Biomedical subjects
Publications and source records attributed to A Harada.
Orosomucoid 1 polymorphism was revealed in human urine by isoelectric focusing and immunoprinting on polyacrylamide gels. The orosomucoid in urine samples was recovered and concentrated by using immobilized allo A lectin. The orosomucoid 1 phenotypes of 60 urine samples correlated with those of the corresponding serum samples.
Curative resection of pancreatic and hepatobiliary tumors is rarely possible because of local invasion, especially into the portal vein. We developed a new antithrombogenic catheter using a heparinized hydrophilic polymer to allow portal vein bypass during resection of tumors invading the portal vein. Pancreatectomy or hepatectomy accompanied by portal vein resection was performed for pancreatic or hepatobiliary cancer, with an intraoperative shunt from the superior mesenteric vein to the femoral vein or from the superior mesenteric vein to the intrahepatic portal vein through the umbilical vein or the hepatic hilar portal vein. Use of the shunt prevented stasis in the superior mesenteric vein and hepatic ischemia even during prolonged occlusion of the portal vein, and portal vein resection was performed in 81 patients with hepatobiliary and pancreatic disease with greater safety and ease.
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Intrapancreatic extension of pancreatic duct cell carcinoma in 19 resectable cases was evaluated by both hematoxylin and eosin (H&E) staining and quick immunostaining using antibodies against cancer-associated antigens--e.g., CA19-9, DUPAN 2, and CEA--on the intraoperative frozen-section biopsy during the course of laparotomy. By H&E staining, malignant lesions were identified in four of 19 cases (21%). By the immunostaining method, however, cancer cells were detected in 10 of 19 cases (52.6%). This immunostaining method was particularly useful in defining cancer cells surrounded by dense fibrous connective tissue, in which cancer cells are usually difficult to evaluate by H&E staining. Thus, the application of such immunostaining, together with H&E staining, on the cryostat sections of biopsy specimens may provide important information for the appropriate operative method. Retrospective study of permanent sections revealed that pathological diagnosis by quick immunostaining was more accurate than diagnosis by H&E staining.
The rapid immunostaining of gastrointestinal cancer-associated antigens, CA19-9, CEA, DUPAN2, and CA50, for intraoperative pathological diagnosis of pancreatic cancer can be completed in only 13 min by applying microwave irradiation, which was used to accelerate the incubation of the primary antibody. Only 3 s irradiation at 500 W for fresh-frozen sections produced specific antigen staining of greater intensity than that obtained with longer incubation, and tissue preservation was satisfactory with minimal non-specific background staining. This rapid immunoperoxidase staining method on the cryostat section of pancreatic biopsy specimens can provide important information for determining an appropriate operative method for pancreatic cancer patients.
Adequate examination of the coronary arteries at forensic autopsy is often difficult. No matter whether the arteries are opened longitudinally or transversely, each method has its disadvantages. A technique making silicone rubber casts of the cardiac vessels providing simultaneous angiography is described. The method allows the three-dimensional precise assessment of stenotic lesions of the coronary arteries. It is concluded that the method examined could be useful for the forensic problems.
Traumatic laceration of a saphenous vein grafted for coronary artery bypass is extremely rare. Successful surgical repair of a laceration was performed in a 64-year-old man who developed retrosternal hematoma from a saphenous vein graft after blunt injury to the chest.
