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Susceptibility to atopic diseases is known to involve genetic factors. The Gly237 allele of a polymorphism (Glu237Gly) of the Fc epsilon R1 beta gene is reportedly associated with atopic asthma in Japanese. To confirm this association, we conducted transmission disequilibrium tests in 76 families identified through atopic asthmatics. A case-control study was also carried out in atopic asthmatic subjects and non-atopic controls. The Gly237 allele was not preferentially transmitted to atopic asthma-affected offspring. Neither the Gly237 allele nor the Gly237/Gly237 + Glu237/Gly237 genotypes were significantly more prevalent in the atopic asthmatics than in the controls. This study failed to confirm a substantial role of the Gly237Glu polymorphism of the Fc epsilon R1 beta gene in the genetic predisposition for atopic asthma in this Japanese population.

In the present study we developed a method for quantifying regional cerebral blood flow (rCBF) using 99mTc-ECD SPECT based on a 3-compartment model. The dynamic SPECT scanning and sequential sampling of arterial blood were performed on 12 subjects with cerebrovascular diseases and etc. We defined brain fractionation index (BFI) as a parameter of rCBF, which was obtained from a single SPECT data and arterial input. The relationship between the values of BFI and rCBF obtained by the 133Xe inhalation method was analyzed by approximation with exponential function. In this method, rCBF was calculated from the values of BFI using the inverse function of the exponential function as a regression curve. The method was applied seven other patients with cerebrovascular diseases and the values of rCBF were compared with those obtained by the 133Xe inhalation method. We observed a good correlation (r = 0.854), and the inclination was approximately 1. This method can be applied to not only large field SPECT cameras but also conventional SPECT cameras.

Atopic dermatitis (AD) is a chronic relapsing dermatitis which belongs to the group of atopy-related diseases as well as asthma and allergic rhinitis. As a probable genetic risk which may contribute to the organ specificity of AD, an association between AD and a genetic variant of the gene encoding mast cell chymase (MCC), which has chymotrypsin-like specificity and is abundant in skin mast cells, has been reported in a Japanese population. We tried to confirm the role of this polymorphism in the development of AD in a Japanese population. A case-control analysis using 100 AD patients and 101 controls did not show a significant difference in the frequency of the BB genotype between the patient and control groups (odds ratio 1. 12, p = 0.81). The haplotype relative-risk analysis using 69 patient-parents trios did not suggest an association (chi2 = 0.177, p = 0.92). Thus, we failed to confirm the association between the polymorphism in the MCC gene and AD in the Japanese population.

The effects of propofol (1 mg.kg-1) and thiopental (2.5 mg.kg-1) on the vascular smooth muscle were investigated in 10 cardiac patients on cardiopulmonary total bypass using a constant perfusion flow. Radial mean arterial blood pressure (mAP) decreased to 85 +/- 5% (P < 0.01, vs preinjection) in the post-injection late phase (2-3 min) after propofol, and to 90 +/- 6% (P < 0.01) after thiopental. MAP after thiopental showed a transient initial increase to 115 +/- 8% (P < 0.01) in the post-injection early phase (-1 min). Skin blood flow (SBF) in the middle finger-tip increased to a peak of 450 +/- 120% (P < 0.01) in the post-injection late phase after propofol, and to 240 +/- 60% (P < 0.05) after thiopental. The increase in SBF after propofol was greater than that after thiopental (P < 0.05). The decrease in mAP and the increase in SBF under the constant perfusion flow are considered mainly due to a direct drug action of dilating peripheral vessels. In conclusion, this in vivo human study suggests that propofol (1 mg.kg-1) causes more potent vasodilatation in the peripheral vessels than thiopental (2.5 mg.kg-1).

A series of ts mutations in the GSP1 gene of Saccharomyces cerevisiae was isolated by error-prone PCR. A total of 25 ts gsp1 strains was obtained. Each of these mutants showed between one and seven different amino acid alterations. In several of these ts gsp1 strains, the same amino acid residues in Gsp1p were repeatedly mutated, indicating that our screen for ts gsp1 mutations was saturating. All of the ts gsp1 strains isolated had a defect in nuclear protein import, but only 16 of the 25 ts gsp1 strains had a defect in mRNA export. Thus, Gsp1p is suggested to be directly involved in nuclear protein import, but not in mRNA export. Following release from alpha-factor arrest, 11 of the ts gsp1 mutants arrested in G1; the remainder did not show any specific cell-cycle arrest, at 37 degrees C, the nonpermissive temperature. While the mutants that are defective in both mRNA export and protein import have a tendency to arrest in G1, there was no clear correlation between the cell cycle phenotype and the defects in mRNA export and nuclear protein import. Based on this, we assume that Ran/Gsp1p GTPase regulates the cell cycle and the nucleus/cytosol exchange of macromolecules through interactions with effectors that were independent of each other, and are differentially affected by mutation.

Background: Susceptibility to the development of asthma and other atopic diseases is known to be associated with genetic components. Several investigator have linked the interleukin-4 (IL-4) gene and nearby markers located on chromosome 5 to atopy and asthma. Recent study has demonstrated that the T allele of a polymorphism in the IL-4 gene promoter region (C-590T) is associated with elevated levels of total serum IgE., Objective: The objective of this study was to evaluate the possible role of this IL-4 polymorphism (C-590T) in modulating the allergic response and asthma in Japanese children., Methods and Results: The study was conducted in two different populations: families ascertained through asthmatic children (asthmatic group, 306 members) in whom linkage of asthma and atopy to chromosome 5q31-33 is suggested and a random general population (control group, 215 members). The IL-4 C-590T polymorphism was genotyped by PCR-restriction fragment length polymorphism analysis. Frequency of the T allele was 0.73 in the asthmatic group and 0.70 in the control group. No significant difference in the levels of total serum IgE and specific IgE to house dust mite was observed between subjects with homozygote for the C allele, a heterozygote and a homozygote for the T allele by intrafamilial and interfamilial comparisons. Using the transmission disequilibrium test, however, we found a significantly frequent transmission of the T allele to asthmatic children (chi2=5.72, P=0.023)., Conclusion: Our data suggest that the IL-4 promoter C-590T polymorphism may be associated with the development of asthma in Japanese children, but not through modulating total serum IgE levels.

