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Biomedical subjects

A Kouno

Publications and source records attributed to A Kouno.

At least 19 recordsLinked to original sources

Fluorometric determination of N-terminal prolyl dipeptides, proline and hydroxyproline in human serum by pre-column high-performance liquid chromatography using 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsufonyl chloride.

A highly sensitive HPLC method for the determination of prolyl dipeptides, Pro and Hyp in serum was developed. After deproteinization of serum and pretreatment with o-phthalaldehyde, the analytes were derivatized with 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride at 70 degrees C for 10 min. The fluorescent derivatives of prolyl dipeptides, Pro and Hyp, were separated on tandem reversed-phase columns by a gradient elution at 55 degrees C and detected by fluorescence measured at 318 nm (excitation) and 392 nm (emission). The detection limits for prolyl dipeptides were 2-5 fmol/injection (S/N=3). Pro-Hyp, Pro-Gly and Pro-Pro were identified as serum prolyl dipeptides. The within-day and between-day relative standard deviations were 1.5-7.9 and 2.4-10.8%, respectively. The recoveries were in the range of 90.8-97.3%. The concentrations of Pro-Hyp, Pro-Gly, Pro-Pro, Pro and Hyp in normal human serum (n = 10) were 0.64+/-0.35, 0.078+/-0.047, 0.022+/-0.016, 177.0+/-43.0 and 11.1+/-3.5 microM, respectively. The concentrations of Pro-Hyp and Pro-Pro in serum of a patient with bone metastases of prostatic cancer were about three times and 50 times, respectively, higher than those in normal human serum.

Chromatography, High Pressure Liquid↗

Development of haemoglobin subtypes and extramedullary haematopoiesis in young rats. Effects of hypercapnic and hypoxic environment.

The influence of repeated hypoxia on the development of haemoglobin (Hb) subtypes and on extramedullary haematopoiesis (EMH) was investigated in young Wistar rats of different ages. The rats were exposed to hypercapnic/hypoxic and to "simple" hypoxic conditions. The results obtained were compared to those of an untreated age-matched control group. Different globin chains were measured using HPLC and time-of-flight (TOF) mass analysis. The number of EMH cells was evaluated by cell counting. By determining the proportions of alpha- and beta-chains, fetal, neonatal and mature types of globin chain composition could be differentiated. The beta-2 chain levels were significantly higher in hypercapnic/hypoxic environments than in the controls and simple hypoxic environments. The numbers of EMH cells in the two groups subjected to hypercapnia/hypoxia decreased significantly more slowly compared to the controls and simple hypoxia groups. Therefore, the development of Hb subtypes and the EMH activity in rats were influenced by both repeated hypercapnia and hypoxia.

Animals↗

Analysis of hemodynamic response with 1/R control on biventricular bypass goat.

A conductance and arterial pressure based method (1/R control) to determine the cardiac output (CO) of a total artificial heart (TAH) was developed to provide a central nervous system with control over the output of TAH. In order to clarify the deference in hemodynamic response between natural heart and 1/R control, biventricular bypass was introduced in the goat. After 2 pneumatically driven sac-type blood pumps were connected to the natural heart, the pulmonary artery was totally clamped to acquire 100% right heart bypass, and the ascending aorta was stenosed to acquire about 60 to 90% left heart bypass; 1/R control was performed substituting the output of the right artificial heart for the CO. The results demonstrated that stable control could be achieved. A discrepancy was often seen between the pulse rate (PR) of the artificial heart and the heart rate (HR) in absolute value. However, the relative changes of PR were quite similar to that of HR for the most part, indicating that the responses of 1/R control were the duplication of natural cardiac responses in normal daily activity.

Animals↗

Development of a miniature undulation pump for the distributed artificial heart.

Research of the distributed artificial heart is important not only to acquire the means of individual organ perfusion but also to clarify the characteristics of the organ and the mechanism of blood distribution. To investigate the distributed artificial heart, the miniature undulation pump was developed. The outer diameter and the thickness of the developed pump were 38 mm and 11 mm, respectively. The priming volume of the pump was 3.2 ml. The total size including the motor unit was 38 mm in diameter and 32 mm in length. The total weight was 67.5 g. The total volume was 27.5 ml. The pump was driven with pulse width modulation by using a 1 chip motor controller. More than 5 L/min of continuous output could be obtained. The results showed that the developed miniature undulation pump system had enough performance for individual organ perfusion.

