Biomedical subjects

A Tanaka

Publications and source records attributed to A Tanaka.

At least 253 records · Page 14Linked to original sources

PubMed2000

Allergen-induced mRNA expression of IL-5, but not of IL-4 and IFN-gamma, in peripheral blood mononuclear cells is a key feature of clinical manifestation of seasonal allergic rhinitis.

OBJECTIVES: To investigate the allergen-induced messenger RNA (mRNA) expression of interleukin (IL) 4, IL-5 and interferon gamma (IFN-gamma) in peripheral blood mononuclear cells from individuals sensitized by Japanese cedar (Cryptomeria japonica) pollens, and to elucidate the clinical role of IL-4, IL-5, and IFN-gamma in the allergen sensitization and clinical manifestation of allergic disorders. DESIGN: This study included 30 patients sensitized to the pollen and 14 nonatopic healthy volunteers. Peripheral blood mononuclear cells (1.0 x 10(6) cells/mL) of each individual were cultured at 37 degrees C for 24 hours in the presence of 10 microg/mL of Cry j 1, a major allergen of the pollens. Total cellular RNA was extracted from the peripheral blood mononuclear cells, and IL-4, IL-5, and IFN-gamma mRNA expression was determined with a reverse transcriptase polymerase chain reaction. RESULTS: From the results of a survey of symptom diary cards and interviews regarding nasal symptoms during the pollen season in 1998, we found that 20 patients (symptomatic group), but not 10 patients (asymptomatic group), had typical symptoms of seasonal allergic rhinitis. Interleukin 4 mRNA was not expressed in the nonatopic subjects but was expressed in 9 asymptomatic patients and in 17 symptomatic patients. Interleukin 5 mRNA was exclusively expressed in the symptomatic patients. Interferon gamma mRNA expression did not differ significantly among the nonatopic subjects, asymptomatic patients, and symptomatic patients. CONCLUSIONS: This study has clearly highlighted an interesting and new concept that IL-4 is implicated in allergen sensitization but not in clinical manifestation, and that IL-5 may not be a feature of atopy in itself but seems to be a hallmark of clinical manifestation of ongoing atopic diseases.

Adolescent↗

PubMed2000

Establishment of a simple system to analyse the molecular interaction in the agglutination of Saccharomyces cerevisiae.

Saccharomyces cerevisiae a-agglutinin, which is involved in mating and covalently anchoring to the cell wall, consists of two components, Aga1p and Aga2p, whose syntheses are individually regulated. To facilitate the analysis of the protein-protein interaction on agglutination between a- and alpha-agglutinins, the construction of a yeast strain (MATa) with the functional protein prepared by genetic fusion of Aga1p- and Aga2p-encoding genes and by the expression system using the UPR-ICL promoter derived from the n-alkane-assimilating yeast, Candida tropicalis, which is functional under the condition of lower glucose concentration was tried and the agglutination ability of the constructed strain was evaluated with a yeast strain (MATa) which expressed AGalpha1 encoding alpha-agglutinin under the control of the same promoter. The genes were integrated into the yeast chromosomes. Cell agglutination between both (MATa) strains was observed microscopically when these two strains were mix-cultured to a glucose-decreased concentration. The agglutination was further confirmed by the sedimentation test and by the quantification using a filter. These results proved that the constructed Aga1p-Aga2p fusion protein was enoughly functional for the interaction with the Agalpha1 protein, and that this phenomenon occurred dependent on glucose concentration, but independent of the peptide pheromones secreted by the cells of the opposite mating types. Using this system, the role of two disulphide linkages between Aga1p and Aga2p on the binding activity between Aga2p and Aga1p was first evaluated. Under the treatment by the SH-compound (dithiothreitol), in which Agalpha2p is easily released into the medium from the intact cell surface, the Aga1p and Aga2p fusion protein was a good tool to make clear the role of the disulphide linkages. As a result, the linkages had a significant effect on not only the assembly but also the binding activity. The novel and simple system described here may further facilitate the study of molecular interaction in agglutination.

Alkanes↗

PubMed2000

Evidence for a locally driven mucosal response and the presence of mitochondrial antigens in saliva in primary biliary cirrhosis.

