Biomedical subjects

K Nakazawa

Publications and source records attributed to K Nakazawa.

At least 73 records · Page 4Linked to original sources

PubMed2001

The efficacy of nafamostat mesilate on the performance of a hybrid-artificial liver using a polyurethane foam/porcine hepatocyte spheroid culture system in human plasma.

Nafamostat mesilate (FUT) is a protease inhibitor of complement activation. The present study investigates whether FUT protects porcine hepatocytes from being injured by human plasma in a multi-capillary polyurethane foam packed-bed culture system (MC-PUF) such as the hybrid-artificial liver (PUF-HAL). Human plasmas with 1 mM of added ammonia were perfused using a small-scale PUF-HAL with porcine hepatocytes. FUT was continuously infused (10 microg/ml, 50 microg/ml). The ammonia detoxification was maintained in human plasma for 24 hours and for 48 hours with FUT which suppressed the rapid increase of asparaginic acid aminotransferase (AST) and alanine aminotransferase (ALT). After 60 hours of perfusion, hepatocyte spheroids completely collapsed in the human plasma, but a small amount of hepatocyte spheroid was maintained by FUT. The effect of FUT was slightly greater at 50 microg/ml than at 10 microg/ml. Our results suggest that FUT has protective effects against porcine hepatocytes in human plasma, and our PUF-HAL using porcine hepatocytes can function in human plasma for about 48 hours with FUT.

Alanine Transaminase↗

PubMed2001

[Basic study about the development of the hybrid-artificial liver support system using human hepatoma cell lines (Hep G2, Huh 7): effects on liver functions by extracellular matrix (type I collagen) in monolayer culture].

The risk of xenozoonosis infections poses the greatest obstacles against the clinical application of hybrid-artificial liver support system (HALSS). To resolve this issue, we used human hepatoma cell lines (Hep G2, Huh 7) in a type I collagen-coated monolayer culture system, and analyzed liver specific functions such as ammonia removal and albumin synthesis capacity. Ammonia removal activity (nmol/10(6) nuclei/hour) and albumin synthesis activity (microgram/10(6) nuclei/day) were upregulated in both Hep G2 and Huh 7 by type I collagen-coated monolayer culture. In particular, Hep G2 cultured in type I collagen-coated monolayer demonstrated relatively high ammonia removal and albumin synthesis capacity. These results indicate the possibility of the application of human hepatocytes to HALSS.

Albumins↗

PubMed2001

[A case of gastric cancer with multiple liver metastases that were completely responsive to hepatic arterial infusion chemotherapy].

A 70-year-old male was hospitalized with a diagnosis of gastric cancer. Multiple liver metastases were detected by CT scan and ultrasonography, but no other metastases were found. After distal gastrectomy with lymphadenectomy, an intra-arterial catheter was inserted and hepatic arterial infusion (HAI) chemotherapy was started with the following regimen: epirubicin, 40 mg (once in 4 weeks); MMC, 4 mg (once in 2 weeks); and 5-FU, 500 mg (once a week). Oral administration of 300 mg/day of tegafur-uracil (UFT) was given concurrently. The metastatic liver tumors were gradually reduced, and then disappeared. The patient has been free from recurrence after HAI. As gastric cancer tends to spread (e.g. peritoneal dissemination) more than colorectal cancer, the indication for HAI in cases of gastric cancer is limited. However, the present case suggests that it may be useful in achieving long-term survival for patients with metastases localized in the liver.

Adenocarcinoma↗

PubMed2001

Mass preparation of primary porcine hepatocytes and the design of a hybrid artificial liver module using spheroid culture for a clinical trial.

