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Measurement of serum 1,25-dihydroxyvitamin D levels is important for diagnosis of various calcium metabolism disorders. Conventional assays for 1,25-dihydroxyvitamin D employed specific 1,25-dihydroxyvitamin D receptor as binding site for the ligand and thus, biologically active 1,25-dihydroxyvitamin D ligand, which is labeled with 3H, was required. Usage of 3H made assays cumbersome works. A new assay which uses specific antibody as the binding site and the radioligand labeled with 125I is now available as a commercial kit. Using these kits, we first studied basically the reproducibility, recovery, cross-reactivity and comparison with conventional assays. All of those results were satisfactory. Secondly, we measured clinically in 111 healthy adults and in patients with various disorders such as renal failure, primary hyperparathyroidism, hypoparathyroidism and sarcoidosis. This newly available kit for measurement of circulating 1,25-dihydroxyvitamin D is proved to be useful in clinical evaluation of calcium metabolic disorders.

Molecular genetic analysis of von Hippel-Lindau tumor suppressor gene (VHL gene) was performed on 38 tissues of human glial tumors (ependymoma, 1; astrocytoma, 6; oligodendroglioma, 1; oligoastrocytoma, 2; anaplastic oligoastrocytoma, 3; anaplastic astrocytoma, 14; glioblastoma multiforme, 11). Somatic DNAs extracted from frozen tumor specimens were examined by single-strand conformational polymorphism analysis and direct sequencing. In addition, loss of heterozygosity (LOH) on chromosome 3p in 15 glial tumor cases, lymphocyte DNAs of which were available, was examined by use of 10 microsatellite probes and two polymorphism markers for the VHL gene. Two cases of low-grade gliomas showed somatic sense mutations in exon 3 of the VHL gene, and 6 of 15 cases (40.0%) showed LOH of chromosome 3p. The VHL gene-mutated cases also showed LOH. The retention of heterozygosity and high pathological grade of glial tumors were correlated significantly. In addition, Kaplan-Meier survival analysis for patients with glial tumors showed that patients with LOH had a significantly longer survival time than those without LOH. These results suggest that somatic mutations on 3p, including the VHL gene, may be involved in tumorigenesis of some low-grade glial tumors.

The Tolosa-Hunt syndrome (THS) consists of a painful ophthalmoplegia related to granulomatous inflammatory process in the cavernous sinus. According to recent concepts, the diagnosis is established only when other causes of painful ophthalmoplegia are ruled out. A typical pattern of response to corticosteroids associated with a benign evolution may reinforce this possibility. Tumors such as lymphoma and meningioma and orbital pseudotumors can make difficult the differential diagnosis because they also may respond to steroids. Thus it is always necessary to make an extensive ancillary investigation. We performed a clinical, laboratory and radiologic study of five patients with THS. Prednisone was used in all, with dosages ranging from 40 to 80 mg/day. In four patients there was a dramatic analgesic effect in less than 48 hours. Improvement of the ophthalmoplegia was not so fast but occurred in all with a complete remission in 4 to 45 days.

This is a report of accessory spleen located within the tail of the pancreas which mimicked a tumor. The correct diagnosis was made noninvasively with 99mTc-HDRBC (heat-damaged red blood cell) SPECT.

N3-Substituted derivatives of arabinofuranosyluracil (1), methyl (2), ethyl (3), propyl (4), butyl (5), allyl (6), benzyl (7), o-, m-, p-xylyls (8, 9, 10) and alpha-phenethyl (11) derivatives, were synthesized and their CNS depressant effects were evaluated by intracerebroventricular (i.c.v.) injection in mice by using hypnotic activity and pentobarbital-induced sleep prolongation as indices. At a dose of 2.0 mumol/mouse, the values of mean sleeping time induced by 7, 8, 9, and 10 were 144, 154, 117, and 33 min, respectively. Although the alkyl (2-6) derivatives did not cause any hypnotic activity, some derivatives tested (5, 7-10) significantly prolonged the pentobarbital-induced sleeping time.

We compared the segmental SMANCS/Lip-TAI and the segmental SMANCS/Lip-TAE and studied the effectiveness of both treatments and the influence and/or the side effects on liver function. In resected cases, we studied histopathologic examination. The response rate of the group treated by TAI was 28.6%, and that of the group treated by TAE was 76.5%. In the group treated by TAE, the therapeutic effects were good in nodular type HCC, using small doses of SMANCS. In both groups, the incidence and degree of side effects showed no significant difference. Hepatic insufficiency occurred in a few cases of the group treated by TAI. In resected cases, viable areas remained below the tumor capsule. In conclusion, segmental SMANCS/Lip-TAE seemed to be an effective treatment without any serious complications.

To identify early prognostic value of brain injury, a comparison was made between computerized tomography (CT) findings, coagulation abnormalities, and clinical features in 51 patients with closed head injury. The patients were divided into three groups according to their plasma level of fibrin-fibrinogen degradation product (FDP): normal group (FDP 10 micrograms/ml or less) in 20 patients; moderately abnormal group (FDP 10-40 micrograms/ml) in 15 patients; and highly abnormal group (FDP 40 micrograms/ ml or more) in 16 patients. Cases with a fatal clinical course were mostly associated with very high FDP level. Mortality rate in the highly abnormal group was 44% and 7% in the moderately abnormal group were dead cases, while no cases in the normal group turned out poor outcome. Injury severity, as assessed by Glasgow Coma Scale (GCS) score, correlated with the increase of plasma FDP level. Although severe head injury (GCS 8 or less) was found in 44% of the highly abnormal group and 13% of the moderately abnormal group, normal group only had one case (5%). Very high FDP concentrations were found to be associated with combined hemorrhagic lesions and mass effect on CT scan, but not with a specific localization of brain damage. In summary, the evaluation of coagulation and fibrinolytic function in patients following closed head injury might have both diagnostic and prognostic value.

