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Aminomethylphenylnorharman (AMPNH) and aminophenylnorharman (APNH) are mutagenic norharman derivatives obtained from o-toluidine and aniline, respectively. APNH is carcinogenic to the urinary bladder of rats and present in urine samples of healthy volunteers, indicating that norharman derivatives may be associated with cancer development in the urinary bladder of humans. To evaluate the possible role of AMPNH and APNH in bladder carcinogenesis, we examined the formation of γ-H2AX, a DNA damage response marker, in the urinary bladder of rats. Seven-week-old male F344 rats were treated with 400 ppm AMPNH or 40 ppm APNH in the diet for 4 weeks. Animals were killed at the end of administration or after 2 weeks of recovery, and immunohistochemistry for γ-H2AX and Ki67, a cell proliferation marker, was performed. At week 4, γ-H2AX formation in bladder epithelial cells was significantly increased by APNH treatment as compared with that in controls. AMPNH also induced upregulation of γ-H2AX formation, although there was no statistical significance. After the recovery period, γ-H2AX-positive cells were reduced but remained significantly higher in AMPNH and APNH groups than in the control group. Ki67-positive cells were significantly increased by AMPNH and APNH at week 4 and reduced to the same level as the control after 2 weeks of recovery. Expression of KRT14, a bladder stem cell marker, was also increased in the basal layer by the two norharman derivatives. Thus, AMPNH and APNH showed in vivo genotoxicity in the bladder epithelium of rats, and APNH may be a potent causative agent of bladder carcinogenesis.

9-(4′-Aminophenyl)-9H-pyrido [3,4-b] indole (aminophenylnorharman, APNH) is a novel mutagenic heterocyclic amine, produced by the reaction of norharman with aniline in the presence of S9 mix. In the present study, the maternal and developmental toxicity of APNH were investigated in ICR mice administered oral doses of 0, 0.625, 1.25, 2.5 or 5 mg/kg/day on gestational days (GD) 6 through 15 or 0, 5, 10, or 20 mg/kg on GD 12. Maternal and foetal parameters were evaluated on day 18 of gestation. Foetuses of dams treated on GD 6–15 were examined for external and skeletal malformations and variations, and foetuses of dams treated on GD 12 were inspected for cleft palate. Maternal death occurred when APNH was administered at 5 mg/kg/day on GD 6 –15. No significant decrease in body weight gain during the administration period was observed at doses of 2.5 mg/kg/day or less when applied on GD 6– 15. Adverse changes in general condition of dams were observed in the groups treated at doses of 2.5 mg/kg/day and above on GD 6 –15, whereas no adverse effects on dams were noted even when APNH was applied at a fairly high dose on GD 12. Intracytoplasmic vacuolation in hepatocytes, necrosis of proximal tubular epithelial cells and desquamation of necrotic epithelial cells in the tubular lumen were observed in dams treated with APNH at 2.5 or 5 mg/kg/day on GD 6 –15. Increased pre-implantation loss was observed at 5 mg/kg/day and post-implantation loss was observed at 2.5 mg/kg/day and above when applied on GD 6–15, or at 20 mg/kg when applied on GD 12. Foetal body weight was decreased by APNH in a dose-dependent manner. The frequency of external malformations (cleft palate) was significantly increased in the group treated with APNH at 2.5 mg/kg/day on GD 6–15 compared to the controls. However, there were no foetuses with cleft palate even when APNH was given at 20 mg/kg on GD 12. No significant increases in skeletally malformed foetuses were found in any APNH-treated group. The frequency of lumbar ribs was increased dose dependently. This study demonstrated the develop-mental toxicity of a mutagenic compound, APNH, in mice at maternally toxic doses, and that cleft palate observed in term foetuses resulted from the adverse effect of APNH on the maternal environment during organogenesis. More detailed studies are warranted to assess the possible risks to pregnant women from exposure to APNH. Human & Experimental Toxicology (2002) 21, 147 – 151.

Objective: To characterize Alexander disease (AxD) phenotypes and determine correlations with age at onset (AAO) and genetic mutation. AxD is an astrogliopathy usually characterized on MRI by leukodystrophy and caused by glial fibrillary acidic protein ( GFAP ) mutations. Methods: We present 30 new cases of AxD and reviewed 185 previously reported cases. We conducted Wilcoxon rank sum tests to identify variables scaling with AAO, survival analysis to identify predictors of mortality, and χ 2 tests to assess the effects of common GFAP mutations. Finally, we performed latent class analysis (LCA) to statistically define AxD subtypes. Results: LCA identified 2 classes of AxD. Type I is characterized by early onset, seizures, macrocephaly, motor delay, encephalopathy, failure to thrive, paroxysmal deterioration, and typical MRI features. Type II is characterized by later onset, autonomic dysfunction, ocular movement abnormalities, bulbar symptoms, and atypical MRI features. Survival analysis predicted a nearly 2-fold increase in mortality among patients with type I AxD relative to those with type II. R79 and R239 GFAP mutations were most common (16.6% and 20.3% of all cases, respectively). These common mutations predicted distinct clinical outcomes, with R239 predicting the most aggressive course. Conclusions: AAO and the GFAP mutation site are important clinical predictors in AxD, with clear correlations to defined patterns of phenotypic expression. We propose revised AxD subtypes, type I and type II, based on analysis of statistically defined patient groups.

Phosphorylated TDP-43 (pTDP-43) is the pathological protein responsible for amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disease. Recently, it has been reported that accumulation of pTDP-43 can occur in the brains of patients with argyrophilic grain disease (AGD), in which phosphorylated 4-repeat tau is the pathological protein. To elucidate the association of ALS with AGD, we examined the brains from 37 consecutively autopsied patients with sporadic ALS (age range 45-84 years, mean 71.5 ± 9.0 years).Sections from the frontotemporal lobe were stained with the Gallyas-Braak method and also immunostained with antibodies against phosphorylated tau, 4-repeat tau and pTDP-43.Fourteen (38%) of the 37 ALS patients were found to have AGD. With regard to staging, 5 of these 14 cases were rated as I, 4 as II and 5 as III. pTDP-43 immunohistochemistry revealed the presence of positive neuronal and glial cytoplasmic inclusions in the affected medial temporal lobe in many cases (93% and 64%, respectively). On the other hand, pTDP-43-positive small structures corresponding to argyrophilic grains were observed only in one case. A significant correlation was found between AGD and the Braak stage for neurofibrillary pathology (stage range 0-V, mean 2.1). However, there were no significant correlations between AGD and any other clinicopathological features, including dementia.The present findings suggest that co-occurrence of AGD in ALS is not uncommon, and in fact comparable with that in a number of diseases belonging to the tauopathies or α-synucleinopathies.

Objective This study aimed to determine the spectrum of pathological involvement of the striatonigral (StrN) and olivopontocerebellar (OPC) systems in Japanese patients with multiple system atrophy (MSA). This study also aimed to compare the pathological spectrum of Japanese MSA patients with the previously reported results in British MSA patients. Methods A semiquantitative pathological analysis of 50 MSA patients9 brains that were referred to the Brain Research Institute, Niigata University, Japan, was performed. The severity of neuronal cell loss was determined as previously described by the study from the Queen Square Brain Bank (QSBB), UK. Results The mean neuronal cell loss score was significantly higher in the OPC area than in the basal ganglia sites examined, except the dorsolateral putamen. The relative prevalence of pathological phenotypes showed that 40% of cases had OPC-predominant pathology, 18% had StrN-predominant pathology and the remaining (42%) had equivalent StrN and OPC pathology. None of the MSA cases had coexistent Lewy bodies in the dorsal motor nucleus of the vagus and the substantia nigra. Conclusions In contrast to the previously reported results involving British patients9 brains from the QSBB (OPC-predominant pathology 17%, StrN-predominant pathology 34%, equivalent StrN and OPC pathology 49%), the results of the present study showed more pathological involvement of the OPC system than of the StrN system. The rarity of Lewy bodies may underlie the phenotypic expression of Japanese MSA. The present observations reflect the disequilibrium in the phenotype distribution between the two populations.

