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Biomedical subjects

K Isono

Publications and source records attributed to K Isono.

At least 109 records · Page 6Linked to original sources

Evidence for transcriptional regulation of plastid photosynthesis genes in Arabidopsis thaliana roots.

Mechanisms underlying suppressed levels of transcripts for plastid photosynthesis genes in nongreen tissues such as roots and calli were analyzed in Arabidopsis thaliana, a plant suitable for further genetic dissection. A region encoding promoters of rbcL, the gene encoding the large subunit of ribulose-1,5-biphosphate carboxylase/oxygenase, and the atpB/E operon for the beta and epsilon subunits of coupling factor one were cloned and sequenced. Transcripts for rbcL, atpB/E, and psbA, the gene for the D1 protein in the photosystem II reaction center, were barely detectable in roots of A. thaliana, whereas 16S rRNA was detected at a low level. The run-on transcription experiment revealed that expression of rbcL, atpB/E, and psbA was regulated at transcription. The copy number of plastid DNA in roots was one-fifth that in green leaves on the basis of total cellular DNA, suggesting that in the latter the DNA copy-number regulation also exists in plastid gene expression. Digestion of DNA with methyl-sensitive and -insensitive isoschizomeric endonucleases and subsequent polymerase chain reaction, as well as in vitro transcription of plastid DNAs with Escherichia coli RNA polymerase, resulted in no evidence of regulation by DNA modification. In spite of predominant suppression of expression of rbcL, atpB/E, and psbA at transcription in roots and calli, 16S rRNA levels were decreased because of low RNA stability.

Arabidopsis↗

No evidence of mutation in the human PTC gene, responsible for nevoid basal cell carcinoma syndrome, in human primary squamous cell carcinomas of the esophagus and lung.

The high frequency of loss of heterozygosity that has been observed on the distal region of the long arm of chromosome 9 in squamous cell carcinomas of esophagus, lung, uterus, and head and neck indicates the presence of a tumor suppressor gene(s) in this region. To investigate the possible role of the PTC gene on chromosome 9q22.3, that was identified as the cause of nevoid basal cell carcinoma syndrome, during carcinogenesis in esophagus and lung, we examined 20 esophageal squamous cell carcinomas and 10 squamous cell carcinomas of the lung for mutations in any coding exon of PTC. Using single-strand conformation polymorphism and direct sequencing, we detected no mutations other than two non-deleterious polymorphisms. Our results suggest that inactivation of some tumor suppressor gene(s) on 9q other than PTC contributes to the development of squamous cell carcinomas in these tissues.

Basal Cell Nevus Syndrome↗

Assessment of energy status and fructose metabolism of liver in obstructive jaundice by 31P magnetic resonance spectroscopy.

The effect of cholestasis on hepatic energy status and fructose metabolism in jaundiced rats and patients was investigated using 31P magnetic resonance spectroscopy. Rats with obstructive jaundice (OJ group) were studied 7 days after bile duct ligation. Drainage rats were studied at 3 days (DR3 group) and 7 days (DR7 group) after the relief of 1 week obstruction of the common bile duct. In the bile duct ligated rat, the beta-adenosine triphosphate (ATP)/Pi (inorganic phosphate) ratio was significantly lower than in sham-operated controls. This ratio recovered rapidly in the DR3 and DR7 groups. The maximum increase in the phosphomonoester peak (PMEmax) after an intravenous bolus of fructose was significantly reduced in both the OJ and DR3 groups, and was accompanied by a decrease in hepatic fructokinase activity. The PMEmax and the fructokinase activity recovered in the DR7 group. In a clinical study, the beta-ATP/Pi ratio in six healthy volunteers was comparable to that of 15 patients with obstructive jaundice, regardless of their biliary drainage status. The PMEmax in all patients (serum bilirubin > or = 5 mg/dL), irrespective of biliary drainage, was significantly lower than in volunteers. Furthermore, the PMEmax in four of the eight patients with biliary drainage (serum bilirubin < 5 mg/dL) was lower than in volunteers. It is concluded that while energy status in jaundiced patients is well maintained, fructose phosphorylation is inhibited and recovery is delayed after the relief of obstruction compared with serum bilirubin. For the non-invasive evaluation of damaged liver function in jaundice, 31P magnetic resonance spectroscopy is a useful technique.

