To elucidate the effects of cytosolic free Ca²⁺ ( [Ca²⁺] _i) on vesicle exocytosis in cholestatic hepatocytes, we established optimal culture conditions for experimental cholestatic hepatocytes. We also investigated the relation between vesicle exocytosis and [Ca²⁺] _i in cholestatic hepatocytes. After the bile duct was ligated for 1 to 7 days, rats exhibited typical features of clinical and pathologic cholestasis. One day after ligation significantly greater amounts of enzymes and lipids were leaked from cultured cholestatic hepatocytes than from normal hepatocytes. These phenomena resembled those in clinical cholestasis. Uptake of horseradish peroxidase (HRP), a marker of vesicle transport, by cholestatic hepatocytes did not differ from that by normal hepatocytes, however, HRP excretion by cholestatic hepatocytes was significantly less than that by norrr.al hepatoctyes. The HRP excretion in the absence of extra-cellular Ca²⁺ was significantly less than that in the presence of Ca²⁺ in both normal and cholestatic hepatocytes. Treatment with nicardipine decreased HRP excretion in normal hepatocytes, but did not affect HRP excretion in cholestatic hepatocytes. Moreover, high K⁺-induced increases in [Ca²⁺] _i in the presence of extracellular Ca²⁺ was observed in normal hepatocytes but not in cholestatic hepatocytes. These results indicate that (1) the hepatocytes isolated 1 day after ligation are a good model cholestasis, (2) cholestatic hepatocytes have impaired [Ca²⁺] _i-dependent vesicle exocytosis, and (3) this impairment of vesicle exocytosis in cholestatic hepatocytes might be related to damage or to smaller numbers of Ca²⁺ channels.

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Using arthroscopy, we studied the status of reduction in developmental dysplasia of the hip. Hips were examined in 2 boys and 15 girls, aged 5 months to 3 years (average, 1 year 4 months) . All joints were irreducible and treated with Pavlik harness or overhead traction. We performed hip arthroscopy using a medial approach with a 2.7-mm arthroscope under general anesthesia and observed the status of reduction during manipulation. We classified the status of reduction into four types: type 1, the femoral head remained outside the acetabular labrum and was irreducible (7 joints) ; type 2, the femoral head was moved on the labrum but was incompletely reducible (4 joints) ; type 3, the top of the femoral head was moved over the labrum but was incongruent and arthroscopically visible (2 joints) ; and type 4, the femoral head was completely reducible and arthroscopically invisible (4 joints) . Type 1 joints were treated with open reduction, and type 2 joints were treated with either open reduction or closed reduction. Joints of types 3 and 4 were treated with closed reduction. Hip arthroscopy is useful for selective screening of irreducible developmental dysplasia of the hip.

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Obstructive sleep apnea has been reported in rheumatoid arthritis (RA) patients who have lesions of the upper cervical vertebrae. We studied the pathophysiology of sleep apnea in RA patients with atlantoaxial subluxation by monitoring movements of the chest wall and abdomen, airflow, and arterial blood oxygen saturation during sleep. Nine female RA patients aged 49 to 80 years took part in this experiment. Five of them were in stage III, and four of them were in stage IV. Anterior subluxation was seen in six patients, and posterior subluxation was seen in three patients. Sleep apnea syndrome, diagnosed when the apnea index was more than 5, was observed in seven of the nine patients. The three patients with posterior atlantoaxial subluxation had high apnea indices (87.7, 48.6, and 12.6) . Central sleep apnea with absence of airflow and chest wall and abdominal wall movements was observed in two patients with posterior atlantoaxial subluxation. In these patients the anterior arc of the atlas was dislocated posteriorly. Compression of the ventral part of the lower brainstem and the upper cervical spinal cord was also observed. These results suggest that central apnea might be induced when the structures in the caudal part of the ventral medulla are blocked. The dysfunction of respiratory structures might occur in RA patients with posterior atlantoaxial subluxation.

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We measured the plasma Interleukin-8 (IL-8) concentration in 240 patients with hematological diseases to investigate the role of IL-8 in hematologic disease. Significantly higher concentrations of IL-8 were observed in some patients with acute myelogenous leukemia (AML), acute lymphoid leukemia, and chronic myelogenous leukemia than in healthy control subjects. We found no correlation between the plasma IL-8 concentration and the number of white blood cells, neutrophils, or monocytes, and the serum C-reactive protein concen-tration. In one patient with acute monocytic leukemia (M5), the high plasma concentrations of IL-8 before treatment fell to control levels after chemotherapy along with a decrease in the number of leukemic monocytes. A positive correla-tion was observed between plasma IL-8 and IL-6, but not between the concen-trations of other cytokines. High concentrations of IL-8 were detected in the supernatant of primary cultures of leukemia cells collected from 10 patients with AML. The reverse transcriptase-polymerase chain reaction showed that these cells expressed IL-8 mRNA. Leukemic cells might therefore be the main source of IL-8 in vivo under some circumstances. IL-8 might participate in the funda-mental disease process of AML, in addition to its role in inflammation.

