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Biomedical subjects

T Funato

Publications and source records attributed to T Funato.

At least 55 records · Page 3Linked to original sources
PubMed1998

[Gene delivery using adeno-associated (AAV) vectors].

Gene transfer vectors based on the replication-defective adeno-associated virus (AAV) are emerging as promising vehicles for gene therapeutic approaches for the neurologic disease, cystic fibrosis and cancers. AAV vectors have been used to efficiently transduce genes into cells in vitro and in vivo. However, transduced recombinant AAV vectors are integrated into human chromosomes. Thus we have discussed that the review about the AAV vectors and transduction, and reported the results used AAV vectors for expression of ribozyme.

Animals↗
PubMed1998

[Quantitative PCR system].

We evaluated a real time quantitative PCR assay using dual-labeled fluorogenic probes for clinical application. Preliminary study using the house-keeping gene, beta-actin confirmed that this method was accurate and reproducible for the quantitative detection of the genes. The system also has merit with regard to the dynamic range of the starting target molecule determination. We then investigated DNA copies of cytomegalovirus (CMV) gene in vivo. The results demonstrated on association between the quantitation of CMV-DNA copies and clinical manifestation associated with CMV infection of immunodeficiency states or infantile hepatitis. It was also successful for quantitative estimation by RT-PCR. Namely, the assay made it possible to discriminate drug-sensitive leukemia cells from resistant cells based on the MDR1 gene and dCK gene. Real time quantitative PCR assay may be useful in a variety of clinical fields.

Cytomegalovirus↗
PubMed1998

[Terminal home care "Ikigai no Yoake--The real aim of our life"--case report].

It is difficult to provide home care especially for terminal cancer patients as their physical conditions deteriorate due to the cancer. It is important to enhance the will of home care providers to make this possible. Although a universal method has been worked out, personal and religious beliefs have made it difficult to create an effective method. We would like to introduce our experience in the paper entitled "Ikigai no Yoake" and subtitled "Influence of scientific study of reincarnation on view of life" by Fumihiko Ida of Fukushima University. It has encouraged home care providers and made it possible to provide home care to terminal patients along with benefits.

Advance Directives↗
PubMed1998

[Benefits and problems of "home care score"--discussion of deviation from home care].

Care ability is the most important factor in providing home care. This ability to care can be classified into family care and community care. The "Home care score" is a system developed through the comprehensive scientific study on long life by the Ministry of Health and Welfare in order to evaluate such care ability. We have scored 71 patients since August 1997 until February 1998, and we found benefits and problems with the system. Here are the benefits: (1) General evaluation can be made on status of patients and home care providers. (2) Comparison of large numbers of patients can be easily made. (3) Helpful to work out practical support. On the other hand, the system has the following problems: (1) Scoring can differ by subjective evaluation. (2) Difficulty to reflect status of disease deterioration and so on. The factors of deviation from home care are: (1) of disease. (2) Loss of desire to provide care. We believe it is important for the home care nurse to intervene so that the will to provide care will not weaken until the end.

Aged↗
PubMed1997

A Turner syndrome woman with a ring X chromosome [45,X/46,X,r(X)(p22.3q27)] whose child also had a ring X chromosome.

OBJECTIVE: To describe a woman with Turner syndrome with ring X chromosome mosaicism who had a child who possessed the same ring X chromosome. DESIGN: Polymorphisms of genes located on the X chromosome from genomic DNA of the mother, father, and the child were evaluated. PATIENT(S): The mother's karyotype was 45,X [48]/46,X,r(X)(p22.3q27) [2], and the child's karyotype was 45,X[33]/46,X,r(X)(p22.3q27) [17]. INTERVENTION(S): Polymerase chain reaction was used to amplify short tandem repeats from the loci of the hypoxanthine phosphoribosyltransferase gene and the androgen receptor gene. RESULT(S): Alleles for both genes in the child originated from both parents in a heterozygous fashion. The alleles originating from the mother originated from the ring X chromosome. However, the amount of amplified DNA was less than that of a normal X chromosome. CONCLUSION(S): The ring X chromosome of the mother was most likely transmitted to the newborn. Thus, an ovum with the ring X chromosome can be fertile and can produce a viable zygote.

Adult↗
PubMed1997

Acute myelogenous leukaemia with t(8;21) translocation of normal cell origin in mosaic Down's syndrome with isochromosome 21q.