Three-dimensional bone scintigraphic images were made and their usefulness and limitations discussed.After usual bone scan procedures, single-photon emission computed tomography (SPECT) data were taken and reconstructed into three-dimensional images. Volume rendering methods were used.Three cases of three-dimensional bone scintigraphy were obtained; one of a normal patient, one of a case of transplanted kidney and incomplete fracture of the left femoral head, and one of a case of degenerative joint disease (DJD) on the left temporomandibular joint (TMJ). The three-dimensional structure of the skeletal system was depicted more clearly by the three-dimensional images than by a conventional bone scan.Three-dimensional bone scintigraphs were thought to provide additional information for better understanding of the nature of bone lesions. Some technical improvements including automated threshold level determination and feature extraction for detecting abnormal high uptake are required before routine use can be envisaged.

To understand age-related changes of whole-body and regional skeletal metabolism, it is important to investigate the mechanisms of age-related bone loss and to develop suitable treatments for it. Bone biopsies show metabolism of the particular site examined while biochemical markers for bone metabolism reflect total skeletal metabolism. Bone scintigraphy is a convenient and simple way to analyze whole-body and regional skeletal metabolism. We attempted to study and understand age-related changes in bone metabolism by quantifying the bone scan and correlating it with biochemical bone metabolic markers.The whole-body skeletal uptake (WBSU) and tracer distribution pattern were studied in men and women by bone scintigraphy using 99mTc-hydroxy-methane-diphosphonate (HMDP). Bone scans were performed using a standard protocol and quantified by setting regions of interest (ROIs) on selected regions. WBSU and the skeletal distribution pattern were compared with simultaneously obtained serum biochemical markers.WBSU showed an increase with age in both sexes, but in women, uptake in the head and legs increased more relatively than in the thoracic region, while in men no such tendency was observed. Increase of WBSU and relative increase of uptakes in the head demonstrated a weak correlation with the serum levels of alkaline phosphatase and type 1 collagen metabolites.These results show an age-related increase of skeletal turnover and sex-dependent regional skeletal metabolism. The age-related changes seen in bone scintigrams might be a sign of progressive bone loss, reflecting changes in local bone metabolism.

Recently, several methods for measurement of bone density have been developed. Among them, dual X-ray absorptiometry (DXA) is now considered to be a standard method in bone densitometry. In most cases, values by DXA measurement reflect bone strength and thus measurements of lumbar spine and femoral neck by DXA are used for diagnosis of osteopenia or osteoporosis. In several clinical trials of new drugs, measurement of DXA proved to be very useful. Other methods such as DXA for os radius or os calcaneus are also useful for screening of general osteopenia and quantitative ultrasound is widely used for mass-screening of osteoporosis because of its convenience. Radiographic absorptiometry is also a convenient method, which can be performed only by taking an X-ray photograph with an aluminum phantom. Quantitative computed tomography (QCT) is a method which enables measurement of trabecular true bone density as well as an analysis of trabecular structure and peripheral QCT, aimed for measurement of peripheral bones, is also expected to be a sensitive method to monitor therapeutic responses. Since several methods are now available, it is important to choose a method which is the most suitable for the purpose of each bone mineral measurement.

Measurements of biochemical markers specific to bone resorption are known to be useful in evaluation of skeletal metastasis. However, most of previous studies were performed cross-sectionally and sensitivity for detection of early bone metastasis was not satisfactory. Since basal levels of bone markers differ significantly individually, longitudinal studies would be preferable to evaluate small metabolic changes such as in early skeletal metastasis. Thus, we performed serial measurements of serum c-terminal telopeptide of type I collagen (ICTP), a bone resorption marker, in patients with malignancy and evaluated its clinical significances for detection of bone metastasis, comparing with measurements of propeptide of type I procollagen (PICP) and tumor markers. In total, 43 patients (41 of them are breast cancer), serial serum samples were obtained during 0.5 to 2.4 years (mean 1.4 years). In all of nine patients, who developed bone metastasis, serum ICTP level increased, and by successful treatment to the metastatic lesions, ICTP level decreased, while in some cases, tumor markers remained to be elevated. PICP was less sensitive and specific for detection of bone metastasis. Thus, serial measurement of ICTP is suggested to be useful for detection and evaluation of therapeutic responses in patients with bone metastasis.