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We studied ischemic injury of the liver and the protective effects of perfusion with a cold solution following total hepatic vascular exclusion (THVE) in a rat model, which was developed in our laboratory. Rats underwent total hepatic ischemia for 20 minutes (group A) or 45 minutes (group B) . In addition each group was further divided into two subgroups : subgroup a, ischemia alone; subgroup b, perfusion with a cold lactated Ringer's solution (4°C) . Serum levels of aspartate transaminase ( AST), alanine transaminase (ALT), and lactate dehydrogenase (LDH), as markers of hepatocellular injury, and serum hyaluronate, as an indicator of damage to endothelial cells, were measured. Liver tissue blood flow was assessed serially after reperfusion of the liver, and the histologic findings in the perfused liver were also evaluated on the 7th day after surgery. Serum levels of AST and ALT were high in the early period in group B, but after recovery there were no differences between the groups. In addition, levels of LDH in the perfusion group were significantly less than in the nonperfusion group. Both postoperative liver tissue blood flow and changes in serum levels of hyaluronate indicated a rather rapid recovery from ischemic damage in perfusion subgroup b after ischemia for 20 or 45 minutes. Histologic examination indicated that hepatocellular damage and derangement of sinusoidal structure were minimal in subgroup b. These findings suggest that perfusion with a cold solution during THVE is a beneficial procedure to protect liver tissue, although serum levels of AST, ALT, and hyaluronate were higher than in the non-perfusion groups at an early stage.

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The effect of an alternating magnetic field on cholesterol metabolism in rabbits was investigated. With four commercial electromagnets of 0.08 T (800 Gauss) in magnetic intensity on the coil surface, a whole-body alternating magnetic field was applied to nine rabbits 4 hours a day for 5 consecutive days. The average magnetic field strength to which the rabbit's liver was exposed was approximately 2 to 7 mT (20 to 70 G) . After exposure to the alternating magnetic field, low-density lipoprotein (LDL) cholesterol decreased (p<0.05 ), and the HDL/LDL cholesterol ratio increased (p<0.05) compared with controls of no magnetic field exposure. However, total cholesterol, HDL cholesterol, and triglyceride levels did not change. We conclude that an alternating magnetic field affects the LDL cholesterol system clearly and decreases the level of LDL cholesterol.

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The aim of the present study was to investigate the role of the pituitary in the delayed-type hypersensitivity (DTH) response in mice. We examined the effect of hypophysectomy on 2, 4, 6-trinitrochlorobenzene (TNCB) -induced DTH, which has been used as a cellular immunity model in vivo. Seven-week-old male BALB/c and C57BL/6 mice were used. Six days after sensitization with TNCB, TNCB was applied to the left ear pinnea and the DTH response was measured as the degree of ear swelling 24 hours after TNCB challenge. The TNCB-DTH reaction was also measured 24 hours after intracisternal injection of α-helical coricotropin-releasing factor (CRF), a CRF inhibitor, at doses of 0.4, 1, 5, and 10 μg. Maximal ear swelling in both hypophysectomized mice and mice that received α-helical CRF was significantly increased. Histologically, adrenocortical atrophy and deposition of lipid were observed in hypophysectomized mice at the time DTH was enhanced. These results suggest that pituitary ACTH plays a suppressive role in the DTH response in mice.

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Phenotypes of apolipoprotein E (apoE) were determined with an isoelectric focusing method in 39 patients with sporadic Alzheimer's disease (AD), 38 patients with dementia from cerebrovascular disease (CVD), and 100 healthy subjects without hyperlipidemia. There was a striking difference in the distribution of apoE phenotypes between patients with AD and healthy subjects (p<0.001) . Such a difference was attributable to different frequencies of phenotype E4/3 and E3/3. The apoE4/ 3 phenotype was detected in 38.5% (15 of 39) patients with AD, more than 3 times higher than in healthy subjects (12.0%, 12 of 100) . In contrast, apoE3 / 3 phenotype, a wild type apoE phenotype, was detected in 81.0% (81 of 100) of healthy subjects but only in 38.5% (15 of 39) of patients with AD. The frequency of apoE-_ε4 allele assessed on the basis of obtained apoE phenotypes in patients with AD (0.269) was significantly higher than that in healthy subjects (0.08, p<0.001) and in patients with dementia from CVD (0.118, p<0.01) . Furthermore, the frequencies of apoE-_ε4 in AD were 0.190 in patients older than 70 years and 0.333 in patients younger than 70 years. These results indicate a strong association between apoE-_ε4 and sporadic AD and suggest that apoE-_ε4 assessed from apoE phenotypes may be a possible risk factor for AD.

