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To investigate the feasibility of repeated gene transfection in suicide gene therapy against human solid tumors by a combination of 5- fluorocytosine (5-FC) and its converting enzyme, cytosine deaminase (CD), we repeatedly transfected the yeast CD gene into the human pancreatic cancer cell line BXPC3 using the hemagglutinating virus of Japan-liposome in a new gene transfer method. The in vivo growth of the s.c. transplanted BXPC3 tumor in nude mice given CD-gene transfection was significantly suppressed by i.p. injection of 5-FC when compared with tumors treated with the control vector. Furthermore, the tumor transfected with the CD gene during a 7-day interval was suppressed much more than that of a single transfection. These results suggest that repeated transfection of the suicide gene together with the combination of 5-FC and the yeast CD gene using the hemagglutinating virus of Japan-liposome gene transfer method may be useful for the treatment of human solid tumors, including pancreatic cancer.

SUMMARYPrimary biliary cirrhosis is an autoimmune disease of the liver in which T helper 1 cytokines predominate over those of T helper 2 in the pathogenesis. Interleukin-18 (IL-18), for which the gene was recently cloned, is a novel T helper 1 cytokine, which augments interferon-gamma production. We designed this study to clarify the role of IL-18 in primary biliary cirrhosis and to examine whether serum IL-18 level can be a prognostic indicator for the disease. Serum IL-18 levels were measured using an enzyme linked immuno sorbent assay with mouse monoclonal antibodies. Twenty-two healthy volunteers, 31 patients with primary biliary cirrhosis (Scheuer's stage I, 13; II, 10; and IV, 8), 20 patients with autoimmune hepatitis, 11 patients with virus-related liver cirrhosis and six patients with obstructive jaundice were enrolled. Significant differences of serum IL-18 levels were observed between patients with Scheuer's stage IV and those with stage I, or II, virus-related liver cirrhosis and obstructive jaundice (P < 0·05). The IL-18 levels in primary biliary cirrhosis increased according to the disease progression, and fell promptly after living-related liver transplantation. Moreover, serum IL-18 levels in primary biliary cirrhosis were correlated with serum bilirubin concentrations and the Risk scores of the Mayo Clinic prognostic model for the disease. The IL-18 levels observed in patients with autoimmune hepatitis were also elevated, and correlated with the activity of the disease. These results indicate that serum interleukin-18 levels reflect the severity of primary biliary cirrhosis, the activity of autoimmune hepatitis, and may be an additive prognostic indicator in primary biliary cirrhosis.

Fibronectin (FN) possesses a wide range of biological functions. However, the role of plasma FN in vivo has not yet been established and there have been no published studies of plasma FN in ALS. The aim of this study was to measure plasma FN in ALS patients.We measured plasma FN levels in 28 ALS patients, 18 control subjects with other neurologic or muscular diseases (control group A) and 21 healthy adults (control group B). The age and sex distributions among the 3 groups were comparable.Plasma FN levels were significantly lower in ALS patients than in control groups A and B. There was also a significant negative correlation between plasma FN levels and duration of illness in ALS patients.These data suggest that a metabolic alteration of FN may take place in ALS and that the measurement of plasma FN may serve as an indicator of clinical progression of this disorder.

The regional toxicity of an anticancer agent for normal liver tissue following hepatic arterial infusion chemotherapy (HAI) was evaluated in terms of morphology, function, and histopathology. Forty-two patients(M:F = 30:12; mean age, 59.9 years) with liver metastases from colorectal cancer were treated with HAI using a totally implantable vascular access port system from July 1994 to March 1999. The regimen used here was so-called weekly high-dose 5-fluorouracil(5-FU) infusion(5-FU, 1,000 mg/m2/week). Volume measurement of the liver demonstrated not only whole liver atrophy including the tumor but also volume reduction of the non-tumorous lobe. Atrophic change of the liver was seen in patients who were administered over 20 g/m2 of 5-FU(p0.01). The CT attenuation values of the liver were examined, and fatty infiltration was seen in six patients. Histologic examination of liver biopsies from the non-tumorous part revealed steatosis and infiltration of inflammatory cells in the portal triad, which were not seen in specimens prior to HAI. On clinical laboratory findings, enzymes representing bile duct, including alkaline phosphatase, leucine amino peptidase, and gamma-glutamyltranspeptidase, were increased in 22 patients. In terms of regional toxicity for long-term HAI, 20 g/m2 of 5-FU, is the key dose at which to consider temporary cessation or dose reduction.