Differential expression of two enzymes, dipeptidyl peptidase IV (CD26/DPP IV) and thyroid peroxidase (TPO), in neoplastic thyroid tissues was studied by Northern blot analysis and histochemical analysis using 31 thyroid tissue specimens of various thyroid diseases. On Northern blot analysis, all 16 differentiated carcinomas (12 papillary and 4 follicular carcinomas) overexpressed CD26/DPP IV mRNA, whereas all 14 benign tissue specimens (4 normal thyroid, 4 Graves' disease, 2 adenomatous goiters and 4 follicular adenomas) showed faint expression of CD26/DPP IV mRNA. All 14 benign tissues expressed high levels of TPO mRNA, whereas all 12 papillary carcinomas strongly underexpressed TPO mRNA. A medullary carcinoma did not show any mRNA expression of either enzyme. TPO mRNA expression in differentiated carcinomas did not always correlate with mRNA expression of thyroglobulin, thyroid stimulating hormone receptor, and thyroid transcription factor-1. Northern blot analysis also revealed that CD26/DPP IV is a more specific marker of differentiated carcinoma than three proto-oncogenes previously reported to increase mRNA expression in thyroid carcinomas: c-met, c-erbB-2, and EGF-R. Histochemically, all 14 benign tissues were CD26/DPP IV negative and strongly TPO positive, while all 12 papillary carcinomas were strongly CD26/DPP IV positive and TPO negative. Three of 4 follicular carcinomas were histochemically positive for the two enzymes. These findings suggest that the differential expression of these two enzymes can be applied to study the thyroid tumorigenesis.

Background and objectives Various measures to inactivate/remove viruses have been implemented for manufacturing plasma-derived products. Here, we examined the heat inactivation ability of an agent of the severe acute respiratory syndrome (SARS), SARS coronavirus (CoV). Materials and methods The Frankfurt-1 strain of SARS-CoV was incorporated in manufacturing processes of several products by using samples collected immediately before liquid heat treatment at 60 degrees C. Results SARS-CoV was easily inactivated by this treatment for 60 min in all in-process samples. However, the different composition of the tested samples affected the heat sensitivity of the virus strain: the infectivity of the virus in Antithrombin III preparation still remained after heating for 30 min at 60 degrees C. Conclusion If by rare chance SARS-CoV contaminates source plasma, there should be no or only minor risk of this virus infection, due to sufficient inactivation by the 60 degrees C 10 h liquid heating step, although we must pay attention to the composition used for blood product preparation.

Although c-kit proto-oncogene product is known to be weakly expressed on normal thyrocytes, its function is unclear. In order to investigate the significance of thyroid c-kit, c-kit gene expression in 37 various thyroid tissues was analyzed by comparing c-kit gene expression with the mRNA expression of three thyroid-specific genes: thyroid peroxidase, thyroglobulin, and thyroid stimulating hormone receptor, c-kit mRNA was hardly detected by the usual northern blot method in 2 of 7 follicular carcinomas, 11 of 12 papillary carcinomas, and a medullary carcinoma. On the other hand, a high level of c-kit mRNA expression was found in all 17 benign thyroid tissues (4 normal thyroid tissues, 4 Graves' disease, 2 adenomatous goiters, and 7 follicular adenomas). This study found that c-kit proto-oncogene is more likely to lose expression in differentiated thyroid carcinoma than any thyroid-specific gene. Decreased c-kit gene expression may serve as an indicator for the de-differentiation of thyrocytes.

Recently the gene responsible for the von Hippel-Lindau (VHL) disease was identified as a tumor suppressor gene. Our ongoing studies on the mutation of the VHL gene in Japanese 28 VHL families with single strand conformational analyses of DNA and Southern blot analyses revealed 6 cases of insertion or deletion, 1 cases of splice site mutation and 9 cases of missense mutation, and 3 possible intragenic deletions. Our analytical findings are essentially similar to those observed in the western countries. The VHL families associated with pheochromocytoma had the same mutational hot spot as those in the western countries. Molecular analyses of the VHL gene in the Japanese VHL disease substantially improved the understanding of this disease and its inheritance character.

To investigate the effect of plaunotol in combination with proton-pump inhibitors on Helicobacter pylori eradication in patients with gastric ulcer.We studied 65 H. pylori-positive gastric ulcer patients. They were randomly assigned to five treatment groups: omeprazole (group I, n = 8), lansoprazole (group II, n = 13), lansoprazole+plaunotol (group III, n = 12), lansoprazole+clarithromycin (group IV, n = 16) and lansoprazole+plaunotol+clarithromycin (group V, n = 16). Ulcer status was diagnosed by endoscopy and H. pylori status by culture, histology, a urease test and a urea breath test at baseline, and after 8 and 12 weeks. The clearance and eradication rates were compared in each group.The healing rates in groups I-V were 100, 92, 100, 100 and 100%, respectively; clearance rates were 0, 23, 42, 50 and 75%, respectively; and eradication rates were 0, 0, 8, 38 and 69%, respectively.Combination therapy with plaunotol is efficacious for the eradication of H. pylori in gastric ulcer patients.

The p53 tumor-suppressor gene is the most commonly altered gene in human cancers. Here we demonstrate that transcripts of the mdm2 gene, which encodes a cellular p53 binding protein, markedly increased in the rat liver within 1 to 3 h, reached a peak at 12 h and returned to the basal level 48 h after the administration of carbon tetrachloride. However, the level of hepatic mdm2 mRNA did not significantly change after partial hepatectomy. This is in contrast to p53 gene expression which increased after either procedure. C-myc transcripts also rapidly increased after the injection of carbon tetrachloride, reaching a maximal level at 3 h. The activity of serum alanine aminotransferase was low within the first 12 h and was maximal 24 h after carbon tetrachloride. These results suggest that the transient hepatic expression of the mdm2 gene prior to the onset of cell death is more likely to reflect events associated with necrosis rather than with cell proliferation.