We studied the accuracy of the method for measuring regional cerebral blood flow (rCBF) loaded with acetazolamide based on the microsphere model using iodine-123-IMP (IMP) SPECT. Two methods were examined, the super-early microsphere method with continuous withdrawal of arterial blood using the SPECT image obtained 5 min after tracer injection and the early microsphere method with one-point arterial sampling using the SPECT image obtained 30 min postinjection. On five subjects, after acetazolamide administration we measured rCBF by the analysis based on the two-compartment model using the data derived from dynamic SPECT scans and the sequential arterial blood sampling after IMP injection. Values of rCBF obtained by both super-early microsphere method and early microsphere method were significantly correlated with those obtained by the two-compartment model analysis (r = 0.982, 0.930, respectively). We conclude that it is possible to use the method based on the microsphere model in measuring rCBF loaded with acetazolamide. The early microsphere method with one-point sampling should be used clinically because of its simplicity and less-invasiveness.

The purpose of the study is to develop a simple and less invasive method for quantifying regional cerebral blood flow (rCBF) at pre- and post Diamox test using split-dose 99mTc-ECD and SPECT. By employing a microsphere model, integral of input function was calculated by the one-point venous sampling method previously reported. The study was performed on 5 subjects with cerebrovascular diseases. A split dose of 99mTc-ECD was injected pre- and post Diamox injection, and rCBF was measured by two SPECT scans and single venous samples, respectively. Mean CBF obtained by the present method was 0.47 +/- 0.07 ml/g/min at the control state, and 0.63 +/- 0.12 ml/g/min loaded with Diamox (mean % increase; 35%), showing good agreement with those obtained by the 133Xe-inhalation method. Since the present method does not require arterial blood sampling, dynamic data acquisition and dose corrections, it is simple, less invasive and useful in clinical SPECT studies.

Susceptibility to the development of asthma and other atopic diseases is known to be associated with genetic components, and several candidate genes have been reported to be linked to atopy and asthma. We have examined a linkage between traits of specific IgE responses to the house dust mite and its purified allergens, high total serum IgE levels and asthma, and the markers at the T-cell receptor (TCR) alpha chain gene (TCR-alpha) and beta chain gene (TCR-beta) in Japanese families ascertained through asthmatic children. Affected sib-pair analyses provided evidence for linkage between TCR-beta and nearby markers (D7S684) on chromosome 7q35 and both IgE and asthmatic phenotypes, but no evidence for linkage of IgE responses nor asthma phenotype to TCR-alpha was detected. The results suggest the presence of major gene(s) controlling atopy and asthma susceptibility in TCR-beta and the nearby region.

The purpose of the study is to develop a simple and less invasive method for quantifying regional cerebral blood flow (rCBF) using 99mTc-ECD and SPECT. By employing a microsphere model, a new method to measure rCBF was developed, which required a single arterial or venous sample instead of continuous withdrawal of arterial blood. Using a regression line, the integral of input function of arterial blood from 0 to 30 min was inferred by activity of arterial blood sampled at time t; A(t), by activity multiplied by its octanol extraction rate; AN(t), by activity of venous blood at time t; V(t), and by activity multiplied by its octanol extraction rate; VN(t). The optimum sampling time of arterial or venous blood was examined when mean % error for inference became minimum. Consequently, minimum error of AN(6 min) was 5.5%, A(3 min) was 8.9%, VN(6 min) was 5.9%, and V(20 min) was 10.0%. Quantitative measurement of rCBF using the value of VN(6 min) was performed on other 6 subjects with dementia etc. To validate the method, 133Xe inhalation SPECT studies were also performed on the same subjects. We found a good agreement between them (r = 0.851). The presented one-point sampling methods were simple and less invasive for quantifying rCBF.

Susceptibility to the development of asthma and other atopic diseases is known to be associated with genetic components, and several candidate genes have been reported to be linked to atopy in Caucasian populations. We conducted a study of linkage between asthma and markers on chromosomes 5q31-q33 and 11q13 in 68 Japanese families (306 members) by affected sib-pair analysis. Families for the linkage study were ascertained through asthmatic children visiting the allergy clinic. The results provide supportive evidence for linkage between asthma and gene markers in or near the interleukin-4 (IL-4) gene, the IL-9 gene, and D5S393 on chromosome 5q31-q33 (p = 0.0013, p = 0.018, and p = 0.0077, respectively). Linkage between atopic phenotype and these genetic markers was also suggested (p = 0.006, p = 0.01, and p < 0.0001 for atopy, respectively). However, we failed to find evidence for linkage of asthma or atopy to the IgE high-affinity receptor gene on 11q13 (p > 0.1). These findings indicate that beyond ethnicity, there are specific loci that contribute to susceptibility to atopy on chromosome 5q31-q33. In addition, our findings suggest that loci on chromosome 5q31-q33 are linked to the development of asthma in childhood.

Using Western blotting and RAST, this study examined IgE reactivities to Dermatophagoides farinae (Df) extract in three groups of children with either asthma, atopic dermatitis (AD) or a combination of both. The Df antigen presented at least nine distinct IgE-binding bands, with apparent molecular weights of 15, 25, 30, 40, 48, 59, 67, 80 and 110 kD. Positive IgE binding reactions to 15-kD proteins were found in more than 85% of subjects in all three groups, and positive reactions to 25-, 30- and 110-kD proteins in 35-45% in each group. The frequencies of positive IgE binding reaction to other components were generally low in all three groups. The pattern of IgE reactivity to Df components did not differ significantly between the three groups of subjects. IgE-binding activities to two major allergens, Der f1 and Der f2, were also examined by RAST, using recombinant antigens. High RAST reactivities to Der f1 and Der f2 were found in three groups, and the average RAST levels for Der f1 and Der f2 did not differ significantly between the three groups. These findings suggest that a difference in IgE responsiveness to house dust mite antigen is not a major discriminative factor in determining the difference between asthma and AD.

