Search PubMed⌕ Search

Biomedical subjects

S Kusunoki

Publications and source records attributed to S Kusunoki.

At least 91 records · Page 5Linked to original sources
PubMed1996

Percutaneous hepatic venous isolation and extracorporeal charcoal hemoperfusion for high-dose intraarterial chemotherapy in patients with colorectal hepatic metastases.

The results of treating 12 consecutive patients with unresectable colorectal hepatic metastases with a hepatic arterial infusion of high-dose Adriamycin, 100-120 mg/m2, using hepatic venous isolation (HVI) and charcoal hemoperfusion (CHP) are reported herein. Adriamycin was administered over 5-15 min under extracorporeal drug elimination by HVI-CHP. HVI was percutaneously accomplished by either the double-balloon technique using a Fogarty occlusion catheter (8/22F) or a balloon-tipped catheter (16F). During the infusion, isolated hepatic venous blood was filtered by CHP and pumped into the left axillary vein. There were no lethal complications, and good hemodynamic tolerance to HVI-CHP was confirmed. Tumor liquefaction accompanied by a sharp decrease in serum carcinoembryonic antigen levels by more than 50% of pretreatment levels was observed in 6 of the 12 patients 1 month after treatment. Apart from chemical hepatitis, which developed in 11 (92%) of the patients, the Adriamycin toxicities were well controlled following the development of nausea and vomiting in 2 patients (17%), leukopenia < 2,000/mm3 in 3 (25%), and gastric ulcer in 1 (8%). These results indicate that this method is a safe and useful procedure for otherwise hazardous high-dose intra-arterial chemotherapy in patients with unresectable hepatic tumors.

Adult↗
PubMed1996

A novel ganglioside, 9-O-acetyl GD1b, is recognized by serum antibodies in Guillain-Barré syndrome.

A hitherto undescribed ganglioside was detected in a crude ganglioside fraction of bovine brain using an IgM M-protein binding to Gal beta 1, 3GalNAc residue. We purified and identified it as 9-O-acetyl GD1b based on results of alkali treatment that yielded GD1b and results of fast atom bombardment-mass and gas chromatography-mass spectrometries. 9-O-acetyl GD1b was also found to be present in human peripheral nerve tissue. The reactivities of the serum antibodies from patients with Guillain-Barré syndrome to 9-O-acetyl GD1b, GD1b, and GM1 were determined by ELISA and TLC immunostaining. Nineteen of 85 serum samples from Guillain-Barré syndrome patients had antibodies that bound to 9-O-acetyl GD1b: 14 of the positive samples also reacted with GM1 and GD1b, three reacted with GM1 but not with GD1b, one with GD1b but not with GM1, and one with neither GM1 nor GD1b. These results show that a subset of patients with Guillain-Barré syndrome had antibodies that react with 9-O-acetyl GD1b; therefore, this ganglioside can serve as a target antigen against the antibodies present in Guillain-Barré syndrome.

Animals↗
PubMed1996

Expression of the beta-galactoside alpha 1,2-fucosyltransferase gene suppresses axonal outgrowth of neuro2a neuroblastoma cells.

The axonal outgrowth of cells of Neuro2a, a mouse neuroblastoma cell line, was suppressed on expression of the beta-galactoside alpha 1,2-fucosyltransferase (alpha 1,2-FT) gene. We recently cloned two types of rabbit alpha 1,2-FT, RFT-I and RFT-II. RFT-I exhibits comparable kinetic properties and structural homology with human H gene alpha 1,2-FT, and RFT-II shows comparable kinetic parameters with human Se gene alpha 1,2-FT. Neuro2a cells expressing RFT-I (N2A-RFT-I) contained a large amount of fucosyl GM1 instead of GM1 and GD1a, major gangliosides in the parent Neuro2a cells, whereas Neuro2a cells expressing RFT-II (N2A-RFT-II) showed a subtle change in the ganglioside pattern. N2A-RFT-II and parent Neuro2a cells showed axonal outgrowth in serum-free medium on the exogenous addition of GM1, whereas N2A-RFT-I cells exhibited multiple neurite sprouts but not axonal outgrowth. This phenotype was fully recovered by N2A-RFT-I cells on the addition of D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol and alpha-L-fucosidase to the culture medium, which resulted in pronounced reduction of fucosyl GM1 expression. These results suggested that expression of H-type alpha1,2-FT, and subsequent incorporation of fucose into glycolipids and glycoproteins, especially the formation of fucosyl GM1, modifies the response of neuronal cells to stimuli that induce axonal extension.

