TRITIUM, HYDROGEN AND OXYGEN ISOTOPE COMPOSITIONS IN MONTHLY PRECIPITATION SAMPLES COLLECTED AT TOKI, JAPAN.

Monthly precipitation samples have been collected at Toki, Japan, from November 2013 to March 2017. In this report, selected data were analysed to identify the regional hydrogen and oxygen isotope compositions. Tritium (3H) concentration in the precipitation ranged from 0.10 to 0.61 Bq L-1 and higher 3H concentrations were observed in spring rather than in other seasons. This range was similar to values reported in Chiba City, Japan. 3H concentration and the ratio d-excess, and δD values were roughly clustered according to each separate season. These regional hydrogen and oxygen isotope compositions will be used for environmental assessments of effects of the deuterium plasma experiments of the large fusion test device.

RECENT TRITIUM CONCENTRATION OF MONTHLY PRECIPITATION IN JAPAN.

To obtain a better understanding of recent tritium concentration and its seasonal cycle in Japan, monthly precipitation samples were collected in Hokkaido, Gifu and Okinawa prefectures from June 2014 to December 2017. The arithmetic mean ( ± standard deviation) of tritium concentrations in precipitation samples from Hokkaido, Gifu and Okinawa were estimated to be 0.62 ± 0.27 Bq L-1, 0.32 ± 0.12 Bq L-1 and 0.13 ± 0.05 Bq L-1, respectively. These results indicate that the concentrations increase with latitude. In addition, the highest and the lowest concentrations appeared in spring and summer, respectively. To clarify the origins and sources of these cycles, further analyses of chemical compositions of precipitation and meteorological conditions are needed.

FWT and OBT concentrations in pine needle samples collected at Toki, Japan (1998-2012).

Free water tritium (FWT) and organically bound tritium (OBT) concentrations in pine needles have been investigated to understand the regional background tritium concentration in Toki City. Samples were regularly collected from pine trees on the National Institute for Fusion Science campus (1998-2012) and the nearby Shiomi Park (SP; 2002-12). FWT and OBT concentrations of the former samples ranged from 0.33 to 0.92 and 0.41 to 1.10 Bq l(-1), respectively, while those of the latter samples ranged from 0.32 to 0.86 and 0.33 to 0.79 Bq l(-1), respectively. Results of both sampling sites were almost the same, and they have been gradually decreased year by year. Concentration level of tritium for Toki City was close to the average background level in Japan. The OBT/FWT ratios were almost 1.0. The apparent half-life of FWT in this period was estimated as almost 10 y, and that of OBT was estimated as almost 12 y; these values were almost the same as the physical half-life.

An atomically controlled Si film formation process at low temperatures using atmospheric-pressure VHF plasma.

To grow epitaxial Si films with atomic- and electronic-level perfection, a high-temperature chemical vapor deposition (CVD) process (>1000 °C) has been generally employed. To reduce the growth temperature below 600 °C but keeping a high deposition rate, other energy sources than thermal heating are required. Atmospheric pressure plasma CVD (AP-PCVD) is considered to be suitable for fabricating high-quality films at high deposition rates due both to the high radical density and to the low ion bombardment against the film surface, because the collision frequency among ions and neutral atoms is high. The present study focuses on the low-temperature growth of epitaxial Si, and experimentally demonstrates that AP-PCVD is capable of growing epitaxial Si films with high perfection applicable for semiconductor devices. It is found that the pre-growth cleaning of the Si surface by H(2) AP plasma is effective to grow high-purity Si films, and that the exposure of a film-growing surface to AP plasma during growth is important to form particle-free and defect-free Si films. From the experimental results and the first-principles molecular dynamics simulations of surface atomic reactions, it can be mentioned that both H atoms in the AP plasma and high-density He atoms having thermal kinetic energy contribute to the reduction of growth temperature by supplying considerable energy to the surface.

Absence of microsatellite instability and germline mutations of E-cadherin, APC and p53 genes in Japanese familial gastric cancer.

To evaluate the genetic factors of familial predisposition to gastric cancer, genetic alterations in the surgically resected stomach samples from gastric-cancer-prone families were investigated. Familial gastric cancer (FGC) was defined as gastric cancer occurring in a family with 3 or more gastric cancer patients over at least two successive generations. We examined replication error (RER) of six microsatellite markers and screened mutations of the 10-(A) repeat sequence in the transforming growth factor-beta receptor type II (TGF-betaRII) gene in individuals from seven unrelated FGC families. Three cases showed RER at one of the six (CA)n microsatellite markers but the other 4 cases showed no RER at any of these loci. No mutation was found in the 10-(A) repeat of the TGF-betaRII gene. Additionally, no germline mutation was found by polymerase chain reaction-single strand conformation polymorphism in exons 1-16 of E-cadherin, exons 5-8 of p53 and in the mutation cluster region of APC. These results indicate that disorders in the DNA mismatch repair system, E-cadherin, p53 and APC may be infrequently involved in the carcinogenesis of Japanese FGC.

