Gene polymorphism and frequencies of the NPC1L1 gene (rs2072183, rs217434 and rs217428) in Japanese patients with dyslipidemia.

WHAT IS KNOWN AND OBJECTIVE: Niemann-Pick C1-Like 1 (NPC1L1) plays a pivotal role in intestinal cholesterol absorption. Ezetimibe is known as an inhibitor for NPC1L1 and decreases concentration of low-density lipoprotein cholesterol (LDL-C) in blood. Responses of the decrease of serum LDL-C levels to ezetimibe have been reported to be different among NPC1L1 variants. However, there are still limited data concerning the genetic variation in the NPC1L1 gene, specifically, in Japanese patients with dyslipidemia. The purpose of this study is to elucidate genotype and allele frequencies of the NPC1L1 gene in Japanese patients with dyslipidemia. METHODS: Written informed consent was obtained from all participants. All patients were administered ezetimibe at the dose of 10 mg for once a day either alone or coadministered with statins. Patient's data were retrospectively obtained from their medical records. Genomic DNA was extracted from whole blood samples and analysed three NPC1L1 SNPs (rs2072183, rs217428 and rs217434) by the direct sequencing method. RESULTS AND DISCUSSION: We found that there is a significant difference of genotype frequencies between healthy Japanese and dyslipidemic subjects in rs2072183. No significant differences were observed in rs217428 and rs217434; however, comparison of our data with literature reports suggests that there are significant differences in the frequencies of rs217428 and rs217434 between Canadian and Japanese dyslipidemic patients. WHAT IS NEW AND CONCLUSION: Our study is the first report concerning the genotype and allele frequencies of the gene coding for NPC1L1 in Japanese patients with dyslipidemia. The most notable result was to demonstrate that there exists a significant difference in rs2072183 variant between healthy Japanese and dyslipidemic subjects and also found that there exists genetic variation of rs2072183 between Japanese and Canadian patients with dyslipidemia. Our results are expected to facilitate research in the proper use of ezetimibe-based mono- or combination therapies. Further studies will be required to evaluate the effects of rs2072183 on the efficacy of LDL cholesterol reduction by ezetimibe.

Abnormal pontine activation in pathological laughing as shown by functional magnetic resonance imaging.

To explore the aetiology of pathological laughing, a 65-year-old woman with pathological laughing was examined by 3-T functional magnetic resonance imaging (fMRI) before and after treatment with drugs. Here, we report that the patient consistently showed exaggerated pontine activation during the performance of three tasks before treatment, whereas abnormal pontine activation was no longer found after successful treatment with the selective serotonin reuptake inhibitor, paroxetine. Our findings in this first fMRI study of pathological laughing suggest that serotonergic replacement decreases the aberrant activity in a circuit that involves the pons.

Amino acid sequence of monkey erythrocyte glycophorin MK. Its amino acid sequence has a striking homology with that of human glycophorin A.

A major sialoglycoprotein, glycophorin MK, was isolated from monkey erythrocyte membranes by extraction with lithium diiodosalicylate and partition in aqueous phenol. Chemical analysis of glycophorin MK revealed that the glycophorin consists of 51% protein and 49% carbohydrate by weight, and contains no N-glycosidic oligosaccharide units. Only N-glycolylneuraminic acid (Neu5Gc) was detected as sialic acid. The amino acid sequence of glycophorin MK was determined, which demonstrated that the glycophorin consists of 144 amino acid residues and 18 oligosaccharide units linked O-glycosidically to the peptide backbone through serine or threonine residues. The molecular weight was estimated to be about 35,000 based on the amino acid residues and carbohydrate content. By comparison of the amino acid sequence with those of human, equine and porcine glycophorins, a striking sequence homology was observed between monkey and human glycophorin. Glycophorin MK demonstrated both M and N blood group activities.

The hemagglutinins of the human influenza viruses A and B recognize different receptor microdomains.

