An in vivo screening system to identify tumorigenic genes.

Screening for oncogenes has mostly been performed by in vitro transformation assays. However, some oncogenes might not exhibit their transforming activities in vitro unless putative essential factors from in vivo microenvironments are adequately supplied. Here, we have developed an in vivo screening system that evaluates the tumorigenicity of target genes. This system uses a retroviral high-efficiency gene transfer technique, a large collection of human cDNA clones corresponding to ~70% of human genes and a luciferase-expressing immortalized mouse mammary epithelial cell line (NMuMG-luc). From 845 genes that were highly expressed in human breast cancer cell lines, we focused on 205 genes encoding membrane proteins and/or kinases as that had the greater possibility of being oncogenes or drug targets. The 205 genes were divided into five subgroups, each containing 34-43 genes, and then introduced them into NMuMG-luc cells. These cells were subcutaneously injected into nude mice and monitored for tumor development by in vivo imaging. Tumors were observed in three subgroups. Using DNA microarray analyses and individual tumorigenic assays, we found that three genes, ADORA2B, PRKACB and LPAR3, were tumorigenic. ADORA2B and LPAR3 encode G-protein-coupled receptors and PRKACB encodes a protein kinase A catalytic subunit. Cells overexpressing ADORA2B, LPAR3 or PRKACB did not show transforming phenotypes in vitro, suggesting that transformation by these genes requires in vivo microenvironments. In addition, several clinical data sets, including one for breast cancer, showed that the expression of these genes correlated with lower overall survival rate.

Enhancement of hepatocarcinogenesis by sorbitan fatty acid ester, a liver pyruvate kinase activity-reducing substance.

Effect on hepatocarcinogenesis of dietary sorbitan fatty acid ester (SorFAE), which had been known to cause decrease in pyruvate kinase (PK) activity, was studied in rats fed a diet containing 0.06% 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) for 6 weeks. The incidence of hyperplastic nodules and/or hepatocellular carcinomas in the rats fed the 3'-Me-DAB diet alone was 45.0% at the end of 51 weeks, whereas the incidence in the rats fed 3'-Me-DAB diet followed by 5 or 10% SorFAE or 0.1% phenobarbital (PB) diet were 76.2, 90.5, and 95.0%, respectively. These incidences were significantly higher compared with the group fed 3'-Me-DAB diet alone (P less than .05). No tumors were observed in rats fed 10% SorFAE diet alone. The results show that SorFAE has an enhancing effect on hepatocarcinogenesis, although the effect was weak compared to that of the effective PB dose. The results seem to confirm our assumption that a chemical that causes decrease in PK activity in rat liver might promote hepatocarcinogenesis.

Segmental anhidrosis in the spinal dermatomes in Sjögren's syndrome-associated neuropathy.

We describe two women with primary Sjögren's syndrome and sensory neuropathy who had anhidrosis segmentally along the dermatomes of the spinal segment, along with sensory loss. Intradermal administration of cholinergic agents elicited no sweat response in the spinal segments with anhidrosis, whereas a normal response was present in the segments with obvious sweating. These features suggest segmental involvement of the postganglionic sympathetic ganglion cells.

Clinicopathological features of chronic inflammatory demyelinating polyradiculoneuropathy in childhood.

The clinical, electrophysiological, and pathological findings, and the therapeutic characteristics in ten children with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), whose onset age was less than 16 years, were evaluated. The clinical progression pattern of the initial phase divided the patients into two groups. One group (six patients) showed a subacute progression for up to 2 months after onset and a subsided progression over 3 months. Three patients in this group had a preceding febrile episode. The other group (four patients) showed a chronic insidious progression for more than 3 months. The former group of patients revealed a favourable response to corticosteroid therapy as compared with the latter group. However, other clinical and laboratory features at the peak impairment were not distinguishable between these two groups. Motor dominant neuropathy was common to all patients, and only three cases showed sensory disturbance on the distal limbs. No cases revealed cranial nerve involvement. Motor and sensory nerve conduction and sural nerve biopsy studies revealed the demyelinating nature of the neuropathy. These clinicopathological features suggest that the subacute progression form frequently associated with prodromal episode and rather favourable corticosteroid response is characteristic in childhood CIDP, while the chronic insidious progression form is indistinguishable from the common adult CIDP.

Change in the flavor of black tea drink during heat processing.