A simple objective parameter for perfusion study of renal transplant.

We proposed a simple parameter, the kidney-to-aorta ratio (KAR), for evaluation of renal transplant perfusion. KAR was calculated from the peak counts of the kidney and the aorta. The calculated values were compared with the visual interpretation of the radionuclide first-pass flow study, percent renal uptake (%RU), and tubular extraction rate (TER) by Bubeck's one point sampling method in 37 studies. KAR correlated well with the visual interpretation of the flow study and the other quantitative parameters. Representative cases, which showed the usefulness of KAR for the objective assessment of the perfusion status of renal transplants, were presented. In conclusion, KAR is a simple and practically useful parameter for objective evaluation and follow-up of renal transplant perfusion.

[Evaluation of postoperative pain relief by infiltration of bupivacaine or epidural block after laparoscopic cholecystectomy].

We compared the analgesic effect of bupivacaine infiltration into surgical wounds with that of epidural block after laparoscopic cholecystectomy (LC). Forty-five patients (ASA physical status I-II) for LC were randomized into three groups (n = 15 in each group). Patients received only general anesthesia (Group C), received infiltration of 0.5% bupivacaine into the surgical wound before surgery combined with general anesthesia (Group L), or received epidural anesthesia combined with general anesthesia (Group E). Postoperative pain was assessed using visual analogue scale (scale: 0-10) at 1, 2, 6 and 12 hours after the operation, the need for additional supplemental analgesics, and the cost of anesthesia. Visual analogue scale in Group C at 1, 2, or 6 hours was significantly greater than that of Group L and E. The number of patients who needed supplemental analgesics was 9 in Group C, 5 in Group L, and 2 in Group E. The cost of pharmaceutical and anesthetic practice of Group E was more expensive than Group L and C. In conclusion, infiltration of bupivacaine combined with general anesthesia is an effective and economical method of postoperative pain relief.

Inactivation of the 14-3-3 sigma gene is associated with 5' CpG island hypermethylation in human cancers.

The cell cycle checkpoint plays an important role in maintaining the integrity of cells. Recently, one of the 14-3-3 protein family members, 14-3-3sigma, was shown to be regulated by p53 and to play a role in the G2-M-phase checkpoint. To determine whether 14-3-3sigma is inactivated in human cancers, the methylation status of the 5' region of 14-3-3sigma was investigated in a series of gastric, colorectal, and hepatocellular cancer cell lines. Of 22 cell lines examined, 6 showed aberrant methylation. The methylation status of 14-3-3sigma was found to be correlated with loss of expression, which was restored by 5-aza-2'-deoxycytidine treatment. Furthermore, normal G2 arrest after DNA damage was not demonstrated in the cell lines with methylation. In primary gastric cancers, 14-3-3sigma hypermethylation was observed frequently in 26 of 60 (43%) cases and observed more frequently in poorly differentiated adenocarcinomas (P = 0.0017). Our findings suggest that 14-3-3sigma is inactivated by aberrant methylation of the 5' region in various human cancers and that it might play an important role in the development of undifferentiated gastric cancers.

Quantitative DNA methylation analysis by fluorescent polymerase chain reaction single-strand conformation polymorphism using an automated DNA sequencer.

A novel DNA methylation assay technique, termed bisulfite single-strand conformation polymorphism (bisulfite-SSCP), is a combination of sodium-bisulfite modification and fluorescence-based polymerase chain reaction (PCR)-SSCP. After bisulfite treatment followed by PCR amplification, methylated and unmethylated alleles can be simultaneously separated in a nondenaturing gel using an automated DNA sequencer. Using bisulfite-SSCP, methylation of hMLH1 was detected in a quantitative manner. This method is not only simple, quick, accurate, and quantitative, but detailed information about methylation is also available with less work. Methylation analysis of large numbers of samples for multiple loci will be facilitated by bisulfite-SSCP.

[Prediction of the clinical efficacy of hepatic arterial chemotherapy for metastatic hepatic cancer by intraarterial infusion of 99mTc-MIBI].

SPECT was performed in 11 patients with metastatic hepatic cancer by intraarterial infusion of 99mTc-MIBI before hepatic arterial chemotherapy was started, and the degree of accumulation and clinical efficacy were compared. Early and delayed SPECT images were obtained and various parameters were calculated, including early ratio (ER), delayed ratio (DR), washout rate (WR), and retention index (RI). Judgement of clinical efficacy was made by CT before and after hepatic arterial chemotherapy and was classified as effective, unchanged, and progressive groups. The mean values of ER and DR in the effective group were higher than those in the progressive group. No relationships were noted among the WR and RI values of the groups. The assessment of ER and DR using 99mTc-MIBI intraarterial SPECT is considered to be useful for prediction of the clinical efficacy of hepatic arterial chemotherapy for metastatic hepatic cancer.

