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Biomedical subjects

J Kobayashi

Publications and source records attributed to J Kobayashi.

At least 145 records · Page 8Linked to original sources

Delayed post-prandial lipid metabolism in subjects with intra-abdominal visceral fat accumulation.

BACKGROUND: Individuals with obesity, in particular those with intra-abdominal visceral fat accumulation, are known to have various complications, such as hyperlipidaemia, impaired glucose tolerance, hyperinsulinaemia and hypertension, leading to the development of coronary heart disease. Post-prandial hyperlipidaemia has repeatedly been shown to be an independent risk factor for coronary heart disease. The aim of the present study was to investigate post-prandial lipoprotein metabolism in subjects with excessive visceral fat accumulation. MATERIALS AND METHODS: Eighty-three patients (52 men, 31 women) [average age 48 +/- 14 years; mean body mass index (BMI) 25 +/- 5 kg m-2] were recruited to the study. Visceral (or subcutaneous) fat accumulation was analysed as areas of fat deposition by computerized tomography at the umbilicus level. After a 12-h overnight fast, oral vitamin A and a fatty meal (40 g m-2 fresh cream containing 50 000 units m-2 vitamin A) were administered to these subjects. The concentration of retinyl palmitate (RP) was measured by high-performance liquid chromatography. RESULTS: The visceral fat area (V) was positively correlated with plasma triglyceride (TG) 0, 2, 4 and 6 h after fat loading and with plasma RP 0, 4 and 6 h after fat loading. The BMI did not show any correlation with plasma TG and RP at any point. The visceral fat area was positively correlated with the RP area under the curve (AUC) in the serum from the subjects [V vs. RP AUC: n = 83, r = 0.327, P = 0.013]. The BMI of the subjects did not show any correlation with the RP AUC (r = 0.021, P = 0.85). CONCLUSION: These results suggest that post-prandial lipid metabolism is impaired in subjects with intra-abdominal visceral fat accumulation, irrespective of BMI, leading to the development and progression of coronary atherosclerosis.

Abdomen↗

Eicosapentaenoic acid modulates arachidonic acid metabolism in rat alveolar macrophages activated by silica.

Eicosapentaenoic acid (EPA) was used to modulate the activation of alveolar macrophages, to examine its potential anti-inflammatory effect in addition to its anti-arteriosclerotic or anti-thrombotic effects. Wistar strain rat alveolar macrophages (2 x 10(6) cell) obtained by bronchoalveolar lavage were preincubated with EPA (0-20 microM), and further incubated with 1 mg of silica for 90 min. Leukotriene (LT) B4 and LTB5 of the supernatant were analyzed by reverse phase HPLC. EPA inhibited the production of LTB4 dose-dependently. The production of LTB5, a metabolite from EPA, was increased at low concentrations of EPA (0-10 microM) and decreased at high concentrations (>10 microM). These results suggest that EPA is competitive with arachidonic acid (AA) at low concentrations, and that EPA may inhibit AA metabolism via inhibition of 5-lipoxygenase or phospholipase A2 at high concentrations.

Animals↗

Modification of lipoprotein lipase catalytic activity by sialic acids.

The role of sialic acid linked with lipoprotein lipase (LPL) in its catalytic activity was studied. When LPL was treated with sialidase, the molecular weight decreased by 2000. The sialidase-treated LPL showed unchanged hydrolyzing activity for tributyrin, a water-soluble substrate of esterase, compared with the untreated LPL. The sialidase-treated LPL also showed similar hydrolyzing activity for triolein emulsified with Triton X-100, phosphatidylcholine and phosphatidylethanolamine, whereas it showed significantly increased hydrolyzing activity for triolein emulsified with phosphatidylserine and cardiolipin (152% and 183%, compared with untreated LPL, respectively). In addition, the sialidase-treated LPL showed significantly increased hydrolyzing activity against triolein incorporated into very low-density lipoproteins and chylomicrons (151% and 186%, compared with the untreated LPL, respectively). These results suggest that the loss of sialic acids does not modify the function of the catalytic site of LPL, but facilitates the interaction of the enzyme with the interface of the surface of substrate lipoproteins.