Bone scintigraphy with 99mTc-phosphate compounds is the most popular examination in clinical nuclear medicine. This was developed more than 20 years ago and its roles in various skeletal disorders are well established. Furthermore, improvement of imaging apparatus and application of SPECT strengthened its value extensively. From scintigram alone, in many cases, differentiation between bone metastasis and other "benign" disorders is easily capable. Further improvement in resolution of scinticamera should strengthen its value more. Other recent developments in skeletal nuclear medicine are those in bone densitometry and in measurement of metabolic bone markers. Bone densitometry using DXA is applied on diagnosis and monitoring of therapeutic effects in various metabolic bone diseases, especially, in osteoporosis. Bone mass measurement combined with assessments of specific bone markers such as bone specific alkaline phosphatase and collagen cross-link metabolites might replace the bone biopsy in evaluating bone metabolism. Treatment of bone metastasis in patients with prostate cancer by administering radiolabeled bone seeking substances is another topics in this field and awaits for more extensive clinical evaluation.

The intestinal absorption efficacy of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G), which has been recently synthesized and characterized as a stable ascorbate (AsA), was determined in guinea pigs by the perfusion technique. Perfusion of AA-2G in isotonic phosphate buffer to the small intestine resulted in a decrease of AA-2G accompanied by an increase of AsA in the perfusate. The results showed that intact AA-2G was not detected in the plasma of the portal vein of guinea pigs at 2 h after perfusion. The disappearance of AA-2G from perfusate was completely inhibited by the addition of castanospermine, a specific alpha-glucosidase inhibitor, or by carbohydrates such as maltose. These results indicate that ascorbic acid released from AA-2G by alpha-glucosidase on the brush border membrane is effectively taken up across the intestinal ascorbate transport channels, into a serosal site, whereas AA-2G permeation was poor via the passive transport system.

Mouse hepatic microsomal enzymes catalyzed the oxidation of o-, m-, and p-tolualdehydes, intermediate metabolites of xylene, to the corresponding toluic acids. Cofactor requirement for the catalytic activity indicates that the microsomes contain NAD- and NADPH-dependent enzymes for this reaction. GC/MS analyses of the carboxylic acids formed by incubation under oxygen-18 gas indicate that the mechanism for this oxidation is an oxygenation and a dehydrogenation for the NADPH- and NAD-dependent reaction. Vmax/Km (nmol/min/mg protein) ratios indicate that the NADPH-dependent activity is more pronounced than the NAD-dependent activity. These results suggest that the NADPH-dependent reaction is mainly responsible for the microsomal oxidation of tolualdehydes. The NADPH-dependent activity was significantly inhibited by SKF 525-A, disulfiram and menadione, inhibitors of cytochrome P450 (P450), suggesting the involvement of P450 in the reaction. In a reconstituted system, P450 MUT-2 (CYP2C29) purified from mouse hepatic microsomes catalyzed the oxidation of o-, m-, and p-tolualdehydes to the carboxylic acids, and the specific activities (nmol/min/nmol P450) were 1.44, 2.81, and 2.32, respectively. Rabbit antibody raised against P450 MUT-2 significantly inhibited the NADPH-dependent oxidation of tolualdehydes to toluic acids by 88% (o-), 63% (m-), and 62% (p-) using mouse hepatic microsomes. The present study demonstrated that a mouse hepatic microsomal aldehyde oxygenase, P450 MUT-2, catalyzed the most of oxidative activity of tolualdehydes to toluic acids in the microsomes.

Hydrolysis activity of phospholipase D from Streptomyces chromofuscus (PLD) was studied in small unilamellar vesicles (SUV) of egg yolk phosphatidylcholine (PC). The enzyme was associated with PC-SUV in a Ca(2+)-dependent manner. Both apparent maximum velocity, Vmax(app), and reciprocal of apparent Michaelis constant, i.e., apparent binding constant, 1/Km(app), increased with Ca2+ concentration, and the maximum values of these kinetic parameters were obtained at about 20 microM Ca2+. Incorporation of 1,2-diacylglycerol (DAG), cholesterol (Chol) or alpha-tocopherol (Toc) into PC-SUV induced shift of the antisymmetric PO2- stretching band of PC to lower frequency. The neutral lipids in SUV brought about increase of the Vmax(app) value (Yamamoto et al. (1993) Biochim. Biophys. Acta 1145, 293-297). On the basis of these findings we discussed the regulation of PLD activity in terms of the Ca(2+)-dependent complex formation of PLD with SUV, and the enhancement of susceptibility of the P-O bond in PC molecule by neutral lipids.

1. ABSTRACT Addition of 50 μM H 2 O 2 to cultured mouse myocytes caused cessation of beating followed by morphological degeneration. On the other hand, cultured quail myocytes did not show any impairment even upon exposure to 100 μM H 2 O 2 . We found that higher intracellular glutathione level ([GSH] i ) in quail myocytes than that in mouse myocytes was the major reason for the high H 2 O 2 resistance of quail myocytes. Both susceptibility to H 2 O 2/ toxicity and [GSH] i of mosaic sheets composed of equal amounts of mouse and quail myocytes were almost the same as those of sheets composed entirely of quail myocytes. Using a radioactive GSH precursor, we showed that GSH and/or its precursor(s) were transmitted from quail to mouse myocytes through gap junctions, and that this protected mouse myocytes in the mosaic sheets from H 2 O 2 toxicity [1].