A cryptically I-active sialylglycoprotein (glycoprotein 2) isolated from bovine erythrocyte membranes as Sendai virus receptor (Suzuki, Y., Suzuki, T. and Matsumoto, M. (1983) J. Biochem. 93, 1621-1633) contains N-glycolylneuraminic acid (NeuGc) as its predominate sialic acid and exhibits poor receptor activity for a variety of influenza viruses. Enzymatic modification of asialoglycoprotein-2 to contain N-acetylneuraminic acid (NeuAc) in the NeuAc alpha 2-3Gal and NeuAc alpha 2-6Gal sequences using specific sialyltransferase resulted in the appearance of receptor activity toward human influenza viruses A and B. The biological responsiveness chicken erythrocytes treated with sialidase and then reconstituted with derivatized glycoprotein 2 showed considerable recovery to influenza virus hemagglutinin-mediated agglutination, low-pH fusion and hemolysis. Specific hemagglutination inhibition activity of derivatized glycoprotein 2 was 5-16-times higher than that of human glycophorin. A/PR/8/34 (H1N1) virus preferentially recognized derivatized glycoprotein 2 containing NeuAc alpha 2-3Gal sequence over that containing NeuAc alpha 2-6Gal while the specificity of A/Aichi/2/68 (H3N2) for the sialyl linkages was reversed. B/Lee virus recognized both sequences almost equally. The biological responsiveness to the viruses of the erythrocytes labeled with the derivatized glycoprotein 2 containing NeuGc was considerably lower than that of derivatized glycoprotein 2 containing NeuAc. The results demonstrate that the hemagglutinins of human isolates of influenza viruses A and B differ in the recognition of microdomains (NeuAc, NeuGc) of the receptors for binding and fusion activities in viral penetration and the sequence to which sialic acid (SA) is attached (SA alpha 2-3Gal, SA alpha 2-6Gal). Inner I-active neolacto-series type II sugar chains may be important in revealing the receptor activity toward the hemagglutinin of both human influenza viruses A and B.

Amino acid sequence and oligosaccharide attachment sites of the glycosylated domain of dog erythrocyte glycophorin.

Glycophorin was prepared from dog erythrocyte membranes by extraction with lithium diiodosalicylate and partition in aqueous phenol. Tryptic and chymotryptic treatments of the glycophorin produced two major glycopeptides labeled T1 and CH1, respectively. The glycopeptides were isolated by gel chromatography followed by ion-exchange chromatography, and subjected to amino acid sequence analysis. Both glycopeptides represented the amino-terminal domain of the major dog glycophorin; T1 of 52 residues and CH1 of 43 residues. The amino-terminal sequence of dog glycophorin does not have significant homology with those of human, horse or porcine glycophorins. This result is in good agreement with our previous proposal that there is no homology in the sequence of the amino-terminal glycosylated domain of glycophorin.

Structural changes of rat liver microsomal membranes induced by the oral administration of carbon tetrachloride. 31P-NMR and spin-label studies.

The acute effects of carbon tetrachloride (CCl4) on the membrane structure of rat liver microsomes were studied using 31P-NMR and spin-labeling techniques. 31P-NMR spectra of rat liver microsomes were not changed appreciably after the oral administration of CCl4, indicating that the surface structures of microsomal membranes probably are not influenced by the oral administration of CCl4. Four different spin-labeled stearic acids, 5-(N-oxyl-4',4'-dimethyloxazolidine)-stearic acid (5SLS), its methyl ester (5SLSM), 12-(N-oxyl-4',4'-dimethyloxazolidine)-stearic acid (12SLS) and its methyl ester (12SLSM), were used for the estimation of membrane fluidity. The apparent rotational correlation time of 12SLS decreased from 4.0 nsec to 3.0 nsec after the oral administration of CCl4, while the order parameter of 5SLS did not change. The results suggest that CCl4 or its metabolites increase the membrane fluidity of liver microsomes primarily at hydrophobic regions rather than at the surface layer. The ESR spectrum of 5SLSM in microsomal membranes comprised two different signals; one was an anisotropic signal and the other was a rather isotropic one. The ratio of the anisotropic signal to the isotropic one decreased markedly after the oral administration of CCl4 and depended on the dose of CCl4. The suitability of this ESR technique with 5SLSM for the estimation of membrane damage is discussed.

Lung fibrosis in hypersensitivity pneumonitis. Association with CD4+ but not CD8+ cell dominant alveolitis and insidious onset.

