord blood-derived cells. The major acidic glycolipid detected was NeuAcalpha2-8NeuAcalpha2-3Galbeta1-4Glcbeta1-1'Cer (GD3). Among peripheral blood leukocytes, only basophils and about 10% of the T cells were labeled with anti-GD3 mAb. Anti-GD3 mAb-conjugated magnetic beads were used to purify mast cells to greater than 90% purity from dispersed skin cells enriched to approximately 12% purity by means of density-dependent sedimentation but were less proficient for dispersed human lung mast cells, most likely because of other cell types that express GD3. CONCLUSION: GD3 is expressed on the surface of developing human mast cells in parallel to tryptase in secretory granules and, like Kit, can serve as a target for their enrichment by immunoaffinity techniques.

[Pulmonary function tests and their clinical significances on outside of textbooks].

Pulmonary functions are classified two kinds of functions which are respiratory and non-respiratory. We explained new clinical significant results by looking at the usual respiratory pulmonary function tests and devices in this paper. We found two new indexes on uneveness of ventilation by analyzed compartments of forced expiratory curves(Tiffneu curves) and at improvement of CO single breathing method on diffusing capacity test. We found new index on bronchial anaphylaxis for bronchial asthma by time constant measured from forced expiratory curves, also. We found new index on ventilatory efficiency at developing the automatic respiratory resistance meter by the forced oscillation method of Du Bois, too.

Calorie restriction reduced blood pressure in obesity hypertensives by improvement of autonomic nerve activity and insulin sensitivity.

Association between obesity and hypertension has been well recognized. A reduction in the body weight of over-weight hypertensive patients is a recommended lifestyle modification. The purpose of our study is to examine the relationship of insulin sensitivity and autonomic nervous activity with reduction of blood pressure by the calorie restriction. We evaluated the heart rate variability, nocturnal change of blood pressure and insulin resistance before and after a short-term low-calorie diet in 12 overweight essential hypertensives. After a week of standard diet (2000 kcal), 2 weeks of low-calorie diet (800 kcal) with normal sodium content induced a significant reduction in body mass index, triglyceride, fasting immunoreactive protein, homeostasis model assessment as an index of insulin resistance, and urinary excretion of sodium and potassium. Systolic blood pressure was significantly reduced both in daytime and night-time after the low-calorie diet (daytime, 134.5+/-6.0 to 122.0+/-4.1 mmHg; night-time, 126.8+/-5.2 to 113.4+/-7.2 mmHg). In daytime, diastolic blood pressure was also reduced (90.3+/-2.1 to 88.1+/-4.8 mmHg). Although heart rate was not significantly reduced, a rise of high frequency in night-time (346+/-82 to 572+/-108 ms2) and a fall of low frequency/high frequency in day-time (3.5+/-0.4 to 2.6+/-0.1) was significant after a low-calorie diet. In conclusion, weight loss by low-calorie diet with a constant intake of sodium, reduced blood pressure in obese hypertensives by improvement of vagal nervous activity and insulin resistance.

[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.

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.

Mechanism of anti-tumor effect of combination of bleomycin and shock waves.

We have previously reported marked enhancement of the cytocidal effect of bleomycin (BLM) on cancer cell suspensions in vitro by the combination with shock waves. In this study, we evaluated the synergistic effects on cancer cell proliferation and apoptosis in solid tumors. A spherical piezo-ceramic element was used as the shock wave source, with a pressure peak of 40 MPa. A human colon cancer cell line, SW480 was implanted onto the back of nude mice. Two thousand shock waves were administered to the tumor immediately following an intravenous injection of BLM at a dose of one-tenth of the LD(50). The tumor was extirpated at 3, 6, 12, 24, 72 h and 1 week following shock exposure. Cell proliferation and apoptosis were detected by Ki-67 using antibody MIB-1 and by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL) method. The lowest percentage (35.7%) of Ki-67-positive cells appeared 24 h following the treatment. The maximum apoptotic index was detected within 6 h following the treatment. Moreover, numerous large cells with enlarged nuclei were detected histologically. These results suggest that shock waves may enhance chemotherapeutic effects by increasing apoptosis and decreasing cell proliferation in the tumor tissue.

[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.

Type 1A dopamine receptor expression in the heart is not altered in spontaneously hypertensive rats.

We have recently demonstrated that type 1A dopamine (D1A) receptor is expressed in the rat heart, but its function still remains unknown. In the present study, we investigated possible changes in the expression level and the distribution of the cardiac D1A receptor in the development of left ventricular hypertrophy in spontaneously hypertensive rats/Izumo strain (SHR/Izm) at the ages of 4, 8, and 20 weeks. We examined D1A receptor protein distribution by immunohistochemistry and gene expression by competitive polymerase chain reaction (competitive PCR). In SHR/Izm, compared with the age-matched Wistar Kyoto rats/Izmo strain (WKY/Izm), blood pressure and heart/body weight ratio were significantly increased at 8 and 20 weeks. By immunohistochemistry, the D1A receptor was localized in cardiomyocytes and vascular smooth muscle cells of coronary arteries, but not in interstitial fibrotic tissue. D1A receptor distribution was not changed either by the strain or the age. Competitive PCR analysis showed that the D1A receptor mRNA level was significantly higher at 4 weeks than at 8 and 20 weeks in both strains of rats and that there was no significant difference in D1A receptor mRNA between SHR/Izm and WKY/Izm at any age (43.2 +/- 10.4 attomol x 10(-3)/L v 43.1 +/- 11.2 attomol x 10(-3)/L at 4 weeks, P = not significant, 3.9 +/- 0.9 attomol x 10(-3)/L v 4.0 +/- 1.3 attomol x 10(-3)/L at 8 weeks, P = not significant, 3.0 +/- 1.2 attomol x 10(-3)/L v 1.9 +/- 1.6 attomol x 10(-3)/L at 20 weeks, P = not significant). These results do not support the hypothesis that changes in D1A receptor expression are associated with the development of left ventricular hypertrophy in SHR.