Dose-volume histogram analysis of external-beam irradiation combined with IORT for unresectable pancreatic cancer.
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Combined Modality Therapy↗
Biomedical subjects
Y Katakura
Publications and source records attributed to Y Katakura.
Fed-batch culture strategy for high yield of baker's yeast with high fermentative activity.
Based on the determination of the operational conditions for batch culture and fed-batch culture of baker's yeast by process analysis and continuous culture, a correlation, which describes the relationship between fermentative activity (FA) and specific growth rate (mu), was obtained. Combining this correlation and the exponential fed-batch culture equation, a fed-batch culture strategy was developed to obtain high yield and high fermentative activity by controlling mu at different stages. The results showed that when the initial and residual sugar concentrations were controlled to be 15-30 g/L and 3-6 g/L, respectively, different stirring speeds were applied at different stages to provide optimal oxygen transfer conditions. When this proposed fed-batch culture strategy was used in a fed-batch culture, the yield of baker's yeast reached 0.432 g/g with high fermentative activity (1180 ml). Thus, the combined bed-batch culture of baker's yeast with high yield and high fermentative activity was realized.
Biotechnology↗
IL-3 augments TCR-mediated responses of type 2 CD4 T cells.
A subset of type 2, but not type 1, CD4 T cell clones expresses IL-3R and can be stimulated by IL-3. Expression of IL-3R on these type 2 T cell clones is induced by TCR stimulation, and subsequent stimulation by IL-3 augmented the proliferation of and IL-4 production by these cells. This augmented response is inhibited by anti-IL-4 mAb, suggesting the involvement of IL-4 in this response. In place of TCR stimulation, treatment of these type 2 CD4 T cell clones with PMA rendered them responsive to further stimulation of proliferation by IL-3, indicating the cooperation between the IL-3R-elicited signals and PKC-mediated signals in stimulating proliferation. Although the augmentation of the TCR-mediated proliferative response by IL-3 was mainly due to the increased production of IL-4, we also demonstrated the presence of IL-4-independent mechanism mediating the response to IL-3. In situ, we found that splenic T cells could be induced to respond to Il-3 by TCR stimulation. Thus, IL-3 can stimulate a specific population of T cells and influence the immune response.
Adjuvants, Immunologic↗
Study of urinary mandelic acid concentration and peripheral nerve conduction among styrene workers.
The relationship between exposure to styrene and nerve conduction velocities was investigated in 32 workers occupationally exposed to styrene. There was a dose-dependent relationship between urinary mandelic acid and ulnar and peroneal motor distal latencies (MDLs). The ulnar and peroneal MDLs of subgroup B (urinary mandelic acid > or = 250 mg/l) were significantly longer than those of subgroup A (urinary mandelic acid < 250 mg/l)(p < 0.05), and the referent group (p < 0.03). After adjusting confounding factors, urinary mandelic acid had a significant positive relationship with ulnar and peroneal MDL. Our study suggests that a low level of styrene, below the 50 ppm exposure limit of the American Conference of Governmental Industrial Hygienists (ACGIH), involves the risk of inducing adverse effects on the peripheral nervous system. The study also indicated that motor distal latency is a sensitive parameter of toxic peripheral neuropathy.
Action Potentials↗
Production of recombinant human monoclonal antibody using ras-amplified BHK-21 cells in a protein-free medium.
A ras oncogene-amplified recombinant BHK-21 cell line (ras-rBHK-IgG) has been established, and was shown to hyperproduce the recombinant IgG chimeric human monoclonal antibody (hMAb) AE6F4, which recognizes lung cancer cells. We found that the ras-rBHK-IgG cell could be easily cultured in a protein-free ERDF medium supplemented with iron(III) nitrate, hydroxyethyliminodiacetic acid, and non-protein synthetic attachment factor as well as in a serum-free ERDF medium supplemented with insulin, transferrin, ethanolamine, and sodium selenite. The productivity of recombinant hMAb from the cells cultured in dishes at high cell densities was higher in protein-free medium than in serum-containing medium. True high density culture of the ras-rBHK-IgG cells was done in protein-free medium using the Tecnomouse, which is a novel hollow fiber bioreactor system. After culture for 30 days in protein-free culture, a total amount of about 14 mg of the recombinant hMAb AE6F4 was obtained, and was shown to be reactive against lung cancer cells in tissues.
Animals↗
Impaired tumor phenotypes in class II major histocompatibility complex antigen-inducible cells originated from human lung adenocarcinoma.
