[Chronic obstructive pulmonary disease].
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Humans↗
Biomedical subjects
S Handa
Publications and source records attributed to S Handa.
New methods using polyvinylidene difluoride membranes to detect enzymes involved in glycosphingolipid metabolism.
Two new methods are described using polyvinylidene difluoride (PVDF) membranes to detect enzymes involved in glycosphingolipid metabolism. One is the detection of enzymes on a PVDF membrane to which glycosphingolipids have been transferred from an HPTLC-plate by TLC blotting. The glycosphingolipids on the membrane were incubated with an enzyme preparation, and the resulting product was detected by immunostaining with a monoclonal antibody directed to the product. IV(3)NeuAc(alpha)Lc(4)Cer that had been transferred to a PVDF membrane was incubated with Clostridium perfringens sialidase. Lc(3)Cer then was transferred to the membrane, and the whole incubated with bovine milk beta1-4 galactosyltransferase, after which the product, nLc(4)Cer, was detected by immunostaining with the monoclonal antibody H11 that recognizes the GAl(beta)1-4GlcNAc(beta)1-3Gal structure of neolactoseries glycosphingolipids. This method detects glycosphingolipid-metabolizing enzymes that produce the same epitope. The second method is the detection of enzymes located on a polyacrylamide gel. A sialidase preparation from C. perfringens was subjected to native polyacrylamide electrophoresis then transferred to a PVDF membrane impregnated with IV(3)NeuAc(alpha)nLc(4)Cer as the enzyme substrate. The membrane then was incubated, and the resulting product, nLc(4)Cer, detected by immunostaining with the monoclonal antibody H11. The area stained shows the location of the sialidase on the polyacrylamide gel. The method is effective for determining the apparent molecular weight(s) of the enzyme(s) in the crude enzyme preparation. This was demonstrated by using crude sialidase preparation from C. perfringens.
Chromatography, Thin Layer↗
Structural analysis of mono- and bis-sulfated glycosphingolipids by negative liquid secondary ion mass spectrometry with high- and low-energy collision-induced dissociation.
Several underivatized mono- and bis-sulfated glycosphingolipids having gangliotriaose or gangliotetraose core structure were analyzed by negative liquid secondary ion mass spectrometry (LSIMS) with high- and low-energy collision-induced dissociation (CID). In the normal negative LSIMS spectra, each mono-sulfated glycolipid gave abundant [M - H]- ions and each bis-sulfated glycolipid gave abundant [M + Na - 2H]- ions as well as the hydrogen sulfate anion [OSO3H]-. In high-energy CID spectra of the deprotonated molecule, only ions containing a sulfate ester were clearly observed. When a sulfate was present on the non-reducing terminal saccharide residue, a series of ions corresponding to sulfated mono- to tetra-saccharides, resulting from sequential cleavage of glycosidic bonds, were observed. If the sulfate was attached to an internal hexose of the sugar chain, the product ions corresponding to the non-sulfated, non-reducing terminal residue were absent. In contrast, the low-energy CID resulted in extremely simple spectra that contained only one or two major product ions characteristic of each sulfated glycolipid. These results provided clear information on the overall sugar and ceramide compositions, and allowed saccharide structures differing in location and number of sulfate esters to be distinguished.
Animals↗
Characterization of blood-group-ABO(H)-active glycosphingolipids in type-AB human erythrocytes.
Neutral glycolipids in Folch's upper phase were isolated from human erythrocyte membranes of 22 individuals with blood type AB. On immunostaining by TLC with anti-A IgG, all reactive glycolipids in type A corresponded to reactive glycolipids in type-AB erythrocytes. With anti-B IgM, all reactive glycolipids in type-B erythrocytes also corresponded to reactive glycolipids in type-AB erythrocytes. By comparison of the reactivity to that of the anti-A and anti-B antibodies, it was found that, in type-AB erythrocytes, all glycolipids reactive with either one of the anti-A or anti-B antibodies were detected in both type-A and type-B erythrocytes, and that A-active glycolipids had higher Rf values than B-active glycolipids on TLC plates. A series of glycolipids reactive with both antibodies were purified from the Folch's upper neutral glycolipid fraction of erythrocyte membranes by column chromatography, and was characterized by TLC-immunostaining and negative secondary-ion mass spectrometry. The results strongly suggested that A-active and B-active carbohydrate chain epitopes existed separately as glycolipid molecules in blood-type-AB erythrocytes. It was also confirmed that these phenotypes observed in erythrocyte membranes were exhibited by blood-group-active glycosphingolipids in the small intestine of blood-type-AB individuals. Furthermore, upon treatment of fractions obtained from silicic acid column chromatography with alpha-N-acetylhexosaminidase or alpha-galactosidase, a branched hybrid-type molecule with both A and B determinants was not detected.