A new highly sensitive thermoluminescent (TL) sheet has been studied as a means of in vivo measurement of spatial dose distribution of radiation therapy. This TL sheet (40 cm x 50 cm x 0.2 mm), which is composed of teflon mixed with BaSO4:Eu doped powder, is very flexible and can be cut to the desired size. The TL sheet was found to have a linear response with a very wide dynamic range of at least 0.002 cGy to 5000 cGy absorbed dose. In addition, this sheet does not need be shielded, because of its insensitivity to room light, and therefore the sheet can detect low-energy electrons decreased by a few millimeter thick air for 3H(maximum beta-ray energy: 18 KeV). The spatial dose distribution was printed out with a newly developed digital readout system. In addition, another high-resolution dosimetry system, which exposes X-ray film with TL photons emitted from the irradiated TL sheet at constant room temperature, is reported. Clinically, the in vivo dose distribution on the surface of the rectal cancer for intracavitary radiation therapy was determined. The applicability of TL sheet for in vivo measurement of dose distribution is discussed.

We reported a case of arachnoid cyst in the craniovertebral junction which was extremely rare. A 36-year-old man presented truncal ataxia and dysesthesia in the right upper extremity. CT and MR images revealed a large cyst in the craniovertebral junction. As for findings of MR images, cystic lesion showed similar intensity as cerebrospinal fluid. Intradural arachnoid cyst with thickened dura was opened to communicate with subarachnoid space. Fluid in the cyst was watery clear. Histological finding of the surgical specimen was arachnoid cyst without inflammatory changes. Arachnoid cyst in the craniovertebral junction is discussed with literature.

The mod genes coding for molybdenum transport system were isolated from Escherichia coli by means of complementation of the tor mutation that causes no synthesis of some molybdenum-containing enzymes. The nucleotide sequence of 3048-base pair showed four open reading frames including the previously reported modC gene. The genes were named modA, modB, modC, and modD. Promoter analysis by lacZ assay and primer extension showed the genes modA and modB individually have their own promoters. No transcriptional regulation of the two genes were observed in the presence of molybdate, nitrate, and oxygen in E. coli strain DH5 alpha.

We investigated the relationship between the severity of Helicobacter pylori-associated gastritis and serum anti-H. pylori antibodies. A total of 158 consecutive patients underwent upper gastroduodenal endoscopy. At the time of endoscopy, three biopsy specimens were taken from the antrum and the body of the stomach for culture of H. pylori, CLO test, and histology. Specimens with a negative rapid urease test and no organisms found by culture were judged to be H. pylori-negative. The GAP test used for the antibody measurement is a kit designed to measure serum IgG, IgA, and IgM antibodies. The severity of gastritis was classified according to the Sydney System. The sensitivity and specificity of the IgG antibody measurement kit was 98.3 and 71.4%, respectively. The sensitivity and specificity of the IgM antibody measurement kit was 14.3 and 60.8%, respectively. The sensitivity and specificity of the IgA antibody measurement kit was 45.1 and 100%, respectively. Serum anti-H. pylori IgG antibody titers were correlated significantly with the severity of inflammation in both the antrum and body. Significant associations were found between serum anti-H. pylori IgG and IgA antibody titers and the development of atrophic gastritis. These results suggest that measurement of serum anti-H. pylori antibody titers is useful in the diagnosis of H. pylori infection and severity of gastritis.

The binding affinity for synaptic membrane from bovine thalamus of N3-phenacyl substituted pyrimidine nucleosides having CNS depressant activity was examined by radio receptor assay. N3-Phenacyl derivatives of uridine, thymidine, deoxyuridine, 6-azauridine and arabinofuranosyluracil inhibited specific [3H]uridine binding and exhibited hypnotic activity. However, N3-phenacyl-2',3'-O-isopropylideneuridine, of which structure was different from sugar moiety of N3-phenacyluridine, showed neither the binding affinity nor the hypnotic activity. The results indicated that CNS depressant effect of pyrimidine nucleoside derivatives might relate to uridine binding affinity, so called uridine receptor.

To evaluate the effect of certain cytotoxic products of Helicobacter pylori on superoxide generation in situ and on the infiltration by polymorphonuclear cells into the gastric mucosa. The agents studied were ammonia, a culture supernatant of H. pylori, and N-formyl-methionyl-leucyl-phenylalanine (FMLP), a chemotactic factor.One or two of these agents were dripped onto rabbit gastric epithelium. The number of polymorphonuclear cells in gastric mucosa were counted histologically. Superoxide generation was measured by a chemiluminescence method, using a Cypridina luciferin analog, 2-methyl-6-phenyl-3,7-dihydroimidazo[1,2-alpha]pyrazin-3-one , a chemiluminescence probe that responds to the generation of superoxides and singlet oxygen radicals. Bovine erythrocyte copper zinc superoxide dismutase was dripped to confirm that the increase in photon counts was a direct consequence of the generation of superoxides.The greatest infiltration of polymorphonuclear cells into the gastric mucosa was observed with the administration of 1 mmol/l FMLP followed by 5.9 mmol/l ammonia, and 10 mmol/l FMLP followed by 59 mmol/l ammonia. There was no increase in the number of polymorphonuclear cells in the other experiments. Increased chemiluminescence was observed in conjunction with the infiltration of polymorphonuclear cells and was inhibited by superoxide dismutase.Acute gastritis and superoxide generation in situ were induced by ammonia after pretreatment with FMLP.