Adenosine Triphosphate↗

Gene identification and classification in the Synechocystis genomic sequence by recursive gene mark analysis.

The GeneMark method has proven to be an efficient gene-finding tool for the analysis of prokaryotic genomic sequence data. We have developed a procedure of deriving and utilizing several GeneMark models in order to get better gene-detection performance. Upon applying this procedure to the 1.0 Mb contiguous DNA sequence of Synechocystis sp. strain PCC6803, we were able to cluster predicted genes into distinct classes and to produce the class-specific GeneMark models reflecting statistical characteristics of each gene class. One gene class apparently includes genes of exogenous origin. Using class-specific models reduces the gene under prediction error rate down to 1.7% in comparison with 8.1% reported in the previous study when only one GeneMark model was used.

Bacterial Proteins↗

[A novel quantitative PCR with fluorogenic probe].

The polymerase chain reaction(PCR) is a powerful tool to amplify small amounts of DNA or RNA for various molecular analysis. However, in these analyses, PCR only provides qualitative results. The availability of quantitative PCR provides valuable additional information in various applications. It is difficult to establish absolute quantitation, because PCR amplification is a complicated reaction process of exponential growth. To trace the amplification process, the initial amount of template and the efficiency of amplification in each cycle, has to be determined. Conventional methods have not achieved absolute quantitative analysis. The ABI PRISM 7700 Sequence Detection System has solved these problems with real-time monitoring of the PCR process. The real-time detection system provides essential information to quantify the initial target copy number, because it can draw an amplification curve. Using the 5' nuclease assay, a specific fluorescent signal is generated and measured at every cycle during a run. This system can perform a variety of applications including, quantitation, allele discrimination, PCR optimization and viral screening. Using the ABI PRISM 7700 Sequence Detection System, the rice genome has been quantitatively analyzed. To monitor maturation of the chloroplast genome from proplastid during germ development, 5' nuclease assay set up for Cab and rbcL genes which are located in the nuclear genome and chloroplast genome, respectively. Cab was used as an internal standard for normalization of cell numbers. The maturation process of chloroplast was estimated using the ratio of gene dosage, [rbcL]/[Cab]. After development of cotyledon, a significant increase in copy numbers of the chloroplast was observed. These results indicate that a light-induced chloroplast maturation process is coupled with an increase in chloroplast genome copy numbers.

Chloroplasts↗

[Effect of heparin on the increase in intracellular calcium caused by inositol 1,4,5-triphosphate in airway epithelial cells].

We studied the effect of heparin on the inositol 1,4,5-triphosphate (IP3)-dependent increase in intracellular calcium (Ca2+) caused by ATP. The Ca2+ concentration ([Ca2+]i) in cultured epithelial cells from cow tracheas was measured by the fura-2 method. ATP (10(-4) M) stimulated IP3 production and caused a biphasic increase in [Ca2+]i a transient response and a subsequent sustained response. Heparin (l 100 U/ml) dose-dependently inhibited the ATP-induced increase in [Ca2+]i but had no effect on IP3 production. Dextran sulfate, a polysaccharide with negative charge density, had a similar inhibitory effect on the ATP-induced increase in [Ca2+]i. These data suggest that heparin inhibits intracellular Ca2+ mobilization by acting at a site "distal to" IP3 production, and that the mechanism of action of heparin may be related to its negative charge.

Adenosine Triphosphate↗

[Surgical treatment and research of esophageal cancer in the past and the present era in our department--for the view point of the future].

We describe the result of esophageal cancer treatment in the past era of the Department of Surgery, Chiba University School of Medicine directed by Professor Seo, Nakayama and Sato and the improved results of the treatment at the present time from the view point of diagnosis (early superficial cancer, lymph node metastasis and adjacent organ invasion) and treatment (three-field lymph node dissection, improved result of operative mortality and morbidity, improvement of long-term survival rate). In addition, we prospect the future through the present study such as mutation of P53 gene and malignancy, immunotherapy using cytokine gene transfer, cancer inhibition therapy by induction of cancer suppressor gene, prodrug therapy, heavy particle iron therapy, and clinical application of virtual reality to the surgical field.