Biomechanical Phenomena↗

Using one rotary blood pump to produce separate pulsatile circulations in the upper and lower halves of the body.

Separate systemic circulations with pulsatile flow were obtained using 1 rotary blood pump as a left ventricular assist device. The outlet of the pump was divided into 2 conduits, 1 connected to the upper half of the body and the other connected to the lower half. An electric actuator that clamped the 2 outlet conduits alternately provided pulsatile flows. An in vitro experiment showed that the pulsatility phases of the upper and lower halves of the body were complementary with pulsatile flow, and an in vivo experiment showed that controlled flow distributions of continuous flows could be obtained.

Animals↗

Implantation of the undulation pump total artificial heart in the goat.

The undulation pump total artificial heart (UPTAH) was developed by using small-size continuous-flow displacement-type blood pumps (undulation pump). To clarify and improve the problems accompanied by the implantation in the chest, 14 animal experiments were performed on goats weighing 41.3-79.2 kg. The UPTAH could be implanted in the chest of all goats and was driven with a modulation pulsatile mode. The first problem was the atrial suction effect. This problem could be prevented to some extent by developing the soft disk and by improving atrial cuffs. An automatic detection and releasing of the atrial suction effect was also tried. The next problem was acute lung edema accompanied by the postural change of the animal. Development of the automatic control of left atrial pressure could prevent this problem. Small blood leakage from a pinpoint hole in the seal membrane was the next problem. Improvement of the manufacturing procedure of the membrane prevented this. With these improvements, a 10 day survival could be obtained with this unique implantable total artificial heart.

Acute Disease↗

Present status of the total artificial heart at the University of Tokyo.

At the University of Tokyo, various types of total artificial heart (TAH) systems have been studied since 1959. At the present time, 2 types of implantable TAH have been developed. One is an undulation pump TAH (UPTAH) and the other is a flow transformed pulsatile TAH (FTPTAH). Using the UPTAH, 14 cases of implantation were performed in goats and 10 days' survival obtained. The new type of FTPTAH is under a prototype study. To prevent ring thrombus, a polyurethane membrane valve, a jellyfish valve, has been developed. The longest in vivo experiences with this valve in the TAH blood pump have been 312 days in the left side blood pump and 414 days in the right side blood pump. Conductance and arterial pressure based control (1/R control) can realize the physiological control of the TAH. Using 1/R control, 532 days of survival could be obtained in a goat with a paracorporeal TAH. The technique required to apply this control method to a implantable TAH is under development. We have proposed a new 5 year research project of the implantable TAH entitled "Comprehensive Basic Research on the Development of a Japanese Original Implantable Total Artificial Heart" to The Ministry of Welfare.

Animals↗

Physiological control of a total artificial heart: conductance- and arterial pressure-based control.

To obtain a physiological response by a total artificial heart (TAH), while eliminating the hemodynamic abnormalities commonly observed with its use, we proposed the use of a conductance- and arterial pressure-based method (1/R control) to determine TAH cardiac output. In this study, we endeavored to make use of a variable more closely tied to central nervous system (CNS) efferents, systemic conductance, to provide the CNS with more direct control over the output of the TAH. The control equation that calculates the target cardiac output of the TAH was constructed on the basis of measurement of blood pressures and TAH flow. The 1/R control method was tested in TAH-recipient goats with an automatic method by using a microcomputer. In 1/R control animals, the typical TAH pathologies, such as mild arterial hypertension and substantial systemic venous hypertension, did not occur. Cardiac output varied according to daily activity level and exercise in a manner similar to that observed in natural heart goats. These results indicate that we have determined a control method for the TAH that avoids hemodynamic abnormalities exhibited by other TAH control systems and that exhibits physiological responses to exercise and daily activities under the conditions tested. The stability of the control and the complete lack of inappropriate excursions in cardiac output is suggestive of CNS involvement in stabilizing the system.

Animals↗

Development of the undulation pump total artificial heart.

The undulation pump is a small size continuous flow displacement type blood pump that has been developed for an artificial heart. Using undulation pumps, 2 types of implantable total artificial hearts (TAHs), the undulation pump TAH (UPTAH) type 1 (UPTAH 1) and UPTAH type 2 (UPTAH 2) were developed. Both UPTAHs were designed to be small enough to implant into the chest of a goat, the experimental animal. UPTAH 1 could be reduced in size to 75 mm in diameter and 78 mm in length. The weight was 520 g. UPTAH 2 could be reduced in size to 75 mm in diameter and 80 mm in length. The weight was 650 g. UPTAH 2 could be tested in an animal experiment using an adult female goat weighing 52.3 kg. The UPTAH 2 could be implanted successfully into the goat's chest with a good fit. The goat stood after the surgery and extubation and survived for 3 h and 40 min; thus, the potential of the UPTAH for a practical implantable TAH was demonstrated.