Heat processing during canning is responsible for the change in flavor of black tea infusion. The quantitative change in the volatile components of the black tea infusion during heat processing is not sufficient for explaining the sensory evaluation. In this study, application of aroma extract dilution analysis using the volatile fraction before and after black tea (Darjeeling) samples were heat processed resulted in the detection of 10 odor-active peaks for which flavor dilution (FD) factors changed. Seven potent odorants were identified from these peaks by gas chromatography-mass spectrometry. Among these components, 3-methylbutanal (stimulus), methional (potato-like), beta-damascenone (sweet), dimethyl trisulfide (putrid), and 2-methoxy-4-vinylphenol (clove-like) showed the highest FD factors after heat processing of the black tea sample. Therefore, these odorants were the most important components involved in changing the black tea odor during heat processing. In addition, the precursor of beta-damascenone in black tea infusion was investigated, and 3-hydroxy-7,8-didehydro-beta-ionol was determined to be one of the beta-damascenone-generating compounds for the first time.

Identification of potent odorants in Japanese green tea (Sen-cha).

Application of aroma extract dilution analysis using the volatile fraction of a Japanese green tea (Sen-cha) sample resulted in the detection of 36 odor-active peaks with flavor dilution (FD) factors between 10 and 5000. Thirty-six potent odorants were identified from 36 odor-active peaks by gas chromatography/mass spectrometry (GC/MS) and/or the multidimensional GC/MS (MDGC/MS) system. Among these components, 4-methoxy-2-methyl-2-butanethiol (meaty), (Z)-1, 5-octadien-3-one (metallic), 4-mercapto-4-methyl-2-pentanone (meaty), (E,E)-2,4-decadienal (fatty), beta-damascone (honey-like), beta-damascenone (honey-like), (Z)-methyl jasmonate (floral), and indole (animal-like) showed the highest FD factors. Therefore, these odorants were the most important components of the Japanese green tea odor. In addition, 4-methoxy-2-methyl-2-butanethiol, 4-mercapto-4-methyl-2-pentanone, methional, 2-ethyl-3, 5-dimethylpyrazine, (Z)-4-decenal, beta-damascone, maltol, 5-octanolide, 2-methoxy-4-vinylphenol, and 2-aminoacetophenone were newly identified compounds in the green tea.

Plasma exchange in chronic inflammatory demyelinating polyradiculoneuropathy; different beneficial effects and their correlation to the clinical features.

We report 7 patients with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) who showed significant clinical improvement by plasma exchange (PE). Their clinical features were extensively varied but fulfilled the diagnostic criteria for CIDP. The patients could be divided into 2 subgroups based on the mode of effectiveness of PE; one group consisted of those who once achieved complete remission, but required highly frequent PEs for long-term maintenance of remission and the others were those who showed persistent and complete remission with several sessions of PE alone. These findings raise issues about the long-term therapeutic goals to be achieved by PE for CIDP patients, particularly on supportive therapy other than PE, as well as the optimization of frequency and duration of PE.

[Postganglionic sudomotor function in multiple system atrophy].

Postganglionic sudomotor function were examined on 12 patients with multiple system atrophy (MSA) consisting of 5 males and 7 females with the clinical duration of 1 to 10 years, and with the age ranged 51 and 70 years. Sixteen healthy volunteers, aged 38 to 75 years were also examined as the control. Local sweating induced by intradermal injection of pilocarpine and nicotine (a concentration of 10(-4) g/ml) was quantitatively measured on the volar surface of the forearm and lower lateral leg using a ventilated capsule method in a climatic chamber at 23 degrees C and 40% of relative humidity. Maximal sweat rate induced by nicotine and pilocarpine was significantly reduced in patients with MSA as compared with controls in both the forearm and lower lateral leg. MSA cases associated with more prominent autonomic dysfunction as well as hyposweating, showed a more remarkable impairment of local sweat responses. Particularly, in 6 cases with Shy-Drager syndrome, there was no sweat response by the injection of both pilocarpine and nicotine. The study of an autopsied case with Shy-Drager syndrome revealed neurons in the para-vertebral sympathetic ganglia were well populated, though neurons in the lateral horns of the lower thoracic spinal cord were almost completely depleted. This substantial discrepancy between the impaired sudomotor function and morphological findings may imply several hypothetical views on the mode of pathology of postganglionic sudomotor nerves. The present results, however, strongly suggested that postganglionic sudomotor functions are more extensively involved in patients with MSA than had ever been believed.(ABSTRACT TRUNCATED AT 250 WORDS)

[Migrating multiple mononeuritis and nonsystemic angitis].