Amyotrophic lateral sclerosis (ALS) is characterized by upper and lower motor neurone involvement with Bunina bodies (BBs) and TDP-43 inclusions. To elucidate the relationship between BBs and TDP-43 inclusions, we examined the spinal cord from 18 patients with ALS.Five serial sections from lumbar cord were first stained with haematoxylin and eosin to detect BBs and subsequently immunostained with anti-TDP-43 antibody. Immunoelectron microscopy was performed on vibratome sections from two cases of ALS.BBs were found in 15 out of 18 cases. TDP-43 inclusions were found in all the cases. The average incidence of anterior horn cells with BBs and TDP-43 inclusions relative to the total number of neurones was 17.1% and 46.4%, respectively. The concurrence of both inclusions in the same neurones was found in 15 cases. The incidence of co-localization of BBs and TDP-43 inclusions was 15.7% of total neurones. The frequency of TDP-43 inclusions was significantly higher in neurones with BBs than in those without. Ultrastructurally, TDP-43-immunoreactive filamentous structures were intermingled with early-stage BBs, but not associated with advanced-stage BBs.These findings suggest that there is a close relationship in the occurrence between BBs and TDP-43 inclusions.

Male Fischer 344 rats were given a single dose of 0.03-30 mg/kg of [2-14C]2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ([14C]PhIP), the radioactivity in urine and feces was determined over 48 h, and the major metabolites were identified and quantified. Dose had little effect on the profile of metabolites in the urine but did influence the profile in the feces. PhIP was more efficiently metabolized at higher doses. In addition, rats were pretreated with Aroclor 1254 (PCB), 3-methylcholanthrene (MC), phenobarbital (PB), PhIP and corn oil prior to a single dose of [14C]PhIP, and compared with a control group receiving [14C]PhIP only. The major metabolites in the urine and feces were quantitated for each group, as well as PhIP binding to serum proteins, hemoglobin and selected tissues. Pretreatment with MC and PCB resulted in an increase in the amount of 4'-hydroxylation of PhIP and a decrease in the amount of N-hydroxylated metabolites in the urine. Pretreatment with PB resulted in an increase in the amount of N-hydroxylated metabolites, but a decrease in 4'-hydroxylation. Pretreatment with either MC or PCB resulted in an increase in PhIP binding to the liver and kidney, while reducing the binding in other tissues. Animals pretreated with PhIP showed few significant differences from the untreated group, while pretreatment with PB in general resulted in a decrease of PhIP binding in tissues.

We conducted ultrasonic decalcification on calcified annulus in patients with aortic stenosis (AS) using an ultrasonic operator, Sonopet (UST 2001) prior to aortic valve replacement (AVR). We studied the reliability of this method.From January 2002 to August 2005, AVR was conducted for AS using the Sonopet in 45 patients, comprising of 18 male and 27 female subjects. The mean age was 73.3 +/- 9.7.Artificial valves were successfully inserted at the intra-annular level in 37 patients and at the supra-annular level in 8 patients without conducting annular enlargement. In the patients with narrow annuli of less than 19 mm (23 patients), the preoperative mean annular diameter was 18.2 +/- 1.0 mm, but significantly larger artificial valves with an average diameter of 19.3 +/- 1.5 mm (p=0.003) were successfully inserted.AVR was proved to be safe and easy by previous ultrasonic decalcification of the annuls using the Sonopet. This method was very useful because it required no enlargement of aortic annulus.

Male F344 5-week-old rats were X-irradiated, and 16 weeks after the first dose, azoxymethane (AOM) was injected or 2-amino-1 -methyl-6-phenylimidazo [4,5-b]pyridine (PhIP) was given by intragastric intubation. Tumors in the pylorus of the glandular stomach were observed in 4 out of the 29 animals receiving X-rays + AOM and in 4 out of the 25 animals receiving X-rays + PhIP, 12 months after administration. No such lesions were found in the chemical or X-ray alone groups. Intestinal metaplasia and some induced tumors were positive for CDX2. It was concluded that the presence of intestinal metaplasia may increase sensitivity to the induction of gastric tumors by colon carcinogens.

Behavioral evidence supports a role for peripheral glutamate receptors in normal nociceptive transmission. In this study, we examined the release of the excitatory amino acids, glutamate and aspartate, in the s.c. perfusate of the rat hind instep by in vivo microdialysis. Antidromic stimulation of the sciatic nerve and noxious stimuli in the form of heat stimulation and local application of capsaicin cream (1%) to the instep caused an increase in excitatory amino acid release. This capsaicin-induced excitatory amino acid release was suppressed by pretreatment with capsaicin. Both systemic (10 mg/kg, i.v.) and local injections (10(-5) M in the perfusate) of morphine inhibited the increase in excitatory amino acid release evoked by local application of capsaicin cream to the instep. This inhibitory effect of morphine was antagonized by naloxone either given systemically (5 mg/kg, i.v.) or locally (10(-5) M). These results suggest that excitatory amino acids are released from small diameter afferent fibers by heat stimulation in the periphery or local application of capsaicin cream, and that activation of opioid receptors, present on the peripheral endings of small-diameter afferent fibers, can regulate noxious stimulus-induced excitatory amino acid release.

Metallothioneins (MTs) are metal-binding proteins that have been regarded as intrinsic factors for protecting cells and tissues from metal toxicity and oxidants. Among the three major classes of MTs, MT-III is different from other MTs because it has neuronal inhibitory activity and is only expressed in the central nervous system. Recent studies, however, have confirmed that MT-III is also expressed in organs other than the brain. These findings not only indicate that MT-III has a much wider tissue distribution than was originally thought, but also suggest that it might have other unknown activities. In the present study, we examined the human salivary and thyroid glands and demonstrated that the MT-III gene is also expressed in the salivary but not in the thyroid gland. While salivary ducts showed intense immuno-reactivity with anti-MT-III, weak immunoreactivity was observed in acinar cells. This, together with the findings that some neuromodulators (i.e. nerve growth factor, etc.) exist in the salivary gland and that MT-III may participate in the transport in renal tubules, suggest that MT-III may have other functions than cytoprotection in the salivary gland.