The class II major histocompatibility complex (MHC) antigens play important roles in T cell activation and are thought to be involved in tumor development. Using anti-class II antibodies with interferon gamma (IFN-gamma), we have here selected class II MHC antigen positive cancer cells from the human lung adenocarcinoma cell line A549, which is originally negative (> 95%) for the antigens. A part of the class II antigen-positive cells presented a flat morphology, which was not observed in the parental A549 cells. Class II antigen expression in these flat cells was IFN-gamma inducible; there was a correlation between the inducibility and phenotypic changes. A class II antigen-inducible flat subline restored the ability of contact inhibition and anchorage-dependent growth as well as lost tumorigenicity in athymic mice.
Adenocarcinoma↗
Molecular cloning of the 31 kDa cytosolic phospholipase A2, as an antigen recognized by the lung cancer-specific human monoclonal antibody, AE6F4.
The human monoclonal antibody AE6F4 specifically reacts with human lung cancer tissues but does not with normal tissues. This monoclonal antibody recognizes a cytosolic 31 kDa antigen in the cancer cells. In a previous study, we elucidated that the 31 kDa antigen belonged to a family of proteins collectively designated as 14-3-3 proteins, which were known as protein kinase-dependent activators of tyrosine/trytophan hydroxylases, or protein kinase C inhibitor proteins. Here we report molecular cloning of the 31 kDa antigen from the human lung adenocarcinoma cell line, A549. Sequencing analysis indicates that the cloned cDNA is identical to that of previously reported human placental cytosolic phospholipase A2 (cPLA2), which is also a member of the 14-3-3 protein family. Western analysis demonstrated that a 31 kDa recombinant cPLA2 expressed in monkey COS cells was recognized by the AE6F4 monoclonal antibody. Binding of the monoclonal antibody to the recombinant cPLA2 was abolished when treated with sodium periodate, suggesting that not only are carbohydrate chains associated with the cPLA2, but they also play a crucial role in antigen recognition by the monoclonal antibody.
14-3-3 Proteins↗
Molecular cloning of yeast cytochrome c-like polypeptide expressed in human lung carcinoma: an antigen recognizable by lung cancer-specific human monoclonal antibody.
We previously determined the amino acid sequence to the epitope (ATLFKTR) of cytochrome c from Candida krusei, which is cross-reactive to the lung cancer-specific human monoclonal antibody HB4C5. Here we report that an antigen messenger RNA, which codes for a structure similar to the cytochrome c epitope, is expressed in the human lung adenocarcinoma A549. Sequencing analysis has revealed that this messenger RNA encodes a novel 190 amino acid polypeptide of 21-kDa containing an amino acid sequence (ALLFFT) similar to the cytochrome c epitope, although the total messenger RNA sequence is apparently different from the cytochrome c messenger RNA. Western analysis indicated that an antibody-recognizable 21-kDa antigen which has the same molecular weight as the predicted polypeptide is expressed in the A549 adenocarcinoma. The in vitro translated product of the antigen messenger RNA and synthesized ALLFFT peptide were both shown to be reactive with the monoclonal antibody, indicating that this protein contains the epitope which enables A549 cells to specifically react with the antibody. The antigen mRNA was not expressed in non-transformed fibroblasts, suggesting that the antigen mRNA expression was associated with cellular transformation. Also in part of the antigen nucleotide sequence, there was a segment that had about 90% homology to the long terminal repeat sequence (no. 297-475) of the human endogenous retrovirus HERV-K10, which was related to the mouse mammary tumor virus.
Adenocarcinoma↗
Epitope in cytochrome c from Candida krusei cross-reacts lung cancer-specific human monoclonal antibody HB4C5.
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Amino Acid Sequence↗
Impaired colour discrimination among workers exposed to styrene: relevance of a urinary metabolite.
OBJECTIVES: To survey the loss of colour vision among Japanese workers who have been exposed to styrene concentrations currently considered low (about 20 ppm). Also to assess the effects of styrene by examination of the nature of the relation between disorder of colour vision and age, alcohol consumption, and other variables. METHODS: Colour discrimination was examined in 64 male workers exposed to styrene (mean age; 38.0, mean exposed years; 7.0) and in 69 controls (mean age; 38.0). A standardised questionnaire was adopted to collect work history, occupational or non-occupational solvent exposure, alcohol consumption, and drug use. Colour vision was evaluated by the Lanthony desaturated panel D-15 test. The results of the test were expressed as the colour confusion index (CCI). RESULTS: The mean atmospheric styrene concentration was about 20 ppm. The mean urinary concentration of mandelic acid was 0.22 g/l. There was a significant difference in CCI between exposed workers and age matched controls. Colour vision of workers whose concentration of urinary mandelic acid was > or = 0.42 g/l was significantly impaired when compared with workers whose concentration was < 0.42 g/l. Multiple linear regression analysis that controlled confounding variables such as age, alcohol consumption, smoking, and educational attainment showed that the CCI was significantly related to the concentration of urinary mandelic acid. In both exposed workers and controls, the types of defects were mostly blue-yellow loss, although a few subjects showed complex loss. No one showed only red-green loss. CONCLUSIONS: These findings suggest that exposure to moderate styrene concentrations can lead to impairment of colour vision, and that there is a significant correlation with the urinary metabolite of styrene.
