Duplex PCR facilitates the identification of immunoglobulin kappa (IGK) gene rearrangements in acute lymphoblastic leukemia.
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Gene Rearrangement↗
Biomedical subjects
M Nakao
Publications and source records attributed to M Nakao.
Rapid and reliable detection of N-ras mutations in acute lymphoblastic leukemia by melting curve analysis using LightCycler technology.
We applied a new strategy for the detection of N-ras gene mutations based on LightCycler technology. We designed two sets of amplimers and internal hybridization probes representing N-ras codons 12/13 and codon 61, respectively. Genomic DNAs from 134 childhood acute lymphoblastic leukemia (ALL) patients (83 common ALL, nine pre-pre-B ALL, 19 pre-B ALL, 23 T-ALL) were amplified, followed by the analysis of the melting temperatures of the PCR products on the LightCycler. PCR products exhibiting an abnormal melting characteristic were directly sequenced. Sequence analyses unravelled nucleotide substitutions at codon 12 in 10 patients, at codon 13 in three, and at codon 61 in one case. The incidence of N-rasmutations (10%) is compatible with previous reports. The LightCycler technology facilitates the rapid analysis of other genes exhibiting hot spot mutations in human malignancies.
Child↗
Tandem duplication of the FLT3 gene in acute lymphoblastic leukemia: a marker for the monitoring of minimal residual disease.
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Amino Acid Sequence↗
Digoxigenin-end labeled oligonucleotides facilitate the detection of minimal residual disease in acute lymphoblastic leukemia.
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Adult↗
Changes in absorptive function of rat intestine injured by methotrexate.
1. Methotrexate (MTX), an anticancer drug, has been shown to induce acute injury in the small intestine. The present study was designed to investigate the in vivo absorptive function of the small intestine injured by MTX using an amino-beta-lactam antibiotic cephalexin (CEX). Time-dependent changes in diamine oxidase (DAO) and alkaline phosphatase (ALP) activity in the small intestine and histopathological findings were also measured in rats treated with MTX (20 mg/kg). 2. Most severe mucosal damage was observed 2 days after MTX treatment and the area under the plasma concentration-time curve of CEX (AUC(CEX)) following oral administration of 20 mg/kg tended to decrease. Thereafter, the AUC(CEX) increased significantly and the histopathological changes diminished within 5 days. 3. Both villus height and mucosal weight followed the same pattern, decreasing in the first 2 or 3 days following treatment, increasing on the 5th day and returning to control levels by the 10th day. Methotrexate-induced changes in the mucosal wet weight/whole intestinal weight ratio were significantly correlated with those of AUC(CEX), but did not correlate with mucosal DAO and ALP activity. 4. These findings provide evidence that the change in the total amount of CEX is an index of the active transport function, probably by intestinal peptide transporter (PEPT1), and is well reflected by histopathological changes in the intestinal mucosa induced by MTX. In addition, there is a possibility that this method could be applied in the clinical setting for diagnosis of intestinal status and absorptive function.
Alkaline Phosphatase↗
Calpain-dependent proteolysis of NF2 protein: involvement in schwannomas and meningiomas.
The neurofibromatosis type 2 (NF2) protein, known as merlin or schwannomin, is a tumor suppressor, and the NF2 gene has been found to be mutated in the majority of schwannomas and meningiomas, including both sporadically occurring and familial NF2 cases. Although the development of these tumors depends on the loss of merlin, the presence of tumors lacking detectable NF2 mutations suggests different mechanisms for inactivating merlin. Recent studies have demonstrated cleavage of merlin by calpain, a calcium-dependent neutral cysteine protease, and marked activation of the calpain system resulting in the degradation of merlin in these tumors. Increased turnover of merlin by calpain in some schwannomas and meningiomas exemplifies tumorigenesis linked to the calpain-mediated proteolytic pathway.
Calpain↗
Relative power contributions of unit discharges simultaneously recorded in the mesencephalic reticular formation.
In order to investigate mutual interactions between neuronal discharges in the mesencepahlic reticular formation, relative power contributions (RPC) were estimated during sleep-wake cycles and serotonin (5HT)-depleted conditions by the administration of parachlorophenylalanine (PCPA). The RPC for most of the neurons dominated in the lower frequency range below 0.4 Hz during rapid eye movement (REM) sleep and the PCPA-treated conditions, which were qualitatively reproduced by the neural network model under global disinhibition. The results supported the hypothesis that the peculiar dynamics of neuronal activities during REM sleep, 1/f fluctuations, were generated by neuronal interactions augmented by the release from the global inhibition of 5HT.
Animals↗
[The bacteriocidal effect of the electrolysed functioning water against Helicobacter pylori].