To clarify the relationship between non-steroidal anti-inflammatory drug (NSAID)-associated gastroduodenal mucosal injury and Helicobacter pylori infection.The incidence of H. pylori infection was determined in a group of patients treated with NSAID for rheumatoid arthritis foror = 3 months and in a control group of patients with mainly abdominal symptoms but without rheumatoid arthritis and not being treated with NSAID. The incidence of H. pylori infection was also determined in patients treated with different NSAID and antirheumatic drugs. In addition, the minimum inhibitory concentration of several NSAID against H. pylori was investigated.The incidence of H. pylori infection in the NSAID group tended to be lower than in the control group, and was significantly lower in patients with gastric ulcers. The incidence of infection did not differ between patients treated with one or with more than one NSAID. Differences in the infection rate were found between individual NSAID, with indomethacin being associated with a particularly low rate. No differences in the infection rate were found between different antirheumatic drugs. The minimum inhibitory concentration of ibuprofen was low.H. pylori appears to have little effect on gastroduodenal mucosal injury associated with long-term NSAID administration.

Radiation therapy holds an important position as one of the multidisciplinary methods of treating lung cancer (non-small cell carcinoma). As a result of the development of platinum preparations such as cisplatin (CDDP) and wide use of digital subtraction angiography (DSA), selective bronchial arterial infusion (BAI) therapy made possible more effective use of anti-lung cancer drugs. The use of radiation therapy in combination with BAI is now recommended as a more effective method. Meanwhile carboplatin (CBDCA) has recently been developed as a second generation platinum preparation with less side effects, and is being used for BAI, too. However, the maximum tolerated dose (MTD) of CB DCA for BAI to be used in combination with radiation therapy is not known yet. We, therefore, carried out a phase-study to determine MTD of CBD CA for combination with radiation therapy. The results show that the MTD of CBDCA is 400 mg/m2, and that clinically recommendable infusion limit is 350 mg/m2. In an angiographic study performed at the same time, a plural number of tumor affected blood vessels were found in 81.3% of the patients with lung cancer. Therefore, infusion of a drug for such patients should be carefully applied.

Hemangioblastoma is one of the benign tumors in the central nervous system. It is often associated with the von Hippel-Lindau (VHL) disease, a well known hereditary tumor syndrome. It is believed that inactivation of both alleles of VHL tumor suppressor gene is essential in the tumorigenic processes in hemangioblastomas associated with VHL disease. The molecular basis for the development of sporadic hemangioblastomas is not known. Here, we analyzed 13 cases of primary sporadic hemangioblastomas for somatic mutations of VHL gene with single strand conformational polymorphism analyses of the tumor DNAs. We detected abnormal single strand conformational polymorphism pattern in 7 tumors (54%). Of these 7 possibly mutated tumors, we successfully characterized 3 tumors by direct sequencing. We were unable to sequence 4 tumors because of the poor quality of DNA obtained from paraffin blocks. Somatic mutations in the 3 tumors were 2 missense mutations and 1 microdeletion. These mutations were observed in 1 tumor in exon 1 and 2 tumors in exon 2. Our results suggest that mutations of VHL tumor suppressor gene are involved in the development of at least 20% of sporadic central nervous system hemangioblastomas.

We report a case of multiple meningioma associated with intraosseous and tentorial meningioma. A 75-year-old woman was admitted to our hospital with blurred vision and exophthalmos in the left eye. Left lower hemianopsia, deterioration of visual acuity and right cerebellar signs were found on neurological examination. CT and MR image revealed an intraosseous tumor of the left sphenoid bone and a right tentorial mass. Cytogenetic analysis failed to reveal any abnormalities of chromosome 22. The intraosseous tumor which was transitional type meningioma and the tentorial mass, which was a fibroblastic meningioma were successfully excised. Postoperatively, the patient's symptoms and signs improved. Intraosseous meningioma associated with multiple meningioma is extremely rare and the histogenesis of meningioma arising from ectopic arachnoid cells is discussed.

Computed X-ray Densitometry (CXD) is a semi-automated method of radiographic densitometry. X-ray photographs of 2nd metacarpal bone, taken with a reference Aluminum phantom, are analyzed by a computed-densitometric machine (Bonalyzer). X-ray films are imaged by a CCD camera and displayed in a screen automatically. By pointing distal and proximal heads of the metacarpal bone, photodensity expressed as Aluminum concentration is computed. Parameters such as metacarpal cortical index (MCI) and cumulated photodensity (sigma Gs/D) at the mid-metacarpal bone are obtained. Precision determined by 5 photographs taken every 5 week, is 2%. Correlation with lumbar BMD assessed by DXA is 0.663 in elderly patients. Using this semi-automated method, radiographic densitometry can be used for clinical purpose, such as screening of patients with osteoporosis or observation of therapeutic effects. Especially, in patients over 70, this method has advantages, since lumbar BMD assessment with DXA at this age is hampered by degenerative diseases.