Animals↗

A general method for rapid purification of soluble versions of glycosylphosphatidylinositol-anchored proteins expressed in insect cells: an application for human tissue-nonspecific alkaline phosphatase.

A soluble form of tissue-nonspecific alkaline phosphatase was purified to apparent homogeneity from the culture media of Sf9 cells which had been infected with recombinant baculoviruses encoding human tissue-nonspecific alkaline phosphatase (TNSALP). To facilitate purification, an oligonucleotide consisting of 6 tandem codons for histidine and a stop codon was engineered into the TNSALP cDNA. The molecular mass of the enzyme purified through a nickel-chelate column was estimated to be 54 kDa by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. That of the native enzyme was 90 kDa as estimated by gel filtration, indicating that the purified soluble TNSALP is dimeric. The enzyme was used for production of antibodies specific for human TNSALP. Immunoblotting analysis showed a single 80-kDa band in the cell homogenate prepared from Saos-2 (human osteosarcoma) cells. However, upon digestion with peptide: N-glycosidase F, the 80-kDa TNSALP of human origin and the soluble enzyme of insect origin migrated to the same position on SDS-polyacrylamide gel, indicating that the size difference between the two enzymes is ascribed to N-linked oligosaccharides. The antibodies prepared against the purified TNSALP were found to be useful also for immunoprecipitation and immunofluorescence studies.

Alkaline Phosphatase↗

Activin from secondary follicles causes small preantral follicles to remain dormant at the resting stage.

The purpose of the present study was to investigate 1) whether activin A can cause primary follicles to become dormant at the resting stage, and 2) the role of the secondary follicle on follicular growth of primary follicles. Preantral follicles (100-120 microm in diameter) harvested from adult mice and cultured in in vitro follicle culture system showed a significant increase in size and estrogen and inhibin secretion in response to FSH, but the administration of activin A blocked the effect of FSH. Withdrawal of activin A not only restored the follicular response to FSH but also enhanced the effect of FSH, indicating that the action of activin A is to cause small preantral follicles to become dormant at the preantral stage. To investigate the role of secondary follicles in early folliculogenesis, small preantral follicles were cocultured with secondary follicle (300-350 microm in diameter) in the presence of FSH. The secondary follicle showed a significant increase in follicular diameter as a result of stimulation by FSH, but the small preantral follicles did not increase in size. After removal of the secondary follicle, however, the small preantral follicles commenced follicular growth, indicating that the growth of small preantral follicles is suppressed by the secondary follicle. Administration of the activin binding protein follistatin caused a significant increase in follicular diameter of both small preantral and secondary follicles as a result of stimulation by FSH. These results have suggested that secondary follicles cause primary follicles to become dormant at the resting stage by secreting activin.

Activins↗

A comparative study on transforming growth factor-beta and activin A for preantral follicles from adult, immature, and diethylstilbestrol-primed immature mice.

Both transformation growth factor-beta (TGFbeta) and activin belong to the TGFbeta superfamily, and each receptor is structurally related. We have shown that the action of activin A on folliculogenesis is different in immature and adult mice, so it is of interest to study whether TGFbeta has such an action on follicular development. The effect of TGFbeta on folliculogenesis was studied in isolated preantral follicles from immature, adult, and diethylstilbestrol (DES)-primed immature mice and was compared with that of activin A. TGFbeta caused a significant increase in follicular diameter and estradiol and immunoreactive inhibin secretion in adult mice in a dose-related manner, but did not affect the size of preantral follicles from immature mice. Activin A, on the other hand, caused a significant increase in the size of follicles from immature mice, but did not change the size of preantral follicles from adult mice. TGFbeta enhanced the effect of FSH, whereas activin A completely blocked the action of FSH on preantral follicles from adult mice. Such a specific action of TGFbeta and activin A was age dependent because preantral follicles obtained from 28-day-old mice, compared with those from 11- and 56-day-old mice, showed an intermediate reaction to TGFbeta and activin A. DES pretreatment of 11- and 28-day-old mice caused an enhanced response to FSH, but this response was completely inhibited by TGFbeta. These results indicate that both TGFbeta and activin A have proliferative action and cytodifferentiative action on granulosa cells, but the action of each is age dependent and opposite in direction. In conclusion, although both TGFbeta and activin A belong to the same family, and each receptor is structurally related, both share a specific role in early folliculogenesis before and after puberty.