The mechanism of cadmium-mediated acute hepatotoxicity has been the subject of numerous investigations and although some uncertainties persist, sufficient evidence has emerged to provide a reasonable account of the toxic process. Acute hepatotoxicity involves two pathways, one for the initial injury produced by direct effects of cadmium and the other for the subsequent injury produced by inflammation. Primary injury appears to be caused by the binding of Cd2+ to sulfhydryl groups on critical molecules in mitochondria. Thiol group inactivation causes oxidative stress, the mitochondrial permeability transition, and mitochondrial dysfunction. Although cadmium may injure hepatocytes directly, there are compelling reasons to believe that hepatocellular injury is produced in vivo as the result of ischemia caused by damage to endothelial cells. Secondary injury from acute cadmium exposure is thought to occur from the activation of Kupffer cells and a cascade of events involving several types of liver cells and a large number of inflammatory and cytotoxic mediators. In this regard, it is clear that Kupffer cell activation and neutrophil infiltration are important events in the toxic process, and the involvement of proinflammatory cytokines and chemokines has also been implicated. The precise roles of the soluble mediators of inflammation warrant further investigation.

Objectives - Type IV collagen (IV-C) abnormalities of skin and serum have been reported in patients with amyotrophic lateral sclerosis (ALS). However, there has been no study of urinary IV-C in ALS. The present study investigates urinary IV-C and the relation to its skin content in patients with ALS. Material and methods - We studied IV-C immunoreactivity of skin and measured urinary levels of IV-C in ALS patients and controls. Results - The basement membrane as well as blood vessels of skin in ALS patients was weakly positive for IV-C as compared with those of controls. Immunostaining became even weaker as ALS progressed. The urinary level of IV-C in ALS patients was significantly decreased as compared to diseased controls (P

Expression of platelet-derived growth factor B-chain and of its specific receptor (β-receptor) was investigated in immature brains with hypoxic/ischemic injury. After the left common carotid arteries of seven-day-old rats were ligated and pups were placed in a hypoxic chamber, the protein and messenger RNA of both B-chain and β-receptor were assessed using immunocytochemistry and northern analysis, respectively. Transcripts for B-chain were localized by in situ hybridization. Faint but definite expression of B-chain and β-receptor was seen in the brains of untreated neonatal controls. Three to 48 h after hypoxia B-chain protein was generally increased above control levels, but focally decreased expression was seen in infarcted areas. Enhanced induction of messenger RNA of B-chain was seen in the both sides of cerebral cortices and hippocampi at 3 h. Strongly increased positivity for B-chain protein and mRNA occurred in the neurons surrounding the infarct. In situ hybridization still showed this up-regulation seven days after hypoxia. Beta-receptor protein expression was enhanced in some neurons immediately surrounding the infarct at 3 h of hypoxia, and marked up-regulation was seen at 16 h. Beta-receptor messenger RNA remained at control levels. Immunocytochemistry showed strong immunoreactivity for the β-receptor on the neurons surrounding the infarct at 72 h. These results indicate that a neonatal hypoxic/ischemic insult induces neuronal up-regulation of the platelet-derived growth factor B-chain as well as β-receptor immediately after hypoxia. While this up-regulation is relatively transient in most neurons, sublethal damage to neurons immediately surrounding an infarct induces sustained up-regulation. Through autocrine and paracrine mechanisms, platelet-derived growth factor B-chain molecules may act as a neuroprotective factor in immature brain experiencing with hypoxic/ischemic injury.

A female patient, who was diagnosed with Leigh syndrome at 15 months of age, developed fulminating severe hypertension and died at 8 years of age. Hypertension has not been reported as an important clinical symptom in Leigh syndrome. Laboratory findings indicated that it was not associated with endocrinopathic diseases such as pheochromocytoma and aldosteronism, or renal diseases. Brain MRI scan showed symmetrical lesions in the basal ganglia and medulla oblongata including the nucleus tractus solitarius. This nucleus is known to play an important role in maintaining blood pressure. Since the medulla oblongata is a vulnerable site, potential development of hypertension should be taken into consideration when managing Leigh syndrome.