To isolate a large number of porcine hepatocytes, we originally developed a mass preparation method that combined the usual collagenase perfusion method of a whole liver with a collagenase redigestion method of tissue fragments after liver perfusion. Using a pig of 10kg, collagenase perfusion only resulted in a yield of 63+/-78 x 10(8) total cells with a viability of 69.2+/-25.3 %, but our combined method had a yield of 167+/-31 x 10(8) total cells with a viability of 87.9+/-4.4% (mean +/- SD). Also, the combined method was applied to two pigs of 10kg body weight at the same time, and isolated 387+/-89 x 10(8) hepatocytes with a viability of 87.1+/-6.9% and a purity of 93.6+/-2.8 % in 11 experiments. We designed a large multi-capillary polyurethane foam (MC-PUF) packed-bed module containing 1 x 10(10) porcine hepatocytes on a clinical trial scale. The porcine hepatocytes in the module formed spherical multicellular aggregates (spheroids) of 200 - 500 microm diameter. Most hepatocytes forming spheroids were viable judged by fluorescein diacetate and ethidium bromide staining. The activities of ammonia removal, albumin secretion and oxygen consumption of the large MC-PUF module were the same as for a small MC-PUF module containing 2 x 10(8) porcine hepatocytes, and were maintained for at least 9 days of culture. These results show that a large MC-PUF module is successfully scaled up 50 times. In conclusion, we succeeded in developing a mass preparation method of porcine hepatocytes and a large hybrid artificial liver module on a clinical trial scale.

Animals↗

PubMed2001

Polyurethane foam/spheroid culture system using human hepatoblastoma cell line (Hep G2) as a possible new hybrid artificial liver.

The risk of xenozoonosis infections poses the greatest obstacle against the clinical application of hybrid artificial liver support system (HALSS). Primary human hepatocytes are an ideal source for HALSS, but the shortage of human livers available for hepatocyte isolation limits this modality. To resolve this issue, we used human hepatocytes with replication capacity (fetal hepatocytes, Hep G2, and Huh 7) in a polyurethane foam (PUF)/spheroid culture system in vitro, and analyzed liver functions such as ammonia removal and albumin synthesis capacity; results were compared to those of porcine hepatocytes. Human fetal hepatocytes, Hep G2, and Huh 7 formed spheroids spontaneously within 24 h in a PUF/spheroid culture system; ammonia removal activity (micromol/10(6) nuclei/h) was upregulated, as was albumin synthesis activity (microg/10(6) nuclei/day). In particular, Hep G2 spheroids demonstrated high ammonia removal and albumin synthesis activities: 85% of the ammonia removal activity and 171.7% of the albumin synthesis activity of porcine hepatocytes in the monolayer culture. These results indicate the possibility of the development of a multicapillary PUF (MC-PUF) packed-bed culture system of hepatocyte spheroids as a HALSS using Hep G2.

Albumins↗

PubMed2001

Effect of long-term bedrest on lower leg muscle activation patterns during quiet standing.

It has been well known that balance instabilities after long-term exposure to microgravity (e.g., Anderson et al. 1986) or bedrest (BR) can be related to alterations and/or adaptations to postural control strategies. Little is known, however, how the reduced muscular activity affects the activation pattern of the lower limb muscles during quiet standing (QS). The purpose of this study was to investigate whether or not any changes in the lower limb muscle activation patterns during QS would occur after BR.

Adult↗

PubMed2001

Reflex response changes during hyper and microgravity.

This paper reports the quantitative evaluation of the H-reflex exhibited by parabolic flight with exposure to micro and high-gravity. With respect to previous findings in parabolic flights and short-term space missions, the analysis focused on reflex activity in weightlessness. The aim of this study was to investigate the effect of gravity on H-reflex and motor evoked potentials (MEP) in soleus muscle (SOL) during parabolic flight.

Adult↗

PubMed2000

Roles of P-selectin in inflammation, neointimal formation, and vascular remodeling in balloon-injured rat carotid arteries.