Primary biliary cirrhosis (PBC) is often considered to be a dry gland disease caused by frequent involvement of salivary and lacrimal glands. Although high titers of antimitochondrial autoantibodies (AMA) have long been recognized in PBC, little is known about the presence of mitochondrial autoantigens in mucosal compartments such as saliva. We investigated saliva and sera in PBC patients and controls for the presence of AMA and mitochondrial antigens. In PBC saliva, AMA were detected in 45 of 49 (92%), with specificity directed against pyruvate dehydrogenase complex (PDC-E2) alone in 22 of 49 (45%), against PDC-E2 and branched-chain 2-oxo-acid dehydrogenase complex E2 (BCOADC-E2) in 4 of 49 (8%), to PDC-E2 and 2-oxoglutarate dehydrogenase complex E2 (OGDC-E2) in 9 of 49 (18%), and to the 3 antigens together in 10 of 49 (20%). Isotyping of the saliva AMA showed that 80% of the patients had immunoglobulin A (IgA) against PDC-E2, 18% had IgM-specific PDC-E2, and 35% had IgG specific PDC-E2. Similar to serum and bile anti-PDC-E2 IgA antibodies, the saliva autoantibodies localized their reactivity to the inner lipoyl domain of PDC-E2. Furthermore, saliva from patients with PBC but not controls inhibited pyruvate dehydrogenase enzyme activity in vitro. In addition, and of particular interest, we detected a molecule with a molecular weight corresponding to PDC-E2 (74 kd) in PBC but not control saliva. These findings make several important points: first, there appears to be localized mucosal immunity in the secretory system of PBC; second, AMA are readily detected in PBC saliva; and third, PDC-E2 may be present in the saliva of PBC.

Antibody Specificity↗

PubMed2000

Fetal microchimerisms in the mother: immunologic implications.

The previously held concept that the fetus is completely separated from the mother, especially by trophoblasts that line the outer layer of the placenta, has recently been questioned. It has recently been shown that fetal cells are detectable not only in the peripheral blood, but also in maternal skin and liver. Although the migration of fetal cells into the maternal circulation has been given a great deal of attention because of its implication in the prenatal diagnosis of genetic diseases, the potential role of such placental transfer of fetal cells in the pathogenesis of autoimmune diseases has only recently been considered. In patients with scleroderma, fetal cell-derived DNA was detected more frequently in the peripheral blood of patients than controls. This finding of a limited number of fetal cells in maternal tissues leading to microchimerism has been proposed to have a role in the induction of scleroderma. Although evidence for microchimerism has also been confirmed in a high frequency of liver tissues from patients with primary biliary cirrhosis (PBC), a similar high frequency was noted in control patients, which suggests that microchimerism by itself is unlikely to fully account for the pathogenesis of PBC. The finding of such a high frequency of fetal microchimerism in the liver suggests that this is a very common event, raising the possibility that the migration of fetal cells may be important in the induction and subsequent maintenance of tolerance against the fetus during pregnancy. In addition, it is clearly possible that such microchimerism could contribute to the pathogenesis of select autoimmune diseases.

Autoimmunity↗

PubMed2000

Search for PET probes for imaging the globus pallidus studied with rat brain ex vivo autoradiography.

We have evaluated the feasibility of using four positron emission tomography (PET) tracers for imaging the globus pallidus by ex vivo autoradiography in rats. The tracers investigated were [11C]KF18446, [11C]SCH 23390 and [11C]raclopride for mapping adenosine A2A, dopamine D1 and dopamine D2 receptors, respectively, and [18F]FDG. The highest uptake by the globus pallidus was found for [11C]SCH 23390, followed by [18F]FDG, [11C]KF18446 and [11C]raclopride. The receptor-specific uptake by the globus pallidus was observed in [11C]KF18446 and [11C]SCH 23390, but not in [11C]raclopride. Uptake ratios of globus pallidus to the striatum for [18F]FDG and [11C]KF18446 were approximately 0.6, which was twice as large as that for [11C]SCH 23390. In a rat model of degeneration of striatopallidal gamma-aminobutyric acid-ergic-enkephalin neurons induced by intrastriatal injection of quinolinic acid, the uptake of [11C]KF18446 by the striatum and globus pallidus was remarkably reduced. To prove the visualization of the globus pallidus by PET with [18F]FDG and [11C]KF18446, PET-MRI registration technique and advances in PET technologies providing high-resolution PET scanner will be required. The metabolic activity of the globus pallidus could then be measured by PET with [18F]FDG, and [11C]KF18446 may be a candidate tracer for imaging the pallidal terminals projecting from the striatum.

Animals↗

PubMed2000

Further characterization of a CNS adenosine A2a receptor ligand [11C]KF18446 with in vitro autoradiography and in vivo tissue uptake.