ABO Blood-Group System↗
Direct mass spectrometric analysis of glycosphingolipid transferred to a polyvinylidene difluoride membrane by thin-layer chromatography blotting.
A simple, rapid method for the analysis of glycosphingolipid that combines "thin-layer chromatography (TLC) blotting" and mass spectrometry is reported. Glycosphingolipids developed by TLC were transferred to a polyvinylidene difluoride membrane by TLC blotting, after which the glycosphingolipid band on the membrane was excised and placed on a mass spectrometer probe tip, and a few microliters of triethanolamine was added as the matrix. The sample was analyzed by secondary ion mass spectrometry. About 1 microgram of glycosphingolipid subjected to TLC can be analyzed. The major advantage of this method is that glycosphingolipid can be analyzed structurally without purification by repeated column chromatography.
Animals↗
Association of hypersensitivity and carriage of dermatophytes in clinically normal sites in patients with Tinea cruris.
Forty nine patients with mycologically confirmed Tinea cruris were investigated for the association of hypersensitivity to trichophytin and the isolation of dermatophytes from clinically normal sites with chronicity and recurrence of infection. At the end of six months following specific therapy, 24 patients returned for follow up and they were similarly studied. Dermatophytes were isolated from clinically asymptomatic sites in 46% patients before treatment and in 21% of the patients on follow up. Immediate weal reaction and increased concentration of IgE antibodies were seen in 73% and 80% of the patients respectively. However, the delayed hypersensitivity reaction was more associated with patients having lesions for more than 6 months (48%) in comparison with patients with a short history (17%). On follow up after 6 months, the different hypersensitivity reactions and IgE antibody concentration maintained more or less the same association. Therefore in persistent or recurrent Tinea cruris infection, besides potential carriage in clinically normal sites, hypersensitivity to antigens of dermatophytes possibly plays an important role in pathogenicity.
Adolescent↗
Ganglioside-like epitopes of lipopolysaccharides from Campylobacter jejuni (PEN 19) in three isolates from patients with Guillain-Barré syndrome.
Sera from patients with Guillain-Barré syndrome (GBS) frequently have anti-GM1 antibody. We earlier showed that an lipopolysaccharides (LPS) from Campylobacter jejuni (PEN 19) isolated from a GBS patient has a GM1 ganglioside-like structure. Aspinall et al. (Biochemistry, 61 (1994) 335-337) reported that OH 4382 has an LPS that bears a CD3 ganglioside-like structure and that OH 4384 has an LPS that bears a GT1a-like structure; both strains were isolated from patients with GBS. They also suggested a GM1-like structure is present in the LPSs from OH 4384, but failed to show the presence in the LPSs from OH 4382. To clarify the pathogenesis of GBS after infection by C. jejuni (PEN 19), we investigated the carbohydrate structures of the three strains by thin-layer chromatography immunostaining with cholera toxin and monoclonal anti-ganglioside antibodies. We found that both OH 4382 and OH 4384 have an LPS with the GM1 epitope as well as one with the GT1a or GD3 epitope.
Antibodies, Bacterial↗
Ganglioside characterization of a cell line displaying motor neuron-like phenotype: GM2 as a possible major ganglioside in motor neurons.
We have examined ganglioside compositions and the presence of sulfated glucuronyl glycolipids of immortalized motor neuron-like cell lines, neuroblastoma-spinal cord (NSC) hybrid cell lines established by fusing mouse neuroblastoma N18TG2 with motor neuron-enriched embryonic spinal cord cells. Among NSC cell lines, only NSC-34 aggregates acetylcholine receptors on co-cultured myotube and expresses a receptor for S-laminin, a neuromuscular junction specific basal lamina protein. GM2, which is only a minor ganglioside component of CNS, was the major component in NSC-34 occupying almost 75% of total gangliosides, whereas GD1a and GM3 were major species in the parental N18TG2, which had only 8.5% GM2. These results indicated that NSC lines have unique ganglioside pattern that is distinctive from other nervous tissues, and this pattern, especially that of NSC-34 cells, might reflect the characteristics of mouse spinal motor neuron gangliosides. Sulfated glucuronyl paragloboside was demonstrated to be present in N18TG2, however, it could not be detected in either of NSC cell lines. Even though the pathogenesis of amyotrophic lateral sclerosis remains unknown, autoimmunological participation has been suggested. Because high-titered antibody against GM2 has been observed in a patient with amyotrophic lateral sclerosis-like disease, GM2 which is possibly expressed on the surface of motor neurons might serve as a potential target antigen in this disorder.