Fas ligand (FasL) belongs to the TNF superfamily. It is induced in activated lymphocytes and eliminates Fas-positive lymphocytes, resulting in the down-regulation of immune responses. FasL has also been detected in tissues other than lymphoid cells. We investigated the expression and function of FasL on human colon cancer cells. FasL mRNA was detected by RT-PCR in all six colon cancer cell lines tested and was not found on fibroblasts. FasL protein was detected in DLD-1, LoVo, HCT-116 and RPMI 4788 cells by immunohistochemical staining. DLD-1, LoVo and WiDr were cytotoxic against mouse T lymphoma cells which were transfected with human Fas receptor cDNA. The cytotoxicity was significantly enhanced by phorbol 12-myristate 13-acetate (PMA) and ionomycin. Our data suggest that the FasL expressed in human colon cancer cells may be regulated by endogenous factors in the microenvironment of the host and facilitates the escape from the host immune system.

Eight patients with uterine cervical cancer received two or three courses of neoadjuvant chemotherapy, including 254-S i.v. and CDDP i.a. Transcatheter arterial embolization (TAE) was added for seven patients. The therapeutic efficacy was evaluated by MR imaging and postoperative histopathological examination. Three patients achieved a complete response (CR) and five others were evaluated as a partial response (PR) on MR imaging. On postoperative histology, three of eight showed CR or PR, which coincided with MR findings. Viable cancer cells were shown in five patients. To detect these viable tumors, dynamic MR imaging was indispensable. However, because of limited spatial resolution, the detection of small residual tumors was not easy using dynamic MR imaging.

To study type IV collagen of skin and serum in patients with ALS.Collagen abnormalities of skin have been reported in ALS patients. However, little is known concerning type IV collagen in ALS.We studied type IV collagen immunoreactivity of skin and measured serum levels of the 7S fragment of the N-terminal domain of type IV collagen (7S collagen) in patients with ALS and control subjects.The basement membrane as well as blood vessels of skin in ALS patients was weakly positive for type IV collagen as compared with those of diseased control subjects. This weak immunostaining became more pronounced as ALS progressed. The optical density for type IV collagen immunoreactivity in ALS patients was significantly lower (p0.001) than in diseased control subjects and was significantly decreased with duration of illness (r = -0.85, p0.01). Serum 7S collagen levels in patients with ALS were significantly decreased (p0.01) as compared with those in diseased and healthy control subjects and were negatively and significantly associated with duration of illness (r = -0.81, p0.001). There was an appreciable positive correlation between concentrations of serum 7S collagen and the density for type IV collagen immunoreactivity in ALS patients (r = 0.81, p0.02).These data suggest that a metabolic alteration of type IV collagen may take place in the skin of ALS patients and that the decreased levels of serum 7S collagen may reflect a decreased type IV collagen immunoreactivity of skin in patients with ALS.

Expression of TrkB receptors were studied in the cerebral cortex of normal rats and young rats with hypoxic/ischemic injury. TrkB expressing cells were present in the piriform cortex at birth and increased in number with age, and were finally present in the entire cerebral cortex. Density of TrkB cells reached adult levels at P30. They were morphologically regarded as pyramidal neurons and interneurons. Hypoxic/ischemic injury induced a tentative increase of full-length TrkB receptors. A novel appearance of TrkB expressing neurons and enhanced immunostaining on both cell soma and dendrites were observed in the peri-infarct areas and increased number of TrkB expressing neurons were detected in the contralateral cortex after carotid artery ligation. This increase was no longer evident after 48 h of hypoxia. Double immunostaining using antiserum against GFA or OX-42 revealed no co-localization of TrkB receptors and these molecules, while there were only slight co-localization of TrkB and calbindin-D28k molecules. The altered levels in responses to injury indicate that TrkB may play a crucial role in the early protective mechanism of the neurons with hypoxic/ischemic injury through ligands BDNF and/or NT-4/5.