BACKGROUND: P-selectin plays key roles in mediating inflammation through promoting adherence of leukocytes to activated platelets and endothelium. This process is one of the initial events in atherosclerosis and restenosis after coronary angioplasty. METHODS AND RESULTS: Using a rat balloon-injury model, we examined the role of P-selectin in vascular inflammatory processes. In the acute phase, immunohistochemistry revealed that P-selectin was intensely expressed on both activated platelets covering the denuded segment and endothelial cells of the inflamed adventitial small vessels. Treatment with an anti-P-selectin monoclonal antibody (MAb) for 8 consecutive days significantly inhibited neointimal formation at day 14 (42% inhibition; P:<0.05), and this effect persisted at day 56 (40% inhibition; P:<0.01) compared with the control group. Vascular shrinking accompanying adventitial fibrosis was also attenuated at day 56. Inhibition of both neointimal formation and vascular shrinking resulted in the lumen area of the anti-P-selectin treatment group being approximately 3 times larger at day 56 than that of the control group. Accumulation of CD45-positive leukocytes in the developing neointima, media, and adventitia at day 8 was significantly inhibited by treatment with the anti-P-selectin MAb. Scanning electron microscopy demonstrated that anti-P-selectin treatment resulted in a less thrombogenic surface of the arterial intima, which featured a pseudoendothelial appearance at day 14 after injury. CONCLUSIONS: These results suggest that inhibition of P-selectin-mediated leukocyte recruitment prevents the development of neointimal formation, adventitial inflammation, and vascular shrinking and promotes pseudoendothelialization by luminal smooth muscle cells. This treatment thus beneficially affects vascular remodeling after balloon injury in rats.

Angioplasty, Balloon↗

PubMed2000

A novel SCN5A mutation associated with idiopathic ventricular fibrillation without typical ECG findings of Brugada syndrome.

Mutations in the human cardiac Na+ channel alpha subunit gene (SCN5A) are responsible for Brugada syndrome, an idiopathic ventricular fibrillation (IVF) subgroup characterized by right bundle branch block and ST elevation on an electrocardiogram (ECG). However, the molecular basis of IVF in subgroups lacking these ECG findings has not been elucidated. We performed genetic screenings of Japanese IVF patients and found a novel SCN5A missense mutation (S1710L) in one symptomatic IVF patient that did not exhibit the typical Brugada ECG. Heterologously expressed S1710L channels showed marked acceleration in the current decay together with a large hyperpolarizing shift of steady-state inactivation and depolarizing shift of activation. These findings suggest that SCN5A is one of the responsible genes for IVF patients who do not show typical ECG manifestations of the Brugada syndrome.

Adult↗

PubMed2000

Extracellular ATP reduces optically monitored electrical signals in hippocampal slices through metabolism to adenosine.

Electrical signals in rat hippocampal slices were optically monitored using a voltage-sensitive dye to determine whether extracellular ATP exhibits direct effects through its own receptors or indirect effects after its hydrolysis to adenosine. The dentate gyrus was stimulated and electrical signals in the CA1 and the CA3 region were analyzed. The signals were divided into two components: a transient component peaking within 10 ms (fast component) and a subsequent sustained component (slow component). ATP (10 to 100 microM) inhibited both the fast and the slow components in the CA1 region by about 30% and 70%, respectively. ADP, AMP and adenosine also inhibited the fast and the slow components. The inhibition by ATP was antagonized by aminophylline and other adenosine receptor antagonists, and by alpha,beta-methylene ADP, an inhibitor of 5'-nucleotidases. These results suggest that extracellular ATP inhibits neuronal electrical signals in hippocampal slices after its metabolism to adenosine.

Adenosine↗

PubMed2000

Effects of changing stance conditions on anticipatory postural adjustment and reaction time to voluntary arm movement in humans.