PET assessment of the adenosine A2a receptors localized in the striatum offers us a potential new diagnostic tool for neurological disorders. In the present study, we carried out in vitro receptor autoradiography of a newly developed PET ligand [11C]KF18446 ([7-methyl-11C]-(E)-8-(3,4,5-trimethoxystyryl)-1,3,7-trimethylxanthin e) with rat brain sections. [11C]KF18446 showed a high striatum/cortex binding ratio (5.0) and low nonspecific binding (<10%), suggesting that [11C]KF18446 has characteristics comparable or slightly superior to [3H]CGS 21680 or [3H]SCH 58261, which are currently available representative A2a receptor ligands. Scatchard analysis indicated a Kd of 9.8 nM and a Bmax of 170 fmol/mm3 tissue in the striatum and a Kd of 16.4 nM and a Bmax of 33 fmol/mm3 tissue in the cortex. Seven xanthine-type and four nonxanthine-type adenosine receptor ligands with an affinity for the adenosine A2a receptors significantly reduced the in vitro binding of [11C]KF18446 to the brain section. The blocking effects were much stronger in the striatum than in the cortex, but did not necessarily parallel their affinity. On the other hand, four xanthine-type ligands and one nonxanthine-type ligand (SCH 58261) of the 11 ligands studied reduced the in vivo uptake of [11C]KF18446 in mice, but other ligands, including A1-selective and nonselective ligands and three nonxanthine-type A2a-selective antagonists did not. We conclude that [11C]KF18446 is a promising adenosine A2a receptor ligand for PET study.

Animals↗

PubMed2000

Cerebral protection selection in aortic arch surgery for patients with preoperative complications of cerebrovascular disease.

OBJECTIVE: Retrograde perfusion is gaining acceptance as a means of cerebral protection, but it remains unclear how long the brain is protected and whether it is effective in patients with preoperative cerebrovascular disease. METHODS: From January 1989 to August 1999, 205 patients--118 male and 87 female patients who ranged 12 to 86 years old, mean: 65.5 years old--underwent surgery at our hospital for aortic arch aneurysm using cerebral protection. We focused on mortality, stroke incidence and perioperative risk factor between 2 groups--selective cerebral and retrograde cerebral perfusion--also studying patients with preoperative cerebrovascular disease that influenced postoperative stroke. RESULTS: The hospital mortality was 11.7% (selective cerebral perfusion group: 12%, retrograde group: 10.9%). Stroke occurred in 11 patients (5.3%), 4.7% in the selective cerebral perfusion group and 7.3% in the retrograde group. Preoperative cerebrovascular disease does not appear to be a risk factor for postoperative brain damage in aortic arch surgery. Regarding total replacement of the aortic arch, the incidence of postoperative brain damage in the retrograde group with preoperative cerebrovascular disease was higher than that in another group (p = 0.072). Cardiopulmonary bypass time and selective cerebral perfusion time in the patients with postoperative stroke were significantly longer than that in non-stroke group. CONCLUSIONS: Preoperative cerebrovascular disease did not appear to be a risk factor in postoperative neurological deficit in the selective cerebral perfusion group. Prolonged selective cerebral perfusion time and cardiopulmonary bypass time may, however, lead to brain edema and cause neurological deficit.

Adolescent↗

PubMed2000

A mutated hygromycin resistance gene is functional in the n-alkane-assimilating yeast Candida tropicalis.

Development of a transformation system in the n-alkane-assimilating diploid yeast Candida tropicalis requires an antibiotic resistance gene in order to establish a selectable marker. The resistance gene for hygromycin B has often been used as a selectable marker in yeast transformation. However, C. tropicalis harboring the hygromycin resistance gene (HYG) was as sensitive to hygromycin B as the wild-type strain. Nine CTG codons were found in the ORF of the HYG gene. This codon has been reported to be translated as serine rather than leucine in Candida species. Analysis of the tRNA gene in C. tropicalis with the anticodon CAG [tRNA(CAG) gene], which is complementary to the codon CTG, showed that the sequence was highly similar to that of the C. maltosa tRNA(CAG) gene. In C. maltosa, the codon CTG is read as serine and not leucine. These results suggested that the HYG gene was not functional due to the nonuniversal usage of the CTG codon. Each of the nine CTG codons in the ORF of the HYG gene was changed to a CTC codon, which is read as leucine, by site-directed mutagenesis. When a plasmid containing the mutated HYG gene (HYG#) was constructed and introduced into C. tropicalis, hygromycin-resistant transformants were successfully obtained. This mutated hygromycin resistance gene may be useful for direct selection of C. tropicalis transformants.