Identification of genes highly expressed in G2-arrested Chinese hamster ovary cells by differential display analysis.

Abnormal cell cycle regulation is believed to be an important step in tumorigenesis. In mammalian cells, DNA damage commonly leads to cell cycle arrest in G2; however, little is known about the detailed biochemical mechanisms underlying the DNA damage-induced G2 arrest. In order to identify genes differentially expressed in association with G2 arrest, differential display analysis was performed between exponentially growing Chinese hamster ovary (CHO) cells and G2-arrested CHO cells induced by etoposide, SN-38, or X-radiation. We identified five cDNA clones whose expression was up-regulated in G2-arrested CHO cells. Sequence analysis revealed that three clones were homologous to known genes: isogene I of translation initiation factor eIF-4A, ribosomal protein L13, and translation repressor NAT1. The remaining two clones showed no homology to known genes. These results indicate that DNA damage can alter the expression of multiple genes, including translational regulators.

[Application of measuring 99mTc-MAG3 plasma clearance based on one-compartment model (MPC method) to renal transplantation].

Measurement of 99mTc-MAG3 plasma clearance (CLmag) based on one-compartment model (MPC method) was applied to renal transplantation and evaluated for the factors which might affect the calculated results, especially concerning renal depth. Correlation coefficient of CLmag between MPC method using real renal depth and Russell or Bubeck single sampling method was good (r = 0.852 or 0.876, respectively). Regression equation between MPC method and Russell method was y = 1.044x - 3.0 and was more closer to y = x than that between MPC method and Bubeck method. CLmag of MPC method calculated by estimated renal depth from the abdominal thickness was also similar to that by real renal depth. Even if the fixed renal depth, 4 cm, was applied, the coefficient and regression equation between MPC method and Russell method were r = 0.884 and y = 1.004x - 10.2. In conclusion, MPC method is applicable to the evaluation of renal transplants. Though measuring renal depth is best, calculation with fixed renal depth of 4 cm might be practically acceptable.

Distinct methylation pattern and microsatellite instability in sporadic gastric cancer.

Aberrant 5' CpG island methylation is an alternative mechanism of gene inactivation during the development of cancer as demonstrated for several tumor-suppressor genes. Also, marked relationship of microsatellite instability (MSI) and DNA methylation has been reported in sporadic colorectal cancer, which is a result of epigenetic inactivation of hMLH1 in association of promoter hypermethylation. In the present study, we investigated the 5' CpG island hypermethylation of hMLH1, E-cadherin and p16 in 61 primary gastric cancers (GCs) by using combined bisulfite restriction analysis (COBRA) and methylation-specific PCR (MSP), and their MSI status. Of 61 GCs investigated, 5 (8.1%) tumors presented hMLH1 methylation, 16 (26.2%) and 25 (40.9%) showed E-cadherin and p16 methylation respectively, and 8 (13.1%) presented high-frequency MSI (MSI-H). Of the 8 MSI-H patients, 5 presented hMLH1 methylation, whereas no low-frequency MSI (MSI-L) and microsatellite stable (MSS) cases exhibited hMLH1 methylation (5/8 vs. 0/43, p < 0.00001). Furthermore, these patients also presented E-cadherin and p16 hypermethylation. Our data showed a significant correlation between hMLH1 methylation and MSI in GC, and suggested that a common mechanism of aberrant de novo methylation can be postulated in these cancers.

Familial gastric cancer in the Japanese population is frequently located at the cardiac region.

The clinical features of familial gastric cancer are still unknown. To approach this question, we investigated the clinicopathological characteristics of 16 cases of familial gastric cancer. In this study the criteria used to define familial gastric cancer was the existence of three or more family members with gastric cancer in at least two successive generations. The clinicopathological characteristics of cases who fulfilled this criteria were studied. This study contained 16 familial gastric cancer probands. Seven cases (44%) of gastric cancer had developed at the cardiac region of the stomach. This frequency was significantly higher than for gastric cancer in the general population in Japan (15.4%, p < 0.01). Undifferentiated types were dominant in familial gastric cancer (69%, p < 0.05). Furthermore, the frequency of disseminated peritoneal (40%) and liver metastases (20%) in familial gastric cancer was also significantly higher than for gastric cancer in the general population in Japan (10.9%, p < 0.01, and 4.4%, p < 0.05, respectively). Familial gastric cancers were frequently located at the cardiac region and appeared to be more aggressive than sporadic gastric cancers. The unique characteristics of familial gastric cancer suggest a genetic background in their etiology.

