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Biomedical subjects

T Baba

Publications and source records attributed to T Baba.

At least 73 records · Page 4Linked to original sources

Construction of a contiguous 874-kb sequence of the Escherichia coli -K12 genome corresponding to 50.0-68.8 min on the linkage map and analysis of its sequence features.

The contiguous 874.423 base pair sequence corresponding to the 50.0-68.8 min region on the genetic map of the Escherichia coli K-12 (W3110) was constructed by the determination of DNA sequences in the 50.0-57.9 min region (360 kb) and two large (100 kb in all) and five short gaps in the 57.9-68.8 min region whose sequences had been registered in the DNA databases. We analyzed its sequence features and found that this region contained at least 894 potential open reading frames (ORFs), of which 346 (38.7%) were previously reported, 158 (17.7%) were homologous to other known genes, 232 (26.0%) were identical or similar to hypothetical genes registered in databases, and the remaining 158 (17.7%) showed no significant similarity to any other genes. A homology search of the ORFs also identified several new gene clusters. Those include two clusters of fimbrial genes, a gene cluster of three genes encoding homologues of the human long chain fatty acid degradation enzyme complex in the mitochondrial membrane, a cluster of at least nine genes involved in the utilization of ethanolamine, a cluster of the secondary set of 11 hyc genes participating in the formate hydrogenlyase reaction and a cluster of five genes coding for the homologues of degradation enzymes for aromatic hydrocarbons in Pseudomonas putida. We also noted a variety of novel genes, including two ORFs, which were homologous to the putative genes encoding xanthine dehydrogenase in the fly and a protein responsible for axonal guidance and outgrowth of the rat, mouse and nematode. An isoleucine tRNA gene, designated ileY, was also newly identified at 60.0 min.

Base Sequence

New assay for glycated lipoproteins by high-performance liquid chromatography.

We developed an automated analytic system for glycated lipoprotein using high-performance liquid chromatography (HPLC) with an affinity boronate column and a gel permeation column. This system can measure glycated lipoprotein (glycated LDL and glycated HDL) in a small (5 microliters) sample of serum in a short time (40 min/sample). Recovery with this system was 92.1%. Therefore a large number of samples can be measured in clinical use. The system should contribute to an elucidation of the role of glycated lipoprotein in atherosclerosis.

Chromatography, High Pressure Liquid

Granuloma formation activity and mycolic acid composition of mycobacterial cord factor.

We analyzed the mycolic acid composition of trehalose 6,6'-dimycolate (TDM) obtained from Mycobacterium, using thin layer chromatography, gas chromatography and gas chromatography-mass spectrometry. Utilizing TDM, whose structure was confirmed, granuloma formation in mice was investigated. All TDM used exhibited considerable granuloma formation activity in the lung and spleen. In particular, TDM from M. bovis showed the greatest activity and toxicity among mycobacterial TDM. We therefore discussed the relationship between the chemical structure and granuloma-forming activity of TDM, especially in relation to the structure of mycolic acid in TDM.

Animals

Dystrophin in rod spherules; submembranous dense regions facing bipolar cell processes.

It is known that the retina contains the protein dystrophin in the ribbon synapse, but the ultrastructural analysis is not yet fully elucidated. Our previous study reported that dystrophin is localized under the rod cell membranes in rat retinas. In the present study, we have investigated the relationship between dystrophin-rich regions of rod cell membranes and other neuronal processes in mouse retinas with a monoclonal antibody raised against the human dystrophin C-terminus. Immunoblotting, immunofluorescence stainings, and immunoelectron microscopy were employed. Immunoblotting analysis indicated that mouse retinas possessed some of the dystrophin isoforms of approximately 260 kDa, 140 kDa, and 70 kDa molecular weight. Confocal images showed a punctate appearance in the outer plexiform layer, as previously described. Immunoelectron microscopy showed that dystrophin immunoreactive products were always observed at submembranous dense regions of the rod spherule abutting bipolar processes. These results suggest that retinal dystrophin may be closely involved in signal transmission from rods to bipolar cells.

Animals

Ubiquitously expressed dynamin-II has a higher intrinsic GTPase activity and a greater propensity for self-assembly than neuronal dynamin-I.