Esophageal Neoplasms↗

[Reasonable lymph node dissection for T2 or T3 midthoracic esophageal cancer with cervical lymph-node metastasis].

In order to determine the reasonable lymph node dissection for T2 or T3 midthoracic esophageal cancer with cervical lymph node metastasis, a retrospective study was carried out on 106 patients receiving resection between 1983 and 1996. Metastasis to cervical lymph node was obtained in 27.4% (29/106) of patients with T2 or T3 midthoracic esophageal cancer. Within 29 patients, metastasis in cervical node only, in two fields and in three fields occupied 17.2%, 41.4% and 41.4%, respectively. And according to the histologic examination of dissected lymph nodes, metastatic sites spreaded from neck to perigastric region. Five-year survival rate of 23 patients receiving curative operation was 33.0%, and that of 13 patients excluding 3-field metastasis was 51.9%. But the main sites of nodal recurrence were cervical or superior mediastinal nodes along the bilateral recurrent laryngeal nerves, and the rate of nodal recurrence was 47.8%. These results of actual state of lymph node metastasis and prognostic benefit of aggressive dissection suggest that 3-field lymph node dissection is mandatory for T2 or T3 midthoracic esophageal cancer with cervical lymph node metastasis. And we should endeavor to upgrade the precise dissection in order to decrease the nodal recurrence.

Esophageal Neoplasms↗

[Arterial infusion chemotherapy in pelvic space for advanced rectal cancer].

Three rectal cancer patients were treated with arterial infusion chemotherapy through the internal iliac artery. Two patients with pelvic recurrences unresectable after APRA were treated with intensive chemotherapy and the other patient with neoadjuvant chemotherapy. In all cases, 5-FU (500 mg/ body/day) was administered continuously for 5 days or 14 days. We attempted three methods for this procedure which were a bilateral catheterization to the internal iliac artery, a single catheterization with an embolization to the other internal iliac artery and a single catheterization without the embolization. As the result of this treatment, in the resected specimen with neoadjuvant case, histological necrosis was found in 50% in the main tumor and the metastatic lymph nodes. One patient with pelvic recurrence showed a partial response in CT imaging, but died one year later of the recurrence around the external iliac artery. The other patient with pelvic recurrence treated with the bilateral catheterization had no efficacy on CT imaging, but his CEA level has decreased at present. It was concluded that arterial infusion chemotherapy was effective for advanced rectal cancer and the pelvic recurrences. However, the efficacy of this treatment is limited to the area to which the drugs are delivered. Thus, it is important that the method and the location of the catheterization are determined adequately for each case.

Antidotes↗

[Evaluation of CTA for arterial infusion chemotherapy for liver metastasis from colorectal cancer].

To compare between the arterial blood supply of metastatic liver tumor and effects of intrahepatic arterial infusion chemotherapy (IHAC), we examined 8 patients with 27 liver metastasis from colorectal cancer. They were treated with ADM/lipiodol/5-FU/LV (19 nodules) or 5-FU/LV (8 nodules). To evaluate the arterial blood supply, CT arteriography (CTA) was performed, which classified tumors into 3 grades. Grade (Gr) 0; almost no enhancement, Gr1: less than one-third of tumor, Gr 2; less than two-thirds; Gr 3 over two-thirds (no case). As a result, 3 of NCs and 3 of PDs were Gr 0, 7 of MRs, 3 of NCs and 2 of PDs were Gr 1, and CR, PR and 7 of MRs were Gr 2. These results suggest that the arterial blood supply is necessary for a better response of IHAC and CTA is effective to forecast the response to IHAC.

Antineoplastic Combined Chemotherapy Protocols↗

A 570-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 28.0-40.1 min region on the linkage map.