The bacteriocidal effect of electrolysed functioning water against Helicobacter pylori was examined by both the culture method (viable count on agar plate) and the bacterial ATP content analysis (biolumminescence assay). The culture method revealed the water's reduced bacteriocidal effect on organic substances such as horse serum and bovine serum albumin. The bacterial ATP content determined by the treatment with lysis solution correlated with the bacterial cell count. The ATP of bacteria treated with electrolysed functioning water, benzalkonium chloride, and chlorhexidine gluconate was still present after contact with these solutions for 30 minutes, while the bacterial growth was completely inhibited by the culture method. Therefore, it was suggested that the bacterial ATP was derived from the coccoid form which was observed morphologically due to the action of these solutions. These results demonstrate that these solutions are not effective bacteriocidally against the coccoid form. Consequently, when we utilize functioning water in the field, we should keep in mind its nonbacteriocidal effect against the coccoid form, as well as its reduced effect under the condition of the contamination by organic substances.
Adenosine Triphosphate↗
[The prevalence of virulence-related genes, eaeA, aggR and astA, of localized and aggregative-adherent Escherichia coli (EPEC and EAggEC) in healthy children and age-matched patients with diarrhea].
The prevalence of virulence-related genes of localized- and aggregated-adherent Escherichia coli (EPEC and EAggEC), such as eaeA, aggR and astA was compared between E. coli isolated from 0 to 5 year old children with and without diarrhea in Saga Prefecture. In the case of eaeA, 233 cases in Aichi Prefecture were included. The subjects were 74 diarrheal patients from which no diarrheagenic bacteria were detected besides E. coli. The control subjects were 304 nursery school children without diarrhea, and E. coli was isolated from 278 children in which 105 strains were of 0-serotype. EaeA-positive E. coli was isolated from nine (12.2%) Saga cases, 19 (8.2%) Aichi cases and 6 (5.7%) control subjects; aggR-positive E. coli was isolated from 10 (13.5%) cases and 6 (5.7%) control subjects and astA-positive E. coli from 10 (13.5%) cases and 14 (13.3%) control subjects. No significant difference (p > 0.05) was observed in the prevalence of eaeA, aggR and astA between healthy and diarrheal children, even in age-matched and 0-serotypable E. coli limited comparisons. The pathogenicity of EPEC and EAggEC should be investigated, considering other known or unidentified factors.
Adhesins, Bacterial↗
Molecular characterization and diagnostic value of Taenia solium low-molecular-weight antigen genes.
Neurocysticercosis (NCC) caused by infection with the larvae of Taenia solium is an important cause of neurological disease worldwide. In order to establish an enzyme-linked immunosorbent assay (ELISA) for this infection using recombinant proteins, we carried out molecular cloning and identified four candidates as diagnostic antigens (designated Ag1, Ag1V1, Ag2, and Ag2V1). Except for Ag2V1, these clones could encode a 7-kDa polypeptide, and Ag2V1 could encode a 10-kDa polypeptide. All of the clones were very similar. Except for Ag2V1, recombinant proteins were successfully expressed using an Escherichia coli expression system. Immunoblot analysis of NCC patient sera detected recombinant proteins, but because reactivity to recombinant Ag1 was too weak, Ag1 was not suitable as an immunodiagnostic antigen. So, Ag1V1 and Ag2 were chosen as ELISA antigens, and the Ag1V1/Ag2 chimeric protein was expressed. Of 49 serum samples from NCC patients confirmed to be seropositive by immunoblot analysis, 44 (89.7%) were positive by ELISA. No assays of serum samples from patients with other parasitic infections recognized the Ag1V1/Ag2 chimeric protein. The Ag1V1/Ag2 chimeric protein obtained in this study had a high value for differential immunodiagnosis.
Amino Acid Sequence↗
Mechanism of transcriptional regulation by methyl-CpG binding protein MBD1.
MBD1 is a mammalian protein that binds symmetrically methylated CpG sequences and regulates gene expression in association with DNA methylation. This protein possesses a conserved sequence, named methyl-CpG binding domain (MBD), among a family of methyl-CpG binding proteins that mediate the biological consequences of the methylation. In addition, MBD1 has at least five isoforms due to alternative splicing events, resulting in the presence of CXXC1, CXXC2, and CXXC3 in MBD1 isoforms v1 (MBD1v1) and MBD1v2, and CXXC1 and CXXC2 in MBD1v3 and -v4. In the present study, we have investigated the significance of MBD, CXXC, and the C-terminal transcriptional repression domain (TRD) in MBD1. A bacterially expressed MBD binds efficiently to densely methylated rather than to sparsely methylated DNAs. In both methylation-deficient Drosophila melanogaster SL2 cells and mammalian CHO-K1 cells, MBD1v1 represses transcription preferentially from both unmethylated and sparsely methylated promoters, while MBD1v3 inhibits densely methylated but not unmethylated promoter activities. The CXXC3 sequence in MBD1v1 is responsible for the ability to bind unmethylated promoter. Furthermore, we have constructed mutant-type MBD1s in which the functionally important residues Arg22, Arg30, Asp32, Tyr34, Arg44, Ser45, and Tyr52 are changed to alanine to investigate the correlation between the structure and function of the MBD in MBD1. Excepting those for Ser45 and Tyr52, none of the recombinant MBD mutants bound to the densely methylated or unmethylated DNAs, and green fluorescent protein-fused MBD1 mutants did not localize properly in the nucleus. All the MBD1v1 and -v3 mutants lost the activity of methylation-dependent gene repression. Based on these findings we have concluded that MBD1 acts as a transcriptional regulator depending on the density of methyl-CpG pairs through the cooperation of MBD, CXXC, and TRD sequences.
