Development of polymorphic microsatellite markers in a tropical tree species, Melaleuca cajuputi (Myrtaceae).
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DNA Primers↗
Biomedical subjects
R Tanaka
Publications and source records attributed to R Tanaka.
Cloning and functional expression of the gene encoding the key enzyme for chlorophyll b biosynthesis (CAO) from Arabidopsis thaliana.
Chlorophyll (Chl) biosynthesis and degradation are the only biochemical processes on Earth that can be directly observed from satellites or other planets. The bulk of the Chls is found in the light-harvesting antenna complexes of photosynthetic organisms. Surprisingly little is known about the biosynthesis of Chl b, which is the second most abundant Chl pigment after Chl a. We describe here the expression and properties of the chlorophyllide a oxygenase gene (CAO) from Arabidopsis thaliana, which is apparently the key enzyme in Chl b biosynthesis. The recombinant enzyme produced in Escherichia coli catalyses an unusual two-step oxygenase reaction that is the 'missing link' in the chlorophyll cycle of higher plants.
Arabidopsis↗
Vascular structure of arteriovenous malformations.
Cerebral arteriovenous malformations (AVMs) are classified angiographically into two types: the arteriovenous fistula (AVF) type and the plexiform type. However, the differences in vascular structure of these two types have not been clarified. The purpose of the present study is to elucidate the vascular structure of plexiform AVMs and to discuss the clinical significance of this classification of AVMs. Specimens of AVMs resected in 8 cases and identified by cerebral angiography as plexiform AVMs were examined. Immediately after their removal, microdissection of the terminal arterial feeder, the nidus, and the venous drainer was performed under a microscope. A histological examination of each element was then conducted. Microdissection of a portion of the vascular mass that formed the nidus made it possible to separate individual vessels of the mass from each other. Many of these individual vessels connected with the feeder on one side, while the other side anastomosed with the drainer, thus exhibiting the morphology of an AVF. From our examination of the AVMs in the present study, we inferred that the plexiform type is fundamentally a conglomeration of AVFs. It is therefore suggested that the vascular structure of this type of AVM is not fundamentally different from that of the AVF type.
Adult↗
Dural arteriovenous malformations associated with cerebral aneurysms.
Only limited information is available concerning dural arteriovenous malformations (AVMs) found in association with cerebral aneurysms. The present report focuses on six such cases, concentrating attention on clinical characteristics and significance. Of a total of 46 dural AVMs encountered over a given period, six (13%) were linked with cerebral aneurysms. Particularly strong associations were noted for dural AVMs in the anterior cranial fossa (three of four cases) and convexity (all of three cases). With dural AVMs in the anterior cranial fossa, subarachnoid haemorrhage is common, so that where this is encountered the possibility of a complicating aneurysm should be considered in order to select an appropriate treatment.
Adolescent↗
Three new lanostane-type triterpene lactones from the stem bark of Abies mariesii.
Three new delta 8-lanostane-type triterpene lactones (1), (2) and (3) were isolated from the stem bark of Abies mariesii M. and unambiguously characterized on the basis of spectroscopic and chemical evidence. Chemical modification of compound 1 showed potent inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA).
Antineoplastic Agents, Phytogenic↗
Bioactive triterpenoids from the stem bark of Picea glehni.
Two new triterpenoids, 3 alpha-methoxyserrat-14-en-21 beta-yl formate (1), and 24-methylcycloartenone (2), were isolated from the stem bark of Picea glehni (Fr. Schm.) Masters together with three known triterpenoids, 3 alpha-methoxyserrat-14-en-21 beta-ol, 3 beta-methoxyserrat-14-en-21 beta-ol, and piceanonol A. Compounds 1, 2, and a synthetic sample, 3 alpha-methoxyserrat-13-en-21 beta-yl formate showed potent inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA).
Antigens, Viral↗
Somatic mutations of the multiple endocrine neoplasia type 1 (MEN1) gene in patients with sporadic, nonfamilial primary hyperparathyroidism.
