[A case of Brunner's gland hamartoma which is consistent of fat tissue and Brunner's gland hyperplasia].
Explore the source record for details and available documents.
Adipose Tissue↗
Biomedical subjects
Y Katakura
Publications and source records attributed to Y Katakura.
[IFOBT-positive gastric cancer].
Among 888 patients who underwent operation or endoscopic resection for gastric cancer (1994-1998), 75 patients, who had no colorectal disease or only small polyps 5 mm or less in diameter, were positive on the immunologic fecal occult blood test (IFOBT) (the positive group). They are compared with the other 813 patients (the negative group) as to the following 6 points: symptoms, presence of anemia, depth of invasion including macroscopic appearance, location, maximum diameter of lesions, and microscopic findings. The rate of positive-IFOBT gastric cancer was 8.4%. The average blood hemoglobin concentration was significantly lower in the positive group than in the negative group. Advanced cancers, especially type 2 and 3, were significantly more frequent in the positive group than in the negative group. The size of the lesions tended to be larger in the positive group than in the negative group. There was no difference between the groups as to symptoms, location, depth of invasion and microscopic findings. In conclusion, IFOBT-positive patients who have no colorectal disease or only small polyps 5 mm or less in diameter should be recommended to undergo upper gastrointestinal endoscopy.
Female↗
[The significance of magnetic resonance cholangiopancreatography in acute cholecystitis].
To clarify the significance of magnetic renonance cholangiopancreatography (MRCP) in the acute phase of acute cholecystitis, MRCP was carried out in forty-five patients with acute cholecystitis in their acute phase. The MR pericholecystic high signal was observed in 38 of the 45 patients (84%). Enlargement of the gallbladder, presence of gallstones, and impacted stones was seen in 71%, 53%, and 18%, respectively. The MR pericholecystic high signal was classified into four categories: type 0, not observed; type 1, a liner high signal; type 2, a band-like high signal; type 3, a radiating high signal. In patients who showed a type 3 MR pericholecystic high signal, 91% required percutaneous transhepatic gallbladder drainage, and most of the gallbladders were diagnosed as necrotic cholecystitis by histology. The accuracy of MRCP for the diagnosis of choledocholithiasis was 96%. It was suggested that MRCP for patients with acute cholecystitis in the acute phase provides useful information for planning the treatment.
Acute Disease↗
Characterization and cDNA cloning of androgenic gland hormone of the terrestrial isopod Armadillidium vulgare.
The sex differentiation in crustaceans is known to be controlled by a peptide hormone called androgenic gland hormone (AGH). AGH was extracted and purified from the androgenic glands (AGs) of the male isopod Armadillidium vulgare by high-performance liquid chromatography. AGH consisted of two peptide chains and their N-terminal amino acid sequences were determined. A cDNA encoding AGH was cloned by PCR and sequenced. The cDNA had an open reading frame of 432 bp, which encoded a preproAGH consisting of a signal peptide (21 residues), B chain (44 residues), C peptide (46 residues), and A chain (29 residues). Through processing, the A and B chains might form a heterodimer interlinked by disulfide bonds. The A chain possessed a putative N-linked glycosylation site. A Northern blot analysis using the cDNA as a probe detected a hybridization signal with 0.8 kb in the RNA preparation only from the AGs.
Amino Acid Sequence↗
Accelerated secretion of mutant beta-lactoglobulin in Saccharomyces cerevisiae resulting from a single amino acid substitution.