1. The effect on reaction time (RT) and anticipatory postural adjustment (APA) of unexpectedly changing stance conditions was studied using a push or pull arm movement task. The aim was to evaluate the modifiability of RT and APA by an external perturbation associated with an automatic compensatory reaction. 2. Subjects standing on a moveable platform were asked to push or pull a rigid handle as quickly and as strongly as possible in response to the 'go-signal', a visual signal from a green or red light-emitting diode. Forward and backward translations of the platform were randomly induced at four time intervals after the go-signal. In some experiments to detect unspecific arousal there were no platform translations but an acoustic signal was given before the go-signal. Surface electromyographic activity (EMG) of upper arm and lower leg muscles was analysed. 3. During the push task both RT and the duration of APA (onset of APA till the force signal indicating hand action) were shorter during backward than during forward translation. During the pull task the effect of platform translations was the reverse. The delay between go-signal and onset of APA remained constant. Consequently, RT and APA became shorter when the platform was translated in the same direction as that in which the upper body was displaced by the push or pull movement, and longer when it was translated in the opposite direction. The effects were maximal when translations were induced 250 ms after the go-signal, but a difference was detected up to 375 ms. 4. Furthermore, with forward and backward platform translations RT was shorter when the translations were induced early rather than late after the go-signal. This was associated with a shortening of the delay between the go-signal and onset of APA, while APA duration remained constant. The shortened RT was in the range of that obtained when an acoustic signal was given just before the go-signal. 5. It is concluded that (i) both the RT and the duration of APA can be modified by a translation of the support surface in a functionally appropriate way by updating the internal representation of the actual stance condition within the central nervous system. Both RT and APA are shortened when the body displacement induced by the push or pull movement and platform translation have the same direction; conversely, an inappropriate translation of the feet requires a greater APA and leads to a longer RT; (ii) both APA and RT are modifiable by platform translation for more than half the time between the go-signal and the focal push or pull movement; (iii) an unspecific effect of platform translation on RT can be identified; it may be mediated by a different neuronal substrate.

Adult↗

PubMed2000

Sound-induced laryngeal and respiratory reflexes originate from vestibular afferents.

To determine whether the auditory or vestibular system causes the sound-induced laryngeal reflex, which has been considered to participate in the auditory feedback control of vocalization, click-induced laryngeal responses were compared before and after sectioning of the cochlear and/or vestibular nerves in cats. The sound-induced reflex modulation of respiratory muscle activity was also investigated, because respiratory movement is important for vocal control. Sectioning of the cochlear nerves had little influence on these responses. In contrast, sectioning of the vestibular nerves abolished these responses. It was concluded that the sound-induced laryngeal and respiratory reflexes are attributed to the vestibular system.

Acoustic Stimulation↗

PubMed2000

Inhaled nitric oxide during partial liquid ventilation shifts pulmonary blood flow to the non-dependent lung regions.

OBJECTIVE: To elucidate the change in pulmonary blood flow brought about by nitric oxide (NO) inhalation during partial liquid ventilation (PLV). DESIGN: Prospective, controlled study. SETTING: A research laboratory at a university medical center. SUBJECTS: Fourteen Japanese white rabbits (3.2 +/- 0.05 kg body weight). INTERVENTIONS: Animals were mechanically ventilated in the right decubitus position. Following saline lung lavage, PLV was started with perflubron (15 ml/kg). In the NO group (n = 7), PLV was supplemented by a 30-min challenge of NO inhalation (10 ppm) from 30 min after the initiation of PLV. In the control group (n = 7), PLV was continued for 60 min. MEASUREMENTS AND MAIN RESULTS: For the pulmonary blood flow analysis, colored microspheres were administered from the right atrium at 30 min (T(PLV1)) and 60 min (T(PLV2)) after the initiation of PLV. The percentage of the left lung blood flow in the total pulmonary blood flow (%Q(L)/Q(T)) was significantly increased by NO inhalation in the NO group (p = 0.0164), while that in the control group was significantly decreased during the same period (p = 0.0107). PaO2 in the NO group was significantly increased by NO inhalation (p = 0.0153), but not in the control group (p = 0.7911). CONCLUSION: Inhaled NO during PLV shifted the pulmonary blood flow to the non-dependent region and improved pulmonary gas exchange. This result suggested that inhaled NO took effect predominantly in the non-dependent region during PLV.