Multiple system atrophy (MSA) is a sporadic neurodegenerative disease characterized by the presence of neuronal and oligodendroglial alpha-synuclein aggregates. To investigate the relationship between the occurrence of neuronal cytoplasmic and intranuclear inclusions (NCIs and NNIs, respectively) and the progression of neuronal degeneration, we performed a quantitative analysis of the pontine and inferior olivary nuclei based on 14 cases of MSA. alpha-Synuclein immunohistochemistry revealed that NCIs and NNIs were present in both brain nuclei in all the cases. The average incidence of NCIs in the pontine and inferior olivary nuclei was 9.1% and 25.8%, respectively, and that of NNIs was 9.2% and 9.0%, respectively. The number of NNIs was strongly correlated with that of neurones in the pontine and inferior olivary nuclei. Although the number of NCIs was not correlated with the neuronal population in both nuclei, the NCI count in patients with moderate MSA was higher than in patients with mild MSA. The NNI count was much higher than the NCI count in the pontine nucleus in four patients, and was the same in the olivary nucleus in three of the four patients. Moreover, the neuronal population in the NNI-predominant cases was significantly higher than in the NCI-predominant cases. These findings suggest that NCI formation is accelerated by the progression of the disease process, and that in MSA, NNI formation is an earlier phenomenon than NCI formation.

The morphologic characteristics of the dentition with maxillary prognathism and reversed occlusion were examined by a computer-assisted dental cast analysing system. Dental casts with normal occlusion, maxillary prognathism and reversed occlusion were selected and measured by a 3D shape measuring system. The dental arches and anteroposterior occlusal curves were approximated numerically by the polynomial expression with a fourth order and second order coefficients, respectively. The coefficients were analysed statistically. Maxillary dental arches with mandibular prognathism showed a more acuminate shape (V-shape) with a significant larger second order coefficient, while arches with reversed occlusion showed more angulate shapes (U-shape) with significantly smaller second order coefficients compared with arches with normal occlusions. However, a notable difference in the shape of the mandibular dental arches was not observed. The mandibular dental arches with maxillary prognathism were positioned backward relative to maxillary dental arches, while the arches with reversed occlusion were positioned forward. The anteroposterior occlusal curves with maxillary prognathism showed larger curvatures in maxilla and mandible, while the curves with reversed occlusion showed smaller curvatures in mandible comparing with the curves with normal occlusion. These results suggest that the present method could offer a useful and objective examination technique for the diagnosis of malocclusion.

The modifying effects of a cyclooxygenase (cox)-2 selective inhibitor nimesulide on tongue carcinogenesis were investigated in male F344 rats initiated with 4-nitroquinoline-1-oxide (4-NQO). The cell proliferation activity measured by proliferating cell nuclear antigen (PCNA)-positive index and apoptotic index, and the immunohistochemical expression of COX-2, and inducible nitric oxide synthase (iNOS) in the tongue mucosa or neoplasms were also examined for mechanistic analysis of modifying effects of nimesulide on tongue carcinogenesis. All animals except those treated with nimesulide alone and untreated rats were given 20 ppm 4-NQO in drinking water for 8 weeks to induce tongue neoplasms. Starting 1 week after the cessation of 4-NQO exposure, rats given 4-NQO were fed the experimental diets containing nimesulide (100 and 400 ppm) for 22 weeks. At week 32, the incidence of tongue squamous cell carcinoma was significantly reduced by feeding of the diet containing 400 ppm nimesulide. Feeding of nimesulide significantly decreased polyamine content and PCNA-labeling index in tongue carcinoma. Apoptotic index in tongue carcinoma was increased by feeding of nimesulide. In addition, nimesulide feeding reduced COX-2 and iNOS expression in the tongue dysplasia and neoplasms. These results suggest that 400 ppm nimesulide in diet, when given during the promotion phase, exerts chemopreventive ability against 4-NQO-induced tongue tumorigenesis through inhibition of cell proliferation activity in conjunction with modification of COX-2 and iNOS expression of the target lesions.

Primary intracranial solitary leptomeningeal gliomas are exceedingly rare. We, therefore, performed a detailed clinical, radiological and pathological analysis to better characterize these tumors in 3 patients (33- and 72-year-old men and a 72-year-old woman). Two of the tumors were located in the frontal region and 1 in the temporal region. Magnetic resonance imaging revealed a well circumscribed large lesion (maximal diameter 4 - 6 cm) with peritumoral edema, mixed low- and isosignal intensity on T1-weighted images, hypersignal intensity on T2-weighted images and non-homogeneous contrast enhancement. External carotid angiography demonstrated a vascular supply to these tumors via branches of the middle meningeal artery. Gross total resection was achieved in all patients. The pathological diagnosis was glioblastoma in 2 patients and oligodendroglioma in 1. The MIB-1 nuclear labeling index ranged from 11.8% - 23.6% (mean 18.2%). Local tumor recurrence was documented in 2 patients after 8 and 11 months, respectively. The other patient with glioblastoma developed a metastasis to the femur 39 months after craniotomy. A definitive diagnosis can be made by careful radiological assessment and histopathological examination.

The purpose of this study was to develop a method to integrate three-dimensional data of dentition and facial morphology measured by a high-speed laser scanner, and visualize them by computer graphics. The accuracy of integration was also examined.The 3-D shapes of upper and lower dental casts and the face of a subject were measured by a high-speed laser scanner with zoom lens facility. As an interface to integrate the coordinates between dentition and face, an extraoral marker plate attached to the occlusal record plate was devised. It was applied between the upper and lower dental casts, and with the subject, and measured. Integration was attempted by coinciding the data of the interface. The accuracy of integration was examined using the conditions of the marker plate such as surface shape and attachment angle.Integration of the data of dentition and face was accomplished. The 3-D relationship between them could be visualized in any direction or section by computer graphics. For optimal accuracy of integration, the marker plate with a mixed shape of flat and sinusoidal wave attached at 45 degrees to the occlusal record plate showed the most appropriate results.An original method to integrate the 3-D data of dentition and face measured by 3-D laser scanner was developed and shown to have acceptable accuracy.

Cyclooxygenases (COXs) and prostanoids play pivotal roles in colon carcinogenesis. This study was designed to determine the chemopreventive effects of ONO-8711, a selective prostaglandin E receptor EP1 antagonist, on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) in male F344 rats and to compare its potential with that of nimesulide, a well-documented selective COX-2 inhibitor.Five-week-old male F344 rats received s.c. injections of AOM (15 mg/kg body weight) or the saline vehicle once weekly for two weeks and were fed the control diet (AIN-76A) or the experimental diets containing 400 or 800 ppm of ONO-8711 or 400 ppm nimesulide for 5 weeks.Administration of ONO-8711 at 800 ppm significantly reduced the total number of ACF/colon and 5-bromodeoxyuridine (BrdUrd) labeling index as compared to the control diet group (by 31% and 66%, respectively). As expected, dietary administration of nimesulide also suppressed the development of ACF and BrdUrd labeling index in the colon, by about 39% and 54%, respectively.Our finding that ONO-8711 significantly suppresses colonic ACF formation and cell proliferation strengthens the hypothesis that the selective prostaglandin E receptor EP1 antagonists possesses chemopreventive activity against colon cancer development.

To produce a safe and effective inactivated poliovirus vaccine (IPV), we have developed S-IPV using Vero cells infected with the Sabin strains in a semi-production scale. All production steps including virus culture on microcarrier beads were highly reproducible. Mean recovery percents of infectious viruses or D-antigens during all processes for concentration, purification and inactivation were 30-50% in the three types. The S-IPV potency was adjusted for D-antigen content as determined by in-house ELISA and was comparable to WHO reference IPV derived from the virulent strains in immunogenicity tests in rats. Antibody development in more than 30 seronegative infant volunteers after two shots of S-IPV at four-week interval were 100% without notable adverse reactions. The mean antibody titres (log2) to Sabin 1, 2 and 3 viruses were 11.1, 8.3 and 8.9, respectively. The antibodies neutralized the Mahoney, MEF-1, and Saukett virulent strains with slightly inferior titres to those of the Sabin strains. D-antigens for each type of S-IPV were stable at 4 degrees C without any significant decrease over more than two years.