Alkanes↗

PubMed2000

Construction of an engineered yeast with glucose-inducible emission of green fluorescence from the cell surface.

An engineered yeast with emission of fluorescence from the cell surface was constructed. Cell surface engineering was applied to display a visible reporter molecule, green fluorescent protein (GFP). A glucose-inducible promoter GAPDH as a model promoter was selected to control the expression of the reporter gene in response to environmental changes. The GFP gene was fused with the gene encoding the C-terminal half of alpha-agglutinin of Saccharomyces cerevisiae having a glycosylphosphatidylinositol anchor attachment signal sequence. A secretion signal sequence of the fungal glucoamylase precursor protein was connected to the N-terminal of GFP. This designed gene was integrated into the TRP1 locus of the chromosome of S. cerevisiae with homologous recombination. Fluorescence microscopy demonstrated that the transformant cells emitted green fluorescence derived from functionally expressed GFP involved in the fusion molecule. The surface display of GFP was further verified by immunofluorescence labeling with a polyclonal antibody (raised in rabbits) against GFP as the first antibody and Rhodamine Red-X-conjugated goat anti-rabbit IgG as the second antibody which cannot penetrate into the cell membrane. The display of GFP on the cell surface was confirmed using a confocal laser scanning microscope and by measuring fluorescence in each cell fraction obtained after the subcellular fractionation. As GFP was proved to be displayed as an active form on the cell surface, selection of promoters will endow yeast cells with abilities to respond to changes in environmental conditions, including nutrient concentrations in the media, through the emission of fluorescence.

Base Sequence↗

PubMed2000

An Aspergillus oryzae CCAAT-binding protein, AoCP, is involved in the high-level expression of the Taka-amylase A gene.

Aspergillus oryzae contains a nuclear protein designated AoCP, which binds specifically to a CCAAT sequence in the promoter region of the A. oryzae Taka-amylase A gene. A gene encoding a homologue of Aspergillus nidulans HAPC, a subunit of the A. nidulans CCAAT binding complex, was isolated from A. oryzae and designated AohapC. AoHAPC comprises 215 amino acids and shows 84% identity to A. nidulans HAPC. Transformation of the A. nidulans hapC deletion strain with the AohapC gene restored the CCAAT binding activity, resulting in both enhancement of taa gene expression and complementation for the poor growth phenotype of this strain. Furthermore, introduction of the AohapC gene also restored the expression of the A. nidulans eglA gene, encoding an endo-beta-1,4-glucanase, in the deletion strain. These results indicate functional interchangeability of the genes from two species.

Amino Acid Sequence↗

PubMed2000

Scalp cavernous angioma presenting as sinus pericranii: diagnostic value of cerebral angiography and magnetic resonance imaging.

OBJECTS: Sinus pericranii is only a symptom complex, and it can have a variety of etiologies. Therefore, it is important to differentiate these etiologies preoperatively by means of radiological examinations. A 5-year-old boy was admitted with a soft and fluctuant tumor in the right parietal region near the midline. The tumor appeared when the child was in a recumbent position, distending noticeably with the Valsalva maneuver and disappearing completely when the patient was in the sitting position. METHODS: Magnetic resonance imaging showed the lesion with honeycomb-like heterogeneous iso- and low-intensity signals on the T1-weighted image and with heterogeneous high- and isointensity signal on the T2-weighted image. Dynamic study with an injection of gadolinium diethylene-triaminopentaacetic acid demonstrated and nodular peripheral enhancement at early phase and subsequent progressive enhancement towards the center of tumor. The internal carotid angiogram was normal. The external carotid angiogram, however, showed a tumor stain fed by the superficial temporal arteries. The stain was retained until the late phase and drained into the scalp veins and into the superior sagittal sinus. Following direct injection of contrast medium into the tumor there was prolonged retention of the medium in the tumor and leakage into scalp veins and the superior sagittal sinus. The mass under the periosteum was totally removed and proved to be a cavernous angioma. CONCLUSIONS: Scalp cavernous angioma is one of the etiologies of sinus pericranii and may be diagnosed preoperatively by cerebral angiography or magnetic resonance imaging. Serial dynamic magnetic resonance imaging will be particularly helpful for this diagnosis.

Cerebral Angiography↗

PubMed2000

Photosynthetic apparatus organization and function in the wild type and a chlorophyll b-less mutant of Chlamydomonas reinhardtii. Dependence on carbon source.