Animals↗
PubMed1996

GM1b is a new member of antigen for serum antibody in Guillain-Barré syndrome.

Serum antibody from some patients with Guillain-Barré syndrome recognized an antigen of a minor component in human brain monosialoganglioside fraction. We purified that antigen, which migrated at a position slightly lower than that of GM1 on a thin-layer chromatogram (TLC), by using Iatrobeads column chromatography and preparative TLC. Structural analyses, including fast atom bombardment mass spectrometry, showed it to be GM1b. An enzyme-linked immunosorbent assay (ELISA) using purified GM1b showed that anti-GM1b antibody was present in 22 of 104 cases tested. No anti-GM1b antibody was present in the sera from control patients with other diseases or from the normal controls. Four sera recognized only GM1b among the 11 ganglioside antigens tested. The other 18 sera had antibodies to other antigens, most of which shared no terminal epitope with GM1b. Eight of nine sera samples with anti-GalNAc-GD1a antibody also had anti-GM1b antibody. Antibody to a minor monosialoganglioside, GM1b, was found to be a useful diagnostic marker for Guillain-Barré syndrome. Further study is needed to determine whether this antibody plays a role in the pathogenetic mechanism of the syndrome.

Carbohydrate Sequence↗
PubMed1995

Molecular cloning and expression of two types of rabbit beta-galactoside alpha 1,2-fucosyltransferase.

Two DNA clones encoding rabbit beta-galactoside alpha 1,2-fucosyltransferase (RFT-I and RFT-II) have been isolated from a rabbit genomic DNA library. The DNA sequences revealed open reading frames coding for 373 (RFT-I) and 354 (RFT-II) amino acids, respectively. The deduced amino acid sequences of RFT-I and RFT-II showed 56% identity with each other, and that of RFT-I showed 80% identity with that of human H blood type alpha 1,2-fucosyltransferase. Northern blot analysis of embryo and adult rabbit tissues revealed that the RFT-I gene was expressed in adult brain, and that the RFT-II gene was expressed in salivary and lactating mammary glands. The identities of these enzymes were confirmed by constructing recombinant fucosyltransferases in which the N-terminal part including the cytoplasmic tail and signal anchor domain was replaced with the immunoglobulin signal peptide sequence. RFT-I expressed in COS-7 cells exhibited similar transferase activity to that of human H blood type alpha 1,2-fucosyltransferase. RFT-II expressed in COS-7 cells showed higher affinity for type 1 (Gal beta 1,3GlcNAc) and type 3 (Gal beta 1,3GalNAc) acceptors than type 2 (Gal beta 1,4GlcNAc) ones, which suggested that RFT-II was a putative secretor-type alpha 1,2-fucosyltransferase.

Amino Acid Sequence↗
PubMed1995

Anti-Gal-C antibody in autoimmune neuropathies subsequent to mycoplasma infection.

Four of 82 patients with Guillain-Barré syndrome (GBS) and 1 of 12 with multifocal motor neuropathy (MMN), who previously had had Mycoplasma pneumoniae infections, had serum antibody to galactocerebroside (Gal-C). Two patients with GBS without mycoplasma infection also had anti-Gal-C antibody, whereas none of the normal or the disease controls had it. As Gal-C is a major glycolipid antigen in myelin, anti-Gal-C antibody may function in the pathogenesis of autoimmune demyelinative neuropathies. Mycoplasma pneumoniae appears to be an important preceding infectious agent in autoimmune neuropathies with anti-Gal-C antibody.