We report a 13-year-old girl with Down's syndrome (DS) having a mosaic karyotype of 46,XX/46,XX, -21,+i(21q), who developed acute myelogenous leukaemia (AML) (FAB M1). The t(8;21) translocation generating a AML1/MTG8 chimaeric gene of her blasts was demonstrated by cytogenetic analysis and reverse transcription-polymerase chain reaction. Interestingly, the leukaemic clone with t(8;21) did not have isochromosome 21q, indicating that the blasts were of normal cell origin. These findings suggest that, in older patients with DS, 21 trisomy cells have no greater predisposition to develop AML than normal karyotypic cells.

Adolescent↗
PubMed1997

[Reversal of drug resistance in human cancer cells by anti-oncogenes].

The demonstration that RNA can be cleaved by cis ribozyme (catalytic RNAs, RNA enzymes) has potentially important therapeutic implications. Ribozymes are effective for modulation of gene expression because of their simple structure, site-specific cleavage activity, and catalytic potential. The targets of ribozyme-mediated gene modulation have ranged from cancer cells to foreign genes that cause infectious diseases. Additional target sites for ribozymes are in initial phases of development and design. Ribozymes have been targeted against myriad genes, including oncogenes (ras, BCR-ABL) and drug resistance genes (MDR-1, c-fos). These ribozymes have cleaved the target RNAs in culture system and developed to the in vivo system. We reported that fos ribozyme has altered the expression of c-fos and DNA repair genes in drug resistance cancer cells, and reversed the sensitivity to cisplatin. Furthermore, we have developed high efficiency by the transfer system in vivo.

Animals↗
PubMed1997

[Circumventing multidrug resistance in human cancer by anti-ribozyme].

The demonstration tha RNA can be cleavaged by cis-ribozyme(catalytic RNAs, RNA enzyme) has potentially important therapeutic implications. Ribozymes are effective for modulation of gene expression because of their simple structure, site-specific cleavage activity and catalytic potential. The targets of ribozyme-mediated gene modulation have ranged from cancer cells to foreign genes that cause infectious diseases. Additional target sites for ribozymes are in initial phases of development and design. Ribozymes have been targeted against myriad genes, including oncogenes (ras, BCR-ABL) and drug resistance genes(MDR-1, c-fos, DHFR). These ribozymes have cleaved the target RNAs in culture system(in vitro) and developed in vivo system. We reported that anti-fos ribozyme has altered the expression of c-fos and DNA repair genes in cisplatin-resistance cancer cells, and reversed the sensitivity to ciaplatin. Furthermore, we have developed high efficiency by the transfer system using an electroporation in vivo.

Animals↗
PubMed1997

[Specific inhibition of anti-DNA antibody production by an anti-DH ribozyme].

In order to develop a new therapy for systemic lupus erythematosus, we designed a ribozyme (catalytic RNA) to specifically cleave the mRNA of anti-DNA-responsible VH genes and tested for their ability to regulate the production of autoantibody by human B cell clone O-81 expressing nephritogenic idiotypes. Based on a preliminary antisense experiments for O-81 VH gene, we determined the regions in O-81 mRNA, which would be the target site for the cleavege. The O-81 ribozyme specifically inhibited the production of anti-DNA antibody by O-81 cones. For application to clinical state, nuclease-resistance ribozyme has been expected. These results indicate to introduce ribozymes as a new method of therapy in autoimmune diseases.

Antibodies, Antinuclear↗
PubMed1996

Trisomy 18 mosaicism associated with secondary amenorrhea: ratios of mosaicism in different samples and complications.

A 28-year-old woman who complained of irregular menstruation was diagnosed as suffering from trisomy 18 mosaicism. She was karyotyped because of her characteristic face, mild mental retardation and aberrant hyperpigmentation of the skin. Her motor function was within normal range. Physical and laboratory examinations, however, revealed obesity, short stature, minor anomalies of the fingers, many areas of hyperpigmentation on the trunk and the hips, hypergonadotropinemia, diabetes mellitus, liver dysfunction, and hyperlipidemia. The ratios of normal/trisomy 18 were 4:135 in blood lymphocytes, 3:11 in a hyperpigmented area of the skin, 20:0 in a normally pigmented area of the skin, and 14:6 in ascitic cells. Laparoscopy revealed that her ovaries contained neither follicles nor germ cells.