The effects of three different inhalation anesthetics (sevoflurane, isoflurane and halothane) delivered with oxygen and nitrous oxide during surgery and of postoperative analgesics on degree %% postoperative sedation immediately following removal of the tracheal tube, in the recovery room and thereafter in the surgical ward, were studied in 74 pediatric patients under 12 years of age. They underwent elective surgical procedures including repair of inguinal hernia, orchidopexy, tonsillectomy, repair of cleft lip and certain orthopedic procedures. The degree of postoperative sedation was assessed using a nine-point behavioral scale (sedation score) designed for the rating of postoperative pain in children. There were no significant differences in sedation score or frequency of use of postoperative analgesics at any time of assessment among the three inhalation anesthetics. Thirty-eight percent of the patients received analgesics such as diclofenac and acetaminophen suppositories, and intravenous pentazocine. There were no significant differences in sedation score between the group of patients who received analgesics and the group of those who did not, and many patients had satisfactory sedation scores in the surgical ward, suggesting that appropriate administration of postoperative analgesics is possible.

A new intraoperative fluid, which is characterized by containing 10 mEq.l-1 of potassium, 140 mEq.l-1 sodium and 1% glucose, was infused into 18 healthy volunteers either at rate of 5, 10 ml.kg-1.h-1 (for 3 hours) or 15 ml.kg-1.h-1 (for 2 hours). Responding to increase in the infusion rate and volume, blood glucose and insulin (IRI) levels increased slightly but remained in the normal range. On the other hand, lipid metabolism was depressed markedly. Although concentrations of both potassium and sodium in serum remained unchanged, sodium and water balance (intake vs. outflow) became absolutely positive. Potassium balance was maintained at zero. Blood renin activity decreased gradually after the infusion was commenced and remained at a lower level for 3 hours after stopping infusion. Blood aldosterone and antidiuretic hormone levels also decreased but more gradually than the decrease of blood renin activity. Noteworthy change was not observed in the variables relating to hepatic and renal functions.

The thoracic aorta was taken from nephrotic rats on the 40th day after a single i.v. injection of daunomycin (10 mg/kg). In the endothelium-intact aorta, the contractions induced by norepinephrine or B-HT 933, an alpha-2 adrenoceptor agonist, were significantly greater in nephrotic rats than in normal animals. However, such a difference was not observed in the KCl- or U46619-induced contractions. The difference in norepinephrine-induced contraction between nephrotic and normal preparations was enhanced by zaprinast, a cyclic GMP phosphodiesterase inhibitor. The contractions elicited by norepinephrine and B-HT 933 were potentiated by either removal of the endothelium or pretreatment with methylene blue, a guanylate cyclase inhibitor. The difference in the contractile response to these agonists between nephrotic and normal preparations was eliminated completely by either treatment. The cyclic GMP level in the endothelium-intact aorta in the presence of zaprinast was significantly lower in nephrotic rats than in normal animals. In the presence of zaprinast, norepinephrine, but not B-HT 933, caused an increase in the cyclic GMP level, which was abolished completely by pretreatment with prazosin, but not by yohimbine. These results suggest that the augmented contractile response to norepinephrine observed in nephrotic aorta may be due to the decrease in the stimulated release of endothelium-derived relaxing factor from the endothelial cells via the stimulation of endothelial alpha-1 adrenoceptors, whereas the augmented response to B-HT 933 may be due, at least in part, to the decrease in spontaneously released endothelium-derived relaxing factor.

Tributyltin (TBT) and triphenyltin (TPT) compounds have been widely used as antifouling paint for ship bottoms and fishery firm nets, and they are known to be aquatic environmental contaminants. A survey of the pollution with organotin compounds in fish and shellfish has been carried out for samples (180 samples from 43 kinds) which were collected in the Hokkaido prefecture during the period from February, 1989 to September, 1992. On the other hand, daily intakes of organotin compounds have been investigated using the Market Basket Method. Relatively high concentrations of TBT and TPT were detected from flat fish and shellfish such as Hirame (bastard halibut), Kurogarei (black plaice), Asari (Japanese littleneck), Hokkigai (Japanese surf clam) and Kaki (oyster). The maximum value of TBT in samples was 0.38 microgram/g (for both Asari and Kaki), and TPT was 0.47 microgram/g (Kurogarei). Estimated daily intakes of DBT, TBT and TPT were 0.45 microgram, 2.40 micrograms and 4.11 micrograms, respectively. The values presented in this study were lower in comparison with the acceptable daily intake. It was revealed that the values of the organotin compounds in this study were not so high as to have any efect on human health at the present.

Structure and properties of various ethylene / α-olefin copolymers were comparatively investigated. Copolymers polymerized with a heterogeneous Ziegler-Natta catalyst showed a wide intermolecular structural distribution, contrary to a narrow distribution of the copolymers obtained with a homogeneous catalyst system. This structural distribution was found to be correspondingly reflected in the distribution on a crystalline level such as a lamella thickness distribution (LTD). Mechanical properties of the copolymers were strongly influenced by the structural distribution; i.e., the narrower the chemical composition distribution and LTD, the stronger the impact strength. This result could be explained from the viewpoints of tie molecular concentration and its orientational homogeniety with deformation.