We have examined the cytoskeletal architecture and its relationship with synaptic vesicles in synapses by quick-freeze deep-etch electron microscopy (QF.DE). The main cytoskeletal elements in the presynaptic terminals (neuromuscular junction, electric organ, and cerebellar cortex) were actin filaments and microtubules. The actin filaments formed a network and frequently were associated closely with the presynaptic plasma membranes and active zones. Short, linking strands approximately 30 nm long were found between actin and synaptic vesicles, between microtubules and synaptic vesicles. Fine strands (30-60 nm) were also found between synaptic vesicles. Frequently spherical structures existed in the middle of the strands between synaptic vesicles. Another kind of strand (approximately 100 nm long, thinner than the actin filaments) between synaptic vesicles and plasma membranes was also observed. We have examined the molecular structure of synapsin 1 and its relationship with actin filaments, microtubules, and synaptic vesicles in vitro using the low angle rotary shadowing technique and QF.DE. The synapsin 1, approximately 47 nm long, was composed of a head (approximately 14 nm diam) and a tail (approximately 33 nm long), having a tadpole-like appearance. The high resolution provided by QF.DE revealed that a single synapsin 1 cross-linked actin filaments and linked actin filaments with synaptic vesicles, forming approximately 30-nm short strands. The head was on the actin and the tail was attached to the synaptic vesicle or actin filament. Microtubules were also cross-linked by a single synapsin 1, which also connected a microtubule to synaptic vesicles, forming approximately 30 nm strands. The spherical head was on the microtubules and the tail was attached to the synaptic vesicles or to microtubules. Synaptic vesicles incubated with synapsin 1 were linked with each other via fine short fibrils and frequently we identified spherical structures from which two or three fibril radiated and cross-linked synaptic vesicles. We have examined the localization of synapsin 1 using ultracryomicrotomy and colloidal gold-immunocytochemistry of anti-synapsin 1 IgG. Synapsin 1 was exclusively localized in the regions occupied by synaptic vesicles. Statistical analyses indicated that synapsin 1 is located mostly at least approximately 30 nm away from the presynaptic membrane. These data derived via three different approaches suggest that synapsin 1 could be a main element of short linkages between actin filaments and synaptic vesicles, and between microtubules and synaptic vesicles, and between synaptic vesicles in the nerve terminals.(ABSTRACT TRUNCATED AT 400 WORDS)
An improved method was devised to purify ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) with high specific activity (2.1 mumol of CO2 fixed/mg protein/min) from Euglena gracilis Z. The purified enzyme stored at -80 degrees C required treatment with dithiothreitol for full activity. The dithiothreitol-treated RuBisCO was activated by 12 mM NaHCO3 and 20 mM MgCl2, and the activated state was stable at least for 60 min in the presence of 4 mM ethylenediaminetetraacetate. The form of inorganic carbon fixed by the Euglena enzyme was CO2, as for the plant enzymes. The carboxylase reaction proceeded linearly with time for at least 8 min. The optimum pH for this reaction was 7.8 to 8.0. The carboxylase activity increased with increasing temperature up to 50 degrees C. The activation energy for the carboxylation reaction was 10.0 kcal/mol. The Michaelis constants of Euglena RuBisCO were 30.9 microM for CO2, 560 microM for O2, and 10.5 microM for ribulose 1,5-bisphosphate. Mathematical comparison between the photosynthesis rate predicted from these enzymatic properties and the observed rate suggested that there is no CO2-concentrating mechanism in E. gracilis.
Clinicohistopathologic and immunohistochemical studies of intrapancreatic development of duct cell carcinoma of the head of the pancreas to the body and tail were done in 34 cases in which total pancreatectomy accompanied by portal vein resection were performed from July 1981 to June 1987. In studies of hematoxylin and eosin (HE) staining, intrapancreatic development from the head to the body or tail was observed in 14 cases of 34 cases (41.1%). Multicentricity or skip development was observed in two of 14 cases. However, by using immunostaining of carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9) and DUPAN2, small cancer nests surrounded by dense fibrous connective tissues could be easily and accurately diagnosed, and finally, in 25 of 34 cases (73.5%), intrapancreatic continuous development from the head to body or tail was observed. The intrapancreatic development correlated with portal invasion and perineural invasion of carcinoma, hardness of body and tail, obstruction of the main pancreatic duct, and irregular pancreaticogram. The intraoperative quick immunostaining on the cryostat sections, together with HE staining, is useful to determine the intrapancreatic development of the carcinoma. The indication of total pancreatectomy or pancreatoduodenectomy for carcinoma of the head of the pancreas can be determined by these results.
A fatal case with an unusual shotgun slug injury caused by an accidental discharge of a shotgun used in hunting bear is reported. The slug is old-fashioned, and is no longer on the market. Some old hunters, however, still possess such shotgun slugs, and the possibility of their use must be borne in mind.
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The observations made under controlled experimental conditions provided us with a clearer understanding of the mechanisms involved in the generation and perpetuation of atrial fibrillation. The magnitude and rapidity of change that occurs in the activation patterns during atrial fibrillation were not appreciated, however, until the arrhythmia was studied in similar detail in humans. These studies provided the scientific basis for devising a surgical treatment for atrial fibrillation that has been successful in three patients. By converting the atrial fibrillation to normal sinus rhythm, all three of the detrimental sequelae of atrial fibrillation have been alleviated in each of these patients. Despite the fact that these clinical results are preliminary at this point, our experience documents that atrial fibrillation can be cured by surgical means. In the absence of other effective forms of therapy and in view of the devastating complications of the arrhythmia, surgical intervention should be considered a viable option for the treatment of atrial fibrillation.