Seventeen cases of hypersensitivity pneumonitis (HP) at a symptomatic phase were categorized into two groups based on computed tomographic (CT) findings and histologic features of transbronchial lung biopsy specimens, HP accompanied by lung fibrosis (fibrosis group), and HP unaccompanied by lung fibrosis (nonfibrosis group). The fibrosis group comprised bird fancier's lung and HP of unknown etiology, whereas the nonfibrosis group mainly comprised summer-type HP. Comparison of results of pulmonary function tests between these two groups confirmed a restrictive impairment in the fibrosis group. Analyses of cellular components of bronchoalveolar lavage (BAL) fluids revealed lymphocytes, especially CD8+ T lymphocytes, were significantly increased in the nonfibrosis group in comparison with the fibrosis group, whereas CD4+ T cells were increased to the same level in the both groups. Analyses of the onset of disease showed that acute onset was observed mainly in nonfibrosis group and strongly correlated with increased CD8+ T lymphocytes in BAL fluids, while insidious onset was related to lung fibrosis and relatively increased CD4+ T lymphocytes in BAL fluids. These findings raise the possibility that highly elevated CD8+ T cells might have a protective effect on pulmonary fibrosis or that relatively increased CD4+ T cells might play an important role in the pathogenesis of pulmonary fibrosis of HP at the chronic phase.

Glycophorins of bovine erythrocyte membranes. Isolation and preliminary characterization of the major component.

Crude glycophorin fraction was prepared from bovine erythrocyte membranes by extraction with lithium diiodosalicylate and partition in aqueous phenol. The crude fraction was further separated into three fractions by gel chromatography and ion-exchange chromatography. The major fraction, designated glycophorin BA, accounts for 80% of the crude fraction. Its carbohydrate content was 79% by weight including galactose, N-acetylgalactosamine, N-acetylglucosamine and N-glycolylneuraminic acid. Iodosobenzoate treatment of glycophorin BA produced two fragments; one is a highly glycosylated segment and the other a hydrophobic peptide. The amino-terminal sequence of the hydrophobic peptide representing the carboxyl-terminal half of glycophorin BA was determined. The other two fractions, IIIa and IIIb, were still heterogeneous when analyzed by gel electrophoresis. Fraction IIIa contained two glycophorins having apparent molecular weights of 46,000 and 42,000, while fraction IIIb contained a glycoprotein having a marked tendency to aggregate.

Isolation and characterization of two glycophorins from horse erythrocyte membranes.

Crude glycophorin fraction was prepared from horse erythrocyte membranes by extraction with lithium diiodosalicylate and partition in aqueous phenol. Two glycophorins, designated glycophorins HA and HB, were isolated by two different techniques: preparative gel electrophoresis in the presence of sodium dodecyl sulfate and ion-exchange chromatography in the presence of the nonionic detergent Ammonyx LO. Each glycophorin formed at least two bands on gel electrophoresis, which corresponded to a dimeric form and a monomeric form. Glycophorin HA, the major component, had a blocked amino-terminus and consisted of 70% protein and 30% carbohydrate. Glycophorin HB, the minor component, had threonine as the amino-terminus and consisted of 80% protein and 20% carbohydrate. Since glycophorin HB showed a chemical composition distinct from that of glycophorin HA, glycophorin HB was not a partially degraded form of glycophorin HA.

Glycophorin-induced differentiation of human myelogenous leukemic cell lines.

Human myelogenous leukemic cell lines, U-937, ML-1, and THP-1, were induced to differentiate into maturing macrophage-like cells upon treatment with any glycophorins from either human, monkey or cow erythrocyte membrane. The differentiated cells expressed lower growth potential, and higher nitroblue tetrazolium-reducing activity, Fc-receptor, phagocytic activity and non-specific esterase activity. The differentiation-inducing activities of the glycophorins were significantly reduced after neuraminidase treatment. On the other hand, sialic acid and colominic acid (a polymer of sialic acid) were inactive. These results suggest that some conformational requirement involving the linkage of sialic acid residues in the glycoproteins is necessary for the observed differentiation-inducing activity.

Role of carbohydrate moiety in granulocyte colony stimulating factor.

Biological activities of two granulocyte-colony stimulating factor (G-CSF) preparations with (Lenograstim) or without (Filgrastim) sugar moiety were compared. Both G-CSF preparations similarly enhanced the N-Formyl-Met-Leu-Phe-induced-migration of human peripheral blood polymorphonuclear cells, but did not significantly affect the proliferation of human oral tumor cell lines (HSC-2, HSG). However, Lenograstim induced cytotoxicity (accompanied by the production of cytoplasmic vacuoles and large DNA fragments) in human promyelocytic leukemic cells HL-60, more potently than Filgrastim. Lenograstim, but not Filgrastim, enhanced the cytotoxic activity of sodium ascorbate. In contrast to Lenograstim, Filgrastim was degraded gradually, but too slowly to explain its lower biological activity. These data suggest that the carbohydrate moiety in G-CSF might confer unique biological activities.