Twenty-four patients were treated with arterial infusion of SMANCS dissolved in Lipiodol. Twenty of these patients had HCC with the main trunks of portal vein occluded by tumor, and four patients had severe cirrhosis and multiple HCC. The actual dose of SMANCS administered each patient ranged from 4 to 6 mg. Side effects occurred in 50%. Severe side effects such as shock and shivering-chilliness were observed in 18%. The differences between the values of hepatic functional serum indexes obtained before and after treatment with SMANCS were small and transient. With regard to the therapeutic response of the arterial infusion of SMANCS, the mean survival time was approximately 2.8 months. It was suggested that the more effective administration of SMANCS was combination of the arterial infusion of SMANCS-Lipiodol with TAE at the level of the right hepatic artery of left hepatic artery for multiple HCC.

Hypersomnia occurs frequently in patients with myotonic dystrophy (MyD). We performed a quantitative immunohistochemical study of serotonin (5-HT)-containing neurons linked to hypersomnia in the dorsal raphe nucleus (DRN) and the superior central nucleus (SCN) in 8 patients with MyD, 5 of whom showed hypersomnia, and in 12 age-matched controls. The densities of 5-HT neurons in the DRN and the SCN were significantly lower in MyD patients with hypersomnia than in MyD patients without hypersomnia and controls. These data suggest that the loss of 5-HT neurons of the DRN and the SCN is associated with the presence of hypersomnia in MyD.

We investigated the cell clonality of 12 cases of female solitary hepatocellular carcinoma (HCC) that were associated with hepatitis virus infection. The clonal origin of HCC could be assessed by the method based on restriction fragment length polymorphism (RFLP) of X-chromosome-linked androgen receptor gene (AR) and phosphoglycerate kinase (PGK) gene, taking advantage of random inactivation of one of two X-chromosomes by methylation in females. We extracted DNA samples from both fresh and paraffin-embedded specimens of the same lesion as a source of DNA sample for polymerase chain reaction (PCR). Consequently, it was possible to use methylation-sensitive restriction enzymes and PCR to study differential methylation patterns among alleles of these genes for both DNA samples. The RFLPs of AR gene and PGK gene were found in eight of 12 cases and five of 12 cases, respectively. There were two cases which had no RFLPs in either AR gene or PGK gene. All cases of HCC which had RFLP in either AR gene or PGK gene demonstrated monoclonal origin of the tumor regardless of their histologic patterns.

Dichlorvos(DDVP)-induced DNA single strand breaks were investigated in isolated rat hepatocytes. In a dose-response study in hepatocytes from PB-treated rats (80 mg/kg i.p., for 3 days), 250 μM DDVP substantially reduced cellular non-protein sulfhydryl (NPSH) content, but had no detectable effect on DNA. At 500 μM, the increase in DNA single strand breaks was significant, with a slight increase in cellular lipid peroxidation. At doses over 1000 μM DDVP, cell death was accompanied with considerable lipid peroxidation, and DNA single strand breaks were evident. When the antioxidant N , N ′-diphenyl- p -phenylene diamine (DPPD) was added or if the hepatocytes were incubated under air instead of 95% O 2 , lipid peroxidation and cell death were attenuated but DNA single strand breaks and reduction in NPSH content were not. On the other hand, ferrous iron-induced DNA single strand breaks, lipid peroxidation, and depletion of NPSH content were all attenuated by DPPD or by incubating the cells under air. With respect to the subcellular lipid peroxidation, DDVP caused a significant increase, mainly in the microsomal fraction, whereas ferrous iron caused rapid and substantial increases in mitochondrial, microsomal, and nuclear fractions. There were more DNA single strand breaks caused by N -nitrosodiethylamine (NDEA), which becomes genotoxic after microsomal metabolism, in hepatocytes from PB-treated rats than in those from control rats. The number of these breaks was reduced by adding the cytochrome P450 inhibitor, metyrapone. On the other hand, the effect of DDVP on DNA was not affected by modification of the cytochrome P450 status. These results suggest that lipid peroxidation induced by DDVP in isolated rat hepatocytes plays a significant role in its cytotoxicity but not in its genotoxicity.

