ODA, H. and FUJITANI, Y. Concentric Electrodes for Produci. Acupuncture-Like Anesthetic Effects. Tohoku J. Exp. Med., 1990, 160 (3), 169-175-We designed concentric electrodes composed of a center electrode and an outer ring electrode. Electrical stimulation with two sets of such electrodes for 15min as conditioning stimuli was given to the left hand of 35 adult subjects to induce acupuncture-like anesthetic effects. The effects immediately after the conditioning were compared between stimulation through a pair of center electrodes alone at 3Hz (conditioning 1) and simultaneous stimulation of 3Hz through a pair of center electrodes and 100Hz through a pair of outer ring electrodes (conditioning 2). In conditioning 2, modulating effects of 100Hz stimuli through a pair of outer ring electrodes made it possible to increase the voltage strength of 3Hz stimuli through a pair of center electrodes with maintaining the minimum perception of pricking sensation. In both conditioning procedures, muscle twitching was not accompanied. It was found that the respective stimulating current thresholds for faint touch sensation and also for pricking sensation at the right forearm could be elevated significantly more by conditioning 2 (1.54 and 1.40times) than by conditioning 1 (1.20 and 1.14times).
MIIDA, T. Apolipoprotein E Phenotypes in Patients with Coronary ArteryDisease. Tohoku. J. Exp. Med., 1990, 160 (3), 177-187-The relationship between apolipoprotein E (Apo E) phenotypes and progression of coronary atheros-clerosis was investigated in 125 patients with coronary artery disease (CAD) proven angiographically (101 males, 24 females). To elucidate the pure effect of Apo E phenotypes on lipoproteins and coronary atherosclerosis, patients with familial hypercholesterolemia were excluded from the subjects. As a control group, 129 norml healthy volunteers (84 males, 45 females) were studied. In the CAD group, VLDL and LDL levels increased and HDL level decreased regardless of Apo E phenotypes in both sexes. The incidence of E4 was higher and that of E2 was slightly lower in the CAD group than in the control group. Two patients with E5/3 who had high LDL-cholesterol levels were found in the male CAD group. LDL-cholesterol level in E3/2 was lower than in E4/3 and E3/3 in the male CAD group. VLDL-cholesterol/triglyceride and VLDL cholesterol/phospholipid ratios in E3/2 were significantly higher than in E4/3 and E3/3 in the male CAD group, but the difference was not so marked as found in typical type III hyperlipidemia. When the male patients with effort angina were examined, coronary score (index of the severity of CAD) was the lowest in E3/2. In addition, the mean age at the onset of CAD was significantly higher in E3/2 than in E4/3. In conclusion, E2 acts protectively against coronary atherosclerosis, while E4 promotes it through the modulation of LDL-cholesterol level.
HANEW, K., SUGAWARA, A., UTSUMI, A., SHIMIZU, Y. and YOSHINAGA, K. GHRH and TRH Tests in Patients with Idiopathic and Secondary GH Deficiency -With Special Reference to GH Response Patterns to GHRH. Tohoku J. Exp. Med., 1990, 160 (3), 189-202-To study whether patients with idiopathic GH deficiency (IGHD) show a delayed GH response pattern to GHRH, 42 patients with IGHD, 14 patients with hypothalamic tumor (2ry GHD), and 23 normal short children (NSC) were examined as to their GH response patterns to GHRH together with their TSH and PRL response patterns to TRH. After TRH injection, the mean time of the TSH peak in IGHD (67.5±6.5min, n=36) and 2ry GHD patients (81.7±14.8min, n=9) was clearly delayed comparing to that of NSC (29.1±2.9 min, n=16; both p<0.01). Similarly, the mean time of the PRL peak in IGHD (38.3±3.6min, n=36) and try GHD patients (39.5±5.8, n=11) was significantly delayed comparing to NSC (22.0±3.5min, both p<0.01). In IGHD patients, the delayed response pattern of TSH and PRL was more remarkable in patients who had breech delivery than in those with normal delivery. In contrast, the mean time of the GH peak was similar in IGHD (62.1±4.0min, n=41), try GHD (64.1±8.1min, n=11) and NSC (58.0±6.1min, n=23). However, the decline from peak GH (120min GH/peak GH) was significantly smaller in IGHD (54.3± 4.2%) and try GHD (60.7±7.3%) than in NSC (39.0±8.1%)(both p<0.05). Further, in IGHD patients plasma GH response was greater in patients with normal delivery than in those with breech and asphyxia delivery. These results seem to indicate: 1) the stimulus-secretion mechanism is different between somatotrophs and thyrotrophs or lactotrophs in man, 2) IGHD patients have hypothalamic lesions as well as pituitary lesions, 3) such hypothalamo-pituitary lesions in IGHD patients are greater in patients with abnormal delivery than in those with normal delivery.