The FLEP regimen (5-FU, LV, ETP and CDDP) is a combination chemotherapy administered regionally and systemically for the control of both local and disseminated disease in intra- and extra-abdominal regions in patients with advanced and recurrent gastric cancer. Sixty-one patients with advanced and recurrent gastric cancer were entered into this study. The treatment regimen consisted of 5-FU at 370 mg/m2 (days 1 to 5, i.v. 24 h); LV at a dose of 30 mg (days 1 to 5, i.v. bolus); and ETP and CDDP each at 70 mg/m2 (days 7 and 21, ia 2 h). This regimen was repeated every four weeks. The overall response rate was 36.1% (22/61) and the 50% and median survival times were 10.23 and 11.80 months, respectively. The adverse events were Grade 3/4 leukocytopenia (18.0%), Grade 3/4 thrombocytopenia (4.9%), Grade 3 nausea and/or vomiting (3.3%) and Grade 3 stomatitis (1.6%). Of the 17 NAC patients, the six curability B patients showed a statistically higher survival rate than the curability C and unresected patients. Based on the encouraging response rate and the improvement in prognosis, we recommend the FLEP regimen for patients with primary gastric cancer. Neoadjuvant chemotherapy using the FLEP regimen should be performed with curative resection as an objective.

We report a case of a 59-year-old man with a severe generalized form of Hailey-Hailey disease that was complicated by fatal liver injury. Erosive lesions were first noted in the axillary and perianal regions at 15 year of age, and Hailey-Hailey disease was diagnosed based on the clinical features and histologic findings in skin biopsy specimens. The patient was treated with at first topical steroids and later a low dose of a corticosteroid, but the skin lesions gradually became generalized. At 45 years of age liver dysfunction was detected after azathioprine and vinblastine treatment for the generalized skin lesions. The liver injury gradually progressed and finally the patient died. The gene responsible for Hailey-Hailey disease was recently identified as ATP2C1, and it encodes a Ca(2+)-transport ATPase with broad expression, including in skin and liver. This finding suggests that mutation of the ATP2C1 gene may give rise to an extracutaneous phenotype, such as the liver dysfunction observed in severe cases, including our own. Further accumulation of cases is necessary to determine whether this is true.

The development of retinal lesions induced by a uniform regimen of interferon-alpha therapy for chronic hepatitis C was prospectively investigated.Eighty-one patients received 6 mega units of natural interferon-alpha intramuscularly daily for 2 weeks and then 3 times a week for 22 weeks. The total dose of interferon-alpha administered was uniformly 478 mega units per patient. Two expert ophthalmologists prospectively examined the patients for retinal lesions before, during and after the therapy.Retinopathy was not found in comparison groups or any of the patients before treatment. In total 34.6% (28/81) of the patients showed cotton-wool spots or minor retinal hemorrhage, or both lesions, during therapy, but these lesions were reversed during or after interferon therapy. The occurrence rates of cotton-wool spots alone, retinal hemorrhage alone and both lesions were 13.6% (11/81), 6.2% (5/81), and 14.8% (12/81), respectively. The appearance of retinopathy did not correlate with patients' background including viral loads and response to the therapy, but was more frequently found in older patients and patients with hypertension and/or diabetes mellitus; disappearance of retinopathy was more prolonged than in patients without these complications. Almost all the lesions appeared 2-4 months after the start of the therapy, and the severity of the lesions did not differ between patients with and without hypertension and/or diabetes mellitus.Although it is not clear if interferon-associated retinopathy occurs in a dose-dependent manner, the present study shows a standard pattern of the occurrence of retinopathy in patients with chronic hepatitis C receiving a uniform dosage of natural interferon-alpha.

Neuronal degeneration in amyotrophic lateral sclerosis (ALS) is associated with ubiquitinated cytoskeletal alterations in the motor neuron system. Ubiquitin-positive inclusions are also seen in the limbic system in ALS with dementia (ALS-D). Recently, similar inclusions were reported to occur in the neostriatum in a case of ALS-D. We, therefore, immunohistochemically examined the neostriatum from 60 patients with ALS and 60 control subjects.Two forms of ubiquitin-positive inclusions were found in the ALS neostriatum: rod-like inclusions in the large neurons and crescent-shaped inclusions in the small neurons. The former were found in 14 ALS and 18 control subjects, whereas the latter were specific to ALS; the crescent-shaped inclusions in small neurons were found in 27 ALS patients, and the immunohistochemical and ultrastructural features were identical to those of the extra-motor inclusions of ALS. Moreover, characteristic temporal lesions consistent with those seen in ALS-D were found in 8 patients, of whom 2 had shown dementia.The present findings strongly suggest that neostriatal small neurons are also involved in the disease process in ALS.

To examine the effect of coffee drinking on serum gamma-glutamyltransferase (GGT) level in relation to alcohol drinking, smoking, and degree of obesity in middle-aged Japanese men.From 1986 to 1994, a total of 7,637 male officials of the Self-Defense Forces of Japan aged 48-59 years received a preretirement health examination. Coffee drinking was ascertained by a self-administered questionnaire, and serum GGT level was measured. After excluding 1,360 men with a possible pathologic condition influencing liver enzyme levels and 182 former alcohol drinkers, effect of coffee drinking on serum GGT was examined by a multiple linear regression model and analysis of variance adjusting for alcohol drinking, smoking, and body mass index (BMI).The adjusted percentage of difference in serum GGT was -4.3 (95% CI = -5.0; -3.5) per cup of coffee. The inverse coffee-GGT relation was most prominent among men drinkingor = 30 ml of ethanol and smokingor = 15 cigarettes daily; and positive associations of alcohol and smoking with GGT were attenuated by coffee drinking, more clearly among men with BMIor = 25.00 kg/m2. Adjusted percentages of difference in serum GGT were -2.6% (p = 0.0003) per cup of brewed coffee, and -5.1% (p = 0.0001) per cup of instant coffee, independently of each other.The present study suggests that coffee consumption may weaken GGT-induction by alcohol, and possibly by smoking. These effect modifications by coffee may differ according to the degree of obesity.

alpha-Synuclein is now known to be a major component of Lewy bodies (LBs) in Parkinson's disease (PD) and diffuse Lewy body disease (DLBD). Recently, cytoplasmic aggregation of alpha-synuclein has also been reported to occur in glial cells in these diseases.We have conducted an immunohistochemical survey to clarify in detail the distribution of alpha-synuclein aggregates in the central nervous system of patients with DLBD. The cerebrum, brainstem, cerebellum and spinal cord of six patients with DLBD were examined immunohistochemically using anti-alpha-synuclein antibodies.In all patients, alpha-synuclein-immunoreactive cytoplasmic inclusions, including those with profiles of typical LBs, were visualized in neurons of the cerebral neocortex, hippocampus, amygdaloid nucleus, hypothalamus, brainstem pigmented nuclei and reticular formation. In some of these patients, similar spherical cytoplasmic inclusions were also detected in neurons of the olfactory bulb, basal ganglia, thalamus, the subthalamic, pontine, inferior olivary and cerebellar dentate nuclei, and in the spinal gray matter. Moreover, alpha-synuclein-immunoreactive cytoplasmic inclusions, which appeared circular or coil-like in shape, were found in glial cells. In four patients with longstanding DLBD, these cytoplasmic inclusions were distributed widely in brain areas, including brainstem, basal ganglia, and cerebral and cerebellar white matter.The widespread occurrence of alpha-synuclein aggregates in both neuronal and glial cells is a pathological feature in patients suffering from DLBD.