Activins↗

Brasilicardin A, a new terpenoid antibiotic from pathogenic Nocardia brasiliensis: fermentation, isolation and biological activity.

A novel tricyclic diterpenoid antibiotic, brasilicardin A, was isolated from the culture broth of Nocardia brasiliensis IFM 0406. The antibiotic exhibited immunosuppressive activity in a mouse mixed lymphocyte reaction (MLR) assay system and its IC50 value was 0.057 microg/ml. Although the inhibitory activity of cyclosporin A (CyA) against IL-2 production was confirmed in the MLR assay system, brasilicardin A did not have the activity. The results of in vitro toxicity testing of brasilicardin A against various human cell lines were compared with those of CyA.

Aminoglycosides↗

[Single-dose and high-volume Bretschneider cardioplegic solution for congenital heart surgery].

Bretschneider cardioplegic solution is used widely in Europe. The aim of this study is to investigate the efficacy of Bretschneider cardioplegic solution for open heart surgery of congenital heart disease in comparison with blood cardioplegia. From June 1995 to July 1997, we treated 32 congenital heart disease patients using Bretschneider cardioplegic solution and 20 patients using blood cardioplegia. Hospital mortality, water balance during operation, percentage of arrhythmia, and intubation time were not significant in both group. Also CPK and CPK-MB were not significant in both group. Bretschneider cardioplegic solution had preserved the heart as same as blood cardioplegia. This is a very convenient method so that we can use only one time infusion.

Cardiac Surgical Procedures↗

[Changes in nitric oxide-related compounds in endotoxemic rats].

NO-related compounds in the blood of septic rats were examined by chemiluminescence method. Rats were treated with 5 mg/kg or 10 mg/kg lipopolysaccharides (LPS) and followed for 30 h. We have determined and evaluated the concentrations of the total NO-related compounds in whole blood (Total-NO), those in plasma (Plasma-NO) and membrane bound-NO (Membranous-NO) in erythrocytes from the LPS-challenged rats. 1) Levels of Plasma-NO after 3 h and Membranous-NO after 7 h in the rats were significantly increased, peaked at 14 h, and then restored to control levels at 30 h. 2) At 7 h in the rats, latter NO levels were exhibited within 10 nM/mg protein. 3) The existence of Membranous-NO can be confirmed by the in vitro experiments of rat erythrocytes incubating with different NO-donors, NOC-3 or SIN-1. 4) Taken together, the determination of Membranous-NO may be an additional marker for pathophysiological states of septic stress.

Animals↗

[Mucosa-associated lymphoid tissue lymphoma with Sjögren's syndrome].

A 68-year-old woman presented with Sjögren's syndrome. Chest X-ray films disclosed consolidated shadows in the right S2 and an infiltration shadow in the right S8 with small nodules. Pathological examination of transbronchial lung biopsy (TBLB) specimens revealed lymphocytic infiltrations that stained positive with UCHL-1 and L 26 in immunohistochemical studies. Lung tissue specimens obtained by video-assisted thoracic surgery showed lympho-epithelial lesions with dense lymphocytic infiltration. Southern blot hybridization and polymerase chain reaction (PCR) assays demonstrated monoclonality and immunoglobulin heavy chain gene rearrangement. These findings yielded a diagnosis of mucosa-associated lymphoid tissue (MALT) lymphoma. The detection of rear-ranged genes encoding for immunoglobulin heavy chains is useful for the diagnosis of primary pulmonary lymphoproliferative disorders, especially malignant lymphomas.