Adult↗
Isolation and structural determination of seminal vesicle-specific peptides of the terrestrial isopod, Armadillidium vulgare.
During the course of purifying the androgenic gland hormone of the terrestrial isopod, Armadillidium vulgare, that induces post-embryonic sex differentiation, four structurally related peptides were obtained and their structures determined by a combination of microsequence and mass spectral analyses. These peptides were found to exist speciffically in the seminal vesicle and vas deferens by a Western blot analysis, therefore being designated as seminal vesicle-specific peptides (SVSPs). They had essentially the same amino acid sequences but differed from one another in the truncation of several residues at the N-terminus and of one residue at the C-terminus, and in the modification of glutamine to pyroglutamate at the N-terminus. The longest peptides, SVSP-4, consisted of 60 amino acid residues with two intramolecular disulfide bridges. There is no significant homology with any other vertebrate or invertebrate peptides.
Amino Acid Sequence↗
Modeling of penicillin G hydrolysis to 6-APA in an immobilized enzyme-ion exchange system.
A composite system of an immobilized enzyme reactor combined with an ion exchange column was employed for hydrolysis of penicillin G to 6-APA with continuous removal of PAA. In this study, a mathematical model of penicillin G hydrolysis to 6-APA in the composite system was developed based on the compartment model, the profile of concentration in the ion exchange column, and the enzyme kinetics in the immobilized enzyme reactor. After checking the simulation values and experimental data, the effects of the resin volume, the flow rate, and the switching time on the time required to reach the desired conversion rate was also examined by computer simulation.
Bacillus megaterium↗
Tryptophan-19 of beta-lactoglobulin, the only residue completely conserved in the lipocalin superfamily, is not essential for binding retinol, but relevant to stabilizing bound retinol and maintaining its structure.
Residue 19 of tryptophan in bovine beta-lactoglobulin (beta-LG) is the only invariant residue throughout the lipocalin superfamily having two characteristic features: binding ability for small hydrophobic molecules and the unique beta-barrel three-dimensional structure. In this study, we investigated whether this strictly conserved Trp-19 of beta-LG would be indispensable for its structure and function such as maintaining the molecular structure and biological activity of beta-LG. Spectroscopic and enzymatic oxidation experiments on retinol bound to W19Y, in which Tyr was substituted for Trp-19, showed that Trp-19 was not critical for this binding, but was important for stably maintaining the environment surrounding retinol and the bound retinol. An using four anti-beta-LG monoclonal antibodies as probes, revealed a structural change in region 20-29, but not in the reverse region of Trp-19. A guanidine hydrochloride-induced unfolding study showed that the conformational stability of W19Y was greatly reduced by 6.9 kcal/mol compared to that of wild-type beta-LG. These facts indicated that Trp-19 is one of the important residues in correctly maintaining the local structure of beta-LG and stably retaining its overall structure, thereby conserving the bound retinol molecule.
Base Sequence↗
Morphological characterization of three phenotypes of the isopod Armadillidium vulgare.
The morphological characteristics and ommochrome quantity in the integument of red, white, and wild type (black-grey) Armadillidium vulgare were studied. The red phenotype was found to possess two kinds of immature ommochrome pigment granules within its pigment cells, in addition to mature pigment granules. The immature granules seemed to contain uniformly distributed fibrilles, or to have an electron-dense central region surrounded by an electron-lucent outer edge. Since these immature pigment granules were typically observed to be distributed along with the mature ones, and were also more easily extractable than the wild type's, it is hypothesized that ommochrome granule maturation in the red phenotype may occur slowly due to a defect in the pigment granule internal process which combines pigments with matrix proteins. Regarding the white phenotype, although its pigment cells were undeveloped, several large-sized vesicles containing a small amount of electron-dense material appeared in the pigment cell cytoplasm. The wild and red type males of A. vulgare were found to have an ommochrome content twice as large as that of the corresponding females, with no ommochrome pigment being detected in the white phenotype. The genetic relationship between the white and red phenotypes was discussed using as a basis the observed pigment granule structure.
Animals↗
[Color vision among workers exposed to styrene].