The corticospinal tract in the rat after neonatal ablation of the unilateral cerebral cortex was studied morphologically using the antegrade horse-radish peroxidase (HRP) tracing method. An aberrant ipsilateral tract was observed 7 days after the operation. Formation of the aberrant neuronal pathway has been confirmed to be contributed mainly by new axons, which ramified at the level of the pyramidal decussation from healthy corticospinal fibers. This ramified axon ran toward the ipsilateral dorsal funiculus. A few fibers also contributed to the aberrant pathway by changing their direction on the way of extension at the level of the pyramidal decussation. These results indicate that the ramification and change of the direction of extending axons play an important role for formation of a new ipsilateral corticospinal tract.

A teratological study was carried out on the plasticizer tri-n-butyl phosphate (TBP). Pregnant Wistar rats were treated orally on days 7–17 of gestation with TBP at 0, 100, 200, 400 or 800 mg/kg/day in the dose-finding study and 0, 62.5, 125, 250 or 500 mg/kg/day in the subsequent teratological study. Caesarean sections were performed on day 20 of gestation. In the dose-finding study, all of the pregnant rats were killed by the treatment with TBP at 800 mg/kg/day. In the teratological study, salivation and depression of body weight gain, adjusted body weight gain and food consumption were observed at the higher doses of TBP. There were no significant differences between the groups in the incidence of dead or resorbed foetuses, the number of living foetuses and the body weights of living foetuses of both sexes. The incidence of rudimentary lumbar rib increased significantly at 500 mg/kg/day. There were two cases of malformation: a foetus with deformity of fore- and hind-limbs at 400 mg/kg/day in the dose-finding study and conjoined twins exhibiting three fore-limbs and four hind-limbs at 125 mg/kg/day in the teratological study. These malformations were rare in the background data of teratology, and the incidence of foetuses with malformations was not increased significantly. Therefore, TBP was considered not to be teratogenic in this study.

Experimental production of congenital hydrocephalus was undertaken by inoculating mumps virus into pregnant hamsters intravenously or intraplacentally. When the mumps virus was inoculated intravenously on the 8th, 10th, 12th, or 14th day of gestation, some fetuses were aborted and those which could come to term did not develop hydrocephalus after birth. Offsprings from the mothers, which had had intraplacental inoculation on the 14th day of gestation, showed ventricular dilatation in about 28%. Histological examination revealed inflammatory infiltration on the surface of ependymal layers, subependymal edema and microglial activation in the underlying ependyma of the aqueduct. These findings were thought to have resulted from ependymitis caused by mumps virus. The transplacental infection of mumps virus is considered to be extremely rare. However, in such conditions as the placental barrier is impaired, mumps virus will possibly pass through the placenta, and will cause hydrocephalus to the infant.

We report a two-year-old boy with clinical Reye syndrome. Brain CT disclosed symmetrical low-density areas in the pons and thalami, a part of which was recognized as being of high-intensity on both T1 and T2 weighted MRI. These lesions were suspected of being caused by vascular involvement, since the methemoglobin induced by bleeding may have contributed to the high-intensity regions. The lesions gradually disappeared within four months after the onset, and clinical symptoms also improved markedly without serious neurologic impairment. This case was characterized by the marked improvement in neurological and radiological findings in spite of the serious neurological involvement at onset.

Antibodies against Japanese encephalitis virus (JEV) were examined in 356 serum samples from individuals visiting Tribhuvan University Teaching Hospital using neutralization test. A total of 13.2% (47/356) seropositivity was observed. Antibody positivity against both Nakayama-NIH and Beijing-1 strains was seen in 7.9% subjects. Infection by Nakayama-NIH strain alone was seen to be 4.2% while 1.1% of study populations were found to be infected only by Beijing-1 strain. High antibody titer was observed in the 20-40 years age-group and did not increase with age. Seropositivity was more common in zone-A (Tarai) area compared to zone-B (Hills) and zone-C (Mountains). Spectrum of antigenicity of JEV in zone-A as shown by neutralization test was identical to Indian isolates (strains). Difference in seropositivity was observed against Nakayama-NIH and Beijing-1 strain in zone-B and C indicating recent spread of JEV in these areas.