Multiple mononeuritis of migrating nature at onset occurs in a variety of disease conditions. "Migrant sensory neuritis" without a systemic underlying disorder described by Wartenberg (1958) is the most distinctive form of this type of neuropathy. Its pathomechanism is not uniform, but angiopathy has been suggested by Matthews et al. (1981). In this report, we describe five cases of sensory-dominant multiple mononeuritis with migrating nature and without systemic visceral involvement. The patients consisted of four females and one male between 26 and 71 years of age. All showed recurrent episodes of sensory involvement along the distal branches of the cutaneous nerve. The patients presented with sudden onset of numbness or pain radiating along the cutaneous branches of the involved nerves often followed by persistent sensory deficit. In mild episodes, the sensory symptom with numbness almost completely subsided. The frequency of these episodes ranged from 7 times in 6 months to 6 times in 9 years. The clinical manifestations were similar or identical to those of migrant sensory neuritis reported by Watenberg and Matthews et al. Cranial nerve involvement and less frequent episodes in the present series differed from those in the previous reports. Laboratory examinations did not disclose any underlying disorder, suggesting systemic collagen vascular disease. Sural nerve biopsy study in two patients revealed vasculitis in small or medium-sized arteries in the epineurium as well as reduced population of large myelinated fibers. Small myelinated fibers were slightly increased in number, probably to regenerating fibers. Sensory action potentials were low or not evoked in the nerves examined.(ABSTRACT TRUNCATED AT 250 WORDS)

[Local effect of calcitonin gene-related peptide on human sweat gland function].

The effect of local administration of calcitonin gene-related peptide (CGRP) on sweating activity was evaluated on normal human volunteers. CGRP and methacholine chloride (MCH) was dissolved in 0.1 ml of 0.9% NaCl solution to a specified concentration, and was injected intradermally at the center of a 1.3 cm2 forearm test area. The sweat rate was recorded continuously by capacitance hygrometry in a relatively cool environment (Ta, 23 degrees C). CGRP did not elicit any sweat secretion when administrated by alone, but significantly increased the sweat rate when it was administrated with MCH. The maximum enhancement of MCH-induced sweating by CGRP was observed at a concentration of 10(-5) g/ml of CGRP. There was clear dose-dependent relationship between the dose of CGRP and its enhancement. Recently, CGRP-like immunoreactivity is demonstrated to be present in cholinergic nerve terminals around the human sweat glands. These observations have strongly suggested that CGRP enhances the cholinergic sweating activity. Although the underlying mechanism is still obscure, CGRP may enhance the sweating as a consequence of vasodilation which has been known to be a major activity of CGRP. As for the evaluation of human sweat gland function, CGRP-induced peptidergic regulation should be considered as well as cholinergic regulation.

[Peptidergic sweating in multiple system atrophy and hereditary spinocerebellar ataxia with special reference to calcitonin gene-related peptide].

Calcitonin gene-related peptide (CGRP)-like immunoreactivity has been demonstrated to be present in nerve terminals around the sweat glands. We have previously demonstrated that CGRP modulates the cholinergic sweating activity. The present study was designed to evaluate the locally administrated CGRP on sweating of patients with multiple system atrophy (MSA) and hereditary spinocerebellar ataxia (HSCA) consisting of 10 males and 11 females. Among 9 HSCA, 3 was diagnosed as Machado-Joseph disease by clinical and DNA diagnostic assessment. CGRP and methacholine chloride (MCH) was dissolved in saline solution to specified concentration, and 0.1ml of which was injected intradermally at the center of a forearm test area. The sweat rate was recorded continuously by the capacitance hygrometry in a relatively cool environment in which the spontaneous sweating was absent. CGRP significantly increased the sweat rate when it was administrated with MCH on normal subjects. There was a clear dose-dependent relationship between the dose of CGRP and its enhancement. This enhancement was significantly reduced in patients with MSA as compared with controls. On the other hand, most of HSCA showed no remarkable impairment of CGRP enhancement. These results suggest that peptidergic sweating is extensively affected in patients with MSA but is not in patients with HSCA, which may be the consequence of peptidergic sweating dysfunction in MSA.