Aged↗

[Maze procedure for a lone atrial fibrillation].

Between 1992. 2 and 1997. 12, the maze procedure for lone fibrillation or flutter (lone af) was performed in 8 patients including 2 patients with a sustained atrial fibrillation, 5 patients with a paroxysmal atrial fibrillation and 1 patient with a sustained atrial flutter. All patients had suffered from a drug resistance lone af which induced sever symptom that is the dyspnea, palpitation and fatigue. Therefore patients requested to receive the maze procedure, and they agreed with the informed concent. After the maze operation, the normal sinus rhythm was recovered in 7 of 8 patients (87.5%). In only 1 patient, a paroxysmal atrial fibrillation remained but his symptoms improved after surgery. So this operation is a good choice of a treatment for a drug resistance lone af.

Adult↗

The effect of Lazaroid U-74389G on extended liver resection with ischemia in dogs.

BACKGROUND: In liver surgery, total clamping of the portal triad (Pringle's procedure) is commonly used, and it sometimes causes liver failure. This study was designed to evaluate the effect of Lazaroid U-74389G (LAZ-G), which inhibits iron-dependent lipid peroxidation, on ischemia-reperfusion injury during liver resection in dogs. METHODS: The experiment animals were divided into 2 groups. The control group was subjected to 60 minutes of warm ischemia by partial inflow occlusion. The LAZ-G-treated group received LAZ-G before ischemia and then underwent liver ischemia. After reperfusion, the nonischemic lobes were resected, and the remnant liver function was evaluated. RESULTS: The LAZ-G-treated group showed a significantly improved animal survival rate. Biochemical analysis and morphologic evaluation by electron microscopy suggest that LAZ-G pretreatment protects both hepatic parenchymal cells and sinusoidal endothelial cells from ischemia-reperfusion injury. Expression of IL-1 beta messenger RNA in hepatic venous blood was measured by a reverse transcriptase-polymerase chain reaction; it was shown to be inhibited in the LAZ-G-treated group after reperfusion. This suggests that LAZ-G decreases the activation of proinflammatory cytokine expression. CONCLUSIONS: Lazaroid U-74389G ameliorates ischemia-reperfusion injury caused by Pringle's procedure during extensive liver resection. This agent may therefore be clinically applicable for extended liver surgery involving vascular isolation.

Animals↗

Plasmodium ovale infections detected by PCR assay in Lao PDR.

Plasmodium ovate infection was demonstrated in 5 out of 143 inhabitants in a village in Lao PDR by blood microscopy and PCR assay. Although the specimen confirmed to be positive for P. ovale by microscopical examination was only one, the target sequences in the 18S rRNA genes of malaria parasite detected in all of the five cases were consisted with those of P. ovale by the PCR assay. This is the first report concerning the presence of so many cases with P. ovale infection in Lao PDR.

Animals↗

Maze procedure for atrial fibrillation associated with atrial septal defect.

BACKGROUND: Atrial fibrillation (AF) is a relatively common complication in elderly patients with an atrial septal defect (ASD). However, use of the simultaneous maze procedure for AF associated with ASD remains controversial. We examined the efficacy and risk of the use of the maze procedure in these patients. METHODS AND RESULTS: Between March 1992 and April 1997, 26 patients underwent the maze procedure as a concomitant operation with ASD closure (maze group). Kosakai's modified maze procedure was performed in 17 patients, the modified Cox maze II or III procedure was performed in 6, and the restrictive right-sided maze procedure was performed in 3. The mean +/- SD age at surgery was 58.2 +/- 9.1 years. The mean +/- SD duration of AF was 7.8 +/- 8.5 years. The mean +/- SD left atrial dimension was 47 +/- 9 mm, and 24 patients (92%) had a larger-than-normal (> 35 mm) left atrium. The mean +/- SD follow-up period was 2.7 +/- 1.7 years. There were no hospital or late deaths. There was no thromboembolic episode in the late follow-up period. Sinus rhythm was regained in all patients who underwent conventional the right- and left-sided maze procedure except for 1 patient. However, 3 patients who underwent the restrictive right-sided maze procedure showed a return to AF rhythm. The atrial A wave was detected with the use of pulsed Doppler study in all patients who had a restored sinus rhythm. During the same period, 45 patients who were > 40 years old and without AF underwent only ASD closure (control group). The incidence of reopening the chest for bleeding was significantly (P = 0.046) higher in the maze group (12%) than in the control group (0%). Paroxysmal AF more frequently (P = 0.023) occurred in the control group (18%) than in the maze group (0%). CONCLUSIONS: These results suggest that the standard maze procedure should be considered in patients with AF associated with ASD. The restrictive right-sided maze procedure was not reliable, probably due to preoperative enlargement of the left atrium.