Animals↗
Preinfarction angina as a major predictor of left ventricular function and long-term prognosis after a first Q wave myocardial infarction.
OBJECTIVES: The purpose of this study was to assess the prognostic significance of preinfarction angina after a first Q wave myocardial infarction. Patients with anterior or inferior myocardial infarction were compared. BACKGROUND: The effect of preinfarction angina on prognosis after anterior and inferior myocardial infarction remains unclear. METHODS: A total of 291 patients with a first Q wave anterior (n = 171) or inferior (n = 120) myocardial infarction were examined to assess the effect of preinfarction angina on short- and long-term prognosis. The relation between predischarge left ventriculographic findings and preinfarction angina was also examined. RESULTS: The presence of preinfarction angina was associated with lower peak creatine kinase activity, a lower in-hospital incidence of sustained ventricular tachycardia and fibrillation and a lower incidence of pump failure and cardiac mortality in patients with either anterior or inferior infarction. Among patients with anterior infarction, preinfarction angina was associated with a lower incidence of cardiac rupture and less need for readmission for heart failure within 1 year after the onset of infarction. In this subgroup it was also associated with a higher ejection fraction, a smaller end-diastolic volume and a lower incidence of aneurysm formation noted on ventriculography during convalescence. In patients with inferior infarction, these variables did not differ significantly in the presence or absence of preinfarction angina. Multivariate analysis confirmed that the presence of preinfarction angina was an independent predictor of development of ventricular aneurysm, late phase heart failure and 1-year cardiac mortality. CONCLUSIONS: The presence of preinfarction angina has a favorable effect on infarct expansion and late phase left ventricular function, especially in patients with anterior myocardial infarction. The mechanisms responsible for this phenomenon are not known but may be secondary to limitations of infarct size through unidentified mechanisms other than collateralization (e.g., ischemic preconditioning).
Adult↗
An alternative limb lead system for electrocardiographs in emergency patients.
It is occasionally difficult to record the standard 12-lead electrocardiograph (ECG) in emergency patients. The aim of this study was to evaluate the influence on electrocardiographic wave form recordings of moving the location of electrodes from the standard limb lead position to the trunk. The participants were 10 normal subjects and 20 patients with heart disease. In the new lead system, the limb electrodes were placed on the anterior acromial region and the anterior superior iliac spine using adhesive electrodes. Conventional 12-lead ECGs were recorded by the standard and the new lead system simultaneously in the supine position. Wave form analysis was done by an automatic analysis program. Motion artifacts in the recordings were less in the new lead system. The R wave amplitude of the new lead system increased in leads II, III and aVF, and decreased in leads I and aVL. However, the amplitudes of each wave obtained by standard electrocardiography and the new lead system correlated well (y = 1.008x + 2.038, r = 0.99, n = 2,880). In 99.6% of all wave forms, the differences in amplitudes were within 5% of the values of standard recordings. The average of differences in the ST-segment was 2.6 +/- 11.4 microV. The frontal plane QRS axis obtained by the new lead system showed a vertical shift of 7.8 +/- 8.5 degrees (y = 0.94911x + 10.346, r = 0.98, n = 30). The recording errors produced by the new lead system were within the permissible range of variation. The new lead system is a reasonable alternative for recording ECGs if application of the standard lead is difficult in an emergency.
Adult↗
Production of monoclonal antibodies directed to Hanganutziu-Deicher active gangliosides, N-glycolylneuraminic acid-containing gangliosides.