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Adolescent↗
Unfolding/refolding studies on bovine beta-lactoglobulin with monoclonal antibodies as probes. Does a renatured protein completely refold?
We investigated whether any local moieties within a protein molecule could completely refold from the denatured state to regain the native conformation. Bovine beta-lactoglobulin (beta-LG) was denatured in the presence of guanidine hydrochloride (GdnHCl) as the denaturant. Renaturation of the denatured beta-LG was attempted by dialyzing to remove GdnHCl. The renatured molecules regained the same retinol binding activity as that of native beta-LG, and physicochemical studies also indicated that refolding of the denatured beta-LG had been almost completely successful. Local structural differences between the native and renatured beta-LG molecules were evaluated by using our panel of four anti-beta-LG monoclonal antibodies (anti-beta-LG mAbs). The structures of the epitope regions in native beta-LG recognized by two of these mAbs were the same as those in renatured beta-LG. However, it is notable that the binding properties of the other two mAbs to native beta-LG indicated a wide structural difference in the epitope regions between the native and renatured beta-LG. These regions unable to completely refold were the same as those that unfolded preferentially to the alpha-helix region, shown in the previous report (Kaminogawa, S., Shimizu, M., Ametani, A., Hattori, M., Ando, O., Hachimura, S., Nakamura, Y., Totsuka, M., and Yamauchi, K. (1989) Biochim. Biophys. Acta 998, 50-56). Complete refolding was never attained by several renaturation conditions such as quicker or slower removal of the denaturant, nor by additional oxidation treatment after reducing the disulfide bonds. These results suggest that some specific moiety(ies) in a protein molecule cannot return to the native conformation from a denatured state, even if the other moieties refold completely, and that such a conformational difference between renatured and native forms has no affect on the biological function of ligand binding.
Amino Acid Sequence↗
Neurobehavioral effects of chronic occupational exposure to organic solvents among Japanese industrial painters.
To investigate the relationship between chronic exposure to organic solvents and changes in central nervous system function, industrial painters were compared with an age- and education-matched referent group of nonexposed workers. Eighty-one male painters completed a symptom questionnaire. Twenty painters underwent both questionnaire and neuropsychological examinations. From the results of pairwise comparisons of the symptoms, dry and scaly skin, being easily depressed without reason, coldness of hands and legs, being easily irritated without reason, loss of appetite, dizziness, and unsteadiness occurred statistically significantly more often among the exposed subjects than among the referents. Performances on the Digit symbol test and vocabulary test scores (synonyms) in exposed subjects were significantly lower than those of controls. In multiple regression models, controlling for age, education, and alcohol intake, a significant relation was found between the duration of the solvent exposure and poor performance in both the Block design and Digit span tests. The relation between toluene exposure and poor performance in both the Santa Ana coordination test and the Benton visual retention test was also significant. The results suggest that a symptom inquiry and some behavioral tests are helpful for detecting the possible effects of exposure to low levels of organic solvents. However, no consistent pattern was observed in regard to the effects of organic solvent exposure on neurobehavioral function, which is coincident with the type I toxic central nervous system disorder as classified by the World Health Organization.
Adult↗
Acute neurobehavioral effects of co-inhalation of toluene and n-hexane on schedule-controlled behavior in rats.
Whether coexposure to toluene and n-hexane had any combined effects on the shock avoidance performance in rats was studied. Eighteen Wistar male rats with an avoidance rate of over 80% were selected and divided to three groups based on performance and body weight: (1) toluene, (2) n-hexane, and (3) toluene + n-hexane. Each group was exposed alternately first to air and then to a particular organic solvent for 4 hr at various concentrations (50, 100, 200, 400, or 800 ppm, in ascending order). The effects of each organic solvent were evaluated by comparing the performance of rats during and after exposure with their own performance under the sham exposure to air by three-way ANOVA. The main results were that (1) 200, 400, or 800 ppm toluene exposures increased lever press rates, (2) 50 ppm n-hexane exposure decreased lever press and avoidance rates in a transitory manner and 800 ppm n-hexane exposure increased the lever press rate, (3) the 50 ppm mixture (25 ppm toluene + 25 ppm n-hexane) decreased lever press and avoidance rates persistently during and after the 4-hr exposure and the 800 ppm mixture (400 ppm toluene + 400 ppm n-hexane) decreased lever press and avoidance rates unpredictably when compared to the results of 400 or 800 ppm of toluene or n-hexane alone. In conclusion, n-hexane showed narcotic effects at 800 ppm and modified the acute neurobehavioral effects of toluene in rats at 400 ppm toward unpredictable results.
Administration, Inhalation↗