Molecular oxygen or cytochrome c has been described as the electron acceptor of the reaction of old yellow enzyme with NADPH. In this study, menadione was found to be a sensitive electron acceptor of the reaction under aerobic as well as anaerobic conditions. The Km value of menadione for old yellow enzyme is as low as 2-3 x 10(-7) M in the presence or absence of superoxide dismutase. The rate enhancement of the cytochrome c reduction of old yellow enzyme with NADPH was about eight times in the presence of menadione. The rate increment was slightly higher under aerobic than anaerobic conditions. The rate enhancement by menadione enabled sensitive determination of the enzyme activity in the assay system, which contained NADPH, cytochrome c, menadione, and old yellow enzyme. In the reaction course, the semiquinone species of menadione was trapped by the reaction with t-butyl-alpha-phenylnitrone. The radical adduct was detected on EPR. The dyestuff, 2,6-dichlorophenolindophenol, was found to be reduced ineffectively even in the presence of menadione; moreover, it was inhibitory in the NADPH consumption reaction. Methylene blue or Lauth's violet, known to be capable of semiquinone formation, also behaved, like menadione, as a mediator of electron transport to cytochrome c. On the basis of the experimental results, the occurrence of the one electron transfer of the old yellow enzyme reaction was emphasized.

In order to elucidate the diagnostic significance of urinary apolipoprotein H (apo H), known as beta 2-glycoprotein I, urinary apo H levels were measured by a non-competitive enzyme immunoassay in diabetics without hypertension and Albustix-positive proteinuria. The relationships between urinary apo H levels and urinary albumin (Alb) levels as well as clinical profiles in diabetics were investigated. The mean urinary apo H level in 45 diabetics was 264.2 +/- 189.5 micrograms/g.cr, and significantly higher than that in 41 healthy subjects (120.6 +/- 84.8 micrograms/g.cr, p0.01). Diabetics were classified into three groups based on urinary Alb levels; 17 group I patients with normoalbuminuria (U-Alb15mg/g.cr), 12 group II patients, whose urinary Alb levels were in between normo- and microalbuminuria (15or = U-Alb30mg/g.cr) and 16 group III patients with microalbuminuria (30U-Albor = 300mg/g.cr). The mean urinary apo H level in group I patients, who were regarded as without nephropathy, was 199.2 +/- 109.0 micrograms/g.cr, significantly higher than that in normal subjects (p0.01). The mean urinary apo H levels in group II and III were 271.4 +/- 177.1 and 327.8 +/- 246.1 micrograms/g.cr, respectively. These values were also higher than in normal subjects (both of p0.01). Urinary apo H levels correlated positively with urinary levels of glycosaminoglycan (r = 0.382, n = 45, p0.01), which was regarded as an indicator of the anion loss from glomerular basement membrane to urine, and with urinary N-acetyl-beta-D-glucosaminidase activities (r = 0.378, n = 37, p0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Pathological examination was carried out of the skeletal muscle of an 8-year-old boy with abetalipoproteinemia. The patient complained of diarrhea, and showed a deficiency of betalipoprotein, decreased fat-soluble vitamins, acanthocytosis and a mild increase in serum creatine kinase. The prominent histochemical finding was punctate deposits of acid phosphatase activity in most fibers. Ultrastructural lesions revealed a number of giant lysosomes. Although these pathological findings seemed to be related to vitamin E deficiency, other pathological findings such as concentric laminated bodies or filamentous bodies were also observed. The clinical course and the changes in the pathological findings in our patient after long-term vitamin E therapy need to be observed.