Administration, Inhalation↗

PubMed2000

Continuous intra-arterial blood gas monitoring during thoracic surgery.

PURPOSE: This study was undertaken to assess the clinical usefulness of a continuous intra-arterial blood gas (CIABG) monitoring system, Paratrend 7, during thoracic surgery. METHODS: A sensor of the CIABG monitoring system was inserted into the radial artery in 50 patients. During one-lung/differential lung ventilation, arterial blood samples for estimation of blood gases (ABG) were taken every 45-90 min or when clinically needed. The PCO(2), PO(2), and pH values displayed by the CIABG monitor were recorded prior to arterial blood sampling and were compared with the results of ABG analysis. The mean (bias) and the standard deviation (precision) of the differences were calculated from the data for each parameter. RESULTS: Ninety-four blood samples were obtained. The correlation between CIABG and ABG measurements was strong for each parameter: r = 0.83 (PCO(2)), 0.89 (PO(2)), 0.74 (pH). The bias +/- precision between the two methods was 0.4 +/- 3.0 mmHg for PCO(2), -6 +/- 47 mmHg for PO(2), -1.2 +/- 27.4% for PO(2) and -0.01 +/- 0.04 for pH. For PO(2) values <150 mmHg, the bias +/- precision was 1 +/- 28 mmHg. CONCLUSION: The agreement between CIABG and ABG measurements was better for PCO(2) and pH than for PO(2). The PO(2) value displayed on the CIABG monitor may not always be reliable during thoracic surgical procedures.

Journal Article↗

PubMed2000

Stability criterion for controlling standing in able-bodied subjects.

A new stability criterion that can be used to assess the standing condition of a subject from center of pressure (CoP) measurements is presented. This criterion can be applied, for example, to control a standing prosthesis, which should allow a paraplegic subject to stand up, sit down and stand safely without using hands for support. Experiments conducted with able-bodied subjects enabled us to establish a relationship between its stability and the subject's CoP position. Four CoP stability zones were identified: high preference, low preference, undesirable and unstable zones. The high preference zone is defined as the area where the CoP is found 99% of the time during quiet standing. The area where the CoP is found during the remaining 1% of the time is called the low preference zone. The undesirable zone is defined as the CoP area where the subject is forced to change posture in order to maintain balance, and the unstable zone is defined as the CoP area in which the subject is forced to step forward, backward or sideways to maintain stability. A general model of the proposed four stability zones was derived, which can be used to compute stability zones a priori for any subject and thus allows one to assess the subject's stability condition from the CoP measurements.

Adult↗

PubMed2000

Differential angle-dependent modulation of the long-latency stretch reflex responses in elbow flexion synergists.

We determined the effect of elbow joint angle on the short-(M1) and long-latency stretch reflex (M2 and M3) responses of the elbow flexion synergists, the brachioradialis (BR), and the biceps brachii (BB), during weak isometric elbow flexion tasks. The elbow joint angle was 35,75 and 115 degrees (full-extension angle was 0 degrees ), and the muscle contraction level was 0,3 and 6% of maximum voluntary contraction (MVC) of the BR. In BR, the M1, M2 and M3 responses were significantly greater at 75 and 115 degrees than at 35 degrees. On the other hand, in BB, the M2 response was significantly greater at 35 degrees than at 75 and 115 degrees, while the M1 and M3 responses were not significantly different among the elbow joint angles. These results indicated that the stretch reflex responses of BR might be dependent on the changes of muscle length in stretch stimulus, while the M2 response of BB might not be dependent on the actual stimulus intensity. Therefore, we concluded that the M2 of BB might be modulated selectively by a higher reflex center in accordance with relationships of the mechanical advantages between synergistic muscles.

Adult↗

PubMed2000

Relationship between airway pressure and the distribution of gas-liquid interface during partial liquid ventilation in the oleic acid lung injury model: fluorine-19 magnetic resonance imaging study.

