Expansion of a paroxysmal nocturnal hemoglobinuria (PNH) clone after cyclosporine therapy for aplastic anemia/PNH syndrome.
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Adult↗
Biomedical subjects
H Takamatsu
Publications and source records attributed to H Takamatsu.
Expression and purification of biologically active human OSF-1 in Escherichia coli.
OSF-1 (also known as pleiotrophin, HB-GAM, HBGF-8 or HBNF) is a heparin-binding, neurotrophic protein. Its tissue-specific expression in rats is developmentally regulated and the protein is highly conserved between species. The protein is believed to be involved in neuronal development. Previous experiments in our laboratory showed that OSF-1 is primarily expressed in brain and bone. The biological function of OSF-1 in bone is unknown. In order to overcome the limited availability of the native protein, we now report on the high-level expression of human OSF-1 in Escherichia coli. The protein is present in the form of inclusion bodies, which were isolated and solubilized. The partially purified protein was refolded and further purified employing heparin sepharose chromatography. N-terminal sequence determination revealed the same amino acid sequence as the natural mature protein. The isolated backfolded recombinant human OSF-1 did promote neurites outgrowth in primary cultures of cortical neurons.
Amino Acid Sequence↗
Treatment for necrotizing enterocolitis perforation in the extremely premature infant (weighing less than 1,000 g).
The frequency of necrotizing enterocolitis (NEC) in the extremely premature infant (less than 1,000 g) is still high and it is very difficult for infants weighing less than 1,000 g with NEC perforation to survive. In our institutes, the management protocol for NEC perforation in infants weighing less than 1,000 g includes peritoneal drainage under local anesthesia, administration of coagulating factor XIII, and the usual treatment for septic shock. During the past 3 years, four infants weighing less than 1,000 g with NEC perforation have survived using this protocol without laparotomy. This management protocol is the treatment of choice in infants in very poor condition or infants weighing less than 1,000 g with NEC perforation.
Diseases in Twins↗
Endorectal pull-through operation for diffuse cavernous hemangiomatosis of the sigmoid colon, rectum and anus.
A 6-year-old girl with diffuse cavernous hemangiomatosis of the sigmoid colon, rectum and anus underwent endorectal pull-through operation for sphincter-saving resection. Rectal mucosa was resected from 4 cm above peritoneal reflection to anal skin margin and the normal sigmoid colon was pulled down through the rectal muscular cuff. Ligation of the superior rectal and left internal iliac arteries at operation achieved satisfactory control of bleeding. Postoperative manometric studies showed almost normal sphincter tone and good response to rectal stimuli by balloon inflation. The endorectal pull-through (Soave-Denda procedure) is a common procedure for Hirschsprung's disease and the best procedure for the sphincter-saving treatment of diffuse cavernous hemangiomatosis of the colon and rectum.
Anal Canal↗
In vivo and in vitro characterization of the secA gene product of Bacillus subtilis.
The putative amino acid sequence from the wild-type Bacillus subtilis div+ gene, which complements the temperature-sensitive div-341 mutation, shares a 50% identity with the sequence from Escherichia coli secA (Y. Sadaie, H. Takamatsu, K. Nakamura, and K. Yamane, Gene 98:101-105, 1991). The B. subtilis div-341 mutant accumulated the precursor proteins of alpha-amylase and beta-lactamase at 45 degrees C as in the case of sec mutants of E. coli. The div-341 mutation is a transition mutation causing an amino acid replacement from Pro to Leu at residue 431 of the putative amino acid sequence. The B. subtilis div+ gene was overexpressed in E. coli under the control of the tac promoter, and its product was purified to homogeneity. The Div protein consists of a homodimer of 94-kDa subunits which possesses ATPase activity, and the first 7 amino acids of the putative Div protein were found to be subjected to limited proteolysis in the purified protein. The antiserum against B. subtilis Div weakly cross-reacted with E. coli SecA. On the other hand, B. subtilis Div could not replace E. coli SecA in an E. coli in vitro protein translocation system. The temperature-sensitive growth of the E. coli secA mutant could not be restored by the introduction of B. subtilis div+, which is expressed under the control of the spac-1 promoter, and vice versa. The B. subtilis div+ gene is the B. subtilis counterpart of E. coli secA, and we propose that the div+ gene be referred to as B. subtilis secA, although Div did not function in the protein translocation system of E. coli.