The activities of N-acetyl-beta-D-glucosaminidase (NAG), gamma-glutamyl-transpeptidase (gamma-GTP) and NAG isoenzyme were measured in the urine of 20 patients with multiple myeloma (IgG/IgA type/Bense Jones type; 15/1/4 cases) and 25 healthy controls to evaluate these activities as indicators of renal disturbance in multiple myeloma. NAG isoenzyme fractions in urine were measured by agarose electrophoresis-m-cresol sulfonphthaleinyl-NAG reaction. Mean urinary NAG activity in the patients with myeloma was significantly higher than that in the controls (20.1 +/- 3.3 vs 4.3 +/- 0.3U/g. cr; p0.001). Urinary NAG activity in these patients correlated positively with the dose (mg/g. cr) of urinary protein (r = 0.755; p0.01), most of which were considered to be light chain protein, but not with creatinine clearance. Each urinary NAG isoenzyme fraction (NAG-1, -2, -3) was higher in the patients than that in the controls, and especially NAG-2 fraction (A form) showed a highly positive correlation with the dose of urinary protein. Urinary gamma-GTP activity in the patients did not differ from that in the controls, but urinary NAG/gamma-GTP ratio was higher in the patients, and reversely correlated with creatinine clearance (r = -0.721; p0.01). It is suggested that the elevation of urinary NAG activity results from the damage of lysosome in proximal tubular cells by urinary light chain protein and its degradation products. Therefore, urinary NAG activity may be a good index for proximal tubular disturbance, and NAG/gamma-GTP ratio may be an index for the extensive damage of nephrons in addition to the damage of tubular cells in multiple myeloma.

The antibody positive rates among Nepalese, in a community, to herpes simplex virus (HSV), varicella-zoster virus (VZV) and cytomegalovirus (CMV) were studied. Immune adherence hemagglutination test (for VZV) and complement fixation test (for HSV and CMV) were used to measure the antibodies. An 80% positive rate of anti-HSV antibodies was found in early childhood (1-4 years) that further increased with age (96.1% positive in greater than 15 years age). Only 25% of children 1-4 years old showed antibodies to VZV but the number of positives increased rapidly with age (82.9% in greater than 15 years age). Antibody against CMV was positive in all the subjects studied.

Rats with huge porencephaly occupying the left cerebral hemisphere were used for this study. Porencephaly was caused experimentally by the ligation of the left common carotid artery and subsequent hypoxic exposure at the age of 7 days. A large band of horseradish peroxidase (HRP) positive fibers was found crossing at the pyramidal decussation to reach the contra-lateral corticospinal tract and the deeper part of the funiculus, in the cervical cord. However, a small band of HRP positive fibers was noticed to reach the ipsilateral corticospinal tract without crossing at the pyramidal decussation. This result indicates that the developing pyramidal neurons in the right hemisphere carry the plasticity even after suffering from considerable hypoxia.

In order to elucidate the clinical significance of urinary apolipoprotein H (Apo H), otherwise known as beta 2-glycoprotein I, we first developed a non-competitive enzyme immunoassay (EIA) to quantify urinary Apo H levels. The measurable range of this assay was about 2 ng/ml to 100 ng/ml. The intra- and inter-assay coefficients of variation were 2.0 to 4.7% and 8.3 to 17.0%, respectively. These results indicated that this assay had high sensitivity and good reproducibility, and that it was useful for clinical study. We then determined urinary Apo H levels in 24 normal subjects and 36 diabetics using this assay. The mean urinary Apo H index (urinary Apo H/cr ratio), in 20 patients without proteinuria, who were regarded as patients without nephropathy, was 569 +/- 560 (X10(-3) mg/g.cr) and significantly higher than in normal subjects (252 +/- 147, p less than 0.01). The mean urinary Apo H index in 16 patients with overt proteinuria was 1,507 +/- 3,701 and also higher than in normal subjects (p less than 0.01). These results taken collectively indicate that urinary Apo H level using the EIA may be a new sensitive marker for detecting minor change of glomerular basement membrane in diabetics.

A case of paroxysmal nocturnal hemoglobinuria (PNH) associated with acute renal failure (ARF) is described. A 57-year-old female, who had been diagnosed as having PNH in 1983 at Kochi Medical School, was admitted to our hospital in April 1989, because of ARF with dark urine after a common cold. Hemodialysis was performed 5 times for ARF, and after almost completely recovering from ARF, she was discharged. The renal biopsy showed the deposition of hemosiderin in the proximal tubular cells. We surveyed fourteen case reports of PNH associated with ARF in Japan including our case. Ten cases developed ARF after infection causing hemolytic attack. Twelve of 14 cases were treated by hemodialysis and 13 cases were reversible. The histopathology of their renal biopsies revealed the deposition of hemosiderin in the proximal tubular cells in six of seven cases and tubular necrosis in three cases. These data showed that hemolytic attack and dehydration related to infection facilitated the induction of ARF.