We have established three kinds of monoclonal antibodies against gangliosides containing N-glycolylneuraminic acid (NeuGc) by immunization of BALB/c mice with the purified gangliosides inserted into liposomes comprising Salmonella minnesota R595 lipopolysaccharides, and fusion of spleen cells with a mouse myeloma cell line. One monoclonal antibody, SHS-1, which was generated by immunizing mice with purified i-active ganglioside(NeuGc), reacted specifically with the i-active ganglioside(NeuGc) used as an immunogen. Structurally related gangliosides, such as GM3(NeuGc), sialosylparagloboside (SPG) (NeuGc), or I-active ganglioside(NeuGc), corresponding gangliosides [GM3 containing N-acetylneuraminic acid (NeuAc), SPG(NeuAc), i-active ganglioside(NeuAc), and I-active ganglioside(NeuAc)], other gangliosides, or neutral glycosphingolipid (GSL) were not recognized by the monoclonal antibody. These findings indicate that the SHS-1 monoclonal antibody may be specific for NeuGc-containing i-active ganglioside. On the other hand, the other two monoclonal antibodies, MSG-1 and SPS-20, which were generated by immunizing mice with purified ganglioside GM3(NeuGc) and SPG(NeuGc), respectively, showed cross-reactivity to structurally related gangliosides. The MSG-1 monoclonal antibody exhibited reactivity to ganglioside GM3(NeuAc). The SPS-20 monoclonal antibody also cross-reacted with SPG(NeuAc), i-active ganglioside(NeuGc), and i-active ganglioside(NeuAc). Neither MSG-1 nor SPS-20 reacted with corresponding gangliosides, other gangliosides, or neutral GSLs tested. Using the SHS-1 antibody specific for i-active ganglioside(NeuGc), we studied the expression of NeuGc-containing antigen in human colon cancer tissue. An NeuGc-containing glycoconjugate was detected in the colon cancer tissue.
Animals↗
Identification of phosphocholine-containing glycoglycerolipids purified from Mycoplasma fermentans-infected human helper T-cell culture as components of M. fermentans.
Previously, we have reported the occurrence of novel phosphocholine-containing glycoglycerolipids (GGPLs: GGPL-I and GGPL-III) in human helper T-cell (MT-4 cell line) (Mustuda et al, Glycoconjugate J. 10:340). However, the GGPLs disappeared from the MT-4 after treatment with an antimycoplasma agent. This disappearance suggested the involvement of microorganisms in the GGPL expression. In this paper, we show that the novel lipids are components of Mycoplasma fermentans itself. The supernatant fluid of the antimycoplasma agent-untreated Mt-4 cell culture produced mycoplasma-like colonies on PPLO agar plates, and PCR and immunological methods revealed the presence of M. fermentans. GGPLs were expressed again in the treated Mt-4 cells after infection with the isolated M. fermentans. The isolated M. fermentans had glycoglycerolipids corresponding to GGPL-I and GGPL-III. Thin-layer chromatography-mass spectrometry and immunological analyses showed that these glycoglycerolipid which were derived from the isolated M. fermentans were identical with GGPL-I and GGPL-III previously obtained. This is the first report that shows mycoplasma has phosphocholine-containing glycoglycerolipids.
Animals↗
Bilateral symmetrical pilar leiomyomas on the breasts.
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Adult↗
Sarcoidosis presenting as cutaneous hypopigmentation.
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Humans↗
Premature thelarche in Möbius syndrome.
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Breast↗
Nevus depigmentosus associated with hemihypertrophy of the limbs.
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Ear, External↗
Syndromic management of genital ulcer disease--a critical appraisal.
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Algorithms↗
Beta-1-selectivity is not essential to achieve therapeutic efficacy with beta-blockade therapy for idiopathic dilated cardiomyopathy.
This study investigated the therapeutic efficacy of two different beta-blockers, metoprolol (beta 1-selective) and nipradilol (nonselective) for the treatment of idiopathic dilated cardiomyopathy (DCM). The New York Heart Association functional class improved in the metoprolol group (n = 9) and the nipradilol group (n = 9), but not in the control group who received conventional therapy (n = 8). The left ventricular ejection fraction increased in both the beta-blocker groups (p < 0.01, p < 0.05). Lymphocyte beta-adrenoceptors were upregulated in the nipradilol group (p < 0.01). Cardiac events were less common in both the beta-blocker groups than in the control group (both p < 0.05). Thus, nipradilol improved symptoms and cardiac function with a favorable effect on sympathoneuronal activity as well as metoprolol in patients with DCM. Therefore, beta 1-selectivity is not essential to achieve therapeutic efficacy with beta-blockade therapy for DCM.
Adrenergic beta-Antagonists↗