Adenosine Triphosphatases↗
Isolation of substance P binding protein from rat brain.
Substance P (SP) binding protein of rat brain was solubilized by digitonin. The solubilized proteins were then purified by sequential gel filtration, concanavalin A lectin Sepharose, and SP-affinity chromatography. The calculated molecular weight of this purified SP binding protein was 76-74 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The rabbits were immunized with the purified protein and resulting polyclonal anti-sera were tested. The immune serum significantly inhibited [3H]SP binding to the 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate solubilized membrane fractions from rat brain, whereas pre-bleed antiserum failed to inhibit the binding. This polyclonal antibody also inhibited the activity of 45Ca influx into astroglioma cells stimulated by SP, but does not inhibit that stimulated by histamine. Furthermore, this polyclonal antibody recognized the 76-74 kDa band as assessed by Western blotting. These data strongly suggest that this polyclonal antibody could recognize a part of the natural SP receptor site.
Animals↗
[Bietti's crystalline retinopathy].
The authors previously reported four cases with crystalline retinopathy and suggested that it be classified into three stages: 1) retinal pigment epithelium (RPE) atrophy in the posterior pole, 2) choriocapillaris atrophy appearing in the enlarged areas of RPE atrophy, and 3) prominent diffuse atrophy of both the RPE and choriocapillaris. We followed up 4 cases, including 2 cases which were previously been reported, and have now evaluated the clinical features of this disease. The results indicated that the above classification was clinically useful. Enlargement of the RPE atrophy from stage 2 to 3 was due to enlargement and confluence of the RPE atrophy between the posterior pole and the periphery. In addition, disturbances in ERG and color vision were thought to correlate with secondary dysfunction at the level of the photoreceptor cell layer within the diseased lesions.
Adult↗
Sequencing reveals similarity of the wild-type div+ gene of Bacillus subtilis to the Escherichia coli secA gene.
We have determined the nucleotide (nt) sequence of the wild-type div+ gene of Bacillus subtilis which complements the temperature-sensitive div-341 mutation and is involved in cell septation, sporulation, secretion of extracellular enzymes, development of competence, autolysis and spore outgrowth. It has an open reading frame encoding 841 amino acids (aa) with homology to the Escherichia coli secA gene, which is involved in protein secretion and cell separation. The deduced aa sequence of the B. subtilis div+ gene shares 50% identity with that of the E. coli secA gene, and highly homologous regions were observed in the N-terminal portions. DNA-DNA hybridization with the E. coli secA gene as the probe showed that the div+ gene could be easily detected by homology and that a single copy of the homologous gene was present in B. subtilis. Since both genes are similar in their functions and deduced aa sequences, we propose that the div+ gene is the counterpart of the secA gene of E. coli.
Amino Acid Sequence↗
Association of hiatus hernia with postero-lateral diaphragmatic hernia (Bochdalek's hernia).
A boy with hiatus hernia following the repair of the left postero-lateral diaphragmatic hernia (Bochdalek's hernia) was reported. At the age of one month, the repair of Bochdalek hernia was performed with transabdominal approach. At that time the stomach was located in the normal position. Eight days after the repair he developed vomiting and hiatus hernia was revealed by barium esophagram. Antireflux surgery was required because there was no response to the conservative management for two months. Esophageal pH study and manometric study were very useful for the diagnosis of hiatus hernia or GER and the evaluation of antireflux surgery.
Abnormalities, Multiple↗
Characterization of the carbohydrate chain on the substance P receptor in the rat brain cortex: effect of lectins on [3H]substance P binding.