In 18 (40%) of 45 patients with CPS, paroxysmal discharges were hard to be found on their interictal EEG examinations. These patients were, however, not different from the other 27 patients with paroxysmal discharges on EEG as for the onset, prognosis for attack, complication of generalized tonic-clonic seizure and febrile convulsion, family history and adaptation to society.

Recently, as a common foundation of various branches in science, thermodynamics is getting fresh notice. One of the reasons are long-range forces that may have far-reaching consequences for the applicability of standard thermodynamics. In this paper we trace these consequences in connection with gravity, particle physics, condensed matter physics, and plasma physics. We point on scenarios (like hot plasmas of elementary particles in the early universe) where usually the applicability of standard thermodynamics is not even questioned. Therefore we wish to attract the reader's attention to such cases and mention some first steps towards non-standard thermodynamics. It is, however, too early to propose final solutions at the present stage. We wish to provoke thoughts about the old thermodynamics from a modern perspective.

We observed a 3-month-old Japanese female infant with severe psychomotor retardation, coarse facial appearance, hepatosplenomegaly, and dysostosis multiplex. Only beta-galactosidase was found to be deficient when the routine lysosomal hydrolase assay was performed on the patient's lymphocytes at 6 months of age. At first GM1-gangliosidosis type 1 seemed the most likely diagnosis. Later, however, additional studies (hydrolase assay in cultured skin fibroblasts, urinary oligosaccharide analysis, genetic complementation study, etc.) revealed that biochemical data of this case were in agreement with those of severe infantile sialidosis. The only important exception was that alpha-neuraminidase in the patient's lymphocytes showed normal activity but abnormal pH dependence toward 4-methylumbellyferyl substrate. In addition, a severely damaged kidney suggested that his case may be classified as a unique type of severe infantile sialidosis (possible nephrosialidosis). These observations stress the importance of careful biochemical diagnosis of a case with GM1-gangliosidosis type 1 phenotype.

A CCD color signal separation IC for solid-state imagers with color filter arrays is described. The device simplifies peripheral circuitry and enhances picture qualities such as resolution, color fidelity, and stability for single-chip color imaging systems by incorporating CCD delay lines, sample-and-hold circuits, and dual clamp circuit. Also described is a color filter array utilizing Bayer geometry, which is used in the separation IC. Color images corresponding to a horizontal resolution of 340 TV lines with no erroneous color have been obtained from the device when used with a 580/spl times/475 element CCD imager.

The stoichiometry of 4-methyl sterol oxidase has been investigated by concurrent assays of rates of oxygen consumption, oxidation of reduced pyridine nucleotide, and formation of steroid 4alpha-oic acid, which is the oxidized product of attack of 4-methyl sterol precursors of cholesterol. The basal, steroid-independent rates of oxidation of alpha-NADH and alpha-NADH-dependent oxygen consumption by rat liver microsomes are about 10 to 15% of the rates observed with beta-NADH. Thus, alpha-NADH is substituted for beta-NADH; alpha-NADH oxidation is observed spectrophotometrically. The slow rate of oxygen consumption is measured accurately with a galvanic oxygen electrode that is attached to an offset amplifier. For maximal velocity, 4alpha-hydroxymethyl-5alpha-cholest-7-en-3beta-ol is the steroid substrate, and oxidase activity is induced 2-fold with a dietary bile acid sequestrant. Under these conditions, accurate measurements are obtained for substrate-dependent increments, which are equal to or greater than basal, substrate-independent rates. For each equivalent of hydroxymethyl group oxidized to carboxylic acid, 2 eq each of oxygen and alpha-NADH are consumed. Thus, the stoichiometry is consistent with that expected for two sequential attacks of the 4alpha-hydroxymethyl group by an external mixed function oxidase. In addition to establishing the stoichiometry of the 4-methyl sterol oxidase, the results further demonstrate that the steroidal 4alpha-carboxylic acid is formed from the hydroxymethyl intermediate by catalysis of a mixed function oxidase rather than dehydrogenases.