Animals↗

Use of a catecholamine sensor in the control of an artificial heart system.

An electrochemical sensor system to allow real-time measurement and feedback of catecholamine concentrations was developed for use in the control of artificial hearts. Electrochemical analyses were carried out using a carbon fiber working electrode, an Ag-AgCl reference electrode, and a potentiostat. The operating parameters of the pneumatically-driven artificial heart system were altered in accordance with the algorithm for changes in the catecholamine concentration. The minimum detectable concentrations of both adrenaline and noradrenaline in a mock circulatory system using a phosphate-buffered solution were approximately 1-2 ng/ml (10(-8) mol/L). An artificial heart control system utilizing this set-up performed satisfactorily without delay, although sensor sensitivity decreased when placed in goat plasma instead of a phosphate-buffered solution, due to the adsorption of various substances such as plasma proteins onto the electrodes. This study demonstrated the future feasibility of a feedback control system for artificial hearts using catecholamine concentrations.

Adsorption↗

Child abuse and neglect in Japan: coin-operated-locker babies.

The coin-operated-locker baby is a type of child abuse that may be unique to Japan. The term refers to newborns who are placed, while alive or dead, in coin-operated lockers. This practice has been decreased by specific measures. It is likely that social and economic variables in Japan account for differences in the frequency and types of child abuse cases when compared to American or European societies as social conditions in Japan change the reported incidence of child abuse may increase in the future. Recently, the government of Osaka organized a group specifically designed to deal with the detection and protection of abused and neglected children. In 1993, they published a manual on how to deal with child abuse, but the Japanese judicial administration still uses old laws for abuse cases. The development of new laws is occurring in parts of the country now. This paper introduces the present status of coping with child abuse and neglect in Japan.

Child↗

Basic study to develop the undulation pump for practical use: antithrombogenicity, hemolysis, and flow patterns inside the pump.

The undulation pump (formerly called the precessional displacement pump) is a continuous flow displacement-type blood pump that is being developed as an implantable total artificial heart. A new undulation pump was developed for chronic use and was examined with animal experiment and flow visualization studies. In the animal experiment using a left ventricular bypass in goats, severe hemolysis occurred. After driving for 12 h, thrombus formation inside the pump was found. In the flow visualization studies, the flow pattern showed that the flow inside the pump was a very complicated turbulent flow. Improvement of hemolysis and thrombus formation is important to realize implantable total artificial hearts using undulation pumps.

Animals↗

Development of a new apparatus to observe microcirculation chronically in continuous flow blood pump research.

To observe microcirculation chronically is an important key to the successful evaluation of the continuous flow blood pump. In this study, we succeeded in developing a new apparatus by which microcirculation could be observed chronically without a microscope in a conscious animal. The apparatus utilizes a charge coupled device (CCD). A thin living tissue, such as mesentery, is put directly on a highly integrated CCD and is lit up through the tissue with a light-emitting diode (LED). The vascular nets in the tissue are projected onto the CCD like a contact photograph, which is then sent to a television screen and which can be used to analyze their motion and function. A 0.5-inch CCD having 250,000 pixels was used in this study. The cover glass of the CCD was removed, and a fiber optic plate was fixed onto the surface of the CCD for the tissue to be able to contact with the apparatus surface without clearance. The CCD as well as the LED were molded with epoxy resin for electrical insulation. The apparatus was 35 mm wide and 12 mm high with a micro stand for an LED, which can be easily implanted into an animal. The apparatus was implanted into a rabbit for 12 h. The configuration of arterioles and venules, tens of micrometers in diameter, and their motions in subcutaneous tissue could be observed.

Animals↗

Percutaneous fiberoptic spinal laser endoscopy.