Amino Acid Motifs↗
Relative locations of the centromere and imprinted SNRPN gene within chromosome 15 territories during the cell cycle in HL60 cells.
Investigations of imprinted regions provide clues that increase our understanding of the regulation of gene functions at higher order chromosomal domains. Here, the relative positions of the chromosome 15 centromere and the imprinted SNRPN gene in interphase nuclei of human myeloid leukemia HL60 cells were compared, because the homologous association of this imprinted chromosomal domain was previously observed in lymphocytes and lymphoblasts. Four targets including the chromosome 15 territory, its centromere, the SNRPN gene on this chromosome, and the nucleus, were visualized simultaneously in three-dimensionally preserved nuclei using multicolor fluorescence in situ hybridization, and the spatial distributions of these probes were analyzed with a cooled CCD camera deconvolution system. We found that preferential association of SNRPN interhomologues did not occur during the cell cycle in HL60 cells, although this gene exhibited asynchronous replication and monoallelic expression in this cells. SNRPN was found to localize at the periphery of the chromosome territories, and it preferentially faced the nuclear membrane, unlike the adjacent centromeric repeat. The SNRPN gene and the centromere were located close to each other late in S phase, reflecting that these DNA segments may be compacted into the same intranuclear subcompartments with the progress of S phase and in course of preparation for the following G(2) phase. Our results suggest that, although an imprinted gene has features similar to those observed with intranuclear localization of other gene coding sequences, the characteristic of mutual recognition of imprinted regions is determined by certain cellular regulation, and it is not necessary for the allele-specific features of an imprinted gene.
Autoantigens↗
Molecular phylogenetic analysis of ixodid ticks based on the ribosomal DNA spacer, internal transcribed spacer 2, sequences.
An internal transcribed spacer (ITS2) sequence between the 5.8S and 28S rRNA genes was used to estimate the phyletic relationships among Ixodes spp. tick vectors of Lyme disease-causing Borrelia spirochetes. Analysis indicates that Borrelia burgdorferi sensu lato species associated with Lyme disease are found mainly in ticks of the Ixodes ricinus species complex. Other closely related tick species are not known to transmit the Borrelia-that cause Lyme disease in humans, but they appear to have a specific association with other closely related Borrelia species. There is a high degree of concordance in the phylogenetics of Borrelia taxa and the phylogenetic relationships among Ixodes ticks.
Animals↗
Collaborative work to evaluate toxicity on male reproductive organs by repeated dose studies in rats 23). A comparative 2- and 4-week repeated oral dose testicular toxicity study of boric acid in rats.
To assess the validity and limitations of 2-week repeated daily dosing to detect toxic effects on male reproductive organs in rodents, a comparative 2- and 4-week oral repeated dosing study of boric acid, a known testicular toxicant, was given to 6- or 8-week-old Crj:Wistar rats at daily levels of 0, 125, 250 and 500 mg/kg. The ages of the rats were selected so that they were all sacrificed at 10 weeks of age. The testes and epididymides were weighed at necropsy; histopathological specimens were prepared in a routine manner and stained with H&E or PAS-H. In addition, the sperm number and motility rates were evaluated. There were no boric acid-induced effects on reproductive organ weights and on gross behavior/appearance in any groups in either the 2- or 4-week studies. The sperm number and motility rate were not decreased in any group after 2 weeks, while both decreased in the 250 and 500 mg/kg groups after 4 weeks. Histopathologically, as evidence of toxicity at the early stage of boric acid exposure, retention of step 19 spermatids of stages IX-XI was observed in the testes of almost all rats treated with 500 mg/kg after both 2 weeks and 4 weeks. Degenerative/necrotic germ cells and multinucleated giant cell formation were observed in 2 weeks, though to a lesser extent than in 4 weeks. On stage analysis of germinal cells in 2 weeks, spermatogonia and spermatids of stage VII were found to be decreased, and pachytene spermatocytes of stage X were increased. In conclusion, the results indicate that if the selection of doses is appropriate, testicular toxicity of boric acid can be detected even after only 2 weeks of repeated daily oral treatment.