The purpose of the study is to validate the split-dose method corrected with dose ratio of 99mTc-ECD for brain perfusion scan. A dose of 600 MBq of 99mTc-ECD was divided into two with various dose ratios from 1:1 to 1:4, and injected to eleven patients with various cerebral diseases. A lesser dose of 99mTc-ECD was injected under a control state for the first SPECT scan, and 15 min SPECT scan was performed 10 min after injection with a triple-head high resolution gamma camera. After the scan, the other dose of 99mTc-ECD was injected under the same control state and the second SPECT scan was performed as same as above. A ratio of the activity of the first scan to the net activity of the second scan corrected by dose ratio, defined as K, was measured in brain regions of each subject. Expected value of K was 1, but the value was distributed with large variations in each subject. The mean % error of the K value was 10.4 +/- 4.9%. Hence it is considered that activity changes by more than 20% from the control values should be required to detect a significant rCBF change in an activation SPECT study. Then, we proposed a new method in which the activity of both two SPECT scans was normalized by cerebellar or occipital activity and compared. The ratio obtained by the proposed method came closer to 1 with less variations and with less mean % error in comparison with those of K value obtained by the dose-correction method. Although the proposed method has a limitation in the use of an activation study loaded with Diamox, it may be useful to evaluate an alteration of rCBF in the study such as postural testing or finger-moving test.

The purpose of this study was to comparatively evaluate the quality of SPECT images for the mapping of rCBF using three tracers, 123I-IMP, 99mTc-HMPAO and 99mTc-ECD. We performed three SPECT studies on seven patients with various cerebral diseases under the same conditions. An effect of Lassen's correction on SPECT images obtained by HMPAO was also evaluated. The same irregular regions of interest were placed on the four transaxial SPECT images. To quantitatively evaluate the pattern of tracer uptake and image contrast, the uptake ratio, regional count/mean count of the cerebrum, and its coefficient variations (CV) were defined, respectively. The order of the value of CV was HMPAO with correction > IMP > ECD > HMPAO without correction. HMPAO with correction showed the best image contrast, but HMPAO without correction was the worst. Uptake ratios of ECD and HMPAO with correction were decreased in the brain stem and thalamus in comparison with those of IMP. Both uptake ratios of ECD and HMPAO without correction were increased in the occipital cortex. IMP provides high quality SPECT images. Images obtained by HMPAO should be modified by Lassen's correction to increase image contrast. ECD or HMPAO should not be used to evaluate patients with spinocerebellar degeneration.

Although asthmatic patients are known to have increased levels of IgG antibody against house dust mite (HDM), it is not clear whether or not the presence of HDM-specific IgG antibody is associated with the etiological mechanism of asthma. To address this problem, we evaluated the relationship between HDM-specific IgG antibody levels and incidence of asthma in a general pediatric population. IgE and IgG antibody levels against Dermatophagoides farinae (Df) were examined by RAST and ELISA in a total of 722 randomly selected schoolchildren including 26 subjects with asthma, and the relative prevalence rates of asthma in this population were evaluated in relation to both Df-specific IgE and IgG levels. The incidence of asthma correlated not only with levels of Df-specific IgE, but also with those of Df-specific IgG. There was a significant correlation between Df-specific IgE and IgG levels both in the total population and in the asthmatic children. Because IgG and IgE responses occurred in parallel in this population, the clinical significance of HDM-specific IgG anti-body remains unclear. However, our findings have suggested that clinical expression of asthma in children is primarily dependent on their capacity to mount a immune response to HDM, which includes both IgE and IgG responses.

Although asthmatic children are often sensitized to the house-dust mite (HDM) during early childhood, it is not clear what antigenic components are associated with the early immune response of these children. To investigate this problem, we evaluated IgE, IgG, and IgG4 antibody responses to Dermatophagoides farinae (Df) by immunoblotting among three groups of children: group I aged 0-4 years, group II aged 5-9 years, and group III aged 10-14 years. In the group I subjects, positive IgE-binding reactions to 15- and 25-kDa components were found in 76% and 44% of sera, respectively. Those to other components were generally low in frequency. In contrast, positive IgG-binding reactions to 15- and 25-kDa components were found in 44% and 24% of sera, and those to 30- and 110-kDa components in 48% and 88% of sera, respectively. Positive IgG4 reactions to 15- and 25-kDa components were found in 48% and 24%, respectively, and those to other components were very low. Positive IgE-binding reactions to these components gradually became more frequent with increasing age in groups II and III, while IgG and IgG4 reactivities were not markedly different in these age groups. These results suggest that the 15- and 25-kDa proteins of Df are important antigens associated with the initial IgE response to HDM in early childhood, while the 30- and 110-kDa proteins are associated with IgG and 15-kDa components with IgG4.

A large volume of liquid nitrogen, (120-150 L) was applied to the houses of 4 asthmatic patients, and the mite densities of the floor dust from these houses were monitored every 2 wk for 4 mo from July to November 1989. The 1st liquid nitrogen treatment was applied in early August and decreased the mite densities to 20-44% of the pretreatment level in all cases, but they returned to the pretreatment level 3-4 wk later (on 3 occasions). The 2nd liquid nitrogen treatment was applied in early September and decreased the mite densities to 6-27% of the pretreatment level within 3 wk in all cases. However, mite numbers again increased to the pretreatment level 4 wk after treatment. The 3rd treatment was applied in mid-October and reduced the mite densities to < 6% of the pretreatment level within 2 wk, and mite numbers remained low thereafter. The results suggested that using liquid nitrogen to freeze houses reduces mite numbers, but that mite recolonization of the houses is an important problem.