Antitumor activity of polysaccharide fractions from pine cone extract of Pinus parviflora Sieb. et Zucc.

Hot water extract of pine cone (PCE) of Pinus parviflora Sieb. et Zucc. dose-dependently suppressed both solid and ascites tumor cells transplanted into various mice. Acidic polysaccharides of PCE significantly increased the survival time of mice bearing ascites tumor cells, and activity increased with acidity. One of the four polysaccharide fractions obtained by NaOH extraction showed the most potent antitumor activity. This fraction significantly suppressed the growth of solid tumor cells, with occasional tumor regression and necrosis, and with little or no cytocidal effect on cultured tumor cells. All acidic polysaccharides were able to activate mouse macrophage-like cell line J774.1. There did not appear to be any correlation between the antitumor activity of these polysaccharides and their content of arabinose (or fucose), mannose, galactose, glucose, or uronic acid.

Abortion in Japanese cows caused by Chlamydia psittaci.

An outbreak of abortion in cows occurring in Niigata Prefecture was shown to be caused by Chlamydia psittaci. Elementary bodies characteristic of Chlamydia were found in the liver of aborted fetuses and C. psittaci antigen was demonstrated by indirect immunofluorescence. Chlamydia was isolated from the liver of aborted fetuses by the yolk sac inoculation of developing chick embryos and by the intraperitoneal inoculation of guinea pigs. Abortion occurred mostly in middle or late pregnancy. Aborted fetuses showed subcutaneous edema and gelatinous infiltration, enlarged liver and spleen, and dark red pleural and ascitic fluid. Focal necrosis was shown in the liver, spleen and lymph nodes. Serological findings and isolation of Chlamydia from fecal specimens indicated a wide dissemination of C. psittaci among cows in the area.

Coronary endothelial damage during off-pump CABG related to coronary-clamping and gas insufflation.

OBJECTIVE: Although off-pump coronary artery bypass grafting (CABG) has been recognized less invasive than conventional CABG on cardiopulmonary bypass, off-pump CABG may be partly invasive especially to the coronary endothelium. The present study was designed to evaluate the adverse effects of coronary snaring with looped sutures and gas insufflation on the coronary endothelium. The protective efficacies on the coronary endothelium of coronary snaring with elastic sutures or humidified gas insufflation with/without heparin and dipyridamole-added were also tested. METHODS: Thirty-six mongrel dogs were used. After systemic heparinization (150 U/kg), a 5 mm longitudinal coronary incision was made with looped non-elastic monofilament sutures or elastic sutures applied proximally and distally. The incised coronary artery was exposed to non-humidified carbon dioxide, humidified carbon dioxide with lactated Ringer solution, or humidified carbon dioxide with heparin and dipyridamole-added lactated Ringer solution for 10 or 20 min in each group. After gas insufflation, the incised coronary artery was repaired, then, the coronary was reperfused. Perfusion-fixation was done for observation of the coronary endothelium by scanning electron microscopy. The adverse effect on the endothelium was graded as follows: grade 1, appeared normal; grade 2, few blood cells deposited; grade 3, many blood cells deposited; grade 4, few endothelial cells delaminated with blood cells deposited; grade 5, many endothelial cells delaminated with blood cells deposited. RESULTS: Non-elastic looping caused much more endothelial tears than elastic looping (P<0.00001). Non-humidified gas blowing for 20 min caused more endothelial cell damage than humidified gas blowing (P=0.00005). Non-humidified gas blowing for 10 min caused less damage than for 20 min (P=0.00326), but still caused more damage than humidified gas blowing (P=0.00253). Heparin and dipyridamole-added humidification reduced coronary endothelial area mottled by the deposited cells when compared with simple humidification (P=0.00120). CONCLUSIONS: Coronary snaring resulted in coronary endothelial injury, which was ameliorated by using elastic sutures instead of non-elastic sutures. Non-humidified gas insufflation made blood cells deposited and endothelial cells delaminated with time. Humidified gas insufflation attenuated these adverse effects. Heparin and dipyridamole-added humidification had potential advantage in terms of reducing deposited blood cells on the endothelium over simple humidification.

Surgery of the dissecting aneurysm involving a right aortic arch.

A dissecting aneurysm in association with a right aortic arch is extremely rare. However, a 50-year-old male was diagnosed as having a dissecting aneurysm (DeBakey IIIa) with a right aortic arch, right descending aorta and an aberrant retro-esophageal left subclavian artery. A graft replacement of the right descending aorta was successfully performed under right thoracotomy and partial cardiopulmonary bypass. Precise anatomical definition and proper surgical procedure permitted a successful surgical result.