BACKGROUND: Multiple endocrine neoplasia type 1 (MEN 1) is a syndrome with tumors of many endocrine tissues. Germline MEN1 gene mutations were found in most patients with familial or sporadic MEN 1. Recently, somatic MEN1 gene mutations were also detected in sporadic non-MEN 1 endocrine tumors. METHODS: We used direct sequence analysis to investigate MEN1 gene mutations in 30 parathyroid tumors obtained from 30 patients with sporadic, nonfamilial primary hyperparathyroidism. RESULTS: Four patients had somatic mutations of the MEN1 gene, comprising 1 small insertion (1091insAGC), one missense mutation (G42S), and 2 non-sense mutations (E388X, R460X). Identical missense and non-sense mutations were found in patients with familial and non-familial MEN 1. There were no differences between clinical features of patients with and without MEN1 gene mutations. CONCLUSIONS: The incidence of somatic MEN1 gene mutations (13.3%) in Japanese patients with sporadic, nonfamilial primary hyperparathyroidism is almost equal to those of such patients in the United States and Sweden. Occasionally, the MEN1 gene mutation sites in sporadic parathyroid tumors are identical to those reported in tumors from patients with familial or sporadic MEN 1.
DNA↗
The effect of encephalo-myo-synangiosis on abnormal collateral vessels in childhood moyamoya disease.
Child patients with Moyamoya disease initially present with ischemic symptoms. However, the long-term risk of intracranial hemorrhage for childhood Moyamoya disease is unknown. Hemodynamic overload to the fragile collateral vessels has been considered to cause hemorrhage. We reviewed angiograms to evaluate the effect of encephalo-myo-synangiosis (EMS) on abnormally dilated collateral vessels in 13 child patients with Moyamoya disease. EMS was performed on 24 sides in 13 patients ranging from 5 to 14 years of age. Post-operative angiography (6-88 months after surgery) revealed good revascularizations through EMS (larger than one-third of the middle cerebral artery (MCA) distribution) in 18 sides (75%) and smaller revascularizations in 6 sides (25%). In cases with a good revascularization through EMS, reduction of the abnormal collateral vessels was observed not only in the basal Moyamoya vessels (94% of sides) but also in the medullary arteries derived from the choroidal arteries (62% of sides), which are considered to cause intraventricular hemorrhages in adult patients. It is suggested that EMS may reduce the hemodynamic load on dilated collateral vessels and, subsequently, the long-term risk of intracranial hemorrhage in childhood Moyamoya disease.
Adolescent↗
Lysosomal cysteine protease, cathepsin B, is targeted to lysosomes by the mannose 6-phosphate-independent pathway in rat hepatocytes: site-specific phosphorylation in oligosaccharides of the proregion.
Cathepsin B, a lysosomal cysteine protease, is synthesized as a glycoprotein with two N-linked oligosaccharide chains, one of which is in the propeptide region while the other is in the mature region. When cultured rat hepatocytes were labeled with [(32)P]phosphate, (32)P-labeled cathepsin B was immunoprecipitated only in the proform from cell lysates and medium. Either Endo H or alkaline phosphatase treatment of (32)P-labeled procathepsin B demonstrated the acquisition of a mannose 6-phosphate (Man 6-P) residue on high mannose type oligosaccharides. To identify the site of phosphorylation, immunoisolated (35)S- or (32)P-labeled procathepsin B was incubated with purified lysosomal cathepsin D, since cathepsin D cleaves 48 amino acid residues from the N-terminus of procathepsin B, in which one N-linked oligosaccharide chain was also included [Kawabata, T. et al. (1993) J. Biochem. 113, 389-394]. Treatment of intracellular (35)S-labeled procathepsin B with a molecular mass of 39-kDa with cathepsin D resulted in the production of the 31-kDa intermediate form, but the (32)P-label incorporated into procathepsin B disappeared after treatment with cathepsin D. These results indicate that the phosphorylation of procathepsin B is restricted to an oligosaccharide chain present in the propeptide region. Interestingly, cathepsin B sorting to lysosomes was not inhibited by NH(4)Cl treatment and about 90% of the intracellular procathepsin B initially phosphorylated was secreted into the medium without being dephosphorylated intracellularly, and did not bind significantly to cation-independent-Man 6-P receptor, suggesting the failure of Man 6-P-dependent transport of procathepsin B to lysosomes. Additionally, about 50% of the newly synthesized (35)S-labeled cathepsin B was retained in the cells in mature forms consisting of a 29-kDa single chain form and a 24-kDa two chain form, while part of the procathepsin B was associated with membranes in a Man 6-P-independent manner. Taken together, these results show that in rat hepatocytes, cathepsin B is targeted to lysosomes by an alternative mechanism(s) other than the Man 6-P-dependent pathway.