Transformed yeasts producing a mutant form of bovine beta-lactoglobulin (beta-LG), W19Y, in which Trp(19) was replaced with Tyr, were shown to secrete 6 times more than those producing wild type beta-LG. Northern blot analysis suggested that the enhanced level of secretion was not the result of upregulated transcription of W19Y. The ratio of the amount of W19Y secreted into the supernatant to the amount of W19Y remaining inside the cells was much larger than that in the case of wild type beta-LG as shown by immunoblot analysis. A pulse/chase experiment revealed that the speed of secretion of W19Y was significantly accelerated, compared to wild type beta-LG. These results indicated that W19Y was more efficiently and rapidly transported in the course of secretion than wild type beta-LG. Our previous study showed that the DeltaG of unfolding of W19Y in water is 6.9 kcal/mol smaller than that of wild type beta-LG. Furthermore, immunoblot analysis of intracellular beta-LG under non-reducing conditions indicated that W19Y as well as wild type beta-LG maintained a specific folded structure inside the yeast cells, whereas other non-secretable mutant beta-LGs with Phe or Ala at position 19 (W19F and W19A, respectively) did not. These data suggest that low molecular stability and the maintenance of a specific folded structure inside the yeast cells are prerequisites for efficient and rapid secretion. W19Y was more efficiently secreted than wild type beta-LG also in transformed ern1 mutant yeast cells expressing only a basal level of BiP which is considered to function in quality control in the endoplasmic reticulum (ER) by playing an important role in determining the secretion efficiency of secretory proteins. Thus, the reason for the enhanced secretion of W19Y is considered to be that the improved folding ability of W19Y can allow the half-life of the W19Y-BiP complex to become shorter than that of the wild type beta-LG-BiP complex, leading to faster translocation of W19Y into transport vesicles, or that W19Y can fold in a BiP-independent manner in the ER of the yeast cells. Our findings demonstrate that the amount of protein secreted can be improved by alteration of a single amino acid residue crucial for its structure.
Amino Acids↗
Effects of prenatal exposure to styrene on neurochemical levels in rat brain.
Styrene was evaluated to determine its neurochemical effects in the offspring of rats exposed during the gestation period. Maternal Wistar rats were exposed to 0, 50 or 300 ppm styrene during gestation days 6 to 20 and the neurochemical effects on their offspring were compared with their pair-feeding and ad lib. feeding controls. The cerebrum weights at birth on day 0 were significantly lower than those for an ad lib. feeding control group. Neurotransmitter analyses showed decreases of neuroamines, especially 5-hydroxytryptamine and homovanillic acid in the cerebrum of newborn offspring of dams receiving a 300 ppm styrene exposure compared with the ad lib. fed control group and homovanillic acid was also decreased compared to the pair-feeding control. On postnatal day 21, the styrene-exposure pups showed a significant decrease of 5-hydroxyindoleacetic acid in the frontal neocortex compared with the ad lib. control group. In the hippocampus a significant decrease of 5-hydroxyindoleacetic acid was observed compared with both control groups. Moreover, the ratio of 5-HIAA/5-HT in the hippocampus was significantly decreased among the styrene-exposure groups. The 50 ppm styrene exposed group induced increase of concentrations of 5-hydroxytryptamine in the striatum. These results suggest that prenatal styrene exposure affects the developing fetal brain in terms of a few signs of neurochemical alteration.
3,4-Dihydroxyphenylacetic Acid↗
Transforming growth factor beta triggers two independent-senescence programs in cancer cells.
Transforming growth factor-beta (TGF-beta)TG has been shown to play a multifunctional role in tumorigenesis. Here we demonstrate that TGF-beta induces a morphological change and expression of senescence-associated beta-galactosidase activity in the human lung adenocarcinoma cell line A549 cells within a week after the addition. These TGF-beta induced phenotypic changes are thought to characterize the rapid onset of senescence. When A549 cells were treated with TGF-beta, cell growth was not completely arrested, but the activity of telomerase was down regulated via transcriptional repression of telomerase reverse transcriptase, which led to a shortening of the telomere during long-term culture and finally resulted in replicative senescence. These results indicate that TGF-beta is able to induce a rapid senescence in A549 cells without significantly inhibiting cell growth and can further direct A549 cells to a replicative senescence state via the suppression of telomerase which culminates in telomere shortening. All these experimental results suggest that TGF-beta transmits several separate and independent signals to shift A549 cells back to a normal senescent cell.
Cell Division↗
Efficient production of recombinant human erythropoietin by replenishment of microcarriers in the hollow fiber culture cassette.
Human erythropoietin (EPO)-producing recombinant BHK cells were cultured in culture medium containing microcarriers, and then microcarriers attached with cells were replenished in the hollow fiber culture cassette. By culture for 14 days, it was possible to produce 450 micrograms of the recombinant EPO, which corresponded to over two-fold of the recombinant EPO production by control hollow fiber culture without microcarriers.
Animals↗
Immortalization by gene transfection.