The 569,750 base pair sequence corresponding to the 28.0-40.1 min region on the genetic map of Escherichia coli K-12 (W3110) was determined. This region includes the replication terminus region and contained at least 549 potential open reading frames. Among them, 160 (29%) were previously reported, 174 (32%) were homologous to other known genes, 102 (18%) were identical or similar to hypothetical genes registered in databases, and the remaining 113 (21%) did not show a significant similarity to any other gene. Of interest was the finding of a large number of genes and gene clusters in and near the replication termination region which had been thought to be genetically silent. Those included a cluster of genes for fatty acid beta-oxidation, the third copy of the pot (spermidine/putrescine transport system) gene cluster, the second dpp (dipeptide transport system) operon, the second dsm (anaerobic dimethyl sulfoxide reductase) operon, a cluster of fim (fimbrial) genes and a DNA helicase-like gene with a high molecular weight. In addition, we found the dnaC- and dnaT-like genes in the cryptic prophage, Rac, and a number of genes originated probably from plasmids.

Chromosome Mapping↗

A 460-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 40.1-50.0 min region on the linkage map.

The 465,813 base pair sequence corresponding to the 40.1-50.0 min region on the genetic map of Escherichia coli K-12 (W3110) was determined. Analysis of the sequence revealed that this region contained at least 466 potential open reading frames, of which 187 (40%) were previously reported, 105 (23%) were homologous to other known genes, 103 (22%) were identical or similar to hypothetical genes registered in databases, and the remaining 71 (15%) did not show a significant similarity to any other gene. At the 45.2-46.0 min region, we found a very large cluster of about 30 genes, whose functions are involved in the biosynthesis of polysaccharides as the components of outer membranes. In addition, we identified a new asn-tRNA gene, designated asnW, between the asnT and asnU genes and a new lysogenic phage attachment site as the cis-element.

Attachment Sites, Microbiological↗

Inhibition of peritoneal dissemination of colon carcinoma in syngeneic mice immunized with interleukin-2-producing cells.

We have examined the antitumor effect of murine colon carcinoma cells engineered to produce human interleukin-2 (IL-2) in syngeneic mice. Subcutaneous inoculation of retrovirally-transduced cells with IL-2 gene formed small tumors, but they became regressed spontaneously. Consequently, the inoculated mice showed prolonged survival. Histological examination of the tumors derived from IL-2-producers revealed predominant infiltration of macrophages around tumor necrotic masses. Thus, inoculation of IL-2-producing cells could protect the mice from subsequent subcutaneous or intraperitoneal challenges with wild-type cells, suggesting the induction of acquired immunity due to the effect of tumor vaccination.

Animals↗

The BCL6 gene is predominantly expressed in keratinocytes at their terminal differentiation stage.

We analyzed the expression of the BCL6 gene in mouse tissues by in situ hybridization. The expression was strong in the upper layer but undetectable in the basal layer of epidermis from adult mice. When human keratinocytes were cultured with a high concentration of calcium ion, these cells stopped their proliferation and differentiated to their terminal stage. In these keratinocytes, BCL6 expression was induced after stimulation and progressively up-regulated. The kinetics was very similar to that of a cyclin dependent kinase inhibitor p21sdil/cip1/WAF1 in these cells. These results suggest that BCL6 plays a role in keratinocytes at terminal differentiation stage.

Animals↗

Expression of taste reception response of fleshfly in Xenopus oocytes.

Functional expression of the gustatory sensitivity to amino acids and sugars was investigated in Xenopus oocytes injected with poly(A+) RNA extracted from fleshfly labellar taste organs. The current induced by the application of amino acids and sugars that stimulate fleshfly sugar receptor cells was recorded under voltage clamp conditions. L-Phenylalanine and L-valine induced a large transient inward current in poly(A+) RNA-injected oocytes but only small responses in uninjected control oocytes. Glucose and fructose also produced inward currents in the RNA-injected oocytes. N-methylated compounds or D-isomers of these amino acids induced either no response or a much smaller response than L-amino acids as they did in the fleshfly sugar receptor cells. The oocyte expression system is a useful tool for characterizing the taste transduction mechanism.

Animals↗