Abnormalities, Multiple↗
PubMed1996

Enhancing the effect of anticancer drugs against the colorectal cancer cell line with electroporation.

Electroporation was applied in vitro and in vivo in the treatment of human colorectal cancer cell lines to study whether it can enhance the effect of bleomycin (BLM), 5-fluorouracil (5-FU) and cis-platinum (CDDP). We used LS174T and Colo320 cells derived from human colon cancer as target cells in this study. When the LS174T cells were used as target cells, the IC50 of BLM decreased to 10(-3) times, while that of 5-FU decreased to only about one fifth with the application of electric current. In the case of the Colo320 cells, the IC50 of BLM and 5-FU were about one hundredth and a half, respectively. The effect of CDDP was not enhanced with electric current. In vivo experiments were also performed using LS174T cells transplanted subcutaneously (s.c.) into nude mice. By treatment with intravenously (i.v.) administered BLM and simultaneous application of the electric current, tumors were markedly decreased in size after three weeks.

Animals↗
PubMed1996

[Molecular diagnosis in hematopoietic malignancy].

Molecular analysis of leukemias has been demonstrated to be useful for understanding the pathogenesis of leukemias or for detection of minimal residual disease. We present studies on the application of molecular diagnosis in malignant lymphomas. Although follicular lymphomas, a part of non-Hodgkin's lymphomas (NHL), often involve a t (14;18) chromosomal translocation, it is difficult to detect the translocation in some cases by conventional chromosomal analysis. We showed the usefulness of amplification of mbr/JH and mcr/JH junction by PCR to detect the translocation. The PCR products of mbr/JH junction were positive in 7 out of 8 cases of follicular lymphomas some of which were negative in tests on the translocation in the malignant cells. However, most NHL do not have cytogenetically specific translocations, indicating the necessity for another way to diagnose the malignant lymphomas. We used IgH gene probes as tumor specific markers in B-cell lineage malignant lymphomas. We determined DNA sequences for complementarity-determining region 3 (CDR3) of IgH in the malignant clone of each case. Then, we prepared a unique 5' primer for PCR or an oligonucleotide probe for detecting the clone-specific VH gene in each case, amplified the DNA by PCR and finally tried to detect minimal disease in bone marrow or peripheral blood, which brought into sequential detection of minimal residual disease in NHL. Thus, molecular diagnosis in hematopoietic malignancy will be more important in diagnosis, staging, and clinical management of patients with NHL.

Base Sequence↗
PubMed1996

[Antioxidant defense capabilities of gastric mucosa induced by water immersion restraint stress of rat].

Lipid peroxidation mediated by free radicals is believed to be one of the important causes of membrane destruction and cell damage. Lipid peroxidation in gastric mucosa induced by the stress is also suggested to cause gastric lesions. However very little is known about the antioxidant mechanisms in gastric mucosa, which is thought to be accelerated by the stress as an adaptive response. So we investigated lipid peroxide (LPO) and the production of lipid hydroperoxides by 1,5-lipoxygenase, which might reflect the antioxidant capabilities in gastric mucosa. The analysis of lipid hydroperoxide was done by high performance liquid chromatography (HPLC) using chemiluminescence which we have established. The production of lipid hydroperoxide by lipoxygenase was done by the condition of Low-Ethanol method. The water immersion restraint stress induced significant rise of gastric mucosal LPO assayed by the thiobarbituric acid-reactive substances method but lipid hydroperoxide was not detected by HPLC. The production of lipid hydroperoxide by lipoxygenase was clearly found in the gastric mucosa before the stress but the stress of 2 or 4 hours depressed the production of lipid hydroperoxides significantly. These findings showed that the stress induced the increase of antioxidant capabilities in the gastric mucosa as an adaptive reaction and the lipid hydroperoxide induced by the stress might be scavenged quickly.

Animals↗
PubMed1996

[Genetic diagnosis of pelvic lymph node metastasis using fine needle aspiration samples in prostate cancer].