The assembly, organization and function of the photosynthetic apparatus was investigated in the wild type and a chlorophyll (Chl) b-less mutant of the unicellular green alga Chlamydomonas reinhardtii, generated via DNA insertional mutagenesis. Comparative analyses were undertaken with cells grown photoheterotrophically (acetate), photomixotrophically (acetate and HCO3-) or photoautotrophically (HCO3-). It is shown that lack of Chl b diminished the photosystem-II (PSII) functional Chl antenna size from 320 Chl (a and b) to about 95 Chl a molecules. However, the functional Chl antenna size of PSI remained fairly constant at about 290 Chl molecules, independent of the presence of Chl b. Western blot and kinetic analyses suggested the presence of inner subunits of the Chl a-b light-harvesting complex of PSII (LHCII) and the entire complement of the Chl a-b light-harvesting complex of PSI (LHCI) in the mutant. It is concluded that Chl a can replace Chl b in the inner subunits of the LHCII and in the entire complement of the LHCI. Growth of cells on acetate as the sole carbon source imposes limitations in the photon-use efficiency and capacity of photosynthesis. These are manifested as a lower quantum yield and lower light-saturated rate of photosynthesis, and as lower variable to maximal (Fv/Fmax) chlorophyll fluorescence yield ratios. This adverse effect probably originates because acetate shifts the oxidation-reduction state of the plastoquinone pool, and also because it causes a decrease in the amount and/or activity of Rubisco in the chloroplast. Such limitations are fully alleviated upon inclusion of an inorganic carbon source (e.g. bicarbonate) in the cell growth medium. Further, the work provides evidence to show that transformation of green algae can be used as a tool by which to generate mutants exhibiting a permanently truncated Chl antenna size and a higher (per Chl) photosynthetic productivity of the cells.

Animals↗

PubMed2000

REAL, an LTR retrotransposon from the plant pathogenic fungus Alternaria alternata.

A retrotransposon was isolated and characterized from strain 15A of the Japanese pear pathotype of Alternaria alternata, which causes black spot disease in certain cultivars of Japanese pear by producing a host-specific toxin known as AK-toxin. The element, which we have named REAL (Retrotransposon of Alternaria alternata), is 6046 bp in size and contains direct long terminal repeats (LTRs) of 218 bp. Target-site duplication of 5 bp was found. REAL contains two long overlapping ORFs. The first ORF shows homology to retroviral gag genes. The second ORF has homology to protease, reverse transcriptase, RNase H and integrase domains of the retroelement pol genes, in that order. Phylogenetic comparison of reverse transcriptase domains from retrotransposons placed REAL in the Ty3/gypsy group of LTR retrotransposons, most closely related to grasshopper from Magnaporthe grisea. Northern analysis detected REAL transcripts of about 2.0 and 6.0 kb. The 6.0-kb species corresponds to a full-length transcript of the element. The element was found by Southern analysis in 12 out of 13 strains of the Japanese pear pathotype, and the banding patterns, copy numbers and signal intensities in these strains were variable. REAL-related elements were also found in some, but not all, of the other strains tested, including nonpathogenic A. alternata and other pathotypes, which cause diseases on different plant species by producing distinct hostspecific toxins. These results suggest that the distribution of REAL in A. alternata is not pathotype specific.

Alternaria↗

PubMed2000

Severe complications of mediastinal pancreatic pseudocyst: report of esophagobronchial fistula and hemothorax.

We report two patients with alcoholic pancreatic pseudocyst which communicated to the mediastinal space through the aortic hiatus, in one patient resulting in hypotensive shock due to hemothorax, and in the other, resulting in esophagobronchial fistula via the mediastinal cyst. The first patient was successfully treated by radical resection of the pancreatic body and tail, and the spleen, with an ultrasonic scalpel, although inflammatory changes caused by pancreatitis were so prominent that the splenic vein was occluded. The second patient could not be treated surgically, because the superior mesenteric vein had been occluded by chronic pancreatitis; he died of respiratory failure and sepsis due to aspiration pneumonia, despite receiving medical treatment.

Adult↗

PubMed2000

Extrahepatic large hepatocellular carcinoma with peritoneal dissemination: multimodal treatment, including four surgical operations.