For percutaneous observation of lesions near the spinal cord and percutaneous spinal laser surgery under the visual field, we studied a method and approach to the spinal cord and intraspinal surgical procedures with laser using a newly developed, small-diameter laser endoscope. The subarachnoid and epidural spaces of a goat were punctured using a 17-gauge epidural needle, and the endoscope (diameter 0.7 mm) was inserted through the needle. The spinal cord and surrounding tissue were observed. Next, an endoscope (diameter 1.5 mm) was inserted into the subarachnoid space, and the quality of the picture and manageability of the endoscope were examined. The Xe:Cl excimer laser was irradiated to a nerve root through a 0.4 mm optical fiber for laser transmission through this endoscope. The spinal cord, nerve roots, and arachnoid were clearly observed through this endoscope. The pulsation of the artery accompanying a root was seen clearly, and the quality of the picture of the endoscope was thought to be sufficient for diagnosis. The nerve root was cut within 30 seconds by excimer laser, and the possibility of laser surgery was demonstrated. Although it was shown that this endoscope could be a useful diagnostic and therapeutic tool, improvement in the manageability of the tip of the endoscope was considered to be important for a clinically usable endoscope.

Animals↗

An epidemiologic and histopathological study of sudden cardiac death in Osaka Medical Examiner's Office.

From 1982 to 1986, 1230 sudden death cases were autopsied in Osaka Medical Examiner's Office. Among them, 810 cases were sudden cardiac deaths (SCD) including coronary heart disease (77%), cardiomyopathy (7%), valvular disease (3%). All SCD cases were dead within 24 hours of the appearance of the fatal symptoms, and most of them (72%) were considered instantaneous death. Many of the fatal symptoms began in bed (31%), at bath (17%), at toilet (8%), or at work (8%). Thirty-four percent of them were thought by themselves or by their families to be healthy before the death. Hypertension (38%), coronary heart disease (13%) and diabetes mellitus (11%) were the major past history recorded. Microscopic observation of the hearts of 200 cases autopsied in 1986 showed various cardiac lesions: hypertrophy, atrophy, degenerations of myocytes, cellular and fatty infiltrations of the interstitium. According to their cardiac lesions and degrees of severity of coronary sclerosis, patients who died suddenly were divided into 8 groups as follows: 1. myocardial infarction (41) 2. myocarditis (6) 3. hypertrophic cardiomyopathy (19) 4. chronic ischemia with severe coronary sclerosis (65) 5. chronic ischemia with moderate coronary sclerosis (27) 6. small vessel disease (18) 7. amyloidosis (1) 8. unknown (23). These results suggest that coronary heart disease and hypertension play an important role in SCD.

Adolescent↗

A statistical study of sudden cardiac death for past five years in Osaka medical, investigated at the Osaka Medical Examiner's Office.

From 1982 to 1986, there were 1230 sudden death cases autopsied in Osaka Medical Examiner's Office. Among them, 810 cases were classified cardiac deaths (SCDs) such as coronary heart disease (77%), cardiomyopathy (7%), valvular disease (3%). All SCD cases were dead within 24 hours after the appearance of the fatal symptoms, and most of them (72%) were supposed to be instantaneous death. Many of the fatal symptoms began in bed (31%), at bath (17%), at toilet (8%) or at work (8%). 34% of them were thought to be healthy by themselves or by their family members before their deaths. Hypertension (38%), coronary heart disease (13%) and diabetes mellitus (11%) were their major past histories recorded. Cardiac lesions such as myocardial hypertrophy, fibrosis and atherosclerosis of coronary artery were frequently admitted in SCD. SCD has been vigorously studied for a long time in various fields including legal medicine, and it has also attracted a great deal of public attention recently. To clarify the causes and the mechanisms of SCD, investigations of the actual situation of SCD cases is essential. Therefore, 1230 autopsy cases of SCD in Osaka Medical Examiner's Office for recent five years were statistically investigated.

Age Factors↗

Quantitative approach to determine the application time of the left ventricular assist device.

This study was designed to establish a method for determining the optimal time of the application of the left ventricular assist device from the viewpoint of the metabolism of the vital organs. Quantitative systemic low flow states were made utilizing goats with total artificial hearts. During these experiments, arterial lactate and pyruvate values were sequentially measured. The behavior of anaerobic metabolism during low cardiac output states and in the recovery process following normalization of the cardiac output was quantitatively described and evaluated. The experimental results are summarized as follows: If a time when the decay of arterial lactate ceases at a level above the normal level by greater than 30 mg/dl is observed despite the use of intraaortic balloon pumping, that time is the exact optimal time for application of the left ventricular assist device. There is a great possibility that circulatory failure will be irreversible if left ventricular assist pumping is not initiated before the arterial lactate value increases to greater than or equal to 140 mg/dl. The method of evaluating the alteration of anaerobic metabolism described in this study is useful for determining the application time of the left ventricular assist device.

Animals↗