IgE reactivities to two recombinant (r) allergens, rDer f1 and rDer f2, synthetized by cDNA clones from Dermatophagoides farinae (Df) were analyzed using sera of Df-sensitive asthmatic patients of varying ages. Positive RAST responses to rDer f1 and rDer f2 were found in 88 and 80%, respectively, with sera from young asthmatic children aged 0-1, positive RAST rates to these antigens increased to 100% up to 5 years of age. There was a significant correlation between RAST levels of rDer f1 and rDer f2 in these young children aged 0-1. IgE reactivities to four mutant proteins of rDer f2, which have at least one amino acid substitution, were similarly examined in the asthmatic sera. IgE reactivity to D7A, in which Asp7 was replaced by Ala, was reduced to 30-60% compared to the rDer f2. IgE binding to C8/119S, in which both Cys8 and Cys119 were replaced by Ser, lacking a disulfide bond between Cys8 and Cys119, was reduced almost to a background level at all ages. In contrast, A72L and A120L, in which Ala72 and Ala120 were substituted for Leu, respectively, almost retained the same IgE binding activity as the rDer f2 at all ages. These results suggest that Der f1 and Der f2 are important antigens associated with early sensitization to house dust mite in young asthmatic children. In addition, the disulfide bond between Cys8 and Cys119 and the N-terminal region including the 7th amino acid residue are considered to maintain IgE epitopes of the Der f2 allergen.

Background: The tsBN462 temperature-sensitive mutant hamster cell line exhibits cell cycle arrest and apoptosis at the restrictive temperature of 39.5 degrees C, due to a point mutation in the CCG1/TAFII250 gene, which encodes a component of the general transcription factor TFIID., Results: We now report that CCG1/TAFII250 persisted as a complex with TBP and associated proteins (TAFs) in tsBN462 cells at the restrictive temperature. FACScan analysis revealed that the tsBN462 mutation resulted in a failure to progress out of G0 into G1. Using two-dimensional gel electrophoresis we observed a decrease in the synthesis of several proteins, starting in the middle of the G1 phase, becoming very pronounced during late G1. The expression of the immediate early genes c-fos, c-jun and c-myc was normally induced by serum treatment of quiescent cells at the restrictive temperature, whereas expression of cyclins A, D1 and D3 was reduced. Expression of the cyclin-dependent kinase (CDK) inhibitor proteins p21 and p27 was enhanced. Consistent with the decreased cyclin D and increased p21/p27 expression, we found that phosphorylation of Rb was decreased at 39.5 degrees C. Cyclin A-, E- and Cdk2-associated histone H1 kinase activity was reduced concomitantly with the increase in p21 protein., Conclusion: Decreased cyclin/Cdk kinase activity and decreased Rb phosphorylation are possible causes of G1 cell cycle arrest in tsBN462 cells at the restrictive temperature.

We validated following five methods to quantitate regional cerebral blood flow (rCBF) using 123I-IMP and SPECT; 1) microsphere method, which is based on a microsphere model using the SPECT image at five minutes after 123I-IMP injection with continuous withdrawal of arterial blood, 2) microsphere+ one-point sampling method, which is the same as the microsphere method except for using one-point sampling instead of continuous withdrawal, 3) conventional microsphere + one-point sampling method, which is the same as the microsphere + one-point sampling method except for using a later SPECT image corrected with the ratio of alteration of measured entire brain activity, 4) Table look-up method, which is based on a two-compartment model (influx; K1 and outflux; k2) using one arterial blood sample taken at 10 min and two SPECT images at 30 min and 180 min post-injection, and 5) functional IMP SPECT, which is based on the two-compartment model using one arterial blood sample taken at 5 min and two SPECT images at 30 min and 60 min post-injection. Those methods were applied to six patients with cerebral infarction and degenerative diseases, and rCBF results were compared with those estimated by non-linear least squares fitting (NLLSF) analysis based on the two-compartment model. The rCBF values obtained by the microsphere method was best correlated with those by NLLSF analysis (r = 0.940), followed by the microsphere+one-point sampling method (r = 0.885) and the functional IMP SPECT (r = 0.882). The table look-up method underestimated rCBF especially at the high flow level, however showed good correlation (r = 0.859). The conventional microsphere + one-point sampling method overestimated rCBF, however showed good correlation (r = 0.849). Distribution volume (Vd = K1/k2) was also estimated by both the table look-up method and the functional IMP SPECT. While values of Vd by the functional IMP SPECT were significantly correlated with those by NLLSF analysis (r = 0.785), the table look-up method overestimated Vd (43.4 +/- 6.6 ml/g) and showed not good correlation (r = 0.287).

To measure distribution volume of 123I-IMP with one-point sampling of arterial blood and 2 times SPECT scans using the Magic square method (Rate constant square method), we introduced a program on a SPECT computer system. In this program four functional images; distribution volume, CBF (K1), k2 and delayed/early ratio, and absolute values with S.D. are obtained on the computer. We have been using the program on daily SPECT studies. In this study we discussed the usefulness of the method. Redistribution phenomenon of 123I-IMP, which is not exactly enough investigated, can be transformed into a functional parameter, distribution volume. Since distribution volume represents an extent of retention of 123I-IMP in brain tissue, it can play an important role to evaluate functional activity in the brain and to diagnose cerebral diseases.

We developed a new method (functional IMP SPECT) to quantitate both regional cerebral blood flow (rCBF) and distribution volume (Vd) using 123I-IMP and SPECT based on a 2- compartment model (influx; K1 =rCBF and outflux; k2). This method requires one arterial blood sampling at 5 min after injection of the tracer and two SPECT scans at 30 min and 60 min. It takes approximately 60 min for total measurements. The integration of arterial input function is estimated by one blood sample taken at 5 min after injection of the tracer without octanol treatment, by using the correlation between one blood sample and the integration, which was obtained from studies on 25 subjects. With the value of integration, K1 and Vd are calculated from the data of two SPECT scans by using the mathematical functions based on the 2-compartment model, which were obtained from studies on 12 subjects. In the practical range of K1 (0.3-0.7 ml/g/min) and Vd (15-35 ml/g) given in the 2-compartment model, statistical errors of K1 and Vd obtained by the functional IMP SPECT were evaluated at approximately 14.0% and 17.2%, respectively. In clinical studies for eight subjects including two healthy volunteers, three patients with cerebral infarction and three patients with degenerative disease, K1 and Vd values estimated by the functional IMP SPECT were significantly correlated with those estimated by non-linear least squares fitting analysis based on the 2-compartment model, and the mean errors of K1 and Vd estimated by the functional IMP SPECT were approximately 8.0% and 11.2% respectively, suggesting the validity of the new method. We conclude that the functional IMP SPECT is clinically useful because of the accuracy, less-invasiveness and convenience.