Cultured cell lines that maintain specific differentiated phenotypes have been indispensable tools in cell biology. Progress in understanding the function of differentiated cells in vivo can be facilitated by creating cell lines via immortalizing gene transduction, if they retain the essential differentiated features of the same cells in vivo. Rodent cells immortalize spontaneously with a frequency of 10(-5) to 10(-6). Thus, it is easy to isolate immortal cells from rodent cell populations even without the transfer of immortalizing genes. Immortalizing genes can be used to increase this frequency to approximately 100%. In contrast, the spontaneous immortalization of human cells is a very rare event; the frequency is thought to be < 10(-12). Immortalizing genes can also be used to increase this frequency. Several genes that promise efficient immortalization of cultured cells have been identified. Immortalizing genes include simian virus 40 large T antigen, papillomaviruses E6 and E7, adenovirus E1A, Epstein-Barr virus, human T-cell leukemia virus, herpesvirus saimiri, oncogenes, and mutant p53 gene. Equally important, innovative means of gene delivery have been developed as well. These immortalizing genes, together with gene transfer methodologies, have provided the means to generate cell lines from cell types that are not abundant or are difficult to obtain in pure form in primary culture, are in short supply as human cells, and/or have brief lifetimes in culture. This chapter focuses primarily on the immortalization method by gene transfection. The chapter is not meant to be comprehensive, but rather to provide an account of the power and usefulness of immortalization methodology.
Animals↗
Cell hybridization, hybridomas, and human hybridomas.
Cell hybridization is one of the most basic cytotechnologies. The hemagglutinating virus of Japan was first used to cause cell fusion; however, polyethylene glycol is widely used now because of simplicity of procedure. This chapter first explains the principles of cell hybridization methods and then describes the practical protocols for preparing mouse hybridomas using polyethylene glycol. So far, lack of an excellent human fusion partner cell line that has high fusion efficiencies and does not produce immunoglobulin has hindered the spread of human-human hybridoma preparation methods. In the authors' laboratory NAT-30 and HO-323, human parent cell lines with high fusion efficiencies, have been established to prepare many hybridoma cell lines producing cancer-specific human monoclonal antibodies. Because NAT-30 and HO-323 cell lines are IgM producers, it is difficult to obtain IgG-producing hybridomas because the types of immunoglobulin produced by hybridomas are strongly affected by the characteristics of parent cells. Thus a nonimmunoglobulin-producing human parent cell line, A4H12, derived from human T lymphoma was established that can efficiently obtain IgG-producing human hybridomas. Another problem with preparing human hybridomas is that it is difficult to obtain B lymphocytes immunized with optional antigens for ethical reasons. To overcome this problem, in vitro immunization methods have been developed that allow exposure of a large number of B lymphocytes to cultured cancer cell or soluble antigens. The section on human hybridomas explains human fusion partners, in vitro immunization methods, and the preparation of human-human hybridomas using an electrofusion method. Finally, the application of human monoclonal antibodies to medical uses and the preparation of supranatural monoclonal antibodies are reviewed. These include multifunctional monoclonal antibodies and altered monoclonal antibodies having increased affinity and specificity by exchanging or modifying light chains.
Animals↗
Radiotherapy combined with transcatheter arterial infusion chemotherapy for locally advanced cervical cancer.
Twenty-four patients with locally advanced cervical cancer were treated with radiation therapy (RT) and transcatheter arterial infusion (TAI) chemotherapy, while 22 patients were treated with RT alone. RT consisted of a combination of external irradiation and high-dose-rate intracavitary brachytherapy. TAI therapy consisted of two sessions using cisplatin and pirarubicin, performed concurrently during the periods of external irradiation. The local-regional control rates at 1 year for the patients treated with RT plus TAI and for those treated with RT alone were 87.5% and 58.3%, respectively (p < 0.05). The 3-year cause-specific survival (CSS) rates for RT plus TAI, and RT alone were 67.1% and 55.9%, respectively (p = n.s.). The 3-year CSS rate for the 14 patients treated with RT and TAI who had well- or moderately differentiated squamous cell carcinoma without pelvic lymph node swelling was 100%, while that for the 19 patients with the same background treated with RT alone was 49% (p < 0.01). Radiation therapy combined with TAI appears to be an effective and safe treatment modality for patients with locally advanced cervical cancer.
Adult↗
An autocatalytic expression system for regulated production of recombinant protein in mammalian cells.
An autocatalytic inducible mammalian expression system was established. This system is composed of two sets of vectors: one carries a selectable marker gene and the other carries a bicistronic expression unit consisting of a target gene, an internal ribosomal entry site, and a tetracycline-controlled transactivator gene. Both the selectable marker and the bicistronic unit are controlled by the tetracycline-responsive promoter (PhCMV*-1). When the two vectors are cotransfected into host cells, only a single selection round in the absence of tetracycline is necessary to generate clones expressing the target gene. The expression level is high and easily regulated by tetracycline. Combination with a gene amplification system may allow further enhance the foreign gene expression. Because the PhCMV*-1 promoter is relatively independent of cellular regulation signals, this system is expected to work in a wide variety of cell types.