Pathology of bovine abortion and newborn calf death caused by dual infection with Chlamydia psittaci and infectious bovine rhinotracheitis virus.

Nine aborted fetuses and one newborn calf, diagnosed as Chlamydia psittaci (C. psittaci) infection, were pathologically examined. The characteristic lesions in the liver were focal necrosis in 9 aborted fetuses and granulomatous necrotic foci in the newborn calf. Moderate numbers of intranuclear inclusion bodies were found in necrotic foci of the liver, spleen, thymus, lymph nodes, adrenal gland, kidney, lung and forestomach. Immunohistologically, a small number of C. psittaci antigens was demonstrated in necrotic foci of the liver and correlated with distribution of elementary bodies. Moderate numbers of infectious bovine rhinotracheitis (IBR) virus antigens were also detected in degenerating and necrotizing parenchymal cells in various organs and correlated with distribution of intranuclear inclusion bodies. Thus, these aborted bovine fetuses and newborn calf were interpreted as being dually infected with C. psittaci and IBR virus during pregnancy.

Pulmonary alveolar proteinosis. Xe-133 scintigraphic findings before and after bronchopulmonary lavage.

A 44-year-old woman with pulmonary alveolar proteinosis was treated by repeated unilateral bronchopulmonary lavages over 6.5 years. The effectiveness of the treatment was assessed by Xe-133 scintigraphy as well as lung function tests and chest roentgenograms. Xe-133 scintigraphy clearly demonstrated improvement of regional ventilation and perfusion, and equalization of the ventilation and perfusion ratio. Therefore, Xe-133 scintigraphy was found to be useful in the analysis of changes in regional aeration, ventilation, and perfusion before and after bronchopulmonary lavage in pulmonary alveolar proteinosis.

[A case of the ascending aorta and aortic arch replacement with thrombo-occlusion of distal arch and descending aorta surround the modified elephant trunk graft].

We report a 65-year-old female who had a extensive thoracic aneurysm from ascending aorta to descending thoracic aorta. The patient underwent a graft replacement of ascending aorta and aortic arch using modified elephant trunk method. The surgery was carried out through median sternotomy with profound hypothermia and selective cerebral perfusion. Postoperatively, the patient was recovered without any complications except bronchial asthma. Postoperative chest computed tomography showed that the surrounding space of the elephant trunk vascular graft inserted into distal arch and descending aneurysm was mostly occupied with thrombus. Therefore, we considered that the second operation on the descending aorta is not necessary at this point and careful attention to the size and shape of the descending aneurysm should be paid.

[Blood concentrations of thrombomodulin in patients with various diseases].

Concentrations of thrombomodulin in blood plasma were measured by a one-step sandwich enzyme immunoassay (EIA). The concentrations in normal healthy subjects were 9.9 +/- 2.9 ng/ml. The concentrations were found to be significantly higher in patients with SLE, RA and other collagen diseases in their active stages than at their non-active stages. The concentrations increased in patients with DIC, and significantly higher levels were observed when DIC was complicated by multiple organ failure. These findings indicate that plasma concentrations of thrombomodulin may be a useful parameter for vascular injuries caused by inflammatory processes or coagulation/fibrinolysis reactions.

[A successful case report of emergency coronary artery bypass grafting for left main trunk obstruction with profound cardiogenic shock: bridge use of balloon pumping and percutaneous cardiopulmonary support system to surgical intervention].

A 73-year-old man was admitted for sudden onset of dyspnea with cardiogenic shock. Chest X-P showed bilateral severe pulmonary edema. Echocardiogram demonstrated diffuse severe hypokinesis of left ventricle. Emergency coronary angiography showed 99% stenosis of left main trunk with delay. After insertion of intraaortic balloon pump (IABP) and percutaneous cardiopulmonary support system (PCPS), vital signs and consciousness were improved. Emergency coronary artery bypass grafting (CABG) was performed, LIMA to left anterior descending artery, saphenous vein grafts to circumflex and right coronary artery. Left ventricular wall motion was improved after bypass grafting and cardiopulmonary bypass was weaned with catecholamine and IABP support. The patient was discharged from hospital 60 days after the operation with good cardiac function. We emphasized a usefulness of combined use of IABP and PCPS to provide systemic organ perfusion and reduce myocardial infarct size and ischemic damages after re-vascularization for coronary insufficiency with profound shock.