TAKEDA, K., NAKAI, H., HAGIWARA, H., TADA, K., SHOWS, T.S., BYERS, M.G. and MYEROWITZ, R. Fine Assignment of β-Hexosaminidase A α-Subunit on15q23-q24 by High Resolution In Situ Hybridization. Tohoku J. Exp. Med., 1990, 160 (3), 203-211-Tay-Sacks disease results from mutation in the gene encoding β-hexosaminidase A α-subunit. Although some reports have suggested the locus on 15q, we tried to determine the finer gene locus using high resolution in situ hybridization. cDNA probe, pβH α-5, containing the full-length sequence for the enzyme subunit, was 3H-labeled within 1-4×107cpm/μg of cDNA by nick-translation. After molecular hybridization and autoradiography, prometaphases were G-banded by Hoechst 33258, UV-exposure and Giemsa. A total of 227 silver grains on chromosomes within 115 prometaphase spreads were analyzed. The region 15q23-q24 had 27 grains, corresponding to 11.9% of the total grains and to 77.1% of the grains on chromosome 15. 20.9% of prometaphases were observed with a grain at 15q23-q24. According to several previous reports, the shortest region of overlap (SRO) of the locus has been 15q22-q25.1. Here we have assigned the gene locus to the narrower region 15q23-q24 by high-resolution in situ hybridization, which is one of the most powerful strategy for the completion of human gene map.
HATAKEYAMA, A., SASAKI, T., MURYOI, T., MURAI, C. and YOSHINAGA, K. Are the Clonotypes of Serum IgG Anti-DNA Antibodies Associated with LupusNephritis in Humans? Tohoku J. Exp. Med., 1990, 160(3), 213-221-We analyzed isoelectrofocusing (IEF) patterns of anti-DNA antibodies originated from sera and the renal eluates of patients with systemic lupus erythematosus (SLE). The spectrotypic patterns of serum anti-DNA-antibodies were heterogenous and bands with single-stranded (ss) and double-stranded (ds) DNA were detected in the PI 5.5-6.5 and PI 8-9.5 regions when SLE sera were tested, whereas healthy subjects failed to form bands even at different saline concentrations. The renal eluates from normal subjects never bound to DNA whereas those from SLE glomeruli showed relatively restricted IEF patterns which were detected mainly in PI 6.0 and PI 8.5, showing that some anti-DNA antibodies may be nephritogenic. However, the spectrotypic patterns of serum anti-DNA antibodies in patients with active lupus nephritis were similar with those in patients lacking renal lesions. The reasons why IEF analysis failed to indentify specific clonotypes of nephritogenic anti-DNA antibodies are discussed in association with pathogenesis of lupus nephritis. This study also suggests that the use of a high concentration of 6M urea in an IEF analysis may be able to expose antigen-binding sites of the circulating immune complex (IC)-derived antibodies.
KANEMITSU, H., SASAKI, S., AKIBA, T. and MARUMO, F. Ureteral ObstructionDecreases ATP Dependent H+-Pump Activity of Rabbit Renal Outer Medulla. Tohoku J. Exp. Med., 1990, 160 (3), 223-229 -Ureteral obstruction has been known to cause a renal acidification defect. To determine the underlying mechanisms of this defect, vacuolar-type H+-pump activity of the membrane vesicles taken from the rabbit renal outermedulla was measured in 24hr obstructed and contralateral kidneys. H+-pump activity measured by acridine orange quenching method was resistant to oligomycin (10μg/ml), vanadate (0.1mM), and the absence of ambient K+, and it was highly sensitive to N-ethylmaleimide (0.5mM) and dicyclohexylcarbodiimide (0.2mM). H+-pump activity was 6.69±1.40 in the obstructed kidney, which was significantly lower than 10.32±1.88% AFU/mg•min in the contralateral kidney. In sham-operated rabbits, H+-pump activities were not different in both kidneys. Passive H+/OH- permeability measured by the rate of dissipation of the imposed pH gradient was not different in obstructed and contralateral kidneys. These results suggest that ureteral obstruction decreases vacuolar-type H+-pump activity of the kidney outermedulla, and this decrease may contribute to the distal acidification defect of obstructed uropathy.