Ammonium Chloride↗
False-positive imaging of Tc-99m pertechnetate Meckel's scintigraphy in a patient with hemorrhagic enteritis.
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Child↗
Assignment of Alteromonas elyakovii KMM 162T and five strains isolated from spot-wounded fronds of Laminaria japonica to Pseudoalteromonas elyakovii comb. nov. and the extended description of the species.
A marine bacterium, Alteromonas elyakovii KMM 162T, which was described recently, and five strains isolated from spot-wounded fronds of Laminaria japonica have been subjected to phylogenetic analysis, and geno- and phenotypic characterization. The phenotypic features of Pseudoalteromonas elyakovii strains were closely related to that of Pseudoalteromonas espejiana IAM 12640T, but utilization of three carbon compounds (D-mannose, L-tyrosine and trehalose) distinguished both species. The G+C content of Pseudoalteromonas elyakovii was between 38.5 and 38.9 mol%. Pseudoalteromonas elyakovii KMM 162T and the five Laminaria isolates constitute a single species different from any other Alteromonas and Pseudoalteromonas species as revealed by DNA-DNA hybridization data, especially Pseudoalteromonas distincta KMM 638T (52.4%), Pseudoalteromonas citrea KMM 216 (49.5%), Pseudoalteromonas carrageenovora NCIMB 302T (46.9%) and Pseudoalteromonas espejiana IAM 12640T (29.9%). All the data indicated that Alteromonas elyakovii KMM 162T should be reclassified as Pseudoalteromonas elyakovii and five strains isolated from Laminaria japonica have to be included in the species. Pseudoalteromonas elyakovii comb. nov. (type strain, KMM 162T = ATCC 700519T) is proposed and a set of phenotypic features which differentiate the Pseudoalteromonas species is described.
Alteromonas↗
Role of magnesium chelatase activity in the early steps of the tetrapyrrole biosynthetic pathway.
Magnesium-protoporphyrin IX chelatase (Mg-chelatase) is located at the branchpoint of tetrapyrrole biosynthesis, at which point protoporphyrin IX is distributed for the synthesis of chlorophyll and heme. We investigated the regulatory contribution of Mg-chelatase to the flow of metabolites. In plants, the enzyme complex consists of three subunits, designated CHL D, CHL I, and CHL H. Transgenic tobacco (Nicotiana tabacum) plants expressing antisense RNA for the Mg-chelatase subunit CHL H were analyzed to elucidate further the role of Mg-chelatase in the distribution of protoporphyrin IX into the branched tetrapyrrolic pathway. The transgenic plants displayed a reduced growth rate and chlorophyll deficiency. Both phenotypical properties were correlated with lower Mg-chelatase activity. Unexpectedly, less protoporphyrin IX and heme accumulated, and a decrease in 5-aminolevulinate (ALA)-synthesizing capacity and ALA dehydratase activity paralleled the progressive reduction in Mg-chelatase activity in the transformants compared with control plants. The reduced activities of the early enzymatic steps corresponded with lower levels of transcripts encoding glutamyl-tRNA reductase and ALA-dehydratase. The decreased expression and activities of early enzymes in the pathway could be explained by a feedback-controlled mechanism in response to lower Mg-chelatase activity. We discuss intercompartmental signaling that synchronizes the activities of the first steps in tetrapyrrolic metabolism with the late steps for the synthesis of end products.