Acupuncture treatment has been used as physiotherapy to treat gastrointestinal disorders in traditional Oriental medicine. In the present study, we selected anesthetized animals as the subjects of this experiment in order to eliminate emotional factors, and investigated the responses to acupuncture stimulation at only a single point by changing animal conditions. 1) There was marked enhancement of gastric acid secretion by electroacupuncture stimulation of the hindlimb based on pH values of the perfusate. However, the enduring response after the momentary initial decrease suggested the existence of humoral factors. 2) The electroacupuncture-specific response was not observed in rats after sciatic nerve denervation. However, the pH values of the perfusate decreased in response to electroacupuncture stimulation of the hindlimb on the sciatic nerve intact side of the same rat. This suggests that the input pathway of acupuncture stimulation was a somatic nerve efferent pathway from the hindlimb. 3) No response to electroacupuncture stimulation was observed in the rats after vagotomy, but a clear enhancement response was seen after sympathectomy. These findings show that the major efferent path of the responses to acupuncture stimulation is the vagus nerve. The results of this experiment suggest that electroacupuncture stimulation of the hindlimb of anesthetized rats increases gastric acid secretion with somatic nerves as the afferent pathway and branches of the vagus nerve to the stomach as the efferent pathway.

To evaluate the accuracy of conventional microsphere method for the measurement of regional cerebral blood flow (rCBF) based on the microsphere model with N-isopropyl-p-[123I]iodoamphetamine (123I-IMP) and SPECT, we performed simulation analysis and clinical studies. Although the microsphere method requires early SPECT scan at a few minutes after injection of the tracer for the accurate measurement of rCBF, the conventional microsphere method, which is generally used, requires more delayed SPECT scan with long scan-duration. In the conventional microsphere method, the delayed SPECT image is corrected to the image at a few minutes after injection of the tracer by using the monitored entire brain activity. By the simulation analysis based on the 2-compartment model (influx; K1 and outflux; k2) using the input function and the entire brain activity obtained from eight subjects respectively, it was found that the conventional microsphere method overestimated the rCBF in the practical range of rCBF and Vd (= K1/k2) given in the 2-compartment model. When the values of rCBF and Vd in the 2-compartment model were given at 0.5 (ml/g/min) and 30 (ml/g) respectively, the rate of overestimation of rCBF by the conventional microsphere method was determined to be 17.3 +/- 0.7% (mean +/- S.D.). Also in clinical studies for eight subjects, the conventional microsphere method overestimated the rCBF compared with those evaluated by non-linear least squares fitting (NLLSF) analysis based on the 2-compartment model. Those results agreed well with the simulation analysis, suggesting the validity of the simulation. The rCBF values clinically estimated by the conventional method were, however, significantly correlated with those calculated by NLLSF analysis, and there were not so much difference between the two quantitative rCBF images obtained by the conventional microsphere method and the microsphere method. Therefore, we conclude that the conventional microsphere method is clinically useful in spite of the overestimation of rCBF.

To determine whether regional cerebral blood flow (rCBF) would change on standing in patients with idiopathic orthostatic hypotension (IOH), Tc-99m HMPAO SPECT studies were performed during postural testing in five patients with IOH. After 10 minutes of quiet rest on a bed, the patients arose quickly and, at the same time, the radiotracer was injected intravenously. SPECT data were obtained with a ring-type SPECT scanner. Another dose of Tc-99m HMPAO was injected with the subjects in the supine position, and SPECT was performed again. Image subtraction was used to evaluate the change in rCBF caused by postural testing. In all patients, the authors observed a decrease of rCBF ni the frontal cortex and basal ganglia. This preliminary study suggests that changes in rCBF occur in patients with IOH on standing, and Tc-99m HMPAO SPECT performed during postural testing may have an important role in evaluating these changes.

CCG1/TAFII250, the largest subunit of the TFIID complex, is mutated in ts cell cycle mutants of BHK21 cells, ts13 and tsBN462, which have a promoter-selective transcriptional defect. A series of deletion mutants of CCG1 cDNA were prepared and transfected into these mutants, in order to identify functional domains of CCG1 required for the complementation of ts 13/BN462 mutation. We determined the minimum size of CCG1:CCG1ME, essential for complementing the ts mutation, which possessed one proline cluster, an HMG1-like domain, and a nuclear localization signal, but which lacked the bromo domains and the acidic phosphorylation sites for casein kinase II common to transcriptional activators. It encodes a protein of 140 kDa. These characteristics of CCG1ME correspond to yeast TAFII145, the yeast homolog of human TAFII250. CCG1ME bound to TBP, creating its own TFIID complex different from that of the endogenous mutated CCG1 in ts+ transformants of tsBN462 cells.

We developed a new non-invasive technique; one-point sampling method, for quantitative measurement of regional cerebral blood flow (rCBF) with N-isopropyl-p-[123I]iodoamphetamine and SPECT. Although the continuous withdrawal of arterial blood and octanol treatment of the blood are required in the conventional microsphere method, the new technique does not require these two procedures. The total activity of 123I-IMP obtained by the continuous withdrawal of arterial blood is inferred by the activity of 123I-IMP obtained by the one point arterial sample using a regression line. To determine when one point sampling time was optimum for inferring integral input function of the continuous withdrawal and whether the treatment of sampled blood for octanol fraction was required, we examined a correlation between the total activity of arterial blood withdrawn from 0 to 5 min after the injection and the activity of one point sample obtained at time t, and calculated a regression line. As the results, the minimum % error for the inference using the regression line was obtained at 6 min after the 123I-IMP injection, moreover, the octanol treatment was not required. Then examining an effect on the values of rCBF when the sampling time was deviated from 6 min, we could correct the values in approximately 3% error when the sample was obtained at 6 +/- 1 min after the injection. The one-point sampling method provides accurate and relatively non-invasive measurement of rCBF without octanol extraction of arterial blood.