To begin to understand mechanistic differences in endocytosis in neurons and nonneuronal cells, we have compared the biochemical properties of the ubiquitously expressed dynamin-II isoform with those of neuron-specific dynamin-I. Like dynamin-I, dynamin-II is specifically localized to and highly concentrated in coated pits on the plasma membrane and can assemble in vitro into rings and helical arrays. As expected, the two closely related isoforms share a similar mechanism for GTP hydrolysis: both are stimulated in vitro by self-assembly and by interaction with microtubules or the SH3 domain-containing protein, grb2. Deletion of the C-terminal proline/arginine-rich domain from either isoform abrogates self-assembly and assembly-dependent increases in GTP hydrolysis. However, dynamin-II exhibits a approximately threefold higher rate of intrinsic GTP hydrolysis and higher affinity for GTP than dynamin-I. Strikingly, the stimulated GTPase activity of dynamin-II can be >40-fold higher than dynamin-I, due principally to its greater propensity for self-assembly and the increased resistance of assembled dynamin-II to GTP-triggered disassembly. These results are consistent with the hypothesis that self-assembly is a major regulator of dynamin GTPase activity and that the intrinsic rate of GTP hydrolysis reflects a dynamic, GTP-dependent equilibrium of assembly and disassembly.

Adaptor Proteins, Signal Transducing

Cerebral ischemic disorders and cerebral oxygen balance during cardiopulmonary bypass surgery: preoperative evaluation using magnetic resonance imaging and angiography.

We compared the preoperative prevalence of small cerebral infarctions and carotid stenosis to jugular venous oxygen saturation (Sjvo2) during coronary artery bypass grafting (CABG). Sjvo2 served as an indicator of whether cerebral oxygen supply meets demand in patients on cardiopulmonary bypass (CPB). The study population consisted of 121 patients who were either older than 65 yr or had a history of cerebrovascular disease. The patients underwent preoperative cerebral magnetic resonance imaging (MRI) and cervical magnetic resonance angiography (MRA) to detect small cerebral infarctions and carotid artery stenosis. Patients with atherosclerosis of the ascending aorta were identified by intraoperative epiaortic ultrasonography. Liberation of emboli from the aorta in these patients was prevented by modification of the standard operation. From preoperative MRI and MRA, 65 patients (54%) had small cerebral infarctions in the white matter or basal ganglia and nine patients (7%) demonstrated moderate or severe stenosis in the carotid arteries. Thirteen patients (11%) had moderate or severe atheromatous disease of the ascending aorta. The severity of aortic atherosclerosis was significantly correlated with the grade of carotid stenosis (P < 0.05). In patients with small infarctions, Sjvo2 was significantly lower than in patients without infarctions (controls) at initiation of CPB, 30 min after aortic cross-clamping, and during the rewarming period of CPB (P < 0.05). Thus, small cerebral infarctions were not uncommon in elderly patients undergoing CABG. Patients with small cerebral infarctions may be at risk for an imbalance in cerebral oxygen supply and demand during the rewarming period because they are unable to deliver the necessary compensatory blood flow.

Aged

Radial artery diameter decreases with increased femoral to radial arterial pressure gradient during cardiopulmonary bypass.

UNLABELLED: A clinically significant femoral to radial artery pressure gradient sometimes develops during cardiopulmonary bypass (CPB), but the mechanism responsible is not clear. We investigated when the pressure gradient developed and what mechanism could be responsible by comparing mean femoral to mean radial artery pressure and radial artery diameter in 75 male patients undergoing coronary artery bypass grafting. A pressure gradient > or =5 mm Hg (High-P) occurred in 38 patients, and the remaining 37 patients had pressure gradients <5 mm Hg (Low-P) at sternal closure. In High-P group, the pressure gradient was significantly greater (4.8 +/- 3.1 vs 1.0 +/- 3.1 mm Hg; P < 0.001) than in Low-P group, and the ratio of radial artery diameter to the diameter after induction of anesthesia was significantly decreased (0.79 +/- 0.12 vs 0.87 +/- 0.14; P = 0.006) at 5 min after aortic clamping. The pressure gradient and the arterial diameter changes persisted until sternal closure. There was a negative linear correlation between the pressure gradient (deltaP) and the radial artery diameter ratio (D) at sternal closure (D = -15.0deltaP + 16.6, r = 0.39, P < 0.001). In a subgroup of 11 High-P patients, palm temperature was significantly lower (P < 0.05) than that of 11 Low-P patients during and after CPB. We conclude that the femoral to radial artery pressure gradient develops by 5 min after aortic clamping during CPB and persists until sternal closure, and that radial artery constriction could be responsible for the pressure gradient. IMPLICATIONS: A femoral to radial pressure gradient has been observed after cardiopulmonary bypass. Arterial vasodilation and vasoconstriction have been considered as causes for this gradient. We measured radial artery diameter using pulsed Doppler ultrasound and examined radial artery vasodilation versus vasoconstriction as possible mechanisms for the pressure gradient.

Aged

Possible migration of harderian gland immunoglobulin A bearing lymphocytes into the caecal tonsil in chickens.