We have studied the behavior of ferritin molecules using a high-speed atomic force microscope (high-speed AFM), which uses a miniaturized cantilever and scan stage to reduce the mechanical response time of the cantilever and to prevent the onset of resonant motion at high scan speeds. Using this instrument, we were able to achieve scan rates of up to 1 frame per second for pure fluids. A conventional AFM can visualize the ferritin molecule-substrate interaction directly at the single-molecule level. The conventional AFM can dynamically visualize the process of the ferritin molecule interacting with the substrate; however, it cannot dynamically visualize the adsorption of ferritin molecules on the substrate, which occurs in a fraction of a second, because of its very low scan speed. In this study, we used a novel high-speed AFM to dynamically visualize the behavior of the ferritin molecule-substrate reaction at the single-ferritin-molecule level by real time.

It is now generally accepted that infection with Helicobacter pylori is the most common cause of active chronic gastritis and duodenal ulceration. Although H. pylori is detected in the majority of patients with gastric ulcer, its pathogenicity and role in gastric ulceration remains unclear. One method to study this relationship is to eradicate H. pylori and observe the recurrence of gastric ulcer. In the present paper we examined the consequence of H. pylori eradication by antibiotics mono therapy with histamine H2 receptor antagonist on the recurrence of gastric ulcer. Two cases treated with proton pump inhibitor plus clarithromycin were reported. It is concluded that eradication of H. pylori infection may diminish the recurrence of gastric ulcers.

We measured the speed of sound (SOS) and broadband ultrasound attenuation (BUA) in the calcaneal bone and compared those values with bone mineral density (BMD) as assessed by single X-ray absorptiometry and dual energy X-ray absorptiometry. Calcaneal speed of sound and broad band ultrasound attenuation showed an earlier decrease with age than lumbar BMD, Calcaneal SOS and BUA correlated well with calcaneal BMD. Calcaneal SOS correlated more closely with neck BMD than lumbar BMD measured by DXA. Furthermore, measurement of calcaneal SOS is more useful than measurement of lumbar BMD to evaluate femoral neck BMD, which might be very important for clinical purposes. Both body weight and height correlated well with calcaneal BMD but not so well with calcaneal SOS or BUA. As ultrasound emits no radiation, we can use it without the restriction of radiation protection.

A radioimmunoassay for circulating levels of the pyridinoline cross-linked carboxy-terminal telopeptide of type 1 collagen (1CTP) was developed and can be available as a kit on a commercial base. Using the kits, we evaluated basically and clinically the assay. The assayed values were reproducible and the assay can detect as low as 0.5 ng/ml of 1CTP. In healthy volunteers, circulating level was high under age 24 and over age 46. In patients with bone metastasis, serum levels elevated even in its early stage and correlated well with clinical status. In other bone diseases, such as primary hyperparathyroidism, hyperthyroidism, post-gastrectomy, hypercalcemia of malignancy and myeloma, serum levels elevated according to their clinical conditions. In patients with chronic renal failure, serum levels were high, suggesting decrease of renal clearance of 1CTP. The circulating 1CTP levels seemed to reflect well clinical bone destructive status. A high correlation between serum 1CTP level and urinary pyridinoline (r = 0.884) was shown, whereas essentially no correlation was observed between bone formation markers such as osteocalcin and alkaline phosphatase. Thus, the measurement of circulating 1CTP seems to be a simple and sensitive method to monitor bone destruction.

A new model of barbiturate tolerance and dependence was developed using i.c.v. infusion of pentobarbital. Male Harlan Sprague-Dawley rats weighing 250 to 300 were implanted with i.c.v. cannulae and infused with sodium pentobarbital (500 micrograms/10 microliters/hour) for 6 days. The pentobarbital-infused group had a shorter duration of pentobarbital-induced loss of righting reflex than the saline-infused group. When i.c.v. pentobarbital- and saline-infused rats were injected with sodium pentobarbital (60 mg/kg i.p.), the time course of pentobarbital levels in the serum and in the brain were not significantly different. The infusion of pentobarbital also did not induce hepatic drug-metabolizing enzymes. The depth of thiopental-induced hypothermia was decreased by i.c.v. pentobarbital infusion. During the course of the infusion, the basal body temperature of the pentobarbital-infused rats did not change. Two days after the infusion was discontinued, the basal body temperature was elevated. The increase in body temperature lasted for 8 days. Twenty-four hours after the infusion was discontinued, the pentobarbital-infused rats had a significantly shorter onset of t-butylbicyclophosphorothionate (TBPS)-induced convulsions. These studies show that i.c.v. infusion can be used to induce pentobarbital tolerance and dependence. This model has the advantage that issues related to induction of hepatic drug-metabolizing enzymes are eliminated, and it may be useful in the study of barbiturate addiction.

We studied the dietary habits of 113 upper urinary tract stone formers and 24-h urine specimens before and after dietary control. Protein intake was decreased in male patients after dietary control but urinary risk factors were not changed significantly. The excretion of urinary risk factors (calcium, uric acid and oxalate) was decreased in the patients who had calcium excretion of more than 250 mg per day or uric acid excretion more than 650 mg per day or oxalate excretion more than 45 mg per day before the control. The daily protein intake was significantly correlated to urinary uric acid, calcium, and phosphate excretion and the daily vitamin C intake to urinary citrate excretion. There was a negative correlation between the daily calcium intake and urinary oxalate. The mean stone episode rate of these patients was significantly decreased from 0.174 to 0.059 stones per year by dietary control (p0.005).