The effect of chest wall vibration on dyspnea at rest was investigated in 15 patients with severe chronic respiratory diseases. The magnitude of their baseline dyspnea was 17.9 +/- SE 3.3 mm on a 150-mm visual analog scale. One hundred hertz out-of-phase vibration (OPV; inspiratory intercostal muscles vibrated during expiration and expiratory intercostal muscles vibrated during inspiration) increased dyspnea an average of 21.9 +/- SE 7.8 mm (p < 0.05). One hundred hertz in-phase vibration (IPV; inspiratory intercostal muscles vibrated during inspiration and expiratory intercostal muscles vibrated during expiration) decreased dyspnea an average of 6.9 +/- SE 2.8 mm (p < 0.05). Changes in the respiratory pattern and arterial blood gas determinations elicited by 5-min IPV were investigated in eight of the 15 patients. Tidal volume was significantly increased in all eight of these patients. Respiratory frequency was decreased in seven patients and increased in one. Functional residual capacity, measured before and during the application of IPV for a period of about 10 breaths, was increased in one patient and decreased in the remaining seven. PaCO2 decreased by 1.3 +/- 1.0 mm Hg (p < 0.05), from 49.6 +/- 8.4 mm Hg; PaO2 increased by 3.4 +/- 2.3 mm Hg (p < 0.05), from 67.7 +/- 12.7 mm Hg. The results indicate that in-phase chest wall vibration decreased pathologic dyspnea in patients with chronic respiratory disease and suggest that the effects on respiratory sensation are mediated by afferent information from chest wall respiratory muscles to supraspinal centers.

Regional cerebral blood flow (rCBF) was measured in 13 patients with various cerebral disease by N-isopropyl-p-[123I]iodoamphetamine (IMP) SPECT with the microsphere model (rCBF (CbST)), which was compared with 133Xe inhalation SPECT (rCBFXe). Continuous withdrawal of arterial blood was performed for 5 minutes and SPECT image was obtained from 25 min to 55 min after the tracer injection. The reconstructed counts at 5 min was corrected using entire cerebral counts collected at 5 min, 20 min, and 60 min after the tracer injection. As compared with rCBFXe, rCBF (CbST) was underestimated in high flow areas and overestimated in low flow areas. Regional CBF was also measured using one minute short time SPECT images at 5 min (rCBF (Cb5)), 20 min (rCBF (Cb20)), and 60 min (rCBF (Cb60)) after the injection. Regional CBF tended to be underestimated with time from the tracer injection due to the wash-out from brain tissue to blood. In comparison with rCBF (Cb5), rCBF (CbST) was underestimated in high flow areas and overestimated in low flow areas. This error was considered to be due to the correction of reconstruct counts using entire cerebral counts, because of the kinetic behavior of 123I-IMP was different in each region of the brain.

The role of the conserved Asp134 residue in Escherichia coli ribonuclease HI, which is located at the center of the alpha V helix and lies close to the active site, was analyzed by means of site-directed random mutagenesis. Mutant rnhA genes encoding proteins with ribonuclease H activities were screened by their ability to suppress the ribonuclease-H-dependent, temperature-sensitive growth phenotype of E. coli strain MIC3001. Based on the DNA sequences, nine mutant proteins were predicted to have ribonuclease H activity in vivo. All of these mutant proteins were purified to homogeneity and examined for enzymic activity and protein stability. Among them, only the mutant proteins [D134H]RNase H and [D134N]RNase H were shown to have considerable ribonuclease H activities. Determination of the kinetic parameters revealed that replacement of Asp134 by amino acid residues other than asparagine and histidine dramatically decreased the enzymic activity without seriously affecting the substrate binding. Determination of the CD spectra indicated that none of the mutations seriously affected secondary and tertiary structure. The protein stability was determined from the thermal denaturation curves. All mutant proteins were more stable than the wild-type protein. Such stabilization effects would be a result of a reduction in the negative charge repulsion between Asp134 and the active-site residues, and/or an enhancement of the stability of the alpha V helix. These results strongly suggest that Asp134 does not contribute to the maintenance of the molecular architecture but the carboxyl oxygen at its delta 1 position impacts catalysis.

To evaluate the regional cerebral blood flow measurement with N-isopropyl-p-[123I]iodoamphetamine (IMP), we performed 123I-IMP SPECT and 133Xe inhalation method on 9 patients with mild cerebrovascular diseases and so on. A tracer dose of 123I-IMP (111 MBq) was injected i.v. and the data were obtained 15 min later. Regional CBF was measured by the microsphere model and arterial blood sampling method. However, mean cerebral blood flow values was not in agreement with both methods. We supposed the reason of the disagreement was due to the difference of PaCO2 when the studies were performed. Hence, CBF measurement with 123I-IMP was performed on the other patient's population with loading of hyperventilation before the IMP injection. Comparing with the control state with normal breathing, we calculated the rate of change of mean CBF between the state with normal breathing and the state with hyperventilation that was 1.02 ml/100 g/min/mmHg PaCO2. After the correction with the rate, we observed a much significant relation between the mean CBF measured with 123I-IMP SPECT and 133Xe inhalation method. When compared CBF values with loading studies or certain therapies, the correction of CBF with the rate of change of PaCO2 is one of the very important factors to be considered. Moreover, 133Xe inhalation method without the correction of PaCO2 can not be a golden standard method for rCBF measurement.

The effects of vehicle and enhancer on the skin permeation of tegafur across excised hairless mouse skin were investigated. Ethanol, water, panasate 800 (tricaprylin), and their mixtures were used as vehicles, and then a series of fatty acids or fatty alcohols was employed as representative enhancers. The skin permeability of tegafur was dramatically increased by the combination of ethanol and panasate 800, whereas the combination of ethanol and water resulted in only a little increase. The greatest permeation ratio (56.8% of dose at 12 h) was obtained with the ethanol/panasate 800 (40/60) binary vehicle. Furthermore, the skin permeability of tegafur could be enhanced by the addition of fatty acids or fatty alcohols to ethanol/panasate 800 (40/60) and ethanol/water (60/40) binary vehicles. The infinite dose studies suggested that panasate 800 decreased the lag time by increasing the diffusion of tegafur, and ethanol increased the permeation rate by increasing the partition of tegafur into skin. The effects of fatty acids or fatty alcohols added to ethanol as a single vehicle were dramatic, but did not appear in the case of panasate 800 as a single vehicle. In conclusion, panasate 800 was an excellent hydrophobic vehicle in that it minimized time lag and promoted tegafur permeability combined with ethanol. This improvement was further promoted when enhancers were added.