This study was designed to evaluate the significance of the Harderian gland (HG) in the chicken IgA-production system by investigating the influence of HG on the increase of surface IgA (sIgA)-positive cells in other lymphoid organs and also the possibility of migration of HG lymphocytes into other organs. Non-contact culture of splenic or caecal tonsil (CT) lymphocytes with HG whole cells did not enhance the expression of sIgA. The experiment of HG lymphocyte transfer showed that the transferred HG lymphocytes migrated mainly into the CT in the 3-week-old recipient, and mainly into the CT and bursa of Fabricius (BF) in the 6-week-old recipient. It was also found that the transferred sIgA-positive HG lymphocytes migrated selectively into CT in both 3-week-old and 6-week-old recipients. These results indicated that chicken HG plays a central role not only in governing the local immunity in the eyes and respiratory tract but also as an organ which supplies precursory IgA-producing cells involved in local immunity in the intestine.

Animals

Immunopotentiating activities of polysaccharide fraction from pine seed shell extract.

The function of polysaccharide (PSE) extracted from pine seed shells as a immunopotentiator was investigated. The phagocytosis and chemotaxis of mouse peritoneal macrophages (MP) were augmented by oral administration of PSE. The incorporation of 3H-thymidine by Con A-stimulated mouse splenic cells was significantly intensified by administration of PSE but the adherent-cells (MP)-eliminated splenic lymphocytes was not increased. It is shown that the existence of activated MP is needed for the activation (proliferation) of T lymphocytes. The responsiveness of mouse T cells to alloantigen was also augmented by administration of PSE. The number of anti-SRBC plaque-forming cells in the spleen of mouse and chicken was also significantly increased. The results indicate that MP are first activated by oral administration of PSE and that the activation of T cells is then initiated indirectly by humoral factors (cytokines) produced by activated MP.

Adjuvants, Immunologic

The method for keeping low perfusion flow weaning from centrifugal pumps: an evaluation of hemolysis in the circulator units.

Currently, the clinical application of extracorporeal and assisted circulation using centrifugal pumps is becoming popular. However, a problem in the use of centrifugal pumps is the difficulty in controlling the flow rate when it is a low range. Although we have been controlling the flow rate with a control unit that simply squeezes the blood outlet tube from the outside, hemolysis resulting from turbulent flow in the tube was discovered. Therefore, this control procedure was evaluated experimentally. Two groups were tested as follows: in Group A (n = 10), the tube was squeezed from the outside using the control unit, and in Group B (n = 10), no control unit was used. Blood in each circuit was circulated at 1,500 rpm, and the flow rate was controlled to 250 ml/min using the control unit for Group A. Time-course changes of lactate dehydrogenase (LDH) and free hemoglobin in the blood were measured during 8 h of circulation. There were no significant differences between the 2 groups in the mean values of LDH and free hemoglobin. Therefore, the present low flow rate control provided by the control unit makes it appear to be a simple and useful apparatus for controlling blood circulation using centrifugal pumps.

Analysis of Variance

Experimental studies on three types of heparin-coated cardiopulmonary bypass circuits.

In cardiovascular operations, we have usually used heparin-coated cardiopulmonary bypass circuits with low systemic heparinization. Three types of heparin-coated cardiopulmonary bypass circuits are available in Japan: 2 of the 3 have covalent heparin bonding, and the other has ionic heparin bonding. We studied these circuits in ex vivo experiments to explore which were the best in terms of biocompatibility. In this study we compared the Carmeda system (Medtronic) and the Capiox system (Terumo) with covalent heparin bonding, and the Duraflo-II (Baxter) with ionic heparin bonding, evaluating them in ex vivo experiments. They were primed with fresh human blood, and we studied and compared the platelet counts, fibrinogen, D-dimmer, beta-thioguanine (TG), thrombin-antithrombin complex (TAT), and C3a and C4a of each of them. Additional research will be presented in the future.

Anticoagulants

Our distal aortic perfusion system in descending thoracic and thoracoabdominal aortic aneurysm repairs.

We have used heparin-bonded partial cardiopulmonary bypass to support distal aortic circulation during aortic cross-clamping. However, there were no cardiotomy reservoirs with fully reliable thromboresistance. To resolve this problem, a short-acting anticoagulant (nafamostat mesilate) was added into a cardiotomy reservoir. The present study was designed to evaluate the efficacy of our distal perfusion system. From May 1995 through the end of May 1996, 27 patients underwent descending thoracic and thoracoabdominal aortic aneurysm repairs with this adjunct, 4 being excluded from the experiment. Twenty patients who had undergone conventional partial cardiopulmonary bypass were defined as the control group. There were no significant differences between the 2 groups in the morbidity, mortality, gas transfer, or transfusion requirements despite the fact that more complicated surgical procedures (shown by a two-fold increase in the prevalence of reoperation) were required in the group that had received the current distal perfusion adjunct the heparin-bonded group. In conclusion, our perfusion system is very effective for descending thoracic and thoracoabdominal aortic aneurysm repairs.