[Chronic recurrent-progressive polyradiculoneuritis--Characteristics of autonomic dysfunction].

We examined autonomic dysfunctions in 12 patients with chronic-recurrent-progressive polyradiculoneuritis (C-R-P-PRN), consisting of 6 males and 6 females with the clinical duration of 8 months to 16 years, and with the age ranged 13 and 71 years. Sixteen healthy volunteers, aged 20 to 70 years, were also examined. Thermal sweat rate was recorded on the palm, forearm, upper arm, anterior chest, lateral thigh and lateral aspect of lower leg using a ventilated capsular method in a climatic chamber at 40 degrees C and 40% of relative humidity. After steady state was attained, thermal sweat rate was measured. Local sweating induced by intradermal injection of pilocarpine and nicotine (a concentration of 10(-4), 0.1 ml) was also measured on the forearm and lower lateral leg at 23 degrees C of room temperature and 40% of relative humidity. Other autonomic functions including skin temperature at rest and after cold loading (15 degrees C, 6 minutes), variation in the R-R interval (CV%), pupillary function (response to 1.25% epinephrine, 2(-5) pilocarpine, 5% tyramine), orthostatic hypotension and bladder dysfunction were also monitored. A decrease in sweat rate and recovery rate of skin temperature was seen more frequently in patients with C-R-P-PRN than normal volunteers. Abnormality in the thermal sweat rate and local sweat response against nicotine and pilocarpine was present more frequently in the forearm and distal leg as compared with the chest and thigh.(ABSTRACT TRUNCATED AT 250 WORDS)

[Autonomic dysfunction in vasculitic neuropathy--special reference to sudomotor function].

We examined autonomic functions in 14 patients with peripheral neuropathy caused by necrotizing vasculitis. These patients consisted of three allergic granulomatous angitis (Churg-Strauss syndrome, AGA), two systemic lupus erythematosus (SLE), two progressive systemic sclerosis (PSS), one mixed connective tissue disease (MCTD), one polyarteritis nodosa (PN) and five nonsystemic vasculitis. All of them were proven to have a vasculitis by sural nerve, muscle or skin biopsy. Sixteen age- and sex-matched healthy volunteers were also examined. Local sweating induced by intradermal injection of pilocarpine and nicotine (a concentration of 10(-4) g/dl, 0.1 ml) was measured with ventilated capsular method on the forearm and lower lateral leg at 23 degrees C of room temperature and 40% of relative humidity. Other autonomic functions including skin temperature at rest and after cold loading (15 degrees C, 6 minutes), variation in the R-R interval of heart beat (CV%), orthostatic hypotension and bladder dysfunction were also monitored. A decrease in sweat rate and recovery rate of skin temperature after cold-loading was seen more frequently in patients with necrotizing vasculitis than normal volunteers. Abnormality in the local sweat response against nicotine and pilocarpine was more frequently present in the involved area of somato-sensory and motor nerves as compared with those in the non-involved area. An occurrence of decrease in recovery rate of skin temperature after cold-loading also well correlated to the region of somato-sensory- and motor involvement. So far other autonomic dysfunction, only one patient had orthostatic hypotension, impotence and bladder dysfunction.(ABSTRACT TRUNCATED AT 250 WORDS)

[Two cases of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) associated with Graves' disease].

We have reported two cases of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) associated with Graves' disease. Case 1: a 45-year-old woman noticed a diffuse goiter, palpitation and emaciation in 1977. Laboratory studies confirmed that she had Graves' disease, and she was treated with antithyroid drug. In 1986, when the hyperthyroidism was subsided, she showed progressive symmetrical weakness and numbness in her limbs, and she was almost in tetraplegia at 1987. Markedly slowed motor and sensory nerve conductions and elevated CSF proteins as well as clinical manifestations confirmed the diagnosis of CIDP. Following corticosteroid-pulse therapy and plasmapheresis resulted in good recovery in both motor and sensory impairment, though two-times of relapses were observed. Case 2: a 33-year-old man first noticed weakness in his legs in 1977, motor and sensory disturbances progressed for 12 years. Slowed nerve conduction, high CSF proteins and two-times of relapses in early phase indicated that the CIDP was the diagnosis. In 1989 he complained general fatigue, hyperhidrosis and body-weight loss. The serum thyroid hormone levels were high, and other laboratory studies confirmed the presence of Graves' disease. The cases with both CIDP and Graves' disease has rarely been reported. The background mechanism of this association is not well understood, but the susceptibility to CIDP and Graves' disease may be related to the HLA antigens and immunoglobulin Gm allotypes of which are the genes linked to the major histocompatibility complex and controlling immune responses. The present two cases commonly shared several HLA-DR antigens, but their significance should be confirmed by examining many cases.