Activation of the beta-catenin/T cell factor-mediated transcription pathway through mutations of the APC or beta-catenin gene is suggested to play an important role in colon carcinogenesis and there is great interest in the target genes. We have described the frequent mutation and an altered cellular localization of beta-catenin in rat colon adenocarcinomas induced by azoxymethane (AOM), along with up-regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. In the present study, the relation between beta-catenin alteration and expression of iNOS and COX-2 in AOM-induced rat colon carcinogenesis was examined in hyperplastic and dysplastic type aberrant crypt, adenoma and adenocarcinoma samples. K-ras gene mutations were also investigated. Mutation analysis by the PCR-single strand conformation polymorphism method and direct sequencing demonstrated the beta-catenin gene to be mutated in two of three dysplastic aberrant crypt foci (ACF), two of six adenomas and 20 of 26 adenocarcinomas, while K-ras was mutated in seven of 10 hyperplastic ACF and seven of 26 adenocarcinomas. Immunohistochemical staining showed an alteration in cellular localization of beta-catenin in all dysplastic ACF, adenomas and adenocarcinomas examined. iNOS expression was also observed in all but one of the lesions in which beta-catenin alterations were observed. Neither iNOS expression nor beta-catenin alterations were observed in any hyperplastic ACF. COX-2 expression in stromal elements was found even in normal colon mucosa and increased in adenomas and adenocarcinomas, while epithelial cells were only positive in large adenocarcinomas. These results show that beta-catenin alterations may be related to induction of iNOS expression, these being early events in AOM-induced colon tumorigenesis which may play important roles in causing dysplastic changes.

The beta-catenin gene is frequently mutated at codons 33, 41 and 45 of the glycogen synthase kinase-3beta phosphorylation motif in human colon cancers in patients without APC mutations. Frequent mutations at codons 32 and 34, as well as 33 and 41, have been detected in rat colon tumors induced by azoxymethane (AOM), with the second G of CTGGA sequences being considered as a mutational hot-spot. In the present study, exon 3 of the beta-catenin gene in mouse colon tumors induced by AOM was amplified by PCR and mutations were detected by the single strand conformation polymorphism method, restriction enzyme fragment length polymorphism and direct sequencing. All 10 colon tumors tested were found to have beta-catenin mutations, four in codon 34, three in codon 33, two in codon 41 and one in codon 37, nine being G:C--A:T transitions. However, no mutations were found in codon 32 of the mouse beta-catenin gene. On immmunostaining, beta-catenin was observed in the cytoplasm and nucleus of the tumor cells. The cytoplasmic staining was homogeneous, while both homogeneous and heterogeneous patterns were noted for the nuclei. Highly frequent mutations of the beta-catenin gene in AOM-induced mouse colon tumors suggest that consequent alterations in the stability and localization of the protein may play an important role in this colon carcinogenesis model.

Although the cyclooxygenase pathway of the arachidonic acid cascade has been suggested to play an important role in colon carcinogenesis, the molecular species of prostanoids and receptors involved have not been fully elucidated yet. We examined the development of aberrant crypt foci (ACFs), putative preneoplastic lesions of the colon, in two lines of knockout mice, each deficient in prostaglandin E receptors, EP1 and EP3, by treatment with the colon carcinogen, azoxymethane. Formation of ACFs was decreased only in the EP1-knockout mice to approximately 60% of the level in wild-type mice. Administration of 250, 500, or 1000 ppm of a novel selective EP1 antagonist, ONO-8711, in the diet to azoxymethane-treated C57BL/6J mice also resulted in a dose-dependent reduction of ACF formation. Moreover, when Min mice, having a nonsense mutation in the adenomatous polyposis coli gene, were given 500 ppm ONO-8711 in the diet, the number of intestinal polyps was significantly reduced to 57% of that in the basal diet group. These results strongly suggest that prostaglandin E2 contributes to colon carcinogenesis to some extent through its action at the EP1 receptor. Thus, EP1 antagonists may be good candidates as chemopreventive agents for colon cancer.

Argyrophilic grain disease (AGD) is a progressive disorder producing dementia in elderly individuals characterized by the presence of numerous AGs in the limbic system. However, the occurrence of AGs has been reported in other neurodegenerative conditions including Alzheimer's disease, Pick's disease, progressive supranuclear palsy and corticobasal degeneration, all of which show tau-positive cytoskeletal abnormalities. We examined the brains of 26 patients with multiple system atrophy (MSA), a neurodegenerative disorder fundamentally lacking tau pathology, histologically and immunocytochemically. Numerous AGs were found in the limbic system in 5 patients, of whom two had shown mild dementia. Immunocytochemically, these AGs were labeled with antibodies against phosphorylation-dependent and -independent tau protein, but not alpha-synuclein, whereas oligodendroglial cytoplasmic inclusions found exclusively in MSA were immunoreactive for alpha-synuclein and, less consistently, for phosphorylation-independent tau but not for phosphorylation-dependent tau protein. Furthermore, many phosphorylation-dependent tau-positive neurons and significant numbers of ballooned neurons (BNs) were found in the limbic system in all of the 5 patients with AGs. These findings suggest that AGs can occur with relatively high frequency in the limbic system of MSA patients, and that as in AGD, they may be accompanied by tau-positive neurons and BNs.

Few studies have investigated the relation between glucose tolerance status and ultrasonographically determined gallstone disease. Using a 75-g oral glucose tolerance test, we examined the association of impaired glucose tolerance (IGT) and non-insulin-dependent diabetes mellitus (NIDDM) with gallstone disease in Japanese men. Subjects were men aged 48 to 59 of the Japan Self-Defense Forces who received a preretirement health examination between October 1986 to December 1994. After exclusion of 12 men under insulin treatment in the consecutive series of 7637 men, 174 were found to have gallstones; 103 were at the state of postcholecystectomy, and 6899 had normal gallbladder. IGT and NIDDM were associated with a modestly increased risk of gallstone disease; adjusted odds ratios were 1.3 (95% confidence interval [CI]: 0.9-1.8) for IGT and 1.3 (95% CI: 0.8-2.0) for NIDDM after adjustment for hospital, rank, smoking, alcohol use, and body mass index. Adjusted odds ratio for IGT and NIDDM combined was 1.3 (95% CI: 1.0-1.7, p=0.08). When prevalent gallstones and postcholecystectomy were considered separately, NIDDM showed a significant, positive association with postcholecystectomy, but not with prevalent gallstones. The findings add to evidence that glucose intolerance is associated with a modest increase in the risk of gallstone disease.

To elucidate the biological functions of poly(ADP-ribose) polymerase (PARP, [EC 2.4.2.30]) in DNA damage responses, genetic and biochemical approaches were undertaken. By disrupting exon 1 of the mouse PARP gene by a homologous recombination, PARP-deficient mouse embryonic stem (ES) cell lines and mice could be produced without demonstrating lethality. PARP-/- ES cells showed complete loss of PARP activity and increased sensitivity to gamma-irradiation and an alkylating agents, indicating a physiological role for PARP in the response to DNA damage. p53, a key molecule in cellular DNA damage response, was found to stimulate PARP activity and became poly(ADP-ribosyl)ated in the presence of damaged DNA. However, PARP-/- ES cells showed p21 and Mdm-2 mRNA induction following gamma-irradiation, indicating that PARP activity is not indispensable for p21 and Mdm-2 mRNA induction in the established p53-cascade. On the other hand, in a reconstituted reaction system, purified PARP from human placenta suppressed the pRB-phosphorylation activity in the presence of NAD and damaged DNA. Human PARP expressed in E. coli showed a similar effect on pRB-phosphorylation activity of cdk2. These findings suggest a direct involvement of PARP in the regulation of cdk activity for cell-cycle arrest.