We developed a new graphic method using N-isopropyl-p-[123I]iodoamphetamine (IMP) and SPECT of the brain, the graph on which all three parameters, cerebral blood flow, distribution volume (Vd) and delayed count to early count ratio (Delayed/Early ratio), were able to be evaluated simultaneously. The kinetics of 123I-IMP in the brain was analyzed by a 2-compartment model, and a standard input function was prepared by averaging the time activity curves of 123I-IMP in arterial blood on 6 patients with small cerebral infarction etc. including 2 normal controls. Being applied this method to the differential diagnosis between Parkinson's disease and progressive supranuclear palsy, we were able to differentiate both with a glance, because the distribution volume of the frontal lobe significantly decreased in Parkinson's disease (Mean +/- SD; 26 +/- 6 ml/g). This method was clinically useful. We think that the distribution volume of 123I-IMP may reflect its retention mechanism in the brain, and the values are related to amine, especially to dopamine receptors and its metabolism.

Examination by thoracoscopy, using flexible fiberoptic bronchoscope, was performed in 43 patients with pleural effusion according to our reported method. In these patients, 14 cases were diagnosed as tuberculous pleuritis. These 14 cases were investigated with respect to clinical and thoracoscopic findings. Their mean age was 38.1 years, and 11 cases were younger than 50 years old (78.5%). The male:female ratio was 2/1. Clinical symptoms recognised were fever (100%), cough (64.2%), chest oppression (50%) and sputum (35.7%). In 5 cases, gastrointestinal symptoms were recognized such as vomiting, abdominal pain, and diarrhea. Tuberculin reaction was positive in all patients with tuberculous pleuritis on admission. With respect to the thoracoscopic findings of tuberculous pleuritis, yellow-white miliary granulomas were observed on the parietal pleura in 12 cases (85.7%), and this characteristic finding was especially observed at the apex. Biopsy specimens, obtained from these miliary granulomas on the parietal pleura showed histological findings of tubercles. In the other 2 cases, generalized reddening of the entire parietal pleura was observed, with no yellow-white miliary granulomas. Biopsy specimens obtained from the reddened lesions on the parietal pleura showed histological findings of tubercles, and these 2 cases were also diagnosed as tuberculous pleuritis. These results indicate that this method may be very useful for the diagnosis of tuberculous pleuritis in patients with pleural effusion.

Crossed cerebellar diaschisis (CCD) is interpreted as a functional deactivation, presumably caused by a loss of excitatory or inhibitory afferent inputs on the corticopontocerebellar pathway and others. A redistribution phenomenon (RD) is usually observed in the contralateral cerebellum with CCD on delayed images of 123I-IMP SPECT. This phenomenon was analyzed in a view point of rCBF measurement in 24 patients with brain tumor, infarction and so forth. Regional CBF was measured by the microsphere method with 123I-IMP and a delayed-to-early counts ratio (D/E ratio) was used. As a result, there was no relation between rCBF and the D/E ratio in the cerebellum, which means that RD is occurred by other factors except for rCBF in the cerebellum. Regional CBF and the D/E ratio in the contralateral and ipsilateral cerebellum was 46.3 ml/100 g/min, 1.01 and 57.0 ml/100 g/min, 0.86, respectively. These results mean that the high activity of IMP gradually decreased in the ipsilateral cerebellum, while, the low activity in the contralateral cerebellum was almost stable, and the difference of both activity reduced after 5 hours and RD was observed on the delayed image. The data indicate that retention mechanism of IMP and vascular permeability are not affected in the cerebellum with CCD.

A case of pulmonary arteriovenous fistulas (PAVF) treated with detachable balloon embolotherapy is reported. A 31-year-old man was admitted to hospital because of polycythemia and hypoxemia. He had a history of dyspnea on exertion and transient cerebral ischemic attacks (TIA). Chest roentgenogram revealed a peripheral lobulated lesion in the right upper lung field and a coin lesion in the left lung field connected by dilated blood vessels to each hilus. Pulmonary angiography and right heart catheterization was performed. Four PAVFs were observed in bilateral lung field, and the intrapulmonary shunt ratio was 25.8%. Because the patient was symptomatic, we embolized each PAVF with a detachable balloon without complications. After embolotherapy, there was improvement in his symptoms. hypoxemia and shunt ratio. Recent reviews have highlighted the need to consider embolotherapy for bilateral multiple PAVF. We conclude that embolotherapy with detachable balloon is a very useful procedure for treatment of bilateral multiple PAVF.

The measurement of serum SLX is thought to be a useful aid in the diagnosis of malignant diseases, particularly adenocarcinoma of the lung. In the present investigation, we measured and compared SLX values in BALF from affected and normal bronchi, obtained from 83 patients. They consisted of 64 males and 19 females, with mean age of 60 years, consisting of 8 normal controls, 19 cases of benign lung disease, and 56 cases of primary lung cancer. SLX value in BALF from normal bronchi was significantly higher in patients with lung cancer than in normal controls, but there was no significant difference in SLX value between lung cancer and benign lung disease. On the other hand, SLX value from affected bronchi was significantly higher in patients with lung cancer than in normal controls and patients with benign lung disease. The rate of elevated SLX in BALF from affected bronchi was significantly higher in patients with lung cancer than in those with benign lung disease. These results suggest that measurement of SLX levels in BALF from affected bronchi may be a useful method for differential diagnosis of primary lung cancer.