The characteristics of the carbohydrate chain on the rat cerebral cortical substance P (SP) receptor were studied. We examined the effects of pretreatment with three lectins (concanavalin A, wheat germ agglutinin, lens culinaris agglutinin) on the [3H]SP binding activities. Each lectin can bind to the specific carbohydrate chain. Among these lectins, only concanavalin A inhibited specific [3H]SP binding by reducing the affinity of the binding sites. The inhibitory action of concanavalin A was dose-dependent and diminished by the addition of alpha-methyl-D-mannoside. The present results suggest that the rat cortical SP receptor has either a biantennary complex-type or a high mannose-type of carbohydrate chain, and that the carbohydrate chain is implicated in the SP binding activity of the SP receptor system.
Animals↗
[Multivitamin preparations for total parenteral nutrition].
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Humans↗
[A case of leiomyoma of the intermediate bronchus].
Leiomyoma of the trachea and bronchus is a rare tumor. Only 45 cases have been reported in the Japanese literature. We report here a case of leiomyoma of the intermediate bronchus which was successfully operated on. A 45-year-old man suffered from a repeated episode of pneumonia in the right lower lobe. Bronchoscopy revealed a polypoid mass on the intermediate bronchus. The diagnosis of leiomyoma was made by bronchoscopic biopsy. Sleeve resection of the intermediate bronchus including the tumor and end-to-end anastomosis was performed. His postoperative course was uneventful and bronchoscopic findings showed clear healing of the anastomotic site. The resected specimen contained a tumor with smooth surface having protrusion into the bronchial lumen with 6 x 5 x 4 mm in size. The final pathological diagnosis was leiomyoma of the intermediate bronchus with no evidence of malignancy.
Anastomosis, Surgical↗
Complementation of the protein transport defect of an Escherichia coli secY mutant (secY24) by Bacillus subtilis secY homologue.
Bacillus subtilis SecY homologue shares 41.3% homology with that of E. coli and remarkably higher homologous regions (more than 80%) are present in the four cytoplasmic regions [(1990) J. Biochem. 107, 603-607]. Based on the formation of the mature form of OmpA in E. coli, we have shown that the protein transport defect of the E. coli secY mutant (secY24) is complemented by the gene product from the B. subtilis secY homologue, which is expressed under the lac promoter control. However, B. subtilis SecY could not restore growth of the E. coli mutant at nonpermissive temperature.
Bacillus subtilis↗
Identification of a bluetongue virus serotype 1-specific ovine helper T-cell determinant in outer capsid protein VP2.
Ovine T-cell lines (including one clone [101A]), which are specific for Bluetongue virus serotype 1 (BTV1), have been established and characterized. Although these T-cell lines react with different isolates of BTV1 (including those from South Africa, Australia, Nigeria, and Cameroon), they do not react with heterologous BTV serotypes. Antigen specificity of these T-cells was studied using purified virus particles, infectious subviral particles (ISVP) and cores, or using individual BTV structural proteins that were either isolated by SDS-PAGE or expressed by recombinant strains of vaccinia virus. The results showed that each of the T-cell lines reacted with outer capsid protein VP2 (the BTV protein exhibiting most serotype-specific variation and the major neutralization antigen). However, all of the uncloned T-cell lines also reacted with either the core structural proteins or the outer capsid protein VP5. In contrast, the T-cell clone 101A only reacted with outer capsid protein VP2. Cell surface marker analysis showed that 101A has a helper T-cell phenotype (CD5+, CD4+, CD8-, T-19-). The T-cell lines and clone 101A all produced large amounts of interleukin 2 (IL-2) when stimulated with purified BTV1 virus particles, or with VP2 (up to 120 IU/ml from 2 x 10(5) T-cells). BTV serotype-specific antigenic sites, for B cells and at least one site for ovine helper T-cells, are therefore located within VP2.
Animals↗
Cloning and characterization of a Bacillus subtilis gene homologous to E. coli secY.