We report a 75-year-old man who presented with parkinsonism, dementia and vertical gaze palsy. Postmortem examination revealed frontoparietal atrophy and nigral depigmentation. The temporal lobe was well preserved. Histologically, the affected cerebral cortex showed vacuolation and neuronal loss in layers 2 and 3. The substantia nigra and globus pallidus were also affected. There was gliosis in the thalamic medial nuclei and midbrain tegmentum. Neither ballooned neurons nor taupositive, argyrophilic inclusions could be found. Moreover, there was no Alzheimer pathology manifested by senile plaques and neurofibrillary tangles. In fact, the clinical and pathological features of the present case were consistent with those observed previously in two other cases reported elsewhere (Kawasaki et al. Acta Neuropathol 91: 140, 1996). In these cases, corticobasal degeneration (CBD) and frontotemporal dementia (FTD) were considered to be the principal entities in the differential diagnosis. However, we believe that these three cases, including the present one, were not examples of CBD or FTD, since neither ballooned neurons nor tau abnormalities were present, and the precentral gyrus was the area most severely involved, the temporal lobes being obviously spared. Therefore we considered that they might represent a hitherto unrecognized new disease entity.

The study aims to examine the relationship between habitual coffee consumption and blood pressure. The subjects were 3336 male self-defense officials aged 48-56 years, who received a preretirement health examination at the Self-Defense Forces Fukuoka Hospital between October 1986 and December 1992. Average coffee intake in the past year was ascertained by a self-administered questionnaire. A significant inverse relation between habitual coffee consumption and blood pressure was found with and without adjustment for alcohol use, cigarette smoking, body mass index, glucose tolerance, and green tea intake. Green tea, another major source of caffeine intake in Japanese, was unrelated to blood pressure. The adjusted mean differences per cup of coffee consumed per day were -0.6 mmHg (95% confident interval [CI]: -0.9 to -0.3, p = 0.0001) in systolic blood pressure and -0.4 mmHg (95% CI: -0.5 to -0.2, p = 0.0002) in diastolic blood pressure. Habitual coffee drinkers had lower blood pressure than non-drinkers at any levels of alcohol use, cigarette smoking, obesity, and glucose intolerance. Our findings consolidate the previous observation that habitual coffee consumption was associated with lower blood pressure.

We describe a 67-year-old man who died 4 days after suffering a subarachnoid hemorrhage. Autopsy revealed a fresh subarachnoid hemorrhage and a ruptured fungal aneurysm near the trifurcation of the right middle cerebral artery. In comparison with 21 previously reported cases in which the fungal aneurysms were proved to be intracranial, the present case had several characteristic features: the causative fungus of the aneurysm was Candida (only one such case has been reported previously). The aneurysm was caused by direct Candida invasion of the arterial wall from the Candida embolus (previously reported aneurysms have been caused by direct invasion of the arterial wall during fungal meningitis). The source of the Candida was endocarditis (the main sources of fungus in previously reported cases have been sinusitis, dental extraction wounds, and some forms of surgery). We describe the features of this rare autopsy case of a ruptured fungal aneurysm caused by Candida originating from endocarditis and review the literature.

The chemopreventive potential of a selective cyclooxygenase-2 inhibitor, nimesulide (NIM), against the development of rat superficial urinary bladder carcinomas after initiation with N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) was examined. Six-week-old Fischer 344 male rats were given 0.05% BBN in their drinking water for 8 weeks, followed by diets supplemented with 0, 100, 200, or 400 ppm NIM for 12 weeks, and they were then sacrificed. NIM decreased, in a dose-dependent manner, the incidence of transitional cell carcinoma (TCC) to 12 of 20 (60.0%), 8 of 16 (50.0%), and 5 of 19 (26.3%) and the multiplicity of TCCs to 0.75 +/- 0.79, 0.56 +/- 0.63, and 0.37 +/- 0.78 per rat at 100, 200, and 400 ppm, respectively, as compared with the BBN alone group values of 18 of 20 (90.0%) and 2.35 +/- 1.23. NIM did not significantly affect the cell differentiation or invasiveness of TCCs. These results indicate clear chemopreventive potential of a selective cyclooxygenase-2 inhibitor against postinitiation development of superficial rat urinary bladder carcinomas.

We investigated the independent associations of total and beverage-specific ethanol consumption with body mass index (BMI) and waist-to-hip ratio (WHR) in middle-aged Japanese males, because of the scarcity of epidemiologic data in Japan. The subjects were 2227 male self-defense officials who received a pre-retirement health examination at the Self-Defense Forces Fukuoka, Kumamoto, and Sapporo Hospitals. Data on alcohol intake, smoking, physical activity, and past medical history were obtained from a self-reported questionnaire. Height, weight, and waist and hip girth measurements were obtained at the examination. Multiple linear regression analysis was employed. Alcohol intake was positively and strongly associated with WHR (p = 0.0001), but not associated with BMI after adjustment for lifestyle variables, including either BMI or WHR. Subjects who consumed 15 ml per day or more of shochu ethanol showed a larger WHR than never drinkers, and a dose-response relationship was found. Dose-response relationships to other beverages were not found. Dose-response relationships to other beverages were not found. These findings suggest that alcohol intake is strongly and independently associated with WHR, but not with BMI. Abdominal obesity was positively associated with shochu ethanol, but not with other types of alcohol.

Prostate tissues obtained from rats given a food-derived carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), at a dose of 400 ppm in the diet for 52 weeks were histopathologically evaluated and found to contain prostate carcinomas limited to the ventral lobe in 18 of 27 cases. Atypical hyperplasias were also detected in the ventral and anterior prostate as well as the seminal vesicles. 32P-Postlabeling analysis of DNA demonstrated that PhIP-DNA adducts are produced in all lobes of the prostate of rats receiving PhIP. The findings indicate that PhIP is carcinogenic to rat prostate in addition to the previously demonstrated targeting of the colon and mammary glands, providing evidence of a possible role of PhIP in human prostate carcinogenesis and highlighting the potential importance of PhIP for man.