During a sparrow-pecking and twisting-needle manipulation to the acupoints BL 23, 24 and 25 for an induction of "Qi", it was found that some transparent materials were binding to the needles after removed from the volunteer's skin. Electron-microscopical analysis of the transparent materials revealed that they corresponded to the injured fascia made up of collagen fibers, elastic fibers, fibroblasts, adipocytes and mast cells. Rarely were nerve fiber-like structures observed in the materials. Immunohistochemically, calcitonin gene-related peptide-positive nerve fibers could be demonstrated in the acupoint BL 24 associated fascia. A possible functional relationship between the needle manipulation and the induction of Qi-sensation is discussed along with the acupoint tissue constitution.

A 66-year-old woman visited our hospital for further investigation of a coin-shaped lesion in the right lower lung field. After admission, CT-scan revealed an anterior mediastinal tumor. Thymoma and lung metastasis from thymoma were suspected. At thoracotomy, an invasive thymoma involving the left phrenic nerve, the left upper lobe, and the pericardium was found, and the diagnosis of lung metastasis was made. Postoperative pathological findings showed a predominantly lymphocytic thymoma with round and oval-shaped epithelial cells, and the metastatic lung lesion revealed the same findings. Postoperative radiotherapy (4000 rads) and chemotherapy with a combination of CDDP, VDS, and CPA were administered. The metastasis of thymoma is not rare, but most often it is found after operation. In this case the diagnosis of metastatic thymoma was made pre-operatively from a solitary lung lesion.

Broncasma Berna (B.B.) prepared from killed polyvalent bacterial vaccine is used as a pleural irritant for pleurodesis on patients with spontaneous pneumothorax. The effects of B.B. on the complement system in vivo after injections into pleural spaces were investigated. Five patients with spontaneous pneumothorax underwent intrapleural injections of B.B. Increased body temperatures with positive C-reactive proteins and increased polymorphonuclear leukocyte counts in peripheral blood were observed in all patients. Serum levels of complement components (Clq, Cls, C4, C3 and B), hemolytic activity of complement (CH50) and plasma levels of C3a were gradually elevated after intrapleural injections of B.B. Analysis of pleural fluids from 4 patients showed that levels of C3a and C5a were increased compared with plasma levels, and an accumulation of polymorphonuclear leukocytes were observed. These findings suggest that intrapleural injections of B.B. activate the complement system and cause inflammatory reactions accompanied by an increase of C3a and C5a and an accumulation of polymorphonuclear leukocytes. Such inflammatory reactions might inversely induce the synthesis of complement components and promote the migration of polymorphonuclear leukocytes from bone marrow. The deposit of fibrin during these inflammatory processes might contribute to the development of pleurodesis. These inflammatory reactions might be mediated through the activation of the classical complement pathway by B.B.

In order to determine the mechanism of bronchoconstriction induced by the inhalation of adenosine in asthmatics, we measured, by the astograph method, cyclic nucleotides, leukotrienes and thromboxane B2 before and after the inhalation, and we studied the relationship between serum theophylline concentration and Rrs cont, Dmin and S Grs/Grs cont of the astogram obtained by inhaled adenosine. Plasma TXB2 level was significantly raised (p less than 0.05) after inhalation, but cAMP, cGMP, LTB4 and LTC4 did not change. In the astogram, Dmin was significantly elevated (p less than 0.01) when plasma theophylline concentration was raised to the therapeutic level, but Rrs cont and S Grs/Grs cont showed no remarkable change. We confirmed that inhaled adenosine caused bronchoconstriction in asthmatic patients, and that Dmin was elevated after the administration of theophylline. These results suggest that TXB2, a metabolic product of arachidonic acid, may be one of the most important mediators in the bronchoconstriction mechanism of inhaled adenosine.

A 25-year-old man was admitted to our hospital because of hemosputum and abnormal shadows on chest X-ray film. The chest film showed a mass shadow located in the right hilum and multiple coin lesions in bilateral lung fields. The chest CT scan showed a bulky mass sized 7 cm X 6 cm at the anterior mediastinum. This case was suspected to be choriocarcinoma because he was young, it was rapidly progressive and also because serum LDH and beta-HCG values were raised. The specimens obtained by surgical biopsy showed HCG-stain positive syncytiotrophoblasts. Cancer chemotherapy was started with CDDP and other anti-cancer drugs, but he succumbed about 3 months after admission. Autopsy findings showed no scar formation of his testes, and the primary lesion was finally diagnosed to be in the anterior mediastinum. Extra-gonadal choriocarcinoma is rare, and beta-HCG is very useful for its diagnosis and the follow-up of the clinical course.

The developmental changes in monoamine oxidase (MAO) activities towards 5-hydroxytryptamine (5-HT) and beta-phenylethylamine (PEA) in chick retina were investigated. The whole pattern of the developmental changes in MAO activity towards 5-HT was similar to that towards PEA, even though the activity towards 5-HT was much lower than that towards PEA during the period from the 12th day of incubation to the 14th day after hatching. The MAO activities on the 12th day of incubation were low and increased remarkably until the 18th day of incubation. Then they remained unchanged until the 7th day after hatching; therefore, they decreased. The activities measured on the 14th day after hatching are on the same level as those of an adult. The multiplicity of MAO in the retinae of a chick on the 12th day of incubation and of an adult was also studied with specific inhibitors. The result showed that the major part of MAO in chick retina is type B enzyme; that type A MAO in the retina of an adult is, however, relatively higher than that in the retina of the embryo on the 12th day of incubation. It was found that an appreciable amount of MAO activity towards 5-HT in chick retina is due to type B MAO.

A simple fluorometric assay for monoamine oxidase (MAO) [EC 1.4.3.4] activity towards beta-phenylethylamine (PEA) was devised. The procedure consists in measuring the disappearance of PEA fluorometrically. The disappearance of PEA was completely inhibited by pargyline, a potent inhibitor of MAO. MAO activity for PEA was linear with 10 mg to 100 mg of liver tissue in 3 ml of reaction mixture for up to 90 min of incubation. Using this method, the V max values and the apparent Km values of MAO for PEA in several rat tissues were determined, and compared with those for benzylamine and 5-hydroxytryptamine (5-HT).