Alpha1-antitrypsin deficiency and toxic shock: a Japanese autopsy case.

A 74-year-old Japanese female presented with the sudden appearance of hemorrhagic purpuric ecchymoses on her lower extremities and with fever and chills, and died on the fifth day of hospitalization. A diagnosis of alpha1-antitrypsin (AT) deficiency was made postmortem. The liver weighed 1260 g. Histological sections from the liver revealed rather severe fatty changes of the hepatocytic parenchyma and partial loss of the normal hepatic architecture with fibrosis. The hepatocytes contained periodic acid-Schiff (PAS)-positive, diastase-resistant and alpha1-AT-positive intracytoplasmic globules. There was markedly increased inflammatory infiltration with severe edema and congestion, accompanied by fibrous, thickened pulmonary alveolar walls with fibrin deposition in the lungs (right, 410 g; left, 280 g), which suggest findings similar to those seen in multiple organ failure. Mild pulmonary emphysema was also present in the upper lobes of the lungs. Histological sections from the hemorrhagic necrotic ecchymoses of the skin showed marked neutrophil infiltration over the subcutaneous tissue with bleeding and blistering. A finding of thrombophlebitis was also found in the subcutaneous tissue. No bacteria were detected in the ecchymoses, the urine or the blood. Plasma protein analysis revealed a lower level (9.5 micromol/L) of alpha1-AT and a higher level (330 U) of anti-streptolysin O (ASO). These findings suggest that the patient died of toxic shock-like syndrome and that alpha1-AT deficiency might have facilitated the development of the toxic shock. To our knowledge, this is the first case of toxic shock associated with alpha1-AT deficiency.

Expression and identification of aberrant c-kit transcripts in human cancer cells.

We have previously cloned and sequenced a novel 3.5 kb c-kit mRNA expressed in a colon carcinoma cell line Colo201. Here we examined the expression of this truncated form of c-kit in 14 gastrointestinal cancer cells and 16 hematopoieic cancer cells by RT-PCR. Expression of the aberrant c-kit transcript was observed in various cancer cell lines. Furthermore, a new transcript which is 78 bp shorter than the transcript previously described was identified and characterized. These results indicate that two kinds of aberrant c-kit transcript produced by alternative promoter in intron 15 are expressed in human cancer cells.

Infrequent alterations of the p16 (MTS-1) gene in human gastric cancer.

p16 (MTS-1, multiple tumor suppressor gene 1), a putative tumor suppressor gene, is one of the cyclin-dependent kinase inhibitors (CDI) and it regulates the G1/S transition of the cell cycle. To clarify the role of p16 in primary gastric cancer, we have investigated somatic mutations of this gene by using the polymerase chain reaction/single strand conformation polymorphism (PCR-SSCP) method. In 23 surgical specimens of primary gastric cancer, none were detected in exon1 and exon 2. Among the 6 human gastric cancer cell lines examined, PCR products were not found in 2, MKN28 and MKN45, suggesting the presence of homozygous deletions. No mutation was found in the other 4 cell lines. Furthermore, decreased expression levels were not observed in 13 gastric cancer tissues by reverse transcription PCR (RT-PCR). Considering the above results of PCR-SSCP and RT-PCR, genetic alterations of the p16 gene are rarely implicated in human gastric cancer tumorigenesis.

Genetic alterations in rat colon tumors induced by heterocyclic amines.

BACKGROUND: In rat colon tumors induced by the cooked food mutagens 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), ras and p53 alterations are rarely detected. To investigate the roles of the APC gene and microsatellite instability (MI) in PhIP-induced colon carcinogenesis, mutations of the APC gene and alterations of microsatellites were examined. METHODS: Complimentary DNA sequence of the rat APC gene were determined by polymerase chain reaction (PCR) using primers based on the human APC sequence. PCR-single strand conformation polymorphism (SSCP) analysis was performed using primers based on sequences of flanking introns and exon 15. Microsatellite alterations were also analyzed using 85 microsatellite sequences dispersed through most of the rat chromosomes. RESULTS: Five mutations in the APC gene were detected in four of eight PhIP-induced rat colon tumors. All five mutations involved deletion of a guanine base in a 5'-GGGA-3' sequence. Only 2 of 13 IQ-induced colon tumors had mutations of the APC gene and these were base substitution mutations. Seven of eight PhIP-induced colon tumors had microsatellite alterations in at least one locus, whereas no alterations were observed in the IQ-induced colon tumors. CONCLUSIONS: The specific 5'-GGGA-3' to 5'-GGA-3' mutation and MI demonstrated in this study are strong evidence of a mutational fingerprint of PhIP.