Animals↗
Bidirectional regulation of telomerase activity in a subline derived from human lung adenocarcinoma.
Telomerase is active in germline cells and tumor cells, but is either not expressed or is repressed in a majority of somatic cells. The regulation mechanism of telomerase activity has lately drawn considerable attention for the possible use of telomerase inhibitors for anti-cancer therapy. We analyzed the regulation mechanism of telomerase activity in the human lung adenocarcinoma cell line A549 and its subline A5DC7 which shows impaired tumor phenotypes. Although A549 and A5DC7 cells have similar growth potentials, when cultured in medium supplemented with 5% fetal bovine serum A5DC7 cells demonstrated a remarkable negative regulation of telomerase activity and telomere shortening. After the long-term culture, A5DC7 cells showed a large senescent cell-like morphology, arrested growth at the 106 population doubling level and entered senescence which was demonstrated by expression of the beta-galactosidase senescence marker. When the serum concentration was raised from 5% to 10% for the senescent A5DC7 cells, telomerase reactivation and telomere lengthening occurred as well as resumption of proliferation. These results demonstrate that the growth arrest seen in senescent A5DC7 cells is reversible and that telomerase activity is also bi-directionally regulated in A5DC7 cells, an event which could not be observed in normal senescent cells. This indicates that stringent telomerase repression and complete growth arrest concomitant with cellular senescence is collapsed in senescent A5DC7 cells in response to exogenous signals. Our study suggests that cellular senescence progresses and/or is regulated at multiple levels.
Adenocarcinoma↗
Electrolyzed-reduced water scavenges active oxygen species and protects DNA from oxidative damage.
Active oxygen species or free radicals are considered to cause extensive oxidative damage to biological macromolecules, which brings about a variety of diseases as well as aging. The ideal scavenger for active oxygen should be 'active hydrogen'. 'Active hydrogen' can be produced in reduced water near the cathode during electrolysis of water. Reduced water exhibits high pH, low dissolved oxygen (DO), extremely high dissolved molecular hydrogen (DH), and extremely negative redox potential (RP) values. Strongly electrolyzed-reduced water, as well as ascorbic acid, (+)-catechin and tannic acid, completely scavenged O.-2 produced by the hypoxanthine-xanthine oxidase (HX-XOD) system in sodium phosphate buffer (pH 7.0). The superoxide dismutase (SOD)-like activity of reduced water is stable at 4 degrees C for over a month and was not lost even after neutralization, repeated freezing and melting, deflation with sonication, vigorous mixing, boiling, repeated filtration, or closed autoclaving, but was lost by opened autoclaving or by closed autoclaving in the presence of tungsten trioxide which efficiently adsorbs active atomic hydrogen. Water bubbled with hydrogen gas exhibited low DO, extremely high DH and extremely low RP values, as does reduced water, but it has no SOD-like activity. These results suggest that the SOD-like activity of reduced water is not due to the dissolved molecular hydrogen but due to the dissolved atomic hydrogen (active hydrogen). Although SOD accumulated H2O2 when added to the HX-XOD system, reduced water decreased the amount of H2O2 produced by XOD. Reduced water, as well as catalase and ascorbic acid, could directly scavenge H2O2. Reduce water suppresses single-strand breakage of DNA b active oxygen species produced by the Cu(II)-catalyzed oxidation of ascorbic acid in a dose-dependent manner, suggesting that reduced water can scavenge not only O2.- and H2O2, but also 1O2 and .OH.
Ascorbic Acid↗
Fluorescence polarization study of a salt bridge between a single-chain Fv and its antigen ribonuclease A.
The interaction between a single-chain Fv (sFv) of the monoclonal antibody 3A21 and its antigen, bovine pancreatic ribonuclease A (RNase A), was studied by site-directed mutagenesis of the hypervariable regions and fluorescence polarization analysis. The affinity constants of wild-type sFv and a mutant sFv D31A (Asp31 of heavy chain was replaced by Ala) for RNase A were found to be 2.7 x 10(7) and 4.7 x 10(6) M-1 in PBS at pH 7.2 and 37 degrees C, respectively. While the affinity constant of D31A is not affected by NaCl concentration, that of wild-type sFv is almost the same as that of D31A in the presence of more than 1 M NaCl. These results demonstrate that Asp31 of the heavy chain interacts electrostatically with a positively charged amino acid residue of RNase A.
