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Biomedical subjects

N Aoki

Publications and source records attributed to N Aoki.

At least 199 records · Page 11Linked to original sources

Molecular cloning of glycoprotein antigens MGP57/53 recognized by monoclonal antibodies raised against bovine milk fat globule membrane.

A cDNA encoding 57 kDa and 53 kDa antigens (MGP57/53) recognized by monoclonal antibodies raised against bovine milk fat globule membrane (MFGM) (Biochim. Biophys. Acta 1199 (1994) 87-95) was cloned from lactating bovine mammary gland by a combination of reverse transcriptase-coupled polymerase chain reaction (RT-PCR) and 3'-rapid amplification of cDNA ends (3'-RACE). The deduced amino-acid sequence showed that mature MGP57/53 consists of 409 amino-acid residues and the calculated molecular weight and isoelectric point are 45,544 and 6.42, respectively. Computer analysis reveals that it has a significant similarity to mouse mammary epithelial cell surface protein, MFG-E8 and a human breast tumor-associated glycoprotein antigen, BA46-1. An N-terminal cysteine-rich domain and a C-terminal tandemly repeated sequence were highly conserved among them, but bovine MGP57/53 lacks 36 amino-acid residues containing a cluster of 5 prolines found in mouse MFG-E8. Northern blot analysis showed that the cDNA hybridized to about 2.0 kb mRNA of lactating bovine mammary gland. These results strongly support our previous report that the two MFGM antigens originate from a single gene and are isoforms with different N-linked sugar chains.

Amino Acid Sequence↗

Characterization of the promoter region of human BCL-6 gene.

The BCL-6 gene encodes a zinc finger-transcription factor and frequently is rearranged in diffuse large cell lymphoma. We have reported here the isolation of the 5'-flanking region of the human BCL-6 gene. Nucleotides sequence of 1.5 Kb promoter region contains TATA box but no CAAT box and a number of potential regulatory elements including CACCC, E-box, and GATA-1 sites. Studies of the promoter activity showed the first 657 nucleotides of the 5'-flanking region significantly drove transcription of the luciferase reporter gene in human Burkitt's lymphoma cell line.

Base Sequence↗

Classification of rice allergenic protein cDNAs belonging to the alpha-amylase/trypsin inhibitor gene family.

Seven cDNA clones encoding rice allergenic proteins were newly isolated. Comparison of the sequences of ten cDNA clones, including the previously isolated three clones results in their classification into four subfamilies. Homologies in the nucleotide sequences among and within subfamilies are 70-85% and above 95%, respectively. A sequence of twenty five amino-acid residues at the C-terminal proximal region is highly conserved among all clones and resembles that of plant lipid transfer proteins.

Allergens↗

Precursor-product relationship between chicken vitellogenin and the yolk proteins: the 40 kDa yolk plasma glycoprotein is derived from the C-terminal cysteine-rich domain of vitellogenin II.

Chicken vitellogenin, a serum lipoprotein specific for laying hens, has been thought to be proteolytically cleaved into the heavy and light chain lipovitellins and phosvitin, the major yolk granule proteins, during or after transportation into oocyte. In this study, another proteolytic product of vitellogenin has newly been isolated from the 'beta-livetin' fraction of yolk plasma. It is a yolk glycoprotein of 40 kDa (YGP40) with asparagine-linked carbohydrate chain(s) recognized by Concanavalin A and castor bean lectin (RCA-I), and it is identified as a C-terminal cysteine-rich fragment of the major vitellogenin (vitellogenin II), the cysteine-rich domain homologous to D2 region of von Willebrand factor. Another yolk plasma glycoprotein of 42 kDa is suggested to be one of the proteolytic products of the minor vitellogenin (vitellogenin I). Both 40 kDa and 42 kDa glycoproteins were shown to be present in growing oocytes but absent in laying hen's serum. Limited proteolysis of vitellogenin II with cathepsin D produced a 40 kDa protein with reactivity to anti-YGP40 antibody. Gel filtration analysis of vitellogenin II digested with cathepsin D showed that YGP40 dissociated from lipovitellin-phosvitin complex after the proteolytic cleavage. These results suggest that after incorporation from serum via a specific receptor vitellogenin II is cleaved in the oocyte into four fragments, heavy and light chain lipovitellins, phosvitin and YGP40, and that YGP40 is released into the yolk plasma before or during compartmentation of lipovitellin-phosvitin complex into the yolk granule.

Amino Acid Sequence↗

A bovine IgG heavy chain contains N-acetylgalactosaminylated N-linked sugar chains.

A 56K protein co-purified with bovine milk fat globule membrane (MFGM) proteins bound to Wisteria floribunda agglutinin (WFA) like most MFGM glycoproteins. Treatment with N-glycanase or beta-N-acetylhexosaminidase abolished the lectin binding to the protein. Amino acid sequence and immunoblot analyses revealed that the 56K protein is an IgG heavy chain. Lectin column chromatography of the oligosaccharides released by hydrazinolysis from the purified IgG heavy chains revealed that 0.08% of the total N-linked sugar chains bind to a WFA-agarose column, suggesting that they contain the beta-N-acetylgalactosaminylated structure.