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We evaluated the effects of 22-oxa-1α, 25-dihydroxyvitamin D₃ (OCT) on the inhibition of growth, the induction of differentiation, and the production of cytokines in human myeloid leukemia (HL-60) cells. While 1α, 25-dihydroxyvitamin D₃ (1, 25 (OH) ₂D₃) induces differentiation and inhibits proliferation of HL-60 cells, it has been found to induce hypercalcemia in clinical trials. However, OCT does not induce hypercalcemia and induces differentiation with a potency similar to that of 1, 25 (OH) ₂D₃. In the present study OCT significantly more strongly inhibited cell proliferation and more effectively induced cytokine production than did 1, 25 (OH) ₂D₃. OCT thus appears to be a promising candidate for treating myeloid leukemia and myelo-dysplastic syndromes.

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Many researchers have been studying the association of monoamine neurotransmitters and their metabolites with brain cell damage after transient cerebral ischemia or in Parkinson's disease. In the present study, we investigated in vitro the antioxidant effect involved in dopamine (DA) metabolism on irondependent and iron-independent lipid peroxidation using frozen striatal tissue, whole brain homogenate and its mitochondrial fraction from rats. Using 7-week-old male Wistar rats, we studied lipid peroxide formation in striatal samples, homogenate and mitochondrial fractions in the presence of various high concentrations of DA. Addition of DA (10^-4 M) to reaction mixtures apparently suppressed lipid peroxidation, as determined with thiobarbituric acid methods, by about 30% to 40% of control. Pargyline, a monoamine oxidase inhibitor, antagonized the decrease in lipid peroxidation in the homogenate and enhanced the effect of DA in the mitochondrial fraction in the presence of iron ion. These effects seem to derive not only from the anti-oxidative effects of DA and their metabolites but also from the process of DA metabolism by monoamine oxidase as a whole.

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The human utricular wall was studied with regard to the distribution of pigment. In general, the distribution of pigment of the superior wall was more dense than that of the inferior wall. The utricular macula and the hemispherical area of the anterolateral portion of the superior wall were devoid of pigment. Unlike the semicircular ducts, the membranous common crus demonstrated dense pigment distribution along the entire wall and less pigment toward the bifurcation of the anterior and posterior semicircular ducts. Masson-Fontana staining, a bleaching method, and elctron microscopic observation revealed the pigment to be melanin. Its significance is discussed from the viewpoint of laser inner ear surgery.

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In recent years, with the changing pattern of cancer death rates among various sites, and the introduction of more effective medical treatments, estimates of future trends of cancer from current death rates are somewhat misleading. Therefore, it is important for policy makers to obtain information on incidence rates when establishing countermeasures against cancers. The Nagasaki Cancer Registry Program was established in 1963 and provides quite accurate data with histologic information. Therefore, in this study, incidence rates of cancer for 2010 were estimated with a linear regression model and the Nagasaki Cancer Registry data from 1985 through 1991. It was estimated that the number of male cancer patients will increase from 3, 373 in 1990 to 5, 624 in 2010 (67% increase) and that of female cancer patients from 2, 734 in 1990 to 4, 426 in 2010 (62% increase) . In both males and females colon cancer is the most common type, followed by stomach cancer; breast cancer is third most common in females. Although incidence rates for stomach cancer are decreasing, because of the aging Japanese population, stomach cancer will remain a major problem in the near future. Discrepancies between the regression coefficient of incidence rates and death rates were found for stomach, colon, and breast cancers. In particular, death rates from breast cancer showed little change in each age group, but incidence rates showed a steep increase in most age groups. Therefore, it is important to analyze incidence rates rather than death rates to determine future trends and to establish effective countermeasures against cancers.

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The Ki-67 labeling index and p53 overexpression determined with immunohistochemistry are both useful indicators of the cellular proliferative potential of brain tumors. We compared the immunohistochemical expression of Ki-67 and p53 proteins in samples of 20 astrocytomas obtained from 19 patients which had been pretreated with three different methods of antigen activation: 95°C heating, 121°C autoclaving, and exposure to target unmasking fluid. The Ki-67 labeling index of specimens pretreated with autoclaving and 95°C heating was positively correlated with the degree of malignancy of astro-cytomas (simple regression analysis, p<0.002 for autoclaving and p<0.03 for 95°C heating) . p53 overexpression was identified in 7 astrocytomas (2 cases of low-grade astrocytomas, 2 cases of anaplastic astrocytomas, and 3 cases of glioblastoma multif ormes) ; however, p53 expression was not correlated with the degree of malignancy of astrocytomas.

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The expression of glial fibrillary acidic protein (GFAP) in the perivascular glia of the developing human retina was studied immunohistochemically in 22 normal eyes. Retinas were stained with anti-GFAP antibody using the labelled streptavidin-biotin method. The GFAP-positive perivascular glial cells were first observed in the human retina at 24 gestational weeks near the optic disc. As the retina matured, the intensity of GFAP immunoreactivity increased until full term. The GFAP immunoreactivity was most intense around large vessels near the optic disc and diminished with decreasing vascular caliber and increasing distance from the optic disc. Our results support the concepts that in the developing human retina, GFAP-positive perivascular glial cells first appear at 24 gestational weeks and that the human retina contains perivascular glial cells.

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