Lyases↗
Surgical management of genital prolapse: is chain cystourethrography useful for evaluating anatomical corrections and urinary symptoms after surgery?
OBJECTIVE: To evaluate the relationship between chain cystourethrography and surgery for genital prolapse and urinary symptoms. METHODS: A retrospective study of women with genital prolapse was conducted from January 1991 through December 1997. To assess the preoperative and postoperative anatomical situations objectively, chain cystourethrography was performed. RESULTS: Fifty-seven patients underwent surgical repairs for genital prolapse. All of them recovered from the genital prolapse both subjectively and objectively after surgery. Chain cystourethrography is useful for evaluating postoperative anatomical corrections. However, 6 patients (12%) suffered from a recurrent feeling of genital prolapse, and 14 patients (28%) had urinary symptoms postoperatively. CONCLUSIONS: Our treatment showed good anatomical corrections based on chain cystourethrography, and there were no prognostic differences among the surgical procedures. However, anatomical correction does not always mean improvement of urinary symptoms. Further studies are needed to clarify what factors contribute to the prognosis in such cases.
Adult↗
Neuropsychological outcome following pallidotomy and thalamotomy for Parkinson's disease.
Cognitive and emotional outcomes were assessed after unilateral posteroventral pallidotomy (PVP) and ventral intermediate nucleus thalamotomy (Vim-Th) in patients suffering from idiopathic Parkinson's disease (PD). PVP was performed on 12 PD patients (8 men and 4 women, mean age 56.4 years, 6 left lesions and 6 right lesions) and Vim-Th was performed on 13 PD patients (5 men and 8 women, mean age 63.2 years, 6 left lesions and 7 right lesions). In both the PVP group and the Vim-Th group, the Hoehn and Yahr staging scores (p < 0.01) and the Unified Parkinson's Disease Rating Scale activities of daily living and motor scores (p < 0.001) improved significantly. No significant changes in cognitive function were observed 4 weeks after unilateral PVP or Vim-Th. PVP produced a significant decline in Hasegawa's Dementia Scale-Revised scores in immediate postoperative assessments (p < 0.05), which resolved 4 weeks after surgery. While the Minnesota Multiphasic Personality Inventory depression and social introversion scores improved significantly in the Vim-Th group (p < 0.01 and p < 0.05, respectively), the hypochondriasis and hypomania scores improved significantly in the PVP group (p < 0.01 and p < 0.05, respectively). We conclude that PVP and Vim-Th influence postoperative emotional status rather than postoperative cognitive status.
Adult↗
Primary malignant lymphoma of the central nervous system presenting with ascites and pleural effusion.
A 70-year-old woman was admitted to our hospital owing to ascites and pleural effusion. Though malignant cells (B-cell type lymphoma) were detected in both the ascites and pleural effusion, neither lymph node swelling nor a tumor was detected upon chest, abdominal and pelvic computed tomography (CT). After weekly THP-COP therapy for 8 weeks, the ascites and pleural effusion completely disappeared. Two years after the first admission, she was re-admitted because of a disturbance of consciousness, and a brain tumor was detected on CT scan. The immunohistological and genetic data for the brain tumor were identical to those of the malignant cells in the pleural effusion and ascites detected 2 years previously. Whereas the symptoms at onset of a primary lymphoma of the central nervous system (CNS) are usually neurological ones, in this rare case of primary CNS lymphoma, the symptoms at onset were the ascites and pleural effusion without neurological symptoms.
Aged↗
Changes in extracellular matrix and tissue viscoelasticity in bleomycin-induced lung fibrosis. Temporal aspects.