We have developed a highly sensitive method to detect pelvic lymph node metastasis using the reverse transcriptase-polymerase chain reaction (RT-PCR) with primers specific for prostate-specific antigen (PSA) gene. Fine needle aspiration biopsy (FNAB) of pelvic lymph nodes was performed in 24 patients with prostate cancer. Each aspirated sample (0.05-0.1 ml) was divided into 2 parts; one for RNA extraction and RT-PCR to detect the fragment of PSA mRNA, and the other to smear on a slide glass for conventional cytology. The PSA gene was detected by RT-PCR in 11 FNAB samples which included not only all 6 cytologically positive and 2 cytologically class III cases but also 3 of 16 cytologically negative cases. The PSA gene was not detected by RT-PCR of FNAB samples in any of the 20 cases of bladder cancer. Thus RT-PCR for detection of the PSA gene in FNAB samples may be useful as a new diagnostic technique for detection of early lymph node metastasis in prostate cancer and an additional tool for cytological diagnosis of prostate cancer.

Aged↗
PubMed1996

[Genetic diagnosis of pelvic lymph node metastasis in prostate cancer using aspiration biopsy samples].

We have developed a highly sensitive method to detect pelvic lymph node(LN) metastasis using reverse transcriptase-polymerase chain reaction(RT-PCR) with the primers specific for prostate-specific antigen(PSA) gene in combination with the fine needle aspiration biopsy(FNAB). The specimens were obtained from pelvic LN from 15 prostate cancer patients and 15 bladder cancer patients. The aspirated samples (0.05 approximately 0.1 ml) were used for detecting the fragment of PSA mRNA by RT-PCR and Southern blot analysis, and the rest of samples were submitted to conventional cytology. Expression of PSA gene was detected in 9 cases of FNAB samples including all 5 cytologically positive and further more 2 cytologically class III cases, and 2 of 8 cytologically negative cases. RT-PCR of FNAB samples from all cases of bladder cancer were negative for the detection of PSA gene. The sensitivity of PSA gene by RT-PCR was very high and could detect 10 degrees cancer cell. In conclusion, our study suggested that RT-PCR for detection of PSA gene in FNAB samples might become a new diagnostic tool for detection of small foci of prostatic cancer metastasis in LN and combination use of RT-PCR and cytology could greatly contribute to accuracy in diagnosis.

Aged↗
PubMed1995

Rapid diagnosis of drug-resistant genes by PCR assay.

This report concerns the utility of the reverse transcription-polymerase chain reaction (RT-PCR) and quantitative PCR (QPCR) assay to detect the drug-resistance of related genes. The expression of some drug-resistance genes was compared with the sensitivity and resistance-acquired cancer cell lines to anti-cancer drugs by Northern blot analysis and PCR assay. The resistance cell lines exhibited an enhanced expression of multi-drug resistance (MDR-1), thymidylate synthase (TS), c-fos and DNA polymerase beta genes. Then these genes that expressed mRNA were quantitated using RT-PCR. The expression of the genes was dependent on their sensitivity (IC50) to anti-cancer drugs. Additionally, the QPCR assay has been developed as a rapid method for the expression of drug-resistance genes and applied to the PCR products amplified by the RT-PCR. Thus the QPCR assay for the expression of genes will allow rapid detection of the drug-resistance to chemotherapy in human cancers.

Blotting, Northern↗
PubMed1995

[Simultaneous measurement of unsaturated fatty acid and lipid peroxide in vivo by high performance liquid chromatography].

A chemiluminescence high performance liquid chromatography (HPLC) method for the simultaneous measurement of unsaturated fatty acid and lipid peroxide is newly described. This method was developed using reversed-phase HPLC which could separate unsaturated fatty acid and lipid peroxide clearly. Linear relationship was observed between these lipid peroxides and their peak areas on the chromatogram. The hydroperoxides were recently suggested to cause some diseases such as gastric lesions due to stress, but convenient method has not been established. The most popular method is the thiobarbituric acid (TBA) test, which has been extensively modified during the last several year. But the major disadvantage in this method is the interference of other thiobarbituric acid reactive substances (TBARS) and it is uncertain whether TBARS reflects the hydroperoxide concentration. This method is a highly sensitive fluorometric method for the quantitation of hydroperoxide at the nanomole level. Chemiluminescence was produced through luminol oxidation during the reaction of the hydroperoxide and cytochrome cheme. It is concluded that reversed phase HPLC can complement each other to serve as a direct and sensitive method for the determination of unsaturated fatty acid and lipid peroxide levels in vivo.

Animals↗