Antibodies↗
PubMed1995

HLA and anti-GQ1b IgG antibody in Miller Fisher syndrome and Guillain-Barré syndrome.

We investigated serological human leukocyte antigen (HLA) types in patients with histories of Miller Fisher syndrome (MFS) and Guillain-Barré syndrome (GBS) with ophthalmoplegia, in whom serum anti-GQ1b IgG antibody was present during the acute phase. We examined class I antigens (A, B and C) in 32 patients and class II antigens (DR and DQ) in 30, but found no association. We conclude that particular serologically defined HLA types are not preferred for the immunoresponse of anti-GQ1b IgG antibody in MFS and GBS.

Autoantibodies↗
PubMed1995

A new genetic assay for rifampicin susceptibility of Mycobacterium tuberculosis.

A new method for testing rifampicin (RFP) susceptibility of Mycobacterium tuberculosis was developed. This method is based on detection of the internal sequence derived from 71-kDa heat shock protein mRNA in tubercle bacilli heat-treated in the presence of RFP. The target sequence was amplified by reverse transcription and PCR, followed by agarose gel electrophoretic analysis. No amplification occurred in one RFP-susceptible strain by exposure to 45 degrees C for 45 min in Middlebrook 7H9 broth containing RFP (10 micrograms/ml) after overnight incubation at 37 degrees C. On the other hand, an amplified 275-bp product was obtained from the RFP-resistant strain MY-129. In a subsequent trial using 65 clinical isolates, this method defined their RFP susceptibility levels as well as the verification of the MICs obtained by the conventional agar dilution method, with the exception of one RFP-susceptible strain. Thus, this method provides a rapid and practical system to determine RFP susceptibility in M. tuberculosis.

Antibiotics, Antitubercular↗
PubMed1995

Evaluation of nested polymerase chain reaction for detecting mycobacterial DNA in pleural fluid.

A protocol based on the polymerase chain reaction (PCR) is the most sensitive method for detecting mycobacteria in clinical samples. However, few studies have assessed the usefulness of this method in the diagnosis of tuberculous effusion. We developed a highly sensitive and specific nested PCR method, that amplifies the bovine tuberculous MPB70 gene and the mycobacterial 16S rRNA gene for use in detecting Mycobacterium tuberculosis (M. tuberculosis) and mycobacteria, respectively, in clinical samples. We determined the sensitivity of this method for detecting mycobacteria in samples containing known amounts of mycobacterial DNA and in DNA extracted from pleural effusions obtained from 10 patients with pulmonary tuberculosis in whom standard microbiological techniques had detected mycobacteria in sputum but not in pleural effusion. The nested PCR method for the bovine tuberculous MPB70 gene and the mycobacterial 16S RNA gene was able to detect M. tuberculosis and mycobacterial genomes only if there were at least 2 copies per sample. Positive results for M. tuberculosis and the mycobacterial genomes were obtained by nested PCR in 2 of 10 and in 3 of 10 samples of pleural fluid, respectively but no mycobacteria were detected in malignant effusions obtained from 9 patients with lung cancer. The nested PCR method represents a rapid means for detecting mycobacteria in some pleural effusions previously found to be negative by culture. We speculate that the reaction of the host against mycobacteria is more important than the mycobacteria themselves in the pathogenesis of pleural effusion in which mycobacteria are not detected.

Base Sequence↗
PubMed1995

[Antiglycolipid antibody in inflammatory neuropathy].