[Double filtration plasmapheresis (DFPP) in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP)].

We studied the therapeutic characteristics of double filtration plasmapheresis (DFPP) in 14 patients with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). The patients were classified into 2 subgroups of the responders (7 patients) and the non-responders (7 patients) to DFPP. The responders to DFPP were designated as those showing the improvement 2 or more grades in measures the activity of daily living by the modified Rankin scale (MRS). All these patients underwent neurological assessment, CSF study, electrophysiological studies at the beginning and end of treatment. Sural nerve biopsy study was performed in 10 cases. Neurological function was assessed serially using a quantitative neurological disability score (NDS). At the beginning of treatment, there were no significant differences in various measurements between the responders and the non-responders except for the frequency of demyelination. In responders, significant improvement was found in mean changes in MRS, NDS, motor nerve conduction velocity, compound muscle action potential, distal motor latency, while in non-responders, all measures remained unchanged or worsened. Muscle wasting was seen in 3/7 responders and 4/7 non-responders, and denervation potentials in needle EMG were seen in 1/7 responders and 3/7 non-responders. Four patients of the responders were classified as chronic relapsing course, and 6 patients of the non-responders as chronic progressive course. We conclude that DFPP was useful for the subgroups of CIDP patients, but the underlying immuno-pathological background that determine the efficacy of plasmapheresis should be elucidated.

[Autonomic dysfunction in sensory ataxic neuropathy with Sjögren's syndrome].

Sjögren's syndrome (SjS) is occasionally associated with chronic progressive sensory neuropathy, and its major pathology is suggested to be dorsal root ganglionitis with the T-lymphocyte invasion. Autonomic dysfunction is frequently accompanied by SjS-associated sensory neuropathy but its underlying pathology is uncertain. We reported four patients with SjS-associated sensory ataxic neuropathy who satisfied the diagnostic criteria of Sjögren's syndrome and also showed a wide variety of autonomic disturbances. The sensory involvement was initially those of multiple mononeuritis in the distal extremities, but in the advanced stage it involved the area distributed along the spinal segmental dermatomes particularly in the trunk. Complete anhidrosis with discrete segmental distribution, as well as absence of response to cholinergic agents, was observed in three cases. Surface skin temperature recorded by a thermoviewer also showed a segmental involvement along the spinal dermatomes in two cases. Adie's pupils were present in two cases. These features suggested that the segmental involvement of the postganglionic sympathetic ganglion cells could be present in the underlying pathological condition. As for the other autonomic dysfunction, two cases showed severe postural hypotension and three cases had an abnormal heart rate variation (R-R interval). Our cases indicated that SjS-associated sensory ataxic neuropathy also manifest variable autonomic disturbances and suggested that similar pathological process to that involving the dorsal root ganglia would be present in the pathology of sympathetic and ciliary ganglion cells.

[Serial MRI findings in patients with CNS cryptococcosis].

We reported the serial magnetic resonance imaging (MRI) findings of two patients with central nervous system (CNS) cryptococcal infection without AIDS. The diagnosis of CNS cryptococcosis was made by visualizing the fungi in the CSF with the India ink test, detecting cryptococcal antigens, and culturing the fungus. Both patients had dilated perivascular Virchow-Robin (V-R) spaces, which were defined as small rounded lesions greater less than 3mm diameter that were hyperintense on T2-weighted images. They were present in the basal ganglia, brainstem and cerebral white matter. Case 1 had bilateral parietal arachnoid cyst which was thought to represent a focal collection of organisms and mucoid material within subarachnoid space. Abnormal optochiasmatic arachnoid enhancement detected in case 2, who had complete loss of vision. With disease progression perivascular V-R increased in size, resulting in the developing cryptococomas which were defined as rounded lesions greater than 3mm diameter, and were hyperintense on T2-weighted images in the basal ganglia, cerebellum and cerebral white matter. In follow-up MRI of those patients, radiological progression was seen despite appropriate treatment and falling CSF cryptococcal antigens. In conclusion, this spectrum of MRI appearances in CNS cryptococcosis reflects the pathological mechanism of invasion by the fungus, and may be relatively specific for cryptococcosis.