Arachnoid cysts very rarely occur within the ventricular system, where no arachnoid tissue exists. We present three cases of intraventricular arachnoid cyst with special reference to its origin. The first patient was a 5-year-old boy who complained of headaches and enlargement of his head. A CT scan revealed obstructive hydrocephalus and a large cystic lesion in the right lateral ventricle. The symptoms resolved after fenestration of the cyst and cystoperitoneal shunt. The second patient was a 49-year-old woman who complained of headache and numbness in her left upper extremity. A CT scan and MRI revealed a large cyst in the trigone of the right lateral ventricle. Fenestration of the cyst wall and cystoperitoneal shunt were performed relieving her complaints. The third patient was a 42-year-old man who complained of frequent seizures and dizziness. A CT scan and MRI demonstrated a moderate size cystic mass in the inferior horn of the right lateral ventricle. The symptoms were improved by partial resection of the cyst wall. Immunohistochemical studies and light microscopy confirmed that the cyst walls were composed of arachnoid membrane, implying that the cysts were arachnoid cysts. CT and MRI in these three cases showed widening of the choroidal fissure bordering the cyst wall, occasionally involving a part of the protruding cyst wall. On enhanced CT and MRI, the choroid plexus in the trigone of ipsilateral lateral ventricle was displaced anterolaterally, implying that the cysts had grown from outside the choroid plexus. Postoperative MRI demonstrated the shrunken cyst wall attached to the choroidal fissure. The surgical findings in case 3 also showed that the cyst wall was attached firmly to the choroid plexus. These findings appeared to indicate that the intraventricular arachnoid cyst originated from the arachnoid layer drawn into the choroidal fissure with choroidal vascular mesenchyme.

We report a 72-year-old man with sporadic amyotrophic lateral sclerosis (ALS) who showed concomitant histopathology of Alzheimer's disease (AD) and incidental Lewy body disease. The patient presented at the age of 70 years with distal upper limb amyotrophy. Thereafter, gait disturbance and respiratory distress progressed. Neuropathological examination showed mild frontal lobe and anterior spinal root atrophy. There was moderate loss of upper and lower motor neurons, and Bunina bodies and skein-like inclusions were present in the spinal anterior horns and facial and hypoglossal nuclei, confirming the pathology of ALS. In addition, however, numerous amyloid plaques were observed throughout the entire cerebral neocortex, nucleus accumbens and amygdaloid body. Many neurofibrillary tangles were also evident in the medial temporal cortex. Moreover, the substantia nigra showed mild degeneration, and Lewy bodies were found in the substantia nigra, locus ceruleus, basal nucleus of Meynert and peripheral autonomic ganglia. Although neither parkinsonism nor dementia was noted during the clinical course, our final neuropathological diagnosis was sporadic ALS, AD and incidental Lewy body disease (or presymptomatic Parkinson's disease). Whether or not the coexistence of these three diseases in the same patient was merely coincidental is of considerable interest.

Norharman, widely distributed in our environment, is alone not mutagenic to Salmonella typhimurium TA98 and TA100 either with or without S9 mix, but becomes mutagenic to S.typhimurium TA98 with S9 mix when non-mutagenic aromatic amines like aniline or o- or m-toluidine are added. Thus norharman has been called a 'co-mutagen'. In the present study we examined whether or not DNA adducts are formed in DNA of S.typhimurium TA98 by treatment with norharman and aromatic amines using 32P-post-labeling analysis under modified adduct intensification conditions. When a sample of norharman (8 mg) and aniline (4 mg) was incubated with 4 ml of overnight culture of S.typhimurium TA98 in the presence of 20 ml S9 mix for 6 h at 37 degrees C, three adduct spots were detected at a total relative adduct labeling (RAL) of 10.8 +/- 2.27/10(8) nucleotides. Under the same conditions, a mixture of norharman (8 mg) and o-toluidine (4 mg) yielded three adduct spots at a RAL of 3.74 +/- 1.71/10(8) nucleotides. With a combination of norharman and m-toluidine, a single adduct spot was seen at a RAL of 0.04 +/- 0.01/10(8) nucleotides. In contrast, norharman with p-toluidine did not produce adduct spots. Furthermore, neither norharman nor the aromatic amines themselves gave any evidence of adducts. Thus DNA adduct formation by norharman with aromatic amines correlates with the co-mutagenic action of norharman in S.typhimurium TA98.

The effects of 1-O-hexyl-2,3,5-trimethylhydroquinone (HTHQ), green tea catechins (GTC), alpha-tocopherol, beta-carotene, chlorophyllin, phenylethylisothiocyanate (PEITC), 3-O-ethylascorbic acid (EAsA), 3-O-dodecylcarbomethyl ascorbic acid (DAsA), n-tritriacontane-16,18-dione (TTAD) and d-limonene on 2-amino-6-methyldipyrido[1,2-a:3',2'-d]imidazole (Glu-P-1)- or dimethylnitrosamine (DMN)-induced hepatocarcinogenesis, and the dose dependence of HTHQ inhibition of Glu-P-1- or 2-amino-3,8-dimethylimidazo [4,5-f]quinoxaline (MeIQx)-influence on lesion development were examined in a rat medium-term liver bioassay system featuring diethylnitrosamine initiation and partial hepatectomy. At the end of week 8, the number and total area of glutathione S-transferase placental form (GST-P) positive liver foci in rats treated with 0.03% Glu-P-1 alone were increased significantly (46.8 +/- 11.0 and 12.0 +/- 5.6 respectively) as compared to the control values (3.8 +/- 1.6 and 0.4 +/- 0.2). Combined treatment with 1% HTHQ remarkably reduced both of these parameters (8.1 +/- 2.1 and 0.6 +/- 0.2). GTC (1%), PEITC (0.1%), beta-carotene (0.1%) and DAsA (1%) also demonstrated inhibition but less than HTHQ. On the other hand, these antioxidants did not influence development of foci initiated by 0.002% DMN. In the dose-response study, up to 0.125% HTHQ significantly reduced the effects of 0.02% Glu-P-1 or 0.03% MeIQx on the number and area of foci. These results indicate that several antioxidants exert chemopreventive effects against heterocyclic amine (HCA)-induced hepatocarcinogenesis, and particularly HTHQ which thus deserves further attention as a chemopreventor in the contest of the environmentally important HCA group of carcinogens.

Effects of transplacental and trans-breast milk exposure to a food-derived mammary and colon carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), were investigated in rats. Female Sprague-Dawley rats were administered PhIP in the diet (100 ppm) for 4 weeks before mating with nontreated males and also during gestation and lactation. As controls, additional females were maintained on the basal diet without PhIP and mated as with the treated animals. The offspring of both groups were subdivided for each sex at weaning into three dietary groups receiving 100, 25, and 0 ppm and were killed at 47 weeks of age. Effects of the transplacental and neonatal exposure to PhIP on mammary carcinogenesis were most evident in females administered 25 ppm PhIP after weaning; the incidence and multiplicity of adenocarcinomas in offspring from the PhIP fed dams (42.9%, 0.62/rat) was significantly higher than the value for offspring from nontreated dams (4.8%, 0.05/rat). Furthermore, in the basal diet groups, the incidence of adenocarcinomas in females was higher, albeit not significantly, in offspring of the PhIP-treated than the nontreated dams (16.7%, 0.22/rat as compared with 3.3%, 0.07/rat). Although the highest incidence of mammary adenocarcinomas was found in the female progeny given 100 ppm PhIP from PhIP-treated dams (70.0%, 1.55/rat), this was only slightly higher than the 61.9% and 0.90/rat of the same dose group from the nontreated dams. In males, no apparent effects of transplacental and neonatal exposures were evident. In a separate experiment, excretion of PhIP into breast milk and transfer of PhIP to fetuses and neonates with resultant hepatic PhIP-DNA adduct formation were demonstrated. Thus, maternal exposure to this food-derived carcinogen may be a critical risk factor for generation of mammary carcinomas.