Acetylgalactosamine↗

Effects of medroxyprogesterone acetate therapy on advanced or recurrent breast cancer and its influences on blood coagulation and the fibrinolytic system.

The effects of medroxyprogesterone acetate (MPA) therapy on advanced or recurrent breast cancer and its influence on blood coagulation and the fibrinolytic system were compared among three different therapy regimens consisting of cyclophosphamide, adriamycin, and 5-fluorouracil (CAF) + MPA and CAF or MPA alone. A clinical response was observed in 42.9% (9/21) of the patients for CAF + MPA, 36.4% (8/22) for CAF and 23.8% (5/21) for MPA alone. No marked thrombosis or its prodromal condition was observed in any group. The effects on the test values for blood coagulation and the fibrinolytic system did not significantly change in the CAF group. However, both AT-III and protein C significantly increased above the normal ranges in the CAF+MPA and MPA groups. Increases in factor X, plasminogen, and alpha 2-plasmin inhibitor/plasmin complex (PIC) and decreases in fibrinogen, tissue plasminogen activator, and D-dimer, were all observed in the MPA and CAF + MPA groups, especially in the MPA group, although these changes remained within the normal ranges. The data indicated that MPA has various influences on blood coagulation and the fibrinolytic system, but these changes did not suggest activation of the blood coagulation system.

Antineoplastic Agents, Hormonal↗

"Head-shaking syndrome" neurological deterioration during continuous head-shaking as an adjunct to cisternal irrigation for clot removal in patients with acute subarachnoid haemorrhage.

To prevent cerebral vasospasm after aneurysmal subarachnoid haemorrhage, cisternal irrigation has been reported to be more effective when combined with continuous head-shaking (head-shaking method). The present study was conducted to evaluate the safety and preventative effect for vasospasm in patients treated with the head-shaking method. Six of 17 patients managed postoperatively by the original head-shaking procedure developed neurological deterioration related to the method: two had intracranial haematoma (one with acute interhemispheric subdural haematoma, and the other with cerebellar haemorrhage), two had acute brain swelling, and two failed to show abnormal findings on computed tomography. These pathological processes may be suitably referred to as "head-shaking syndrome". Delayed ischaemic neurological deficits associated with low-density lesions on computed tomography were demonstrated in five patients (29%). From these observations, the head-shaking method may not be as safe as described in the original articles, and is critically evaluated in terms of its preventative effect for cerebral vasospasm.

Aged↗

Vasospasm after resection of skull base tumors.

Vasospasm after resection of skull base tumors is a rare complication that often produces serious ischemic sequelae. In four of the authors' recent cases, vasospasm complicated the patient's postoperative course. A review of the literature produced a number of cases that can help determine possible causes of vasospasm after tumor resection, ways to prevent it, and methods to evaluate it when it occurs. The cause appears to be multifactorial and the surgical approach may contribute to the pathogenesis of vasospasm. Physicians must have a high degree of suspicion to detect vasospasm at an early stage of skull base surgery. Cerebral blood flow measurement and transcranial Doppler are useful monitoring tools.

Adenocarcinoma↗

Differing kinase activity of the c-yes and c-src gene proteins in TPA-induced megakaryocytic differentiation of T-33 and K562 cell lines.

We examined the protein kinase (PK) activity of the c-yes and c-src gene proteins (c-YES, c-SRC) at an early phase of 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced megakaryocytic differentiation of T-33 and K562 cells with use of immunoprecipitation and in vitro kinase assay. We found that c-SRC PK activity of TPA-treated T-33 and K562 cell lines had been enhanced compared with the untreated ones, but in contrast, no enhancement of c-YES PK activity by the TPA treatment was observed in these cell lines. We also examined PK activity in TPA-induced monocytic differentiation of U937 monoblastic cells that exhibited no megakaryocytic markers and found that both the c-YES and c-SRC PK activity was enhanced by the TPA treatment. Our data suggest that c-YES and c-SRC play different and unique roles in TPA-induced megakaryocytic differentiation in T-33 and K562 cells.

Cell Differentiation↗

Enzymic method for the amperometric determination of nicotinic acid in meat products.

An enzymic method for the determination of nicotinic acid is described, based on the indirect electrochemical monitoring of nicotinic acid via its reaction with oxygen in the presence of nicotinic acid hydroxylase. Derivative amperometric signals due to oxygen depletion enable a one-point kinetic analysis to be carried out. Nicotinic acid hydroxylase catalyses the hydroxylation of nicotinic acid to produce 6-hydroxynicotinic acid, which is accompanied by a stoichiometric consumption of oxygen. The method was applied successfully to the analysis of real samples.

Catalysis↗

Using biopsy materials to detect mutations in gastrointestinal tumors.