We report a patient with extrahepatically growing large hepatocellular carcinoma (HCC) associated with disseminated intraabdominal tumor and spontaneous tumor bleeding who was treated with four operations, transcatheter arterial embolization, systemic chemotherapy, and hyperthermia. It took 12 months for the multimodal treatment to normalize the alpha-fetoprotein (AFP) level, and remission continued for 6 months. We performed the fourth surgical treatment for a recurrent abdominal tumor involving the small intestine and mesentery, but the patient died 26 months after the first admission. Multimodal treatment, including repeat surgical treatments, for such advanced HCC should be encouraged, to prolong life and to maintain quality of life.

Antineoplastic Combined Chemotherapy Protocols↗

PubMed2000

At least four non-env factors that reside in the LTR, in the 5'-non-coding region, in gag and in part of pol affect neuropathogenicity of PVC-441 murine leukemia virus (MuLV).

PVC-441 murine leukemia virus (MuLV) is neuropathogenic in F344 rats. Recently, an infectious DNA clone was isolated and its nucleotide sequence was determined (J. Virol. 72: 3423-3426. 1998). To identify the viral determinants of neuropathogenicity of the molecularly cloned PVC-441 MuLV, chimeras were constructed between PVC-441 MuLV and F-MuLV clones at appropriate restriction enzyme sites that divide the viral genome approximately in LTR-non-coding, gag-, pol-, and env-gene regions. Results indicated that the LTR-non-coding and the gag-gene regions of PVC-441 MuLV affected independently the neuropathogenicity in combination with the env gene region as evidenced clinically and pathologically. Studies on the distribution of vacuolar degeneration suggested that the pons and cervical spinal cord areas were the primary targets and the large brain was the latest target of PVC-441 MuLV. Further studies with chimeric viruses that were formed in the LTR-non-coding and the gag gene regions revealed that at least four factors affected the neuropathogenicity of PVC-441 MuLV. Two factors were found in the U3, and R-U5-5'-non-coding regions, and at least two factors in the gag gene region that contained the N-terminal part of the pol gene. Among these factors, at least two factors seemed to be 'cis-acting' from each other

5' Untranslated Regions↗

PubMed2000

Genetic immobilization of proteins on the yeast cell surface.

A genetic system has been exploited to immobilize proteins in their active and functional forms on the cell surface of yeast, Saccharomyces cerevisiae. DNAs encoding proteins with a secretion signal peptide were fused with the genes encoding yeast agglutinins, a- and alpha-type proteins involved in mating. The fusion gene was introduced into S. cerevisiae and expressed under the control of several promoters. Appearance of the fused proteins expressed on the cell surface was demonstrated biochemically and by immunofluorescence and immunoelectron microscopy techniques. Alpha-galactosidase from Cyamopsis tetragonoloba seeds, peptide libraries including scFv and variable regions of the T cell receptor from mammalian cells have been successfully immobilized on the yeast cell wall in the active form. Recently, surface-engineered yeasts have been constructed by immobilizing the enzymes and a functional protein, for example, green fluorescent protein (GFP) from Aequorea victoria. The yeasts were termed 'arming yeasts' with biocatalysts or functional proteins. Such arming cells displaying glucoamylase from Rhizopus oryzae and alpha-amylase from Bacillus stearothermophilus, or carboxymethylcellulase and beta-glucosidase from Aspergillus acleatus, could assimilate starch or cellooligosaccharides as the sole carbon source, although S. cerevisiae cannot intrinsically assimilate these substrates. GFP-arming cells can emit green fluorescence from the cell surface in response to the environmental conditions. The approach described in this review will enable us to endow living cells, including yeast cells, with novel additional abilities and to open new dimensions in the field of biotechnology.

Journal Article↗

PubMed2000

Automatic synthesis of L-[beta-11C]amino acids using an immobilized enzyme column.

We have developed a system for the automatic synthesis of L-[beta-11C]amino acids for i.v. injection by means of enzyme-mediated reactions from 11CO2 via 11CH3I and D,L-[beta-11C]alanine as labeled intermediates. This system, which incorporates an ultrafilter cartridge sterilized by electron beam irradiation and a column packed with immobilized enzymes, was effective for eliminating enzymes and endotoxins that may contaminate the product. Using this system, 1.3 +/- 0.5 GBq of 5-hydroxy-L-[beta-11C]tryptophan with a radiochemical purity of 97.1 +/- 0.6% and a specific activity of 39.6 +/- 8 GBq/mumol a pH value of 4 could be obtained in about 32 min (n = 3, at EOS). No endotoxin, enzyme, or bacteria was detected in the product. L-[beta-11C]dihydroxyphenylalanine (L-[beta-11C]DOPA) was also synthesized using this system.

Alanine↗