Administration, Oral↗
Effectiveness of interferon therapy for reducing the incidence of hepatocellular carcinoma among patients with type C chronic hepatitis.
STUDY PURPOSE: To evaluate the effect of interferon treatment for reducing the incidence of hepatocellular carcinoma among patients with type C chronic hepatitis. METHODS: Retrospective cohort study was conducted on 923 patients with type C chronic hepatitis, who were identified through databases of Osaka Medical Center for Cancer and Cardiovascular Diseases. Two hundred and twenty-four of those had undergone interferon treatment, while the other 699 patients had not. Kaplan-Meier method and the proportional hazards model were used for statistical analysis. RESULTS: Five-years' cumulative incidence of hepatocellular carcinoma was 2.2% among the interferon treated patients, while 9.5% among the interferon untreated. Difference between the 2 curves of the cumulative incidence was statistically significant (p=0.0015). After adjustment for possible confounders, hazard rate ratio of hepatocellular carcinoma was 0.31 in the interferon treated group, significantly lower than that in the untreated (p=0.015). Hazard rate ratio for death from causes other than hepatocellular carcinoma and liver diseases was also lower among the interferon treated group than that among the untreated, although not significant. CONCLUSIONS: Interferon treatment is suggested to reduce the risk of hepatocellular carcinoma among patients with type C chronic hepatitis, and not to increase the risk for death from causes other than hepatocellular carcinoma and liver diseases.
Antiviral Agents↗
[Factors influencing noncompliance with medication regimens in the elderly].
Poor medication compliance is a major issue in the health care of older patients. To identify risk factors for medication noncompliance in the elderly, inpatients aged 65 years and older at Nagoya University Hospital and at Chubu National Hospital underwent a comprehensive geriatric assessment and tests for the assessment of medication compliance. The dependency of medication assistance by a caregiver is associated with low physical function activity, cognitive impairment, depression and communication inability. Medication noncompliance was not associated with the score of any component of comprehensive geriatric assessment. There was a good relationship between patient's knowledge of medications and the frequency of dosage interval, both of which were associated with the score of instrumental ADL, cognitive function and communication ability. The knowledge was also associated with the medication compliance at Nagoya University Hospital but not at Chubu National Hospital. These results may suggest that the elderly patient's understanding of a medication regimen is important but that other factors are also required to maintain their treatment regimen.
Aged↗
[Epidemiology of alveolar hydatid disease (AHD) and estimation of infected period of AHD in Rebun Island, Hokkaido].
An epidemiological study was performed of endemic alveolar hydatid disease (AHD, multilocular echinococcosis), Rebun Island, Hokkaido and the period of AHD infection of patients was estimated. Death certificates of the residents of the island were analyzed, and 74 deaths (43 males and 31 females) by AHD were found out of the 3,126 deaths that occurred during the period from 1948 to 1990. The red fox population of the island was estimated on the basis of past researchs. The deaths due to AHD distributed around a major peak (n = 67) between 1948-1975 and there were 7 sporadic cases between 1976-90. The red fox population on the island had been estimated to be largest in 1935. The mean infection period from initial AHD infection to death was estimated to be 26 +/- 7 years (x +/- SD) on the basis of the period between the year in which the peak red fox population was observed (1935) and the major peak of patient death (1962). The mean symptomatic period was 5 +/- 5 years, and the mean latent period from infection to the onset of AHD was 21 +/- 7 years. Sex ratio (M/F = 28/13 = 2.15) was higher (P < 0.05) at the age groups below 10 and 26-45 years than the other age groups (15/18 = 0.83), and playing outdoors during childhood and working outdoors in the prime of life were assumed to be the causes of infection.
Adolescent↗
[Salvage chemotherapy in refractory testicular cancer].
A treatment strategy for patients with refractory testicular cancer who failed in the initial therapy has not been established. Patients with metastatic testicular cancer are treated with BEP (BLM, etoposide, CDDP) therapy, which is recognized as the standard first line chemotherapy regimen. About 80% of patients attain complete remission (CR) with BEP therapy and following salvage surgery. The patients who fail to achieve CR or have recurrences during the period of follow-up will be candidates for salvage chemotherapy. Salvage chemotherapies include VIP (ETP, ifosfamide, CDDP) therapy or high dose chemotherapy with peripheral blood stem cell auto-transplantation (PBSCT); however, the effectiveness of these therapies is limited. Those who fail in these salvage therapies are treated with irinotecan hydrochloride (CPT-11) in Japan. Clinical trials with paclitaxel and gemcitabine have recently been started in the United States and Europe. Further investigations are necessary to develop more useful regimens with these novel anticancer agents for refractory testicular cancer.
Germinoma↗