Bleomycin-induced lung fibrosis results in changes in tissue mechanical properties due to alterations in the extracellular matrix (ECM). How oscillatory mechanics and changes in the matrix evolve over time has not been addressed. Sprague-Dawley rats were instilled with bleomycin sulfate (BM) (1.5 U) intratracheally; control animals (C) received saline. At 7, 14, and 28 d after BM, parenchymal strips (7 x 2 x 2 mm) were obtained and strips suspended in a Krebs-filled organ bath. One end of the strip was attached to a force (F) transducer and the other to a lever arm that effected sinusoidal length (L) oscillations. Strips were oscillated at varying amplitudes (1, 3, and 10% of resting L) and frequencies (f = 0.3, 1, 3, and 10 Hz) at an operating stress of 2 kPa. Resistance (R) and elastance (E) were estimated by fitting changes in F and L to the equation of motion. Hysteresivity (eta) was calculated as eta = (R/E) 2pif. Strips were then fixed for morphological study of collagen, elastic fibers, and the small proteoglycans (PGs), biglycan and fibromodulin (FM). R and E were significantly greater and eta significantly less in BM versus C strips (p < 0.001). The increase in R and E peaked at 14 d after BM; the decrement in eta was maximal at Day 7. Biglycan was increased in BM lung strips at all time points, FM and elastic fibers were increased at 14 and 28 d, and collagen was increased at 28 d only. Hence, changes in mechanics were maximal before collagen content had increased. In addition, we demonstrated a significant correlation between biglycan and all mechanical parameters. These data suggest that changes in PGs may be critical in determining changes in lung tissue viscoelastic behavior in this fibrosis model
Animals↗
Expansion of human NOD/SCID-repopulating cells by stem cell factor, Flk2/Flt3 ligand, thrombopoietin, IL-6, and soluble IL-6 receptor.
Here, we demonstrate a significant ex vivo expansion of human hematopoietic stem cells capable of repopulating in NOD/SCID mice. Using a combination of stem cell factor (SCF), Flk2/Flt3 ligand (FL), thrombopoietin (TPO), and a complex of IL-6 and soluble IL-6 receptor (IL-6/sIL-6R), we cultured cord blood CD34(+) cells for 7 days and transplanted these cells into NOD/SCID mice. Bone marrow engraftment was judged successful when recipient animals contained measurable numbers of human CD45(+) cells 10-12 weeks after transplantation. When cells were cultured with SCF+FL+TPO+IL-6/sIL-6R, 13 of 16 recipients were successfully engrafted, and CD45(+) cells represented 11.5% of bone marrow cells in engrafted recipients. Cells cultured with a subset of these factors were less efficiently engrafted, both as measured by frequency of successful transplantations and prevalence of CD45(+) cells. In animals receiving cells cultured with all 4 factors, human CD45(+) cells represented various lineages, including a large number of CD34(+) cells. The proportion of CD45(+) cells in recipient marrow was 10 times higher in animals receiving these cultured cells than in those receiving comparable numbers of fresh CD34(+) cells, and the expansion rate was estimated at 4.2-fold by a limiting dilution method. Addition of IL-3 to the cytokine combination abrogated the repopulating ability of the expanded cells. The present study may provide a novel culture method for the expansion of human transplantable hematopoietic stem cells suitable for clinical applications.
Animals↗
Lysosomal cysteine protease, cathepsin H, is targeted to lysosomes by the mannose 6-phosphate-independent system in rat hepatocytes.
The contribution of mannose 6-phosphate (Man 6-P)-dependent and -independent systems to lysosomal targeting of cathepsin H, a lysosomal cysteine protease, was investigated by metabolic labeling with [32P]phosphate and [35S]methionine/cysteine in primary cultures of rat hepatocytes. Metabolic labeling experiments with [32]phosphate revealed that only the proform of cathepsin H acquired Man 6-P residues on its high mannose type oligosaccharide, and that most of the phosphorylated procathepsin H was secreted into the medium without having undergone significant intracellular dephosphorylation. Thus, acquisition of Man 6-P residues did not correlate with targeting of cathepsin H to lysosomes. Pulse-chase experiments with [35S]methionine/cysteine showed that only about 10% of the newly synthesized cathepsin H was secreted as a proform while the remainder was retained intracellularly in processed mature form. These results indicate that the majority of newly synthesized cathepsin H is targeted to lysosomes by a Man 6-P-independent mechanism, at least in rat hepatocytes.
Animals↗