To obtain basic data concerning the availability of medical care for intractable diseases in a large city and its environs, we analyzed the data of Saitama Prefecture. These data were selected from a nationwide survey conducted by the Epidemiology of Intractable Diseases Research Committee of the Ministry of Health and Welfare in Japan. The characteristics of medical institutions visited by patients with 26 intractable diseases were analyzed. The patients were receiving financial aid for treatment. The subjects were 4234 patients in 1984 and 6804 patients in 1988. In addition, we compared the data of 1984 with those of 1988. The results are as follows: 1) In both years, the characteristics of medical institutions which were visited varied in terms of the individual disease. The proportion of patients who visited medical institutions in the same medical service area and in the same prefecture were both very low. The percentage of patients who visited medical institutions in Tokyo was more than 30% of the total, and was more than 50% for the disease with the highest proportion of patients. The dependency on Tokyo for medical institutions was inversely proportional to the distance from Tokyo. Most patients were highly dependent on a large hospital, but the proportion of patients with SMON and ulcerative colitis who visited a large hospital was markedly lower than that for other diseases. 2) A comparison of the data of 1984 with those of 1988 showed that in most of medical service areas, the proportion of patients who visited medical institutions in the same medical service area and in Saitama Prefecture increased, but in Tokyo Prefecture the proportion decreased. We continuously observed a high dependency on a university hospital located about 60km distance from downtown Tokyo, but the dependency slightly decreased in 1988. When a new branch of a university hospital opened, many intractable disease patients then depended on that branch. These results suggest that the self-sufficiency levels of medical services for intractable diseases gradually rose in Saitama Prefecture. The illness behavior of intractable disease patients in Saitama Prefecture did not show any remarkable changes, therefore those results in Saitama Prefecture might indicate a universal characteristic of medical institutions visited by intractable disease patients who live in the envirous of a large city (Tokyo).

To investigate whether waist-hip ratio (WHR) influences glucose tolerance, blood pressure, and serum lipids independently of body mass index (BMI).Cross-sectional study.2,228 male self-defense officials aged 49-55 years in Japan.BMI, WHR, serum lipids, systolic and diastolic blood pressure, and glucose tolerance status according to World Health Organization (WHO) criteria.Adjustment was made for smoking, alcohol use, physical activity, and inter-hospital variation in statistical analysis. Adjusted odds ratios of non-insulin-dependent diabetes mellitus (NIDDM) were progressively increased with increasing levels of WHR, but not BMI. Both BMI and WHR were independently positively associated with impaired glucose tolerance to almost the same degree. Adjusted means of blood pressure, total cholesterol, and triglycerides were almost linearly increased with increasing levels of both BMI and WHR, and high-density lipoprotein cholesterol was progressively decreased. Although the associations of BMI and WHR with blood pressure and serum lipids attenuated after mutual adjustment for each, BMI was less influenced by WHR than vice versa, except for triglycerides.WHR is an important contributor not only to NIDDM but also to impaired glucose tolerance, blood pressure, and serum lipids, although the associations of WHR with blood pressure and serum lipids were generally weaker than those of BMI, except for triglycerides.

Spontaneously hypercholesterolemic (SHC) rats become hypercholesterolemic on normal diets. SHC rats, especially males, exhibit renal lesions similar to those found in focal segmental hyalinosis/sclerosis (FGS). We presented here a detailed natural history of serum lipid, renal function and pathological changes in male SHC rats from 5 to 40 weeks of age. Increased urinary protein excretion and glomerular injury were apparent before the detection of lipid or immune deposits, indicating that the renal lesions were not caused by these deposits. Serum total cholesterol levels, already high at 5 weeks, abruptly increased in concurrence with increased urinary protein excretion, suggesting that the severe hyperlipidemia of this strain is modified by a nephrotic syndrome. By 30 weeks of age, glomerular sclerosis was evident in more than half of the glomeruli and tubular dilatation was prominent, with an abrupt increase of BUN, SCr, and urinary volume; these findings indicate a rapid progression to renal failure. Our results suggest that SHC rats would be a very useful model to investigate the process leading to glomerular sclerotic lesion and renal failure, as well as the effect of hyperlipidemia on glomerular injury.

Microsatellite instability in rat colon tumors induced by heterocyclic amines was examined by studies on the lengths of 85 microsatellite sequences, covering most of the rat chromosomes in tumors and normal tissues. Seven of eight colon tumors induced by 2-amino-1-methyl-6- phenylimidazo-[4,5-b]pyridine showed alterations at least at one locus of microsatellite sequences, whereas no mutations were observed in colon tumors induced by 2-amino-3-methylimidazo[4,5-f]quinoline. Three 2-amino-1-methyl-6-phenylimidazo-[4,5-b]pyridine-induced colon tumors had mutations in more than one microsatellite, their mutation rates being 2 of 85, 2 of 85, and 3 of 85 allele/mircrosatellite sequence, respectively. These data suggest that rat colon adenocarcinomas induced by 2-amino-1-methyl-6- phenylimidazo-[4,5-b]pyridine but not 2-amino-3-methylimidazo[4,5-f] quinoline show a trait of microsatellite instability. This is the first systematic study of microsatellite instability in experimental animal models of carcinogenesis.

A total of 8,993 cases of gastric cancer which were undergone gastrectomy at Cancer Institute Hospital, Tokyo were subjected to the multi-variate analysis of prognostic factors in gastric cancer. Relative ratio of hazard risk indicated the rank of importance in prognostic factors as the following order: Hepatic metastasis, radicality of surgery, macroscopic tumor type, serosal involvement, number of involved lymph node, and grade of lymph node metastasis. Difference in the results of analysis was observed between mono- and multi-variate analysis in the following categories: site of primary lesion, type of resection, and chemotherapy. These results suggested the importance of multimodality therapy to achieve the absolute curative surgery for advanced cancer, and also the importance of multi-variate analysis for the proper evaluation of prognostic factors.

The recurrence rate of resected gastric cancer was 31.9%. The rate was in creasingly higher in proportion to the lack of radicality. The most common type of recurrence was peritoneal dissemination, followed by hematogeneous metastasis, local recurrence, and distant lymph node metastasis. The treatments for recurrence were surgery (9.3%) and chemotherapy (16.1%). Systemic chemotherapy (intravenous iv) was common, and local chemotherapy (intraarterial ia, intraperitoneal ip) was rate. The efficacy of systemic chemotherapy (iv) for recurrent gastric cancer without local recurrence was suggested. The efficacy of local chemotherapy (ia) was suggested for peritoneal dissemination and local recurrence.

To evaluate the clinical significance of the surgical procedure of left upper abdominal evisceration (LUAE), we analysed the prognosis of patients who received LUAE in the Cancer Institute Hospital from 1980 to 1990 using pair matching analysis. Survival benefits were observed in the patients with the tumor located in the upper or middle part of the stomach, and especially in those with Borrmann type 4 gastric cancer. Even in cases of peritoneal dissemination, patients who received LUAE survived longer than those with conventional total gastrectomy. These results indicated that LUAE should be a standard surgical approach for Bormann type 4 gastric cancer.

A 16-year-old female patient with gastric cancer was recently treated at our institute. She had macroscopically curative surgery, but relapsed to die 7 months after surgery due to peritoneal dissemination. We experienced three teen-agers over past 45 years, who were 0.019% of all cases treated in our institute. Histological type of these cases was undifferentiated adenocarcinoma, exposing to serosa. Though they had suffered abdominal discomfort for long time, proper diagnosis was not done due to their young age. Even the teen-ager who has abdominal complaints should be subjected to a thorough examination of gastrointestinal tract. Oncogene analysis should be done for the familial strain of gastric cancer.