Antiglycolipid antibody is frequently detected in the acute phase sera from patients with acute inflammatory demyelinating polyneuropathy (Guillain-Barré syndrome, GBS). The titer is highest in the serum sample taken first after the neurological onset, and decreases with clinical improvement. Antiglycolipid antibody may play a role in the pathogenetic mechanism of GBS. GM1 and GD1b are the antigens most commonly recognized. Monoclonal anti-GD1b antibody specifically bound to the paranodal myelin of the peripheral nervous system. Serum anti-GD1b antibody may cause demyelinative neuropathy by binding to the paranodal myelin of the peripheral nervous system. Anti-GQ1b IgG antibody is specifically raised in almost all the sera from Fisher syndrome and GBS with ophthalmoplegia. Anti-GQ1b monoclonal antibody immunostained specifically the paranodal myelin of the extramedullary portion of oculomotor, trochlear and abducens nerves, but no such staining was observed in the other peripheral nerves. Anti-GQ1b antibody may cause conduction block in the cranial nerves innervating the muscles for extraocular movement by binding to the paranodal myelin of those nerves. Anti-GalNAc-GD1a antibody is detected in the patients with GBS with very low or inexcitable compound muscle action potentials. The sera from patients with GBS subsequent to mycoplasma infection had antigalactocerebroside antibody. Further study on antiglycolipid antibody is needed for understanding the pathogenetic mechanism of GBS.

Animals↗
PubMed1994

Unique localization of fucosyl GM1 in rabbit spinal cord and peripheral nerve: immunohistochemical study using monoclonal anti-fucosyl GM1 antibody CRD73-6.

We generated a murine monoclonal antibody (CRD73-6) for ganglioside fucosyl GM1 using a homogenate of rabbit dorsal root ganglia as an immunogen. CRD73-6 immunostained a subset of neurons and the satellite cells surrounding them in rabbit dorsal root ganglia, unmyelinated fibers in the dorsal root, and dorsal horn of the spinal cord. After the tissue sections had been treated with acetone, CRD73-6 also immunostained large neurons and the perineuronal area around them in the ventral horn, and axons and myelin of the small myelinated fibers in the dorsal root. Unique localization of fucosyl GM1 implicates its possible role in cell adhesion and recognition in the nervous system.

Acetone↗
PubMed1994

N-acetylgalactosaminyl GD1a is a target molecule for serum antibody in Guillain-Barré syndrome.

Serum antibodies against such major glycolipids as GM1, GD1b, and LM1 have been reported in patients in the acute phase of Guillain-Barré syndrome (GBS). Because minor unidentified glycolipids also may be targets of antibodies in GBS sera, we assayed serum antibody against a crude ganglioside fraction using thin-layer chromatogram immunostaining. Antibody activity was detected against a band that migrated just below GD1a in 6 of the 50 patients with GBS tested. Antibody titer, as determined by enzyme-linked immunosorbent assay, decreased during the course of the disease. All 6 patients had suffered gastrointestinal infection before the neurological onset of GBS and showed low amplitudes for the compound muscle action potentials and normal or only slightly decreased nerve conduction velocities. Thin-layer chromatogram immunostaining did not show this antibody activity in any of the 16 normal and 119 disease controls. The unidentified glycolipid was isolated by DEAE-Sephadex A-25 column chromatography, sialidase treatment, and Iatrobeads column chromatography. Fast atom bombardment-mass spectra showed it to be N-acetyl-galactosaminyl GD1a.

Adult↗
PubMed1994

Cerebellar ataxia and polyneuropathy in a patient with IgM M-protein specific to the Gal(beta 1-3)GalNAc epitope.

A 79-year-old man with sensory dominant polyneuropathy, cerebellar ataxia, and palatal myoclonus had serum IgM M-protein that specifically bound to GM1, GD1b, and asialo-GM1. IgM with the same specificity was detected in his cerebrospinal fluid. Results of immunohistochemical studies showed specific binding of this monoclonal IgM to the cerebellar granular layer, dentate nucleus, inferior olive, and gray matter of the cerebrum and spinal cord. Monoclonal antibody GGR12, monospecific to GD1b, had an immunostaining distribution similar to that of the patient's IgM M-protein. The binding of M-protein may be associated with the development of cerebellar ataxia and palatal myoclonus in this patient.