OBJECTIVE: To elucidate the relationship between airway pressure (Paw) and the distribution of gas-liquid interface d ring partial liquid ventilation (PLV). DESIGN: Prospective, controlled study. SETTING: A research laboratory at a university medical center. SUBJECTS: Ten Japanese white rabbits. INTERVENTIONS: Ten rabbits were tracheostomized and mechanically ventilated (F(IO2) = 1.0, tidal volume = 40 mL, respiratory rate = 25 breaths/min) under general anesthesia. In the oleic acid lung injury group (n = 5), oleic acid (0.10-0.12 mL/kg) was intravenously administered to make a lung injury model. When PaO2 became <100 torr (13.3 kPa), 15 mL/kg of perflubron was instilled into the endotracheal tube, and PLV was performed for 60 mins. In the control group, PLV was performed for 60 mins without any insult to the lung. After animals were killed under deep anesthesia, Paw was set at 0, 5, 10, 15, 20, 25, 30 cm H2O and fluorine-19 magnetic resonance imaging was performed. Pressure-volume relationship was investigated using proton magnetic resonance imaging. MEASUREMENTS AND MAIN RESULTS: Gas-liquid interface reduces fluorine-19 magnetic resonance signal. The total lung fluorine-19 signal intensity was reduced by the Paw above the lower inflection point on the pressure-volume curve, and severe lung injury interfered with establishing gas-liquid interface, especially in the dependent region, even during PLV. CONCLUSION: Paw above the lower inflection point on the pressure-volume curve established gas-liquid interface dose dependently during PLV, and severe lung injury with low compliance could cause difficulty in the establishment of gas-perflubron interface, especially in the dependent lung region.

Animals↗

PubMed2000

Outbreak of cryptosporidiosis after contamination of the public water supply in Saitama Prefecture, Japan, in 1996.

An outbreak of cryptosporidiosis occurred in Ogose Town, Saitama Prefecture. Japan, in June 1996. Of 12,345 respondents to a questionnaire sent to households in the town (population; 13,809), 8,812 (71.4%) reported an acute gastrointestinal illness some time between May and July. In addition, 274 traceable visitors at local inns, golf courses, and the like during this period and 54 employees from out of town were infected. Cases of cryptosporidiosis were estimated to 9,140. Of these, 2,856 subjects were treated at outpatient clinics and 24 subjects were hospitalized (some subjects counted twice). No deaths were attributed to the outbreak. Among the visitors to Ogose who were traced, 7 persons who stayed only one day during the outbreak and drank half a glass to 2 glasses (100 to 360 ml) of tap water had cryptosporidiosis confirmed by laboratory tests. The median incubation period for the 14 persons for whom this calculation was possible was 6.4 days (range, 5 to 8 days). Of 469 pupils reporting details of their fever and diarrhea, abdominal cramps, or these combined signs and symptoms, the median maximum body temperature was 37.8 degrees C (range, 36.7 to 40.3 degrees C). The duration of illness, reported by 608 of the pupils, was 5.2 days (range, 1 to 15 days), and that reported by 187 employees was 4.8 days (range, 1 to 18 days). The longest known time for discharge of oocysts after onset was 44 days. Blood was not found in the 609 stool specimens examined. The outbreak was caused by contamination of the town's potable water by Cryptosporidium parvum oocysts. The town's water treatment plant treated river water by coagulation, sedimentation, sand filtration, and chlorination. Contamination arose because of various natural and artificial factors: one was that the monthly precipitation in May was much lower than average, causing the river water level to drop. Another factor was heavy rainfall one night in May that increased water turbidity. The amounts of the coagulant added seemed to be insufficient. There are two inns, three public lavatories, and two small-scale wastewater treatment plants upstream 400 m and 1,200 m of the intake point of the town's water treatment plant. However, there are no farms with livestock in the area. We suggest that the location of the water treatment facilities was inappropriate, and that oocysts had circulated from the potable water to humans to sewage to the river and back to the potable water.

Adolescent↗