Orosomucoid 1 phenotypes were detected in seminal plasma by isoelectric focusing and immunoprinting. The orosomucoid 1 phenotypes in seminal plasma correlated with the types found in the corresponding serum specimens. Semen stains stored for ten days could be typed for orosomucoid 1. The present work revealed that orosomucoid 1 is a useful genetic marker for the medicolegal grouping of semen stains.
In case of an arterial infusion chemoembolization therapy for primary or metastatic liver cancer, gradual release of the anti-cancer drug from lipiodol is a very important factor for a higher drug concentration in the tumor and for longer contact. We studied basic points about what kind of drug form has a gradual drug release. We prepared 3 forms of drugs. (1) Powder form: Powder of ADM, MMC and CDDP was suspended in lipiodol with ultrasonic suspender. (2) URO form: ADM and MMC were dissolved with Urografin and mixed with lipiodol. (3) Surfactant form: ADM and MMC were dissolved with water and then mixed with lipiodol using surfactant. We put lipiodol suspension into physiological saline and then stirred water at 100 rpm with the paddle method, measuring drug release from the suspension or emulsion. Powder form had a lowest drug release. In clinical trials, we administered intra-arterially (1) ADM, MMC dissolved with physiological saline water as usually used (physiological saline water form) (2) Powder form; (3) URO from; (1) CDDP solution as usually used was administered; (2) Powder form. Then we studied the changes of serum concentration of ADM, MMC and CDDP. The results indicated that powder form had the lowest drug release. Thus, the water-soluble anti-cancer drugs ADM, MMC and CDDP should be used in powder form.
A recently developed computer program is capable of rapidly (less than 5 minutes) constructing a series of potential-distribution maps (PDMs) for every msec of a 4-second window of ventricular tachycardia (VT). This study was performed to assess the ability of a series of PDMs to localize the site of earliest activation of VT originating in the interventricular septum. In 12 dogs, 13 morphologies of VT were initiated with programmed electrical stimulation 3-6 days after anterior septal coronary artery infarction. VT was mapped with endocardial and epicardial unipolar electrodes with a multipoint, computer-assisted mapping system. PDMs were compared with activation-time maps, and the former correctly identified the site of earliest activation of all 13 VT morphologies. When PDMs were viewed in sequence on a computer monitor, the site of earliest activation was signaled by abrupt development of a negative potential of less than -3.0 mV. The initial negative point subsequently expanded, and the spread of this negative-potential field correlated with activation sequence. PDMs provide an accurate, unambiguous, rapid means of analyzing large numbers of electrograms acquired with multipoint, computer-assisted mapping systems.
The patient with the anterior-paraseptal type WPW syndrome was operated upon with closed heart technique (so-called epicardial approach) of cryosurgery. Following intraoperative electrophysiologic studies, with ultrasonic aspirator, the epiannular fat-pad of the anterior right atrium was completely dissected to the epicardial side of the tricuspid annulus. Then the anterior paraseptal accessory pathway on the tricuspid annulus was ablated by cryosurgery (-65 degrees C, 3 min, total 9 trials). With this method, cardiotomy and establishment of cardiopulmonary bypass were not necessary to divide the accessory pathway, and significant hemodynamic instability was not caused. But in the case of accessory pathway near interatrial septum (for example, anterior paraseptal or posteroseptal) WPW syndrome, although cryoprobe was applied just adjacent to the AV node, careful ECG monitoring throughout cryosurgery obviated the injury to the AV node.
Effects of open-heart surgery on renal function were studied in 38 patients who had uneventful postoperative course with no sign of acute renal failure. Of these, 19 underwent aorto-coronary bypass grafting (Group 1) and 19 underwent valve replacement (Group 2). During cardiopulmonary bypass (CPB), renal plasma flow (RPF) and glomerular filtration rate (GFR) significantly decreased in both groups as compared to the preoperative values. The decreased GFR remained significantly low on the day of operation in both groups and returned to the preoperative level on the 1st postoperative day (POD) in group 1 and on the 3rd POD in group 2. On the other hand, plasma antidiuretic hormone (ADH) level markedly increased during CPB and remained significantly higher than the preoperative level through the 1st POD. Despite the decreased GFR and increased ADH, however, urinary flow markedly increased during CPB in both groups and remained at significantly higher level than the preoperative values through the 3rd POD in group 1 and through the 5th POD in group 2. Fractional excretion of sodium significantly increased during CPB in both groups and remained high through the 1st POD in group 1 and through the 3rd POD in group 2. These data demonstrate that the renal function of urine concentration is disturbed in the early phase following open-heart surgery even in the patients who have uneventful postoperative course.