Aged↗
PubMed1994

[Detection of Mycobacterium avium, M. intracellulare and M. kansasii in sputum by two-step PCR].

A DNA amplification assay using PCR, which consists of amplification of genus specific mycobacterial 16S rRNA gene as the 1st step and reamplification of the amplicon with species specific primers as the next step, could detect M. avium, M. intracellular and M. kansasii in the sputum. The results with type or standard strains showed that M. avium PCR and M. intracellular PCR were highly specific for identification of each species but M. kansasii PCR detected M. gastri besides M. kansasii. Among 22 clinical samples which were positive by PCR, the 17 results were confirmed by culture. The PCR detected 27 (7.5%) of nontuberculous mycobacteria from 360 sputum and showed that the 27 of NTM consists of 9 M. avium, 8 M. intracellulare, 5 M. kansasii, 1 reacted both M. avium and M. intracellulare and 4 unidentified.

Base Sequence↗
PubMed1994

Induction of cytotoxic T lymphocytes from splenocytes after orthotopic penetrating keratoplasty in the rat.

We used a rat model of orthotopic penetrating keratoplasty to study splenic cell cytotoxicity in the host. DA (RT1avl) rats received grafts of Fischer (F344, RT1lv1) rat corneas. Cytotoxic T lymphocyte (CTL) activity was measured by the cell-mediated lymphocytotoxicity assay using 4/4R.M.-4 cells as target cells. CTL activity became detectable in vitro 2 weeks after grafting and peaked at 3 weeks. This simple method of quantifying CTL activity in the orthotopic penetrating keratoplasty model should provide a reliable tool for studying the CTL response in corneal transplantation.

Animals↗
PubMed1994

[Fisher syndrome and anti-GQ1b antibody].

Serum anti-GQ1b IgG antibody is specifically detected in almost all the patients with Fisher syndrome and those with Guillain-Barré syndrome with ophthalmoplegia in the acute phase. An anti-GQ1b mouse monoclonal antibody immunostained the paranodal regions of the extramedullary portion of the human oculomotor, trochlear and abducens nerves. Such immunostaining was not observed in the other cranial and peripheral nerves. Anti-GQ1b IgG antibody may be involved in the pathogenetic mechanism of ophthalmoplegia in Fisher syndrome and Guillain-Barré syndrome by binding to the paranodal regions of the oculomotor, trochlear and abducens nerves. Investigation of serum anti-GQ1b antibody is considered to be a useful diagnostic marker in Fisher syndrome and Guillain-Barré syndrome with ophthalmoplegia.

Autoantibodies↗
PubMed1993

Generation of a monoclonal antibody specific for a new class of minor ganglioside antigens, GQ1b alpha and GT1a alpha: its binding to dorsal and lateral horn of human thoracic cord.

We have established a monoclonal antibody, GGR41, specific for a new class of minor gangliosides, such as GQ1b alpha and GT1a alpha, by immunizing mice with a GQ1b-rich ganglioside fraction extracted from bovine brain. Each of those minor gangliosides has been reported to be one of the cholinergic-specific gangliosides (Chol-1). Careful examination of binding specificity of the antibody by both an enzyme-linked immunosorbent assay and immunostaining on thin-layer chromatograms showed that the antibody recognizes three sialyl residues separately attaching to the gangliotetraosyl backbone structure. Immunohistochemical analysis revealed that GGR41 immunostained lamina I and III of dorsal horn and lateral horn of human thoracic cord but motor neurons were not immunostained. Except for negative staining of motor neurons, this distribution is similar to the distribution pattern of staining as reported in rats and humans using a polyclonal antibody against Chol-1. Thus, the antibody obtained in this study should be a useful reagent to study the function of a unique new class of the minor gangliosides.

Animals↗