Aged↗

9-Methyl-7-bromoeudistomin D induces Ca2+ release from cardiac sarcoplasmic reticulum.

9-Methyl-7-bromoeudistomin D (MBED), the most powerful caffeine-like releaser of Ca2+ from skeletal muscle sarcoplasmic reticulum, induced Ca2+ release from the cardiac sarcoplasmic reticulum. MBED (5 microM) and caffeine (1 mM) caused rapid Ca2+ release from the fragmented cardiac sarcoplasmic reticulum in a Ca2+ electrode experiment. [3H]MBED bound to a single class of high-affinity binding sites in cardiac sarcoplasmic reticulum membranes (Kd = 150 nM). These results suggest that MBED binds to a specific binding site on cardiac sarcoplasmic reticulum membranes to induce Ca2+ release from the cardiac sarcoplasmic reticulum. Thus, MBED is a useful probe for characterizing Ca2+ release the channels not only in skeletal sarcoplasmic reticulum but also in cardiac sarcoplasmic reticulum.

Animals↗

The optical activity of lysozyme crystals.

The components of the gyration tensor of the enzyme lysozyme were measured by using the HAUP method: g11 = -0.90 x 10(-5) and g33 = 1.05 x 10(-5) at 303.4 K and a wavelength of 4880 A. The optical rotatory powers along the a and c axes in the same conditions were calculated: rho a = -21.3 and rho c = -24.8 degrees cm-1. The optically active property of lysozyme is strange in that, although it contains a considerable quantity of alpha-helices (about 30%), the rotatory powers are unexpectedly small in magnitude, one order of magnitude less than those of quartz and with very large anisotropy. A conceptual consideration of this phenomenon is given. In order to assess the difference between the structures in both crystalline and solution states, the chirality index r was calculated to be 0.16. This value indicates that the structural change of lysozyme from the solution into the crystalline state is expressed by an increase of 19% in optical activity. From the NMR results [Smith et al. (1993), J. Mol. Biol. 229, 930-944], it is anticipated that the r value reflects the increased constraint in atomic motion in the side chains of exposed amino acid residues in the crystalline state.

Animals↗

Nucleotide sequence of Bombyx mori cytoplasmic polyhedrosis virus segment 8.

The segments 8 (S8) of the 10 double-stranded RNA genomes from Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) strains I and H were converted into cDNAs, amplified by PCR, and cloned. The nucleotide sequence analysis of the two full-length S8 cDNAs showed that the segments consist of 1328 nucleotides encoding putative proteins (p44) of 390 amino acids with molecular masses of about 44 kDa, which have glutamic acid-rich and proline-rich domains in their central regions. They had quite high identity with each other: about 98% in nucleotide and amino acid sequences. The recombinant p44 expressed in BmN4 cells using the baculovirus vector was detected by immunoblot analysis. p44 was also confirmed with the same antiserum to be present in BmCPV-infected midgut cells, but not in polyhedra, virus virions and uninfected midgut cells, indicating that p44 is expressed as a nonstructural protein of BmCPV.

Amino Acid Sequence↗