Analysis of Variance

Clinical, pathological, and etiologic aspects of acquired dermal melanocytosis.

To study the pathogenesis of acquired dermal melanocytosis (ADM), we reviewed the clinical, immunohistochemical, and ultrastructural features of 34 cases (female, 33, and male, 1) of ADM. The patients' ages at onset ranged from 8 to 51 years and averaged 26.8 +/- 12.7 years. There was a positive family history. Gray-brown macules were mostly recognized on the face. Not only active dermal melanocytes but also non-pigmented c-KIT- and TRP-2-positive immature melanocytes were detected in the dermis. Taken together those clinical and histological findings, activation of pre-existing immature melanocytes by sunlight, estrogen, and/or progesterone, and some other factors, may be the most likely mode of the development of ADM. Moreover, using cultured murine neural crest cells as a model of c-KIT-positive immature melanocytes, we confirmed that endothelin-1, which is produced and secreted by keratinocytes after UV-irradiation, affects melanocytes and accelerated melanogenesis.

Adult

DAX-1 gene mutations and deletions in Japanese patients with adrenal hypoplasia congenita and hypogonadotropic hypogonadism.

Abnormality of the DAX-1 gene accounts for many instances of congenital adrenal hypoplasia. In the present study, we performed molecular genetic analysis of DAX-1 in 4 unrelated Japanese patients with adrenal hypoplasia congenita and hypogonadotropic hypogonadism. A double-point mutation for V126M and W171X was identified in 1 family and a complex de novo insertion-deletion mutation was identified in a second. The DAX-1 gene was entirely deleted in a 3rd patient as well as in a 4th with the additional feature of glycerol kinase deficiency.

Adolescent

Pubertal changes in testicular 3 beta-hydroxysteroid dehydrogenase activity in a male with classical 3 beta-hydroxysteroid dehydrogenase deficiency showing spontaneous secondary sexual maturation.

Males with classical 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) deficiency manifest appropriate secondary sexual maturation with an elevation in serum testosterone levels at pubertal age. To define the origin of serum testosterone, we evaluated a male patient with classical 3 beta-HSD who showed pubertal development. High values of testosterone and a ratio of delta(5) to delta(4) steroids in the spermatic vein indicated direct production of considerable amounts of testosterone and a persistent defect of 3 beta-HSD activity in the gonad. Immunohistochemical analysis showed distinct immunoreactivity in the Leydig cells of the patient. The patient was homozygous for a nonsense mutation in the type-II 3 beta-HSD gene. We propose that gonadal type-I 3 beta-HSD could be expressed by gonadotropin stimulation at pubertal age, and delta(4)-steroid precursors would convert to testosterone.

3-Hydroxysteroid Dehydrogenases

Effects of adenosine triphosphate on ventriculoatrial conduction--usefulness and problems in assessment of catheter ablation of accessory pathways.

The effects of adenosine triphosphate (ATP) on ventriculoatrial (VA) conduction were examined before and after accessory pathway (AP) ablation, with emphasis on assessment of the complication of dual atrioventricular (AV) node pathway. By evaluating the differences in the response to ATP of APs and other pathways, we assessed the usefulness and problems of this method. Of 59 patients who underwent AP ablation, 31 showed pre-excitation and 28 had concealed APs. A dual AV node pathway was found in 9 patients (15.3%) before ablation. After ablation, a dual AV node pathway was newly found in 9 patients. Thus, the total number of patients with a dual AV node pathway was 18 (30.5%). VA conduction over APs was not blocked in 26 of 29 patients, but the remaining 3 APs were blocked transiently by ATP. ATP caused VA block over the AV node in 15 of 16 patients and a dual AV node pathway in all 11 patients. In contrast, VA conduction over the retrograde fast pathway was blocked in 9 of 14 patients with AV node re-entrant tachycardia. ATP has little effect on APs, so observation of the response to ATP provides a more reliable and useful means of evaluating successful ablation. With this method, however, it is important to consider the possibility of the presence of ATP-sensitive APs and ATP-resistant retrograde fast pathways. The influence of ablation-induced injury has not been fully clarified. It is therefore essential to take into account various data, including the comparison between data obtained before and after ablation.

Adenosine Triphosphate