SAITO, R., KONDO, T., FUJIMURA, S., IIANDA, M., ICHINOSE, T. and SHIRAISHI, Y. Detection of Peripheral Blood and Bronchoalveolar Lavage Fluid Lymphocytesin Rat Lung Transplantation for Early Diagnosis of Rejection. Tohoku J. Exp. Med., 1990, 160 (3), 231-249-Left lung transplantation was performed in two combinations of rat strains. In group 1, lung grafts were rejected within 7 day postoperatively, and in group 2, grafts were rejected within 18 day postoperatively. The histological appearance of rejection was classified into 4 stages, and lymphocytes from peripheral blood (PB) and bronchoalveolar lavage fluid (BALF) were collected at each stage. In group 1, a significant decrease in the number of lymphocytes in PB was obserbed as the rejection progressed, whereas the number of lymphocytes in PB increased slightly in group 2. The number of total cells in BALF increased significantly as the rejection progressed in both groups. A marked increase in the value of spontaneous blastogenesis (SB) was observed in stage 2 in BALF lymphocytes, whereas that in PB lymphocytes was found to increase after stage 3 in both groups. The ratio of T-helper/T-nonhelper did not change significantly in PB lymphocytes in both groups. On the other hand, a significant decrease in the value of T-helper/T-nonhelper (<1.0) was observed in BALF at stage 3 and 4 in both groups. These results show that the studies of BALF lymphocytes were very useful for early diagnosis of rejection in lung transplantation.
SASAHARA, M., MATSUO, M., KAKIZAKI, G., AIKAWA, T., SATO, T., UEJIMA, K., YAMADA, A., SAITO, M. and NAKAMURA, K. The Effect of Long Term EthanolIntake on the Parotid Gland in Rats. Tohoku J. Exp. Med., 1990, 160 (3), 251-275-To elucidate the effects of long term ethanol intake on the parotid gland, light and electron microscopic observations of the parotid glands from male Wistar rats that had been given 20% ethanol ad lib instead of water were carried out. The findings were compared with the changes in the pancreas previously reported (Kakizaki et al. 1987). The results were as follows: (1) Severe lesions involving lobular units of the parotid gland were found in three rats exposed to ethanol. (2) From the early stages of ethanol ingestion, the lateral interdigitations of acinar cells widened irregularly. In the striated ducts, basal enfoldings became also irregular and short, while the mitochondria (which were a slender rod shape and arranged parallel to the basal enfoldings in control rats) became swollen, developed a dark matrix, a decrease in the cristae, and showed random arrangement. (3) Later, both tortous proliferation of the intercalated ducts and destruction as well as irregular lamination of the basal lamina of the striated ducts were observed frequently. (4) The animals developing severe morphological changes in the pancreas following ethanol ingestion were different from the animals developing severe changes in the parotid gland.
TANITA, T., KOIKE, K., FUJIMURA, S. and STAUB, N.C. A New Method forEstimating Filtration Variables in Isolated Zone 1 Rat Lung. Tohoku J. Exp. Med., 1990, 160 (3), 277-284-The filtration variables, K (filtration coefficient), Ppmv (perimicrovascular pressure) and σ (reflection coefficient), were estimated independently in previous reports using the Starling equation or the micropuncture method. We used matrix algebra to estimate these variables simultaneously. We measured filtration rate (Q) by a gravimetric method in isolated rat lung lobes in zone 1 conditions (alveolar pressure=20cmH2O) at two vascular pressures, Pvasc=15 or 18cm H2O and perfused the lobes with plasma containing a low or a high concentration of protein. By extrapolating the log of the rate of weight gain to t=0, we obtain the initial filtration rate before any of the pressure variables (microvascular and perimicrovascular hydrostatic pressures) in the Starling equation changed. Assuming that protein filtered into perimicrovascular space only by convection, we substituted it into the Starling equation as follows: Q=K[(Pmv-Ppmv)-σ2 (Πmv)], where Pmv and Πmv are microvascular and perimicrovascular plasma protein osmotic pressures. Πmv was estimated by Yamada's equation (Yamada et al. 1985). For the matrix algebra, we used three values, we omitted the value for the high protein, low vascular pressure experiment. We obtained K=26.3 [mg/(min×cmH2O×g wet weight)], Ppmv=6.2cmH2O and σ=0.46. These values agree with values from previous reports. Since these 3 filtration variables are interrelated, this new method for simultaneous measurement is more accurate than independent measurements are. The chief advantage of this method is that it does not require a separate estimate of isogravimetric pressure or a direct measurement of interstitial pressure, and all variables are obtained simultaneously.
SAWAI, T., MURAKAMI, K., OHTANI, Y., KURKINNEN, M., KYOGOKU, M. and HAYASHI, M. Stromelysin Synthesizing Cells in the Synovial Tissues ofRheumatoid Arthritis Demonstrated by In Situ Hybridization and Immunohisto-chemical Methods. Tohoku J. Exp. Med., 1990, 160 (3), 285-286-We identified the cells synthesizing stromelysin to be synovial cells in the synovial tissue from four biopsy cases with rheumatoid arthritis (RA) by in situ hybridization and immunohistochemical studies. In the cases which showed severe inflammation of synovia such as well developed lymphoid follicle and diffuse inflammatory infiltration, synovial cells located only in the superficial layer of synovial tissues showed abundant mRNA and enzymic protein of stromelysin.