[Phenotypic heterogeneity in Japanese Charcot-Marie-Tooth disease type 1A patients with PMP-22 gene duplication].

We studied phenotypic heterogeneity in 18 Japanese patients with Charcot-Marie-Tooth disease type 1A (CMT1A) with PMP-22 gene duplication, together with heterogeneity of duplication size. In order to detect the duplication of PMP-22 gene region, the PMP-22 cDNA and a polymorphic marker VAW409R3 were used as probes for Southern blot analysis. As the clinical phenotypes, we assessed the degree of foot deformity, muscular weakness and atrophy, tendon reflexes, sensory impairment and electrophysiologic and sural nerve biopsy findings. Although the degree of muscular weakness and atrophy was slightly more severe in the advanced age, there was a patient with calf hypertrophy in the older age or a patient with marked muscular atrophy of the leg in the younger age. The incidence of foot deformities and sensory impairment was high and these phenotypes did not relate to aging. Diminished or absent tendon reflexes and slowing of motor conduction velocities were commonly seen, but the motor conduction velocity varied greatly among the patients. The clinical phenotypes were extensively variable among the CMT 1A patients with the same gene mutation of PMP-22 gene duplication, suggesting that there is a factor other than PMP-22 gene duplication, which influences phenotypic manifestation in CMT 1A.

[Abnormality of PMP-22 gene in Japanese patients with Charcot-Marie-Tooth disease--comparison between Southern blot and polymerase chain reaction analysis in the detection of PMP-22 gene duplication].

We investigated the duplication of the PMP-22 gene in 23 Japanese patients with Charcot-Marie-Tooth disease (CMT) by Southern blot and PCR analysis. To detect duplication of the PMP-22 gene region, PMP-22 cDNA and a polymorphic marker, VAW409R3, located in the region flanking the PMP-22 gene, were used as probes for Southern blot analysis. A marker, 6G1, also located in the PMP-22 flanking region, was amplified using the quantitative PCR method. The signal intensity of the 2.8-kb and 2.7-kb bands of MspI digests probed with VAW409R3 was different in patients with duplication. The ratio of these two bands, measured by densitometry, ranged from 1.75 to 2.13 in the patients with duplication and from 1.01 to 1.15 in those without duplication and in normal controls. The signal ratio of PMP-22 to the reference marker SF85 of BamHI digests ranged from 1.15 to 1.33 in the patients with duplication and from 0.96 to 1.04 in those without duplication, when compared with normal controls assigned a value of 1.0. The ranges of the intensity in these two groups were narrow, but did not overlap. The signal ratio of the PCR products of 6G1 to reference marker D1S80 on quantitative PCR analysis was also measured. The ratio ranged between 1.67 and 2.23 with duplication and between 1.29 and 1.74 without duplication according to the results of Southern blot analysis. However, some patients exhibited overlapping signal intensity in the duplication and non-duplication ranges. Thus, these three methods each has its own advantages and disadvantages in regard to detecting duplication.(ABSTRACT TRUNCATED AT 250 WORDS)

[Evaluation of correlation between the disappearance rate of indocyanine green and the maximum removal rate].

The disappearance rate of indocyanine green (K.ICG) and the maximum removal rate (Rmax) usually correlate with each other. However, in some cases it was shown there was a dissociation between them. We investigated the relationship between the two rates in 146 subjects. K.ICG and Rmax correlated strongly with a correlation coefficient of 0.749 (p less than 0.001). Sixty-six cases were included in the limits of 95% confidence, and the other 80 cases outside the limits were defined as dissociated cases. Among them a lower Rmax rate as compared to the K.ICG rate was found in many cases of obstructive jaundice. Particularly a lower K.ICG rate compared to the Rmax rate was found in many cases of liver cirrhosis accompanied by esophageal varices and idiopathic portal hypertension. On the other hands, we performed multiple regression analysis on 12 other liver function tests. K.ICG was strongly related to platelet count, circulatory blood volume, and albumin, all factors relating to portal hypertension. Rmax largely depended on LCAT, A/G ratio, and cholinesterase, which are Therefore, the dissociation between K.ICG and Rmax was caused by differences in the characteristic of each disease.