Alanine↗
Development of DNA diagnostic methods for the detection of new fish iridoviral diseases.
A new disease of epidemic proportions caused by fish viruses within the Iridoviridae family inflicts serious damage on red sea breams (Pagrus major) and striped jack (Caranx delicatissimus) populations grown in aquacultures in Japan. A partial segment of the fish iridoviral DNA was directly amplified using the polymerase chain reaction (PCR) with synthetic primers designed from well conserved nucleotide sequences between the frog virus 3 (Ranavirus) and the silkworm iridescent virus type 6. The deduced amino acid sequence from the nucleotide sequence of the PCR fragment demonstrates a high correlation with a partial sequence from the frog virus 3. Using the PCR method with specific primers, we could detect three of four different known types of fish iridoviruses in diseased fishes. To construct more reliable detection methods specific for this viral family, DNA fragments which can specifically hybridize with all of the four known iridoviridae viral DNAs were screened from the genomic library of one iridoviridae strain. The hybridization assay, using a specific fragment which contains regions which are highly homologous with a characterized partial sequence from the frog virus 3, proved to be a reliable diagnostic tool for fish iridoviral diseases.
Animals↗
Structure and function of the recombinant fifth domain of human beta 2-glycoprotein I: effects of specific cleavage between Lys77 and Thr78.
In order to elucidate the mechanism of binding of beta 2-glycoprotein I (beta 2-GPI) to cardiolipin (CL), we constructed a high-level expression system for the C-terminal domain (Domain V) of beta 2-GPI using Pichia pastoris and studied its conformation and liposome-binding activity. Purified Domain V was found to have the native disulfide bonds. It had a compactly folded conformation, judging from the circular dichroism spectrum, and exhibited a cooperative unfolding transition induced by pH or urea. Also, it bound liposomes containing CL. Commercially available human beta 2-GPI is known to be selectively cleaved between Lys 317 and Thr 318. We found that bovine factor Xa weakly but specifically cleaves the corresponding site of recombinant Domain V, i.e., the peptide bond between Lys 77 and Thr 78. The conformation of the "nicked" Domain V, which was cleaved at this site, was examined by circular dichroism and fluorescence measurements, and concluded to be similar to that of the intact protein. The stability of the nicked Domain V to urea was slightly lower than that of the intact protein. Although both Domains V bound to liposomes containing CL, the affinity of the nicked Domain V was greatly reduced in comparison with the intact protein, indicating that the cleavage of the peptide bond between Lys 77 and Thr 78 controls the binding to CL. In addition, analysis of the fluorescence spectra in the presence and absence of CL liposomes indicated that Trp 76 is involved in the binding site. These results suggest that the region including Trp 76, Lys 77, and Thr 78 has a critical role in binding to CL.
Amino Acid Sequence↗
Genetic and biochemical studies on ommochrome genesis in an albino strain of a terrestrial isopod, Armadillidium vulgare.
Genetic studies and quantitative determination of levels of 3-hydroxykynurenine and kynurenine were performed in an albino strain of a terrestrial isopod Armadillidium vulgare. From the results of matings between the albino and the albino, the red, the dark red, or the wild type individuals, the albino A. vulgare seems to be regulated by an autosomal gene(s) recessive to its wild allele. Litter mating of F1 progenies obtained by crossing the albino and the red mutant or the albino and the dark red mutant yielded progenies at a ratio of 3:6:3:4 for the red, the dark red, the wild, and the albino phenotypes, respectively. The albino gene(s) seems not to be allelic but to be epistatic to the red gene(s) with respect to ommochrome biosynthesis. Quantitative determination of 3-hydroxykynurenine carried out by high-performance liquid chromatography with electrochemical detection revealed that the 3-hydroxykynurenine content in the albino was significantly lower than that in the wild or the red type. The whole content of 3-hydroxykynurenine after enzymatic conversion of kynurenine to 3-hydroxykynurenine was still considerably lower than that found in the wild type, even though it increased after the conversion. The albino gene(s) seems to be associated with a blockage at distinct level(s) of ommochrome biosynthesis.
Albinism↗