Because endoscopic examinations are a crucial first step in the diagnosis of carcinomas, we have analyzed point mutations of the c-K-ras and p53 genes using biopsied samples. There is no previous report of such an approach, using only biopsied materials. We analyzed point mutations of the c-K-ras and p53 genes using 60 colorectal and 31 gastric tumor biopsy specimens. None of the gastric and seven of the colorectal tumors had mutations of the c-K-ras gene. p53 gene mutations were detected in six gastric carcinomas [two out of 11 intramucosal carcinomas (18.2%) and four out of 11 invasive carcinomas (36.4%)]. Eight of 15 invasive colorectal carcinomas (53.3%) showed point mutations in the p53 gene. Although point mutation frequencies in this study were relatively low when compared with previous reports in which surgical samples were used, our study shows that biopsied specimens are adequate and useful for analysis of gene point mutations.

Adenocarcinoma↗

Epidemiological investigation of insulin resistance syndrome (syndrome X) in a city in Japan.

1. In order to study the prevalence of insulin resistance syndrome (syndrome X) in Japanese subjects, inhabitants aged above 40 years living in Osaka-Sayama city from September 1992 through December 1993 were investigated. The population-based study was performed on 2498 subjects (661 males and 1837 females) constituting 10.9% of the total population aged above 40 years. 2. The prevalence of glucose intolerance was 8.7% (n = 218) in 2498 subjects. The prevalence of hypertension was 36.9% (n = 923) and that of hypertriglyceridaemia was 19.0% (n = 475). The prevalence of syndrome X as characterized by an association of glucose intolerance, hypertension and hypertriglyceridaemia was 1.6% (n = 39) in all subjects examined and 17.4% in subjects showing glucose intolerance. 3. Fasting serum insulin levels were significantly higher in patients with syndrome X than in normal subjects. Furthermore, the levels were significantly correlated with blood levels of frucutosamine, fasting glucose and triglyceride, and with body mass index as well. 4. In conclusion, insulin resistance syndrome (syndrome X) is also found among the larger Japanese population, and fasting serum insulin levels can be a useful marker of this metabolic disorder.

Adult↗

[A trial of clinical application of sparfloxacin for treating mycobacterial infections].

Sparfloxacin seems to be a good candidate for antimycobacterial treatment. However, there have been no clinical studies. We experienced 2 SPFX-treated cases, who could not use other antimycobacterial agents because of side effect, we tried SPFX-treatment on these cases. Good results were obtained, however, for long time use to prevent side effects, we tried SPFX every second day and monitored the serum levels of SPFX. SPFX-every second day treatment gave good clinical results and adequate serum levels of SPFX were observed.

Aged↗

Spontaneous regeneration of the foramen magnum after decompressive suboccipital craniectomy in Chiari malformation: case report.

A 21-year-old woman, who had undergone foramen magnum decompression for her symptomatic Chiari malformation (Type 1) 7 years before, presented with recurrence of the symptoms. Neuroradiological examinations demonstrated regeneration of the foramen magnum that was caused by new bone formation, as confirmed at the second surgery. Neurological improvement was obtained after the removal of the regenerated foramen magnum. This observation, though rare, deserves to be kept in mind during the postoperative follow-up period in young patients with Chiari malformation.

Adult↗

Four rice seed cDNA clones belonging to the alpha-amylase/trypsin inhibitor gene family encode potential rice allergens.

Four rice seed proteins encoded by cDNAs belonging to the alpha-amylase/trypsin inhibitor gene family were overexpressed as TrpE-fusion proteins in E. coli. The expressed rice proteins were detected by SDS-PAGE as major proteins in bacterial cell lysates. Western blot analyses showed that all the recombinant proteins were immunologically reactive to rabbit polyclonal antibodies and to a mouse monoclonal antibody (25B9) specific for a previously isolated rice allergen of 16 kDa. Some truncated proteins from deletion mutants of the cDNAs retained their reactivity to the specific antibodies. These results suggest that the cDNAs encode potential rice allergens and that some epitopes of the recombinant proteins are still immunoreactive when they are expressed as their fragments.

Allergens↗

Isolation and culture of bovine mammary epithelial cells and establishment of gene transfection conditions in the cells.

Bovine mammary epithelial cells (BMEC) were isolated as acinous fragments from a mammary gland of a lactating cow. They grew well on plastic substratum, showed the characteristic cobblestone morphology of epithelial cells, and secreted alpha s1-, beta-, and kappa-caseins even when grown on plastic substratum. A plasmid containing the bacterial chloramphenicol acetyl transferase (CAT) gene was transfected to the isolated BMEC by calcium phosphate precipitation and electroporation methods. The transfection efficiency of BMEC by the calcium phosphate method was greatly improved by post-transfection osmotic shock with glycerol or polyethylene glycol. An about 700 bp DNA fragment containing 5'-flanking sequence of bovine alpha s1-casein gene showed promoter activity in the transfected BMEC. The primary culture of BMEC might be useful for studies on regulation of bovine milk-protein gene expression.

Animals↗