A 3.5-kb HindIII DNA fragment containing the secY gene of Bacillus subtilis has been cloned into plasmid pUC13 using the Escherichia coli secY gene as a probe. The complete nucleotide sequence of the cloned DNA indicated that it contained five open reading frames, and their order in the region, given by the gene product, was suggested to be L30-L15-SecY-Adk-Map by their similarity to the products of the E. coli genes. The region was similar to a part of the spc operon of the E. coli chromosome, although the genes for Adk and Map were not included. The gene product of the B. subtilis secY homologue was composed of 423 amino acids and its molecular weight was calculated to be 46,300. The distribution of hydrophobic amino acids in the gene product suggested that the protein is a membrane integrated protein with ten transmembrane segments. The total deduced amino acid sequence of the B. subtilis SecY homologue shows 41.3% homology with that of E. coli SecY, but remarkably higher homologous regions (more than 80% identity) are present in the four cytoplasmic domains.
Adenylate Kinase↗
Characterization of long-term cultured bovine CD4-positive and CD8-positive T-cell lines and clones.
Long-term cultured CD4+ or CD8+ bovine T-cell lines and clones were established. The CD8+ T-cell line and clones had a strong lectin-dependent cytotoxicity, whereas the CD4+ T-cell line did not. Both phenotype cell lines grew in an interleukin-2 (IL-2)-dependent manner and expressed 50,000-55,000 MW and 65,000-75,000 MW proteins associated with a putative IL-2 receptor (IL-2R), as demonstrated by the cross-linking of radioiodinated recombinant human IL-2 (rhIL-2). Additional molecules of 13,000 and 27,000 MW were also observed on CD8+ T cells. The binding of rhIL-2 was blocked by crude bovine IL-2, and Scatchard plot analysis of the binding data showed that both phenotype cells expressed two different affinity IL-2R that had equilibrium dissociation constants of 12-20 pm (3000-6000 sites/cell) and 146-490 pM (16,000-25,000 sites/well). Only IL-2 stimulated DNA synthesis in these cell lines, whereas mitochondrial enzymes activity, protein synthesis and protein secretion were enhanced by IL-2, mitogens and phorbol myristate acetate. The supernatant from mitogen-stimulated CD4+ cells was unable to enhance the DNA synthesis of either the CD4+ or CD8+ lines, whereas both freshly prepared Con A blasts and anti-immunoglobulin-treated bovine B cells showed elevated DNA synthesis under the same conditions.
Animals↗
Cultivation of bluetongue virus-specific ovine T cells and their cross-reactivity with different serotype viruses.
Bluetongue (BT) virus-specific ovine T-cell lines were prepared from BT virus-infected sheep by three cycles of alternate stimulation and resting culture in vitro. In antigen-specific proliferation assays and/or cytotoxicity assays, most of these T-cell lines responded not only to homologous serotype virus but also heterologous serotype viruses. This cross-reactivity did not correlate with the relatedness of serotypes as defined by cross-neutralizing antibodies. One cell line, 58-014, has grown continuously for more than 7 months without loss of antigen specificity. However, most of the cell lines lost their antigen specificity 2-4 months after cultivation. Certain BT virus-specific T-cell lines were able to reduce BT virus replication in autologous skin fibroblast cell culture.
Animals↗
Inhibition of in vitro immunocyte function by sera from cattle with bovine leukosis.
Most sera from leukaemic cattle inhibited phagocytic activity of normal bovine peripheral polymorphonuclear leukocytes, growth of interleukin 2-dependent bovine T cells and mitogen-induced (phytohaemagglutinin, concanavalin A, pokeweed mitogen, lipopolysaccharide and protein A) blastogenesis of normal bovine lymphocytes. By contrast, antibody-dependent, and spontaneous cell-mediated cytotoxicity were suppressed by only a few sera. The antibody titer against bovine leukaemia virus in these sera correlated with the percent inhibition of lymphocyte blastogenesis. These leukotic sera had no direct cellular cytotoxicity and the inhibitory activity was not lost by dialysis or heat inactivation at 62 degrees C for 30 min. However, the activity was reduced by heating at 80 degrees C for 30 min. Neither the concanavalin A sepharose 4B effluent fraction nor 3.5% polyethyleneglycol-treated serum was found to contain significant lymphocyte-inhibitory activity. Blastogenic transformation of lymphocytes prepared from leukaemic cattle was hardly detectable; however, the mitogen responsiveness of these lymphocytes was improved by a 37 